Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

623

Mycobacteria in water, soil, plants and air: a review

K. Hruska, M. Kaevska

Veterinary Research Institute, Brno, Czech Republic

ABSTRACT: Amazingly, despite the 24 143 papers on mycobacteria, indexed in the Web of Science database during the last six years, published by 67 008 authors from 13 128 organizations located in 166 countries or territories, interna-tionally accepted legal directives on how to control the public health risk associated with environmental mycobacteria have yet to be developed. Mycobacteria are human and animal pathogens, causing not only tuberculosis and leprosy, but mycobacterioses of skin, soft tissues and lung. Due to their cell wall composition and their adaptability mycobac-teria can survive in different habitats for years. Their immunomodulatory ability has been recognised for more than 50 years and hundreds of papers published during the last two decades have demonstrated that small chemical products derived from mycobacterial cells participate in inflammatory pathways involved the pathogenesis of important human diseases like Crohn’s disease, asthma, type 1 diabetes mellitus, psoriasis, arthrosis, Blau syndrom, sarcoidosis, autism etc. Mycobacteria can influence inflammatory pathways not only as live organisms, but also by means of components derived from dead cells. Pasteurisation or cooking does not affect this ability. Hence, how many mycobacterial cells are ingested, what factors play a role concurrently, and how long the harmful effect persists become important questions. This paper presents only a short review based on selected papers about mycobacteria in water, soil, plants and air with the aim of attracting attention to this significant global problem and of making the first steps towards protection of people. Selected bibliographic references of published data from 2007 to 2012 are presented in easy-to-navigate tables.

Keywords: Mycobacterium; water; soil; plant; vegetables; air; biofilm; sediment; determination; zoonoses; food safety

List of abbreviations

CFU = colony forming units; DGGE = denaturation gradient gel electrophoresis; IMS = immuno-magnetic separation; IS = insertion sequence; ITS = intergenic transcribed spacer; MAC = Mycobacterium avium complex; MAP/Map = Mycobacterium avium subsp. paratuberculosis; MTC = Mycobacterium tuberculosis complex; MWF = metal working fluids; nPCR = nested PCR; NTM = non-tuberculous mycobacteria; PCR = polymerase chain reaction; PPM = poten-tially pathogenic mycobacteria; qPCR = quantitative real time PCR; RFLP = restriction fragment length polymorphism

Contents

1. Introduction1.1. The database used1.2. The format used

2. Selected review articles3. Mycobacteria in water 4. Mycobacteria in soil5. Mycobacteria in plants6. Mycobacteria in air7. Identification of mycobacteria8. Acknowledgements9. References

Supported by the Ministry of Education, Youth and Sports, Czech Republic (AdmireVet; Grant No. CZ 1.05/2.1.00/01.0006-ED 0006/01/01) and the Ministry of Agriculture of the Czech Republic (Grant No. MZE 0002716202).

Tables

Table 1. Search profiles used and numbers of results retrieved

Table 2. Selected review articlesTable 3. Mycobacteria in water Table 4. Mycobacteria in soilTable 5. Mycobacteria in plantsTable 6. Mycobacteria in airTable 7. Identification of mycobacteria

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1. Introduction

Potentially pathogenic mycobacteria, also re-ferred to as non-tuberculous mycobacteria, are known pathogens of animals and can cause diseases also in humans, especially in immunocompromised persons. Mycobacterioses differ depending upon the species and hosts involved and upon ways of in-fection, and may present as pulmonary, skin or soft tissue lesions (Wagner and Young 2004; Griffith et al. 2007; Jarzembowski and Young 2008). Some hosts can develop a generalised mycobacteriosis. The immunomodulatory potential of mycobacteria is in the spotlight as a consequence of the compo-sition of the mycobacterial cell wall. Bacterial cell wall components have a high immunomodulatory potential. Dead mycobacteria have been used in the complete Freund adjuvans for more than 50 years. Muramyldipeptides were discovered as the mini-mal structures responsible for the improved reac-tion to antigens (Ellouz et al. 1974; Traub et al. 2006; Coulombe et al. 2009). This ability has been proven by experiments which showed that syn-thetic molecules have the same effects. Coulombe et al. (2009) reported that N-glycolyl MDP has a greater NOD2-stimulating activity than N-acetyl MDP, consistent with the historical observation attributing exceptional immunogenic activity to mycobacterial cells. N-glycolyl MDP is produced by degradation of mycobacterial peptidoglycans. The importance of a lipid antigen in the molecu-lar pathogenesis of ruminant paratuberculosis and human inflammatory bowel diseases are subjects of recently published data (Momotani et al. 2012; Mori and De Libero 2012). It is evident that under specific conditions mycobacteria can be zoonotic or environmental pathogens for humans and an agents that participate in foodborne autoimmune or autoinflammatory human diseases. Crohn’s disease, type 1 diabetes mellitus, psoriasis, mul-tiple sclerosis, asthma, arthrosis, autism, Blau syndrome and sarcoidosis are the most frequently mentioned diseases with respect to bacterial trig-gers. Nevertheless, mycobacteria are not unique in their ability to act as bacterial triggers. Some known pathogens are possible sources of compo-nents that trigger inflammatory processes as a con-sequence of their intensive replication during the primary infection. Non-tuberculous mycobacteria were not a focus of interest for a long time because their participation in pathogenesis need not follow the Koch’s postulates completely or unequivocally.

Those who cannot accept the term “pathogen” for cells unable to replicate can describe the harmful microorganism as an immunomodulator, bacterial trigger or allergen-like factor.

The difficult diagnosis by culture of slow or non-growing mycobacteria has also contributed to an underestimation of mycobacteria as a public health risk. However, the current understanding of the molecular pathogenesis of autoimmune or allergic diseases, the recognition of genetic or epi-genetic components in the pathogenesis of many diseases, the expanding use of molecular biol-ogy in research on mycobacteria, and the rapidly growing number of publications and data on the distribution of mycobacteria in the environment, namely in water, air and soil, have all contrib-uted to the evolution of a new understanding of the role of mycobacteria. Mycobacterium avium subsp. paratuberculosis plays an important role in this paradigm. Paratuberculosis (Johne’s disease) in cattle and sheep was for a long time considered unimportant both for animal breeding and food safety and remained uncontrolled with regard to milk and meat contamination and in animal trade and mobility. Thus, the herd incidence increased in countries with intensive cattle and sheep industry enormously, up to an estimated 70% to 90% of all herds. The infectious agent is very resistant, can survive for a long time in water and liquid dung and can survive and replicate in amoebae. The number of mycobacteria in faeces can reach 108 per gram, in milk and meat 104 per gram and in water 104 per ml. It is therefore evident that humans are not abso-lutely protected against exposure to mycobacteria and their components. The important factors in this exposure are the numbers of mycobacteria and the age and dispositions of the hosts. Obviously some sensitisation can occur inapparently and an interval of many years can exist between the first contact and development of the clinical form of disease. The unknown sources of mycobacteria and the creeping development of health problems make the understanding of possible consequences rather difficult.

The risk of direct transmission of live tuberculous mycobacteria between humans or animals takes the form of droplet infection in open forms of pulmo-nary tuberculosis (M. tuberculosis, M. bovis and M. caprae). The risk of contracting human tuber-culosis increases with the time of sharing a small room with the mycobacterial shedder, e.g. in a class room, pub or nearest neighbour during intercon-

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tinental flight. Non-tuberculous mycobacteria can be transmitted in raw milk or insufficiently heat-treated meat. Water and soil are frequent sources of mycobacterial infections either in the form of direct contacts for aquarists or gardeners or by means of aerosols in showers or indoor swimming pools. Water in hospitals and dental units or metal working fluids (Falkinham 1996, 2009a,b; Primm et al. 2004; van Ingen et al. 2009) have been a recent focus of attention. A recently published book was devoted to the ecology of mycobacteria and their impact on human and animal health (Kazda et al. 2009).

Readers should pay special attention to biofilms, aerosols, resistance to disinfectants, and myco-bacterioses as professional diseases. Several spe-cific phenomenona are typical for mycobacteria: Isolation of mycobacteria from the environment is hampered by their slow or limited growth in vitro.

Mycobacteria are frequently overgrown by other microorganisms present in the sample. To over-come this obstacle, different decontamination methods have proven to be effective, although with negative consequences for the sensitivity of the culture.

As is the case for other microorganisms myco-bacteria can be detected and identified directly and quantitatively using different molecular methods.

Mycobacteria survive for a long time in the en-vironment and can be found in great numbers in rivers that collect water from pastures, in river and lake sediments and in soil.

The hydrophobic character of the mycobacterial cell wall is responsible for their easy aerosolisation over swimming pools and river water, by sea break-ers as well as in the shower bath.

1.1. The database used

The publications on mycobacteria were retrieved from the Web of Science® (Thomson Reuters) data-base using the search profiles described in Table 1, and directed to water, soil, plants and air. The num-bers of results retrieved from the complete data-base Science Citation Index Expanded (timespan: 1945 to 2012) are mentioned only to demonstrate the huge number of sources available. We acknowl-edge that the key words used for searching are too general and also that many inappropriate papers have been omitted. The most important papers published from 2007 to 2012 have been selected for

this review using abstracts or full papers. However, certain important references published before 2007 were also included. The utility “Analyse results” was used for the selection of review articles.

1.2. The format used

The review follows the format of our recently published reviews (Eyer and Hruska 2012; Hruska and Franek 2012). Selected papers are presented in tables with the basic key words in the first column, full or shortened abstract in the second column and the link to the List of References in the third column. This format is easy-to-navigate, supplies readers with more information and minimises the misinterpretation of papers through a subjective wording by the authors of the review. The text in the tables contains several format imperfections, which exist in the Web of Science® database and are caused by transmission and copying of data between various information sources.

2. Selected review articles (Table 2)

Many reviews are devoted to mycobacteria in the environment, namely in water (Falkinham 1996, 2002, 2009a,b, 2010; Whiley et al. 2012). Attention has been also paid to non-tuberculous mycobacteria that cause human disease (Set and Shastri 2011), and guidelines on how to control its transmission is also an area of focus. M. avium complex members and non-tuberculous mycobac-teria cause pulmonary or lung diseases and their diagnosis and treatment have been reviewed by Kasperbauer and Daley (2008) and McGrath et al. (2010). Catheter-related infections (Adekambi 2009) and nosocomial outbreaks of mycobacterial infections (Garcia-Martos and Garcia-Agudo 2012) are other fields of interest. Dental units and their waterlines are reviewed by Szymanska et al. (2008) and Szymanska and Sitkowska (2012). M. avium subsp. paratuberculosis which causes paratubercu-losis in animals, namely cattle and small ruminants, should be regarded also as a bacterial trigger for Crohn’s disease (Carbone et al. 2005), and as a food safety risk factor (Skovgaard 2007; Gill et al. 2011). A small number of reviews present new methods for the detection of mycobacteria using biosensors (Nayak et al. 2009), or fluorescence in-situ hybridi-sation (Cerqueira et al. 2008).

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3. Mycobacteria in water (Table 3)

Mycobacteria are present in most natural wa-ters and piped water supplies. The main features of mycobacteria as a public health risk have been characterized already in 1984 as evident from the sub-headings of a review published by Collins et al. (1984):Resistance of mycobacteria to chlorination Access, persistence and colonization in piped sup-

plies Is water the natural habitat of free-living myco-

bacteria? Water as a vector for mycobacterial infections Immune response to environmental mycobacteria Mycobacteria as indicators of pollution

Most of the bacteria in drinking water distribu-tion systems are associated with biofilms. M. avium has been described to survive in biofilms for more than two to four weeks in culturable forms. Lehtola et al. (2006) studied the survival of M. avium in drinking water biofilms after the spiking of the wa-ter using fluorescent in situ hybridization (FISH) with an rRNA-targeted PNA probe. They conclud-ed that culture examination seriously underesti-mates the occurrence of M. avium in biofilms and water. The study performed by Lehtola et al. (2007) clearly proved that pathogenic bacteria entering water distribution systems can survive in biofilms for at least several weeks, even under conditions of high-shear turbulent flow, and may be a risk to water consumers. In order to understand microbial communities in drinking water biofilms, Liu et al. (2012) sequenced 16S rRNA in three faucet bio-films using 454-pyrosequencing. They found that the abundance of Legionella and Mycobacterium was affected by the residual chlorine in the water.

Most of the non-tuberculous mycobacteria not only survive in water for a long time, but can grow there as well (Kazda et al. 2009). Water, regardless of origin and quality, can be contaminated by my-cobacteria and, under specific conditions, can jeop-ardise the users (Falkinham III 2003; Falkinham 2009a,b). Pickup et al. (2005) reported that M. avium subsp. paratuberculosis can be present in high con-centrations in the river water in the catchments area of pastures. Data presented in this paper bring evidence of a higher incidence of Crohn’s disease in districts bordering rivers. Exposure to waters whose catchments include heavily grazed pastures was associated with conspicuous clusters of Crohn’s disease. The first of these involved a rural com-

munity of about 2000 people in England, in which 12 people developed Crohn’s disease between 1960 and 1983 (Allan et al. 1986). The village, which had its own water supply from local springs, lay in a hol-low surrounded by upland pastures grazed by cattle in which clinical paratuberculosis (Johne’s disease) was evident. A further suspicious cluster of seven cases of Crohn’s disease amongst 285 graduates of the Mankato West High School class of 1980 was reported by Van Kruiningen and Freda (2001). All seven students had been swimming in local ponds and lakes.

A novel study into the diversity of mycobacteria with regard to the physical and chemical charac-teristics of the water in a coastal lagoon was per-formed by Jacobs et al. (2009). The abundance of mycobacteria was high; their presence was detected in 96% of the stations sampled. There was a positive correlation between the number of mycobacteria and elevated temperatures, turbidity, nitrogen and phosphorus components, whereas negative cor-relations existed for the dissolved oxygen content, depth and salinity.

A high hydrophobicity of mycobacteria leads to their enrichment in natural ejected droplets and transfer from water to air (Blanchard and Syzdek 1970). The enrichment factor for transfer of myco-bacteria from water to air ranged from 68 to 15 000 in M. intracellulare (Parker et al. 1983). Obviously, communal water poses a risk. Two case control epidemiological studies carried out independently in the United Kingdom each unexpectedly iden-tified the availability of fixed hot water supplies in the early childhood home as a significant risk factor for the subsequent development of Crohn’s disease (Gent et al. 1994; Duggan et al. 1998). An urban cluster of Crohn’s disease possibly linked to fully treated drinking water has been described by Pierce (2009). Mycobacteria were found in 15% of bottled water in Greece, in 4% of cases at a concen-tration greater than 103 CFU/l (Papapetropoulou et al. 1997).

4. Mycobacteria in soil (Table 4)

M. avium subsp. paratuberculosis present on pastures or barns is the most common non-tuber-culous mycobacteria detected in soil (Eisenberg et al. 2009; Pribylova et al. 2011). Soil is easily con-taminated by fertilisation with manure or liquid dung or by water contaminated by animal faeces.

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Survival of mycobacteria in soil for as long as one year or longer is associated with amoebae or other protozoa or with the shedding of mycobacteria by wild ruminants, wild board, hairs, rabbits and other animals. Mycobacteria from river sediments can be transferred to soil by floods or by the ejection of mi-cro droplets forming aerosols. Any of these transfer mechanisms can explain the finding of M. avium subsp. paratuberculosis in amoebae from fields not used for grazing. (White et al. 2010).

Other M. avium subspecies were studied to de-termine sources of infection for patients (Kaevska et al. 2011). A small number of studies were con-cerned with the detection of M. bovis (Young et al. 2005) or M. leprae in soil. The association was observed between endemicity of leprosy in Africa and India, the distribution of mycobacteria in soil and water with respect to dry or wet season and geographical distribution. The mycobacterial iso-lates from soil were identified as M. fortuitum, whereas the uncultured sequences obtained from soil DNA fell into a few closely related groups, ei-ther M. fortuitum or other fast-growing mycobac-teria, like M. tokaiense, or M. austroafricanum and M. heidelbergense. However, the method described in this study based on the sequencing of a 473 bp region of the 16SrRNA gene, cannot be used to dis-criminate many species that are human and animal pathogens, i.e., M. tuberculosis, M. avium, M. bovis and M. leprae, although sequences belonging to this group were identified (Chilima et al. 2006; Lavania et al. 2008; Turankar et al. 2012).

With regard to mycobacterial diversity in poly-cyclic aromatic hydrocarbon-contaminated soils, investigations have revealed the presence of certain species typical for that environment. Cheung and Kinkle (2001) studied the diversity of mycobacte-ria in petroleum-contaminated soils. 16S rRNA sequences were amplified and subjected to tem-perature gradient gel electrophoresis analysis. All of the sequences belonged to fast-growing myco-bacteria, some of them similar to M. monascense and M. chlorophenolicum. A similar study was con-ducted by Leys et al. (2005). The sequences detect-ed in the contaminated soil belonged to the species M. frederiksbergense, M. austroafricanum, M. pe-troleophilum and M. tusciae. In a study conducted on heavily contaminated soil in Southern Finland, Denaturation gradient gel electrophoresis revealed that 30% of the clone library of the contaminated soil belonged to the genus Mycobacterium (Bjorklof et al. 2009).

5. Mycobacteria in plants (Table 5)

The presence of mycobacteria in plant tissues has been a concern owing to possible transmission to animals and humans (Kazda et al. 2009). The contamination of food of plant origin with myco-bacteria has been reported already several dec-ades ago (Nassal et al. 1974). Mycobacteria were found in fruits and vegetables, such as strawber-ries, radish, cucumbers etc. mainly in edible parts which were close to, or beneath the soil surface. Crucially, mycobacteria were present, although in smaller numbers, even after the washing of the fruits. In the same study, the first experiments demonstrating the presence of bacteria inside fruits were reported. This finding was explained by root uptake of bacteria. In the past couple of decades, the numbers of papers which have con-nected mycobacteria to food contamination and which have recognised its impact on animal and human health have been increasing (Kaevska and Hruska 2010). Mycobacteria were detected also in seven out of 121 vegetable samples which posed a danger to HIV-infected individuals (Yajko et al. 1995). A later study compared the genotypes of M. avium isolates from patients and foods and demonstrated a link between them (Yoder et al. 1999). Mycobacteria (predominantly M. avium) were isolated from 46 samples of salads, leak, let-tuce, mushrooms, and other vegetables as well as apple juice and twenty nine isolates were tested (Argueta et al. 2000). Zwielehner et al. (2008) studied the microbial communities present in the phylosphere of lettuce leaves. After denaturing gradient gel electrophoresis and sequencing anal-yses, sequences from the genus Mycobacterium were found on leaves as well as soil samples. The sequence obtained from conventionally grown let-tuce was most similar to M. alvei. The adoption of the routine use of molecular biology methods, i.e., DNA isolation and PCR/real time PCR rep-resents a major breakthrough in the detection of mycobacteria in general. These techniques enable a considerably more rapid and sensitive detection of mycobacteria, with the possibility of quantifi-cation. Most of the methods developed so far are used for the detection of M. a. paratuberculosis (MAP). Pribylova et al. (2011) detected MAP in grass samples using IS900 real time quantitative PCR. M. avium subsp. paratuberculosis-specific DNA was detected in 13 out of 19 samples exam-ined (approx. 102 copies/g).

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6. Mycobacteria in air (Table 6)

Mycobacteria in air are associated with dust or particles originating from water or soil. Tuberculous mycobacteria can be spread by dried sputum or ex-crements. The transmission of M. tuberculosis and M. bovis in droplets ejected by patients suffering from open forms of pulmonary tuberculosis is a special risk. The time of exposure, quantity and virulence of the pathogen, frequency and intensity of coughing, air exchange rate in the room and many factors related to the endangered person sharing the same room play a role in the dissemination of tuberculosis. It is obvious that species and con-centration of mycobacteria in the air depend on many factors. House dust samples was collected from vacuum cleaners, homogenised by vigorous shaking, and sieved. Mycobacteria were found with both qPCR and traditional culture methods in all 20 dust samples tested. The median cell count was 106 cells/g and the median colony count was 103 CFU/g. Identification of samples was not possi-ble by qPCR, but the species isolated by culture were M. nonchromogenicum, M. kumamotonense, M. ter-rae, M. avium complex and M. gordonae (Torvinen et al. 2010). The contamination of air with mycobac-teria in a peat moss processing plant was assessed by Cayer et al. (2007). A fragment of the 16S rRNA gene was amplified, cloned and sequenced. Forty-nine mycobacterial clones were obtained, most of which were M. intracellulare species. The other detected mycobacteria were M. graecum, M. inter-jectum, M. bohemicum and M. smegmatis. M. avium subsp. paratuberculosis was also detected in dust on dairy farms (Eisenberg et al. 2009).

Mycobacteria in dust do not pose a unique risk of harm for humans and animals. They only supple-ment the other microorganisms, allergens, mites, pesticides and other foreign bodies which may have an adverse effect and are disseminated by means of dust. Appropriate house and street cleaning tech-nology should be thoroughly defined and required. Vacuum cleaners must be tested for efficiency and the sweeping of streets using hand held blowers should be prohibited.

Mycobacteria were detected in the air of a hos-pital therapy pool environment (Angenent et al. 2005). Among the indoor air sequences, there were a total of 77 belonging to mycobacterial rRNA genes. No mycobacteria were detected in the out-side air sample. Perkins et al. (2009) sampled water and aerosol samples from showers in a stem cell

transplantation unit. From the sequences obtained and analysed, the most notable potential pathogen identified was M. mucogenicum.

7. Identification of mycobacteria (Table 7)

Classical culture using solid or liquid media with the identification of colonies using different meth-ods was a standard method for more than 100 years. The sample has to be decontaminated to prevent an overgrow by the other microorganisms. Waiting for results for several weeks or months and the inability to determine the concentrations of mycobacteria in a sample means that the popularity of culture has waned. Nowadays, sophisticated, instrumental ana-lytical methods based on DNA or RNA specificity or determination of specific proteins is preferred. A description of these methods is outside of the scope of this review and the reader is directed elsewhere for this information (Cerqueira et al. 2008; Nayak et al. 2009).

The identification of mycobacteria in environ-mental samples can be achieved using different ap-proaches. In several studies isolated mycobacterial DNA has been subjected to sequencing. Using this method, the mycobacterial diversity in different environments can be assessed. The most commonly used gene, 16S rRNA, has variable and conserved regions within the genus. For sequencing of the genus Mycobacterium, hsp65, dnaJ, or rpoB genes have been used (Mendum et al. 2000; Angenent et al. 2005; Feazel et al. 2009). Next generation sequencing based on pyrosequencing techniques has also been used for the identification of bacte-rial and mycobacterial diversity (Liu et al. 2012; Gomez-Alvarez et al. 2012). The discovery of inser-tion sequences which are specific for certain my-cobacterial species or complexes has been crucial for their direct detection using PCR or real time PCR. IS900 is specific for M. avium subsp. para-tuberculosis and is the most widely used sequence for its detection (Pickup et al. 2005, 2006; Whan et al. 2005; Torvinen et al. 2010). For detection of M. avium subsp. avium and M. avium subsp. homi-nissuis IS901 and IS1245 are used most commonly (Kaevska et al. 2011). For direct detection of M. ul-cerans and M. marinum, PCR and real time PCR system specific for the insertion sequences IS2404 and IS2606 has been described (Fyfe et al. 2007). For total mycobacterial DNA analysis in soil, dif-ferent attempts have been made at DNA isolation.

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Humic acids and other organic material in soil have been the biggest obstacle for extracting microbial DNA due to their inhibitory effects. The diversity of mycobacteria in soil was most often assessed using Denaturation gradient gel electrophoresis or T-RFLP followed by cloning and sequencing (Mendum et al. 2000; Niva et al. 2006; Kopecky et al. 2011).

Many other methods including hybridisation as-says, flow cytometry or MALDI-TOF have been employed for the identification of mycobacteria, though they are so far restricted to bacterial isolates or clinical material. A suitable method should be selected according to the specific aims, the matrix to be analysed, specificity and sensitivity required, accuracy needed, time available for the determina-tion, etc.

Table 1. Search profiles used and numbers of the results retrieved

Web of Science databases = SCI-EXPANDED, SSCI, A&HCI, Lemmatization = On

Timespan All years 2007–2012WaterTopic = (mycobact* AND water AND (drinking OR potable OR tap OR surface OR river OR swimming OR plumbing OR household OR tub))Results 532 231Selected 83Cited in Table 3 55Review articles in Table 2 12SoilTopic = (mycobact* AND soil)Results 803 349Selected 92Cited in Table 4 23Review articles in Table 2 10PlantsTopic = (mycobact* AND (plant* OR vegetable*))Results 1099 612Selected 29Cited in Table 5 12Review articles in Table 2 1AirTopic = (mycobact* AND (air OR aerosol))Results 1136 503Selected 35Cited in Table 6 18Review articles in Table 2 6 Ta

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nts.

A re

view

of

the

case

s of

M. m

ucog

enic

um c

ompl

ex in

fect

ion

in th

e lit

erat

ure

is a

ddre

ssed

her

e, a

s w

ell a

s tw

o ad

ditio

nal c

ases

of t

he

clos

ely

rela

ted

spec

ies

Myc

obac

teri

um a

ubag

nens

e.

Ade

kam

bi

2009

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

630

Met

al w

orki

ng fl

uids

Pote

ntia

l dem

ogra

phic

risk

fact

ors f

or o

utbr

eaks

of r

espi

rato

ry d

isea

se d

ue to

wat

er-b

ased

met

alw

orki

ng fl

uids

(MW

Fs) w

ere

inve

stig

ated

thro

ugh

syst

emat

ic re

view

of p

ublis

hed

outb

reak

inve

stig

atio

ns. S

earc

h te

rms w

ere

sele

cted

by

a m

ultid

isci

plin

ary

team

, ass

iste

d by

an

expe

rien

ced

libra

ry in

form

atio

n se

rvic

e. S

ever

al c

ompu

teri

zed

liter

atur

e da

taba

ses w

ere

sear

ched

for a

rtic

les

publ

ishe

d be

twee

n Ja

nuar

y 19

90 a

nd O

ctob

er 2

011,

rela

ting

to il

l hea

lth o

utbr

eaks

due

to M

WFs

. Pap

ers m

eetin

g th

e se

arch

cri

teri

a w

ere

revi

ewed

in d

etai

l, an

d th

eir r

efer

ence

s che

cked

for a

dditi

onal

art

icle

s. St

udy

desi

gn a

nd d

emog

raph

ic d

etai

ls of

the

outb

reak

w

ere

extr

acte

d fr

om th

e se

lect

ed a

rtic

les a

nd e

nter

ed in

to st

anda

rdiz

ed e

vide

nce

tabl

es. Th

irty

-five

art

icle

s rel

atin

g to

inve

stig

atio

ns

of 2

7 ou

tbre

aks o

f res

pira

tory

ill h

ealth

att

ribu

ted

to M

WF

expo

sure

wer

e id

entifi

ed. Th

e m

ajor

ity o

f rep

orts

wer

e ca

se se

ries

of d

is-ea

se o

r obs

erva

tiona

l cro

ss-s

ectio

nal s

tudi

es o

f sym

ptom

s and

hyg

iene

mea

sure

men

ts. E

ight

of t

he o

utbr

eak

inve

stig

atio

ns in

clud

ed

an e

lem

ent o

f cas

e-co

ntro

l ana

lysi

s. M

ost o

utbr

eaks

wer

e fr

om th

e U

SA, h

ad o

ccur

red

in la

rge

car-

or a

eron

autic

al-m

anuf

actu

ring

pl

ants

, and

wer

e as

soci

ated

with

the

use

of c

entr

al sh

ared

sum

ps. H

ygie

ne st

udie

s hav

e no

t dem

onst

rate

d co

nsis

tent

risk

fact

ors f

or

resp

irato

ry o

utbr

eaks

, in

term

s of t

he ty

pe o

f MW

F ut

ilize

d, d

egre

e of

mic

robi

al c

onta

min

atio

n, o

r lev

els o

f per

sona

l exp

osur

e. S

ix

stud

ies w

ere

iden

tified

that

foun

d w

orke

rs w

ith M

WF

expo

sure

dur

ing

outb

reak

s wer

e m

ore

likel

y to

repo

rt re

spira

tory

or s

yste

mic

sy

mpt

oms t

han

unex

pose

d co

ntro

l wor

kers

. Six

cas

e-co

ntro

l ana

lyse

s wer

e al

so id

entifi

ed th

at fo

und

wor

kers

with

ext

rins

ic a

llerg

ic

alve

oliti

s (EA

A) w

ere

mor

e lik

ely

to d

emon

stra

te c

erta

in im

mun

e re

spon

ses t

o m

icro

bial

con

tam

inan

ts a

nd/o

r use

d M

WFs

than

w

orke

rs w

ithou

t EA

A. D

espi

te a

num

ber o

f det

aile

d w

orkp

lace

and

imm

unol

ogic

al st

udie

s of a

sthm

a an

d al

veol

itis o

utbr

eaks

in

MW

F-ex

pose

d w

orkf

orce

s, ou

r und

erst

andi

ng o

f the

ir ae

tiolo

gy re

mai

ns li

mite

d.

Burt

on e

t al

. 201

2

Fluo

resc

ence

in

sit

u hy

brid

izat

ion,

pe

ptid

e nu

clei

c ac

ids,

lo

cked

nuc

leic

aci

ds

Fluo

resc

ence

in s

itu h

ybri

diza

tion

(FIS

H) i

s a

wel

l-es

tabl

ishe

d te

chni

que

that

is u

sed

for

a va

riet

y of

pur

pose

s, r

angi

ng fr

om

path

ogen

det

ectio

n in

clin

ical

dia

gnos

tics

to th

e de

term

inat

ion

of c

hrom

osom

al s

tabi

lity

in s

tem

cel

l res

earc

h. T

he k

ey s

tep

of F

ISH

invo

lves

the

dete

ctio

n of

a n

ucle

ic a

cid

regi

on a

nd a

s su

ch, D

NA

mol

ecul

es h

ave

typi

cally

bee

n us

ed to

pro

be fo

r th

e se

quen

ces

of in

tere

st. H

owev

er, s

ince

the

turn

of t

he c

entu

ry, a

n in

crea

sing

num

ber

of la

bora

tori

es h

ave

star

ted

to m

ove

on

to th

e m

ore

robu

st D

NA

mim

ics

met

hods

, mos

t not

ably

pep

tide

and

lock

ed n

ucle

ic a

cids

(PN

A a

nd L

NA

). In

this

revi

ew, w

e w

ill c

over

the

stat

e-of

-the

-art

of t

he d

iffer

ent D

NA

mim

ics

in re

gard

to th

eir

appl

icat

ion

as e

ffic

ient

mar

kers

for

the

pres

ence

of

indi

vidu

al m

icro

bial

cel

ls, a

nd c

onsi

der

thei

r po

tent

ial a

dvan

tage

s an

d pi

tfal

ls. A

vaila

ble

PNA

pro

bes

are

then

reas

sess

ed

in te

rms

of s

ensi

tivity

and

spe

cific

ity u

sing

rRN

A d

atab

ases

. In

addi

tion,

we

also

att

empt

to p

redi

ct th

e ap

plic

abili

ty o

f DN

A

mim

ics

in w

ell-

know

n te

chni

ques

att

empt

ing

to d

etec

t in

situ

low

num

ber

of c

opie

s of

spe

cific

nuc

leic

aci

d se

quen

ces

such

as

cata

lyze

d re

port

er d

epos

ition

(CA

RD) a

nd re

cogn

ition

of i

ndiv

idua

l gen

es (R

ING

) FIS

H.

Cer

quei

ra

et a

l. 20

08

Envi

ronm

enta

l m

ycob

acte

ria

Alth

ough

the

envi

ronm

enta

l myc

obac

teria

are

slow

gro

win

g re

lativ

e to

oth

er m

icro

orga

nism

s in

wat

er a

nd so

il w

hich

wou

ld su

g-ge

st th

at th

ey a

re p

oor c

ompe

titor

s, co

mpe

nsat

ing

fact

ors p

erm

it su

rviv

al, g

row

th a

nd p

ersis

tenc

e in

nat

ural

and

hum

an-e

ngin

eere

d en

viro

nmen

ts. F

acto

rs su

ch a

s the

hyd

roph

obic

, lip

id-r

ich

impe

rmea

ble

enve

lope

, bio

film

form

atio

n, a

cid

resis

tanc

e, a

naer

obic

surv

ival

an

d m

etab

olism

of r

ecal

citr

ant c

arbo

n co

mpo

unds

per

mit

surv

ival

and

gro

wth

of t

he e

nviro

nmen

tal m

ycob

acte

ria in

a w

ide

rang

e of

na

tura

l and

hum

an-e

ngin

eere

d ha

bita

ts. H

igh

num

bers

of e

nviro

nmen

tal m

ycob

acte

ria a

re fo

und

in c

oast

al sw

amps

and

est

uarie

s and

bo

real

, pea

t-ric

h fo

rest

soils

and

wat

ers.

The

hydr

opho

bic

surf

ace

resu

lts in

con

cent

ratio

n of

the

envi

ronm

enta

l myc

obac

teria

at i

nter

-fa

ces (

air-

wat

er a

nd su

rfac

e-w

ater

) and

in a

eros

oliz

ed d

ropl

ets e

ject

ed fr

om w

ater

. The

surv

ival

and

gro

wth

in p

roto

zoa

and

amoe

bae

perm

it en

viro

nmen

tal m

ycob

acte

ria to

per

sist i

n ha

bita

ts su

bjec

t to

pred

atio

n an

d lik

ely

led

to su

rviv

al a

nd g

row

th in

pha

gocy

tic c

ells

of a

nim

als.

Fina

lly, s

low

gro

wth

allo

ws t

ime

for m

ycob

acte

rial c

ells

to a

dapt

to c

hang

ing

cond

ition

s bef

ore

loss

of v

iabi

lity.

Falk

in-

ham

20

09b

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

631

Ecol

ogy

of m

ycob

acte

ria

A m

ajor

ity o

f the

Myc

obac

teri

um sp

ecie

s, c

alle

d th

e no

ntub

ercu

lous

myc

obac

teri

a (N

TM

), ar

e na

tura

l inh

abita

nts

of n

atur

al

wat

ers,

eng

inee

red

wat

er s

yste

ms,

and

soi

ls. A

s a

cons

eque

nce

of th

eir

ubiq

uito

us d

istr

ibut

ion,

hum

ans

are

surr

ound

ed b

y th

ese

oppo

rtun

istic

pat

hoge

ns. A

car

dina

l fea

ture

of m

ycob

acte

rial

cel

ls is

the

pres

ence

of a

hyd

roph

obic

, lip

id-r

ich

oute

r m

embr

ane.

The

hyd

roph

obic

ity o

f NT

M is

a m

ajor

det

erm

inan

t of a

eros

oliz

atio

n, s

urfa

ce a

dher

ence

, bio

film

-for

mat

ion,

an

d di

sinf

ecta

nt-a

nd a

ntib

iotic

resi

stan

ce. T

he N

TM

are

olig

otro

phs,

abl

e to

gro

w a

t low

car

bon

leve

ls [>

50

µg a

ssim

ila-

ble

orga

nic

carb

on (A

OC

) l-1

)], m

akin

g th

em e

ffect

ive

com

petit

ors

in lo

w n

utri

ent,

and

disi

nfec

ted

envi

ronm

ents

(dri

nkin

g w

ater

). Bi

ofilm

form

atio

n an

d ol

igot

roph

y le

ad to

sur

viva

l, pe

rsis

tenc

e, a

nd g

row

th in

dri

nkin

g w

ater

dis

trib

utio

n sy

stem

s. In

ad

ditio

n to

thei

r ro

le a

s hu

man

and

ani

mal

pat

hoge

ns, t

he w

ides

prea

d di

stri

butio

n of

NT

M in

the

envi

ronm

ent,

coup

led

with

th

eir

abili

ty to

deg

rade

and

met

abol

ize

a va

riet

y of

com

plex

hyd

roca

rbon

s in

clud

ing

pollu

tant

s, s

ugge

sts

that

NT

M m

ay b

e ag

ents

of n

utri

ent c

yclin

g.

Falk

in-

ham

20

09a

Wat

er a

nd m

ycob

acte

ria

Non

tube

rcul

ous

myc

obac

teri

a (N

TM

) are

env

iron

men

tal o

ppor

tuni

stic

pat

hoge

ns o

f hum

ans

and

anim

als.

The

y ar

e fo

und

in

a w

ide

vari

ety

of h

abita

ts to

whi

ch h

uman

s ar

e ex

pose

d, in

clud

ing

drin

king

wat

er d

istr

ibut

ion

syst

ems

and

hous

ehol

d w

ater

an

d pl

umbi

ng. I

n th

at re

gard

, the

y ar

e di

stin

ct fr

om th

eir

oblig

ate

path

ogen

ic re

lativ

es, t

he m

embe

rs o

f the

Myc

obac

teri

um

tube

rcul

osis

com

plex

. Ow

ing

to th

e pr

esen

ce o

f NT

M in

the

hum

an e

nvir

onm

ent,

hum

an a

ctiv

ities

hav

e ha

d di

rect

impa

cts

on th

eir

ecol

ogy

and

ther

eby

thei

r ep

idem

iolo

gy. N

TM

are

olig

otro

phic

, abl

e to

gro

w a

t low

org

anic

mat

ter

conc

entr

atio

ns

and

over

a w

ide

rang

e of

tem

pera

ture

s, a

nd e

ven

at lo

w o

xyge

n co

ncen

trat

ions

. Thu

s, N

TM

are

nor

mal

inha

bita

nts

of n

atu-

ral w

ater

s an

d dr

inki

ng w

ater

s. D

isco

very

of t

he p

rese

nce

of N

TM

-pol

lute

d so

ils is

not

sur

pris

ing

in li

ght o

f the

abi

lity

of

NT

M to

deg

rade

a v

arie

ty o

f hyd

roca

rbon

pol

luta

nts.

A m

ajor

hum

an a

ctiv

ity s

elec

ting

for

the

grow

th a

nd p

redo

min

ance

of

myc

obac

teri

a in

hab

itats

is d

isin

fect

ion.

In c

ompa

riso

n to

oth

er b

acte

ria,

NT

M a

re d

isin

fect

ant,

heav

y m

etal

and

ant

ibio

tic

resi

stan

t. T

here

fore

, the

use

of a

ny a

ntim

icro

bial

age

nt s

elec

ts fo

r m

ycob

acte

ria.

Use

of d

isin

fect

ant i

n dr

inki

ng w

ater

trea

t-m

ent s

elec

ts fo

r m

ycob

acte

ria

that

can

gro

w a

nd c

ome

to p

rolif

erat

e in

dri

nkin

g w

ater

dis

trib

utio

n sy

stem

s in

the

abse

nce

of

disi

nfec

tant

-sen

sitiv

e co

mpe

ting

mic

roor

gani

sms.

NT

M s

elec

tion

may

als

o oc

cur

as a

con

sequ

ence

of a

ntib

iotic

s in

dri

nkin

g w

ater

sou

rces

.

Falk

in-

ham

201

0

Rap

idly

gro

win

g

myc

obac

teri

a in

fect

ion

and

trea

tmen

t

Rapi

dly

grow

ing

myc

obac

teri

a (R

GM

) are

ubi

quito

us in

nat

ure

and

wid

ely

dist

ribu

ted

in w

ater

, soi

l and

ani

mal

s. D

urin

g th

e pa

st

thre

e de

cade

s we

have

obs

erve

d a

nota

ble

incr

emen

t of i

nfec

tions

cau

sed

by R

GM

, bot

h lo

caliz

ed a

nd d

isse

min

ated

, as w

ell a

s nos

o-co

mia

l out

brea

ks o

f con

tam

inat

ed m

edic

al e

quip

men

t. Th

e m

icro

biol

ogic

al d

iagn

osis

of R

GM

infe

ctio

ns in

clud

es d

irect

mic

rosc

opic

ob

serv

atio

n an

d cu

lture

. The

taxo

nom

ic id

entifi

catio

n is

per

form

ed b

y ph

enot

ypic

, bio

chem

ical

, chr

omat

ogra

phic

and

mol

ecul

ar

biol

ogy

tech

niqu

es. Th

e tr

eatm

ent d

iffer

s fro

m th

at o

f oth

er m

ycob

acte

rios

is li

ke tu

berc

ulos

is, o

win

g to

the

vari

able

in v

itro

susc

ep-

tibili

ty o

f the

spec

ies o

f thi

s gro

up. Th

e RG

M a

re re

sist

ant t

o co

nven

tiona

l ant

itube

rcul

ous d

rugs

, but

can

be

susc

eptib

le to

bro

ad

spec

trum

ant

imic

robi

al a

gent

s. In

this

stud

y w

e co

mm

ent o

n th

e si

gnifi

cant

asp

ects

of h

uman

infe

ctio

ns b

y RG

M, i

nclu

ding

thei

r bi

olog

y, ep

idem

iolo

gy, p

atho

logy

, mic

robi

olog

ical

dia

gnos

is, t

axon

omic

iden

tifica

tion,

ant

imic

robi

al su

scep

tibili

ty a

nd tr

eatm

ent.

Gar

cia-

Mar

tos

and

Gar

cia-

Agu

do

2012

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

632

Wat

er, m

ilk a

nd m

eat

M. a

. par

atub

ercu

losi

sM

ycob

acte

rium

avi

um su

bsp.

par

atub

ercu

losis

(Map

) is t

he c

ause

of J

ohne

’s di

seas

e, a

chr

onic

infe

ctio

n of

the

gut,

in ru

min

ant

anim

als t

hat p

rovi

de m

ilk a

nd/o

r mea

t for

hum

an c

onsu

mpt

ion.

Map

als

o m

ay b

e in

volv

ed in

Cro

hn’s

dise

ase

and

type

I di

abet

es in

hu

man

s. A

lthou

gh th

e ro

le o

f Map

in h

uman

dis

ease

s has

not

bee

n es

tabl

ishe

d, m

inim

izin

g th

e ex

posu

re o

f hum

ans t

o th

e or

gan-

ism

is c

onsi

dere

d de

sira

ble

as a

pre

caut

iona

ry m

easu

re. I

nfec

ted

anim

als c

an sh

ed M

ap in

fece

s and

milk

, and

the

orga

nism

can

be

com

e di

ssem

inat

ed in

tiss

ues r

emot

e fr

om th

e gu

t and

its a

ssoc

iate

d ly

mph

nod

es. Th

e pr

esen

ce o

f at l

east

som

e M

ap in

raw

milk

an

d m

eat a

nd in

nat

ural

wat

ers i

s lik

ely,

but t

he n

umbe

rs o

f Map

in th

ose

food

s and

wat

ers s

houl

d be

redu

ced

thro

ugh

cook

ing

or

puri

ficat

ion.

The

avai

labl

e in

form

atio

n re

latin

g to

Map

in m

ilk a

nd d

airy

pro

duct

s, m

eats

, and

dri

nkin

g w

ater

is re

view

ed h

ere

for

asse

ssm

ent o

f the

risk

s of e

xpos

ure

to M

ap fr

om c

onsu

mpt

ion

of su

ch fo

ods a

nd w

ater

.

Gill

et a

l. 20

11

Wat

er, f

ood

and

feed

Pape

rs o

n m

ycob

acte

ria

in fo

od, f

eed

and

wat

er, p

ublis

hed

betw

een

1945

and

201

0 an

d in

dexe

d in

the

data

base

Web

of S

cien

ce (R

) (Th

omso

n Re

uter

s) w

ere

rank

ed a

ccor

ding

to a

utho

rs, i

nstit

utio

ns, c

ount

ries

and

sour

ce ti

tles.

The

tota

l num

ber o

f pap

ers o

n m

yco-

bact

eria

and

food

and

myc

obac

teri

a an

d w

ater

wer

e 14

86 a

nd 1

419,

resp

ectiv

ely.

Mor

e th

an 4

0% o

f pap

ers h

ave

been

pub

lishe

d in

th

e la

st fi

ve y

ears

. In

addi

tion

to p

ublic

atio

ns in

pee

r rev

iew

ed jo

urna

ls th

e ar

chiv

es o

f Pro

MED

-mai

l and

the

Rapi

d A

lert

Sys

tem

for

Food

and

Fee

d of

the

Euro

pean

Uni

on w

ere

also

sear

ched

. It i

s evi

dent

that

muc

h at

tent

ion

is b

eing

pai

d to

myc

obac

teri

a in

food

, fe

ed a

nd w

ater

as t

hey

likel

y po

se a

pub

lic h

ealth

risk

.

Kae

vska

an

d H

rusk

a 20

10

Pulm

onar

y di

seas

esM

. avi

um c

ompl

exM

ycob

acte

rium

avi

um c

ompl

ex (M

AC

) con

sist

s of n

ontu

berc

ulou

s myc

obac

teri

a th

at c

ause

dis

ease

in im

mun

ocom

prom

ised

and

im

mun

ocom

pete

nt h

osts

. The

orga

nism

s are

ubi

quito

us in

the

envi

ronm

ent,

and

acqu

isiti

on o

ccur

s thr

ough

inge

stio

n or

inha

latio

n of

aer

osol

s fro

m so

il, w

ater

, or b

iofil

ms.

Dis

ease

may

man

ifest

as d

isse

min

ated

infe

ctio

n, so

ft tis

sue

infe

ctio

n, c

hron

ic p

neum

onia

, or

hyp

erse

nsiti

vity

pne

umon

itis.

Non

tube

rcul

ous m

ycob

acte

ria

are

incr

easi

ngly

ass

ocia

ted

with

pul

mon

ary

dise

ase,

with

MA

C

bein

g th

e m

ost c

omm

on n

ontu

berc

ulou

s myc

obac

teri

a to

cau

se p

ulm

onar

y di

seas

e in

the

Uni

ted

Stat

es. P

ulm

onar

y sy

mpt

oms,

nodu

lar o

r cav

itary

opa

citie

s on

a ch

est r

adio

grap

h or

hig

h-re

solu

tion

com

pute

d to

mog

raph

ic sc

an w

ith m

ultif

ocal

bro

nchi

ecta

sis

and

mul

tiple

smal

l nod

ules

, plu

s pos

itive

cul

ture

resu

lts fr

om tw

o sp

utum

spec

imen

s or o

ne b

ronc

hosc

opic

spec

imen

are

con

sist

ent

with

MA

C p

ulm

onar

y di

seas

e. T

reat

men

t con

sist

s of a

mac

rolid

e, ri

fam

ycin

, and

eth

ambu

tol g

iven

thre

e tim

es w

eekl

y fo

r non

cavi

-ta

ry d

isea

se a

nd d

aily

with

or w

ithou

t an

amin

ogly

cosi

de fo

r cav

itary

dis

ease

.

Kas

per-

baue

r and

D

aley

20

08

Lung

dis

ease

cau

sed

by

myc

obac

teri

aN

ontu

berc

ulou

s myc

obac

teri

a (N

TM

) are

resi

lient

bac

teri

a th

at g

row

in v

irtu

ally

any

env

ironm

ent,

espe

cial

ly th

ose

whe

re c

ompe

t-in

g m

icro

orga

nism

s are

des

troy

ed, s

uch

as in

chl

orin

ated

wat

er. Th

ey h

ave

been

dis

cove

red

in so

il, d

ust,

food

, wat

er, a

nd d

omes

tic

and

wild

ani

mal

s. N

ontu

berc

ulou

s myc

obac

teri

a te

nd to

infe

ct in

divi

dual

s with

loca

l (e.

g., d

amag

ed sk

in o

r lun

g) o

r sys

tem

ic (e

.g.,

HIV

, dru

gs, m

alig

nanc

y) d

efec

ts in

hos

t def

ence

, and

thei

r inc

iden

ce a

nd p

reva

lenc

e ha

ve c

onsi

sten

tly in

crea

sed

in th

e la

st d

ecad

e.

Diffi

culty

may

ari

se in

det

erm

inin

g w

heth

er a

n is

olat

ed N

TM

from

a m

icro

biol

ogic

al sa

mpl

e is

in fa

ct a

con

tam

inan

t or a

pat

ho-

geni

c or

gani

sm. I

n th

is re

view

, we

disc

uss t

he im

port

ant m

ycob

acte

ria

invo

lved

in lu

ng d

isea

se, f

acto

rs th

at p

redi

spos

e in

divi

dual

s to

infe

ctio

n, a

nd th

eir d

iagn

osis

and

trea

tmen

t acc

ordi

ng to

upd

ated

gui

delin

es. E

nglis

h la

ngua

ge p

ublic

atio

ns in

MED

LIN

E an

d re

fere

nces

from

rele

vant

art

icle

s fro

m Ja

nuar

y 1,

199

0 to

June

28,

200

9 w

ere

revi

ewed

. Key

wor

ds se

arch

ed w

ere

“non

tube

rcul

ous”,

“m

ycob

acte

ria”,

“dia

gnos

is”, a

nd “t

reat

men

t”.

McG

rath

et

al.

2010

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

633

Con

tact

trac

ing

in p

ublic

tr

ansp

ort

Whi

le g

uide

lines

on

cont

act t

raci

ng (C

T) a

fter

exp

osur

e to

cer

tain

infe

ctio

us p

atho

gens

dur

ing

air

trav

el e

xist

, no

guid

ance

do

cum

ents

are

ava

ilabl

e on

CT

in re

spon

se to

pot

entia

l exp

osur

e on

pub

lic g

roun

d tr

ansp

ort.

We

revi

ewed

sci

entif

ic a

nd

non-

scie

ntifi

c lit

erat

ure

on tr

ansm

issi

on o

f air

born

e pa

thog

ens

in p

ublic

gro

und

tran

spor

t and

on

fact

ors

pote

ntia

lly in

flu-

enci

ng tr

ansm

issi

on. W

e id

entif

ied

32 re

leva

nt p

ublic

atio

ns (1

5 sc

ient

ific

and

17 n

on-s

cien

tific

). M

ost o

f the

sel

ecte

d st

udie

s de

alt w

ith tr

ansm

issi

on o

f tub

ercu

losi

s. H

owev

er, t

he re

latio

n be

twee

n tr

avel

dur

atio

n, p

roxi

mity

to th

e in

dex

case

and

env

i-ro

nmen

tal f

acto

rs, s

uch

as v

entil

atio

n, o

n di

seas

e tr

ansm

issi

on in

pub

lic g

roun

d tr

ansp

ort i

s po

orly

und

erst

ood.

Con

side

ring

th

e di

ffic

ulty

and

pro

babl

y lim

ited

effe

ctiv

enes

s of

CT

in g

roun

d tr

ansp

ort,

our

resu

lts s

ugge

st th

at o

nly

exce

ptio

nal c

ircu

m-

stan

ces

wou

ld ju

stify

CT.

Thi

s co

ntra

sts

with

the

high

leve

l of a

tten

tion

CT

in a

ir tr

avel

see

ms

to re

ceiv

e in

inte

rnat

iona

l re

gula

tions

and

reco

mm

enda

tions

. We

ques

tion

whe

ther

the

indi

catio

n fo

r C

T s

houl

d be

revi

site

d af

ter

a ri

sk-b

enef

it as

sess

-m

ent t

hat t

akes

into

acc

ount

exp

osur

e in

bot

h gr

ound

and

air

tran

spor

t.

Moh

r et

al. 2

012

Det

ecti

on o

f mic

roor

gan-

ism

s us

ing

bios

enso

rsA

long

with

use

ful m

icro

orga

nism

s, th

ere

are

som

e th

at c

ause

pot

entia

l dam

age

to th

e an

imal

s an

d pl

ants

. Det

ectio

n an

d id

entif

icat

ion

of th

ese

harm

ful o

rgan

ism

s in

a c

ost a

nd ti

me

effe

ctiv

e w

ay is

a c

halle

nge

for

the

rese

arch

ers.

The

futu

re o

f de

tect

ion

met

hods

for

mic

roor

gani

sms

shal

l be

guid

ed b

y bi

osen

sor,

whi

ch h

as a

lrea

dy c

ontr

ibut

ed e

norm

ousl

y in

sen

sing

an

d de

tect

ion

tech

nolo

gy. H

ere,

we

aim

to re

view

the

use

of v

ario

us b

iose

nsor

s, d

evel

oped

by

inte

grat

ing

the

biol

ogic

al a

nd

phys

icoc

hem

ical

/mec

hani

cal p

rope

rtie

s (o

f tra

nduc

ers)

, whi

ch c

an h

ave

enor

mou

s im

plic

atio

n in

hea

lthca

re, f

ood,

agr

icul

ture

an

d bi

odef

ence

. We

have

als

o hi

ghlig

hted

the

way

s to

impr

ove

the

func

tioni

ng o

f the

bio

sens

or.

Nay

ak e

t al

. 200

9

Free

-liv

ing

amoe

bae

Myc

obac

teri

um s

peci

es e

volv

ed fr

om a

n en

viro

nmen

tal r

ecen

t com

mon

anc

esto

r by

redu

ctiv

e ev

olut

ion

and

late

ral g

ene

tran

sfer

. Str

ateg

ies

sele

cted

thro

ugh

evol

utio

n an

d de

velo

ped

by m

ycob

acte

ria

resu

lted

in re

sist

ance

to p

reda

tion

by e

nvi-

ronm

enta

l uni

cellu

lar

prot

ists

, inc

ludi

ng fr

ee-l

ivin

g am

oeba

e. In

deed

, myc

obac

teri

a ar

e is

olat

ed fr

om th

e sa

me

soil

and

wat

er e

nvir

onm

ents

as

are

amoe

bae,

and

exp

erim

enta

l mod

els

usin

g A

cant

ham

oeba

spp

. and

Dic

tyos

teliu

m d

isco

ideu

m w

ere

expl

oite

d to

ana

lyse

the

mec

hani

sms

for

intr

acel

lula

r su

rviv

al. M

ost o

f the

se m

echa

nism

s ha

ve b

een

furt

her

repr

oduc

ed in

m

acro

phag

es fo

r m

ycob

acte

ria

rega

rded

as

oppo

rtun

istic

and

obl

igat

e pa

thog

ens.

Am

oeba

l cys

ts m

ay p

rote

ct in

trac

ellu

lar

myc

obac

teri

a ag

ains

t adv

erse

con

ditio

ns a

nd m

ay a

ct a

s a

vect

or fo

r m

ycob

acte

ria.

The

latt

er h

ypot

hesi

s w

arra

nts

furt

her

envi

ronm

enta

l and

clin

ical

stu

dies

to b

ette

r as

sess

the

role

of f

ree-

livin

g am

oeba

e in

the

epid

emio

logy

of i

nfec

tions

cau

sed

by

myc

obac

teri

a.

Sala

h et

al

. 200

9

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

634

Free

-liv

ing

amoe

bae

Aca

ntha

moe

bae

are

free

-liv

ing

amoe

bae

dist

ribu

ted

wor

ldw

ide.

The

y ar

e am

ong

the

mos

t pre

vale

nt p

roto

zoa

foun

d in

the

envi

ronm

ent,

and

have

bee

n is

olat

ed fr

om a

wid

e va

riet

y of

pub

lic w

ater

sup

plie

s, s

wim

min

g po

ols,

bot

tled

wat

er, v

entil

atio

n du

cts,

soi

l, ai

r, su

rgic

al in

stru

men

ts, c

onta

ct le

nses

, den

tal t

reat

men

t uni

ts a

nd h

ospi

tals

. Aca

ntha

moe

bae

feed

on

bact

eria

by

pha

gocy

tosi

s, b

ut s

ome

bact

eria

are

abl

e to

sur

vive

and

som

etim

es m

ultip

ly in

the

host

, res

ultin

g in

new

pro

pert

ies

of

the

bact

eria

. The

intr

acel

lula

r gr

owth

of b

acte

ria

has

been

ass

ocia

ted

with

enh

ance

d en

viro

nmen

tal s

urvi

val o

f the

bac

teri

a,

incr

ease

d vi

rule

nce

and

incr

ease

d re

sist

ance

aga

inst

ant

ibio

tic s

ubst

ance

s. T

he a

dvan

tage

of u

tilis

ing

free

-liv

ing

amoe

bae

is

that

rese

arch

can

be

carr

ied

out o

n no

n-m

amm

alia

n ce

lls a

s a

mod

el b

ased

on

natu

ral r

ealit

y to

stu

dy b

acte

rial

vir

ulen

ce a

nd

path

ogen

icity

. Am

oeba

e ar

e ea

sy to

han

dle

expe

rim

enta

lly c

ompa

red

with

mam

mal

ian

cells

and

allo

w s

tudi

es o

n ho

st fa

ctor

s fo

r ho

st-p

aras

ite in

tera

ctio

ns. B

acte

ria

are

easi

ly m

anip

ulat

ed g

enet

ical

ly, w

hich

cre

ates

the

poss

ibili

ty o

f res

earc

h on

mut

ants

to

stu

dy b

acte

ria-

host

inte

ract

ions

. Thu

s ut

ilisi

ng th

is n

on-m

amm

alia

n m

odel

can

resu

lt in

bet

ter

unde

rsta

ndin

g of

inte

rac-

tions

bet

wee

n pr

okar

yotic

and

euk

aryo

tic c

ells

and

ass

ist i

n th

e de

velo

pmen

t of n

ew th

erap

eutic

age

nts

to re

cogn

ise

and

trea

t in

fect

ions

.

Sand

-st

rom

et

al. 2

011

Rap

idly

gro

win

g m

yco-

bact

eria

The

pat

hoge

nic

pote

ntia

l of t

he r

apid

ly g

row

ing

myc

obac

teri

a (R

GM

) has

sta

rted

bei

ng re

cogn

ized

. Thi

s is

due

to m

ore

sens

i-tiv

e an

d sp

ecifi

c te

chni

ques

in th

e la

bora

tory

. The

RG

M a

re g

ener

ally

def

ined

as

nont

uber

culo

us s

peci

es o

f myc

obac

teri

a th

at

show

vis

ible

gro

wth

on

agar

med

ia w

ithin

7 d

ays.

RG

M a

re w

idel

y di

stri

bute

d in

nat

ure

and

have

bee

n is

olat

ed fr

om n

atur

al

wat

er, t

ap w

ater

, and

soi

l. Se

vera

l bio

chem

ical

test

s, h

igh

perf

orm

ance

liqu

id c

hrom

atog

raph

y, a

nd m

olec

ular

tech

niqu

es h

ave

been

dev

elop

ed fo

r ra

pid

iden

tific

atio

n of

thes

e sp

ecie

s. T

he A

mer

ican

Tho

raci

c So

ciet

y an

d th

e In

fect

ious

Dis

ease

Soc

iety

of

Am

eric

a re

com

men

d th

at R

GM

sho

uld

be id

entif

ied

to th

e sp

ecie

s le

vel u

sing

a re

cogn

ized

acc

epta

ble

met

hodo

logy

suc

h as

pol

ymer

ase

chai

n re

actio

n re

stri

ctio

n en

zym

e an

alys

is o

r bi

oche

mic

al te

stin

g an

d ro

utin

e su

scep

tibili

ty te

stin

g of

RG

M

shou

ld in

clud

e am

ikac

in, i

mip

enem

, dox

ycyc

line,

the

fluor

inat

ed q

uino

lone

s, a

sul

phon

amid

e or

trim

etho

prim

-sul

pham

eth-

oxaz

ole,

cef

oxiti

n, c

lari

thro

myc

in, l

inez

olid

, and

tobr

amyc

in. T

he d

isea

ses

caus

ed b

y th

ese

orga

nism

s ha

ve v

arie

d m

anife

sta-

tions

. The

y ha

ve b

een

resp

onsi

ble

for

a nu

mbe

r of

hea

lthca

re-a

ssoc

iate

d ou

tbre

aks

and

pseu

do-o

utbr

eaks

. For

reco

gniti

on o

f ou

tbre

aks,

it is

impo

rtan

t to

be fa

mili

ar w

ith th

e ca

usat

ive

orga

nism

s lik

e RG

M w

hich

are

mos

t fre

quen

tly in

volv

ed in

hea

lth-

care

-ass

ocia

ted

outb

reak

s an

d ps

eudo

out

brea

ks. I

t is

esse

ntia

l to

inte

rven

e as

soo

n as

pos

sibl

e to

inte

rrup

t thi

s tr

ansm

issi

on.

Larg

e ga

ps s

till e

xist

in o

ur k

now

ledg

e of

RG

M. U

nque

stio

nabl

y m

ore

stud

ies

are

requ

ired

. Thr

ough

this

revi

ew, w

e w

ish

to

emph

asiz

e th

at re

port

ing

of R

GM

from

clin

ical

set

tings

alo

ng w

ith th

eir

sens

itivi

ty p

atte

rns

is a

n ab

solu

te n

eed

of th

e ho

ur.

Set a

nd

Shas

tri

2011

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

635

Food

bor

ne p

atho

gens

The

em

erge

nce

of p

atho

gens

is th

e re

sult

of a

num

ber

of im

pact

in a

ll pa

rts

of th

e fo

od c

hain

. The

em

ergi

ng te

chno

logi

es in

fo

od p

rodu

ctio

n ex

plai

n ho

w n

ew p

atho

gens

can

est

ablis

h th

emse

lves

in th

e fo

od c

hain

and

com

prom

ise

food

saf

ety.

The

im

pact

of t

he fo

od te

chno

logy

is a

naly

sed

for s

ever

al b

acte

ria,

such

as Y

ersin

ia, C

ampy

loba

cter

, Arc

obac

ter,

Hel

icob

acte

r pul

lo-

rum

, Ent

erob

acte

r sak

azak

ii, M

ycob

acte

rium

avi

um sp

p. p

arat

uber

culo

sis,

prio

ns re

late

d to

vC

JD a

nd o

ther

s. Th

e im

port

ance

of

the

abili

ty o

f man

y m

icro

bes t

o fo

rm V

BNC

form

s is e

labo

rate

d on

. Res

earc

h on

cul

ture

inde

pend

ent m

etho

ds m

ay a

ddre

ss th

is

outs

tand

ing

issu

e to

the

bett

er u

nder

stan

ding

of e

mer

ging

pat

hoge

ns. Th

e “d

emer

ging

” of p

atho

gens

als

o oc

cur,

and

exam

ples

of

this

are

exp

lain

ed. Th

e re

actio

n of

bac

teri

a to

stre

sses

and

subl

etha

l tre

atm

ents

, and

how

exp

osur

e to

one

stre

ss fa

ctor

can

con

fer

resi

stan

ce to

oth

er st

ress

es, l

itera

lly sp

eaki

ng c

ausi

ng c

onta

giou

s res

ista

nce,

are

exp

lain

ed. Th

e im

plic

atio

n of

this

e.g

. in

mod

em

appr

oach

es o

f foo

d pr

eser

vatio

n, su

ch a

s Min

imal

ly p

roce

ssed

Foo

ds, i

s con

side

rabl

e. In

test

inal

col

oniz

atio

n of

EH

EC m

ay b

e re

gula

ted

by Q

uoru

m se

nsin

g, a

nd th

is a

bilit

y of

mic

robe

s pla

ys a

n im

port

ant r

ole

in th

e co

loni

zatio

n of

mic

robe

s in

food

and

on

food

pro

cess

ing

equi

pmen

t, an

impo

rtan

t fac

tor i

n th

e em

erge

nce

of p

atho

gens

. The

emer

genc

e of

Sac

char

omyc

es c

erev

isia

e,

as a

n op

port

unis

tic h

uman

pat

hoge

n, u

sed

for c

entu

ries

for f

ood

and

prod

uctio

n of

alc

ohol

ic b

ever

ages

, cal

ls fo

r res

earc

h in

m

olec

ular

tool

s to

dist

ingu

ish

betw

een

prob

iotic

and

clin

ical

stra

ins.

Cyc

losp

ora

caye

tane

nsis

and

Nor

ovir

us o

utbr

eaks

can

no

long

er b

e de

sign

ated

as e

mer

ging

pat

hoge

ns, t

hey

shar

e ho

wev

er o

ne c

hara

cter

istic

in th

e ep

idem

iolo

gy o

f em

ergi

ng n

atur

e, th

e im

port

ance

of t

he h

ygie

ne in

the

prim

ary

prod

uctio

n st

age,

incl

udin

g su

pply

of p

otab

le w

ater

, and

the

appl

icat

ion

of G

MP

and

the

HA

CC

P pr

inci

ples

in th

e be

ginn

ing

of th

e fo

od c

hain

. Hep

atiti

s E v

irus

is a

pot

entia

l em

ergi

ng fo

od b

orne

pat

hoge

n an

d sw

ine

may

serv

e as

a so

urce

of i

nfec

tion

in h

uman

, a m

ost c

halle

ngin

g is

sue

in g

reat

er p

art o

f the

wor

ld ra

isin

g pi

gs. T

ick-

born

e en

ceph

aliti

s vir

us in

fect

ion,

eith

er th

ick

born

e or

cau

sed

by c

onsu

mpt

ion

of ra

w m

ilk, i

s an

incr

easi

ng tr

end

in th

e in

dust

rial

-iz

ed p

art o

f the

wor

ld. C

onsu

mer

aw

aren

ess,

eth

ics o

f foo

d, su

stai

nabi

lity

in fo

od p

rodu

ctio

n, a

nd tr

ust i

n fo

ods,

are

of g

row

ing

impo

rtan

ce to

the

cons

umer

. The

reac

tion

of th

e co

nsum

er to

new

tech

nolo

gy, s

uch

as n

anot

echn

olog

y, is

unp

redi

ctab

le. M

any

effor

ts sh

ould

be

devo

ted

to c

omm

unic

atio

n of

non

-bia

sed

info

rmat

ion

to b

oth

the

food

pro

duce

rs a

s wel

l as t

he c

onsu

mer

.

Skov

gaar

d 20

07

Den

tal u

nit w

ater

lines

T

he s

peci

fic s

truc

ture

of d

enta

l uni

ts fa

vour

s th

e pr

esen

ce o

f bio

film

and

mic

robi

al c

onta

min

atio

n of

the

dent

al u

nit w

ater

-lin

es (D

UW

L) w

ater

. The

abi

lity

of b

acte

ria

to c

olon

ize

surf

aces

and

to fo

rm b

iofil

m in

wat

er s

uppl

y tu

bes,

incl

udin

g D

UW

L,

is a

com

mon

phe

nom

enon

, whi

ch h

as b

een

wel

l doc

umen

ted,

just

as

with

diff

icul

ties

in b

iofil

m re

mov

al a

nd p

reve

ntio

n of

its

regr

owth

. Mic

roor

gani

sms

from

con

tam

inat

ed D

UW

L ar

e tr

ansm

itted

with

aer

osol

and

spl

atte

r, ge

nera

ted

by w

orki

ng

unit

hand

piec

es. O

n th

e ba

sis

of th

e de

taile

d lit

erat

ure

revi

ew, t

he s

tate

-of-

the

art k

now

ledg

e of

the

mic

roflo

ra o

f den

tal u

nit

wat

erlin

es is

pre

sent

ed. M

ost o

f the

mic

roor

gani

sms

isol

ated

from

DU

WL

are

of lo

w p

atho

geni

city

. Nev

erth

eles

s, th

e pu

blic

he

alth

sig

nific

ance

of m

any

of th

e m

icro

orga

nism

s fo

und

in D

UW

L is

unk

now

n. A

ccor

ding

to c

urre

nt k

now

ledg

e, it

is n

ot th

e m

ere

pres

ence

of b

acte

ria

that

is im

port

ant i

n D

UW

L co

ntam

inat

ion

mon

itori

ng, b

ut th

eir

num

ber,

the

pres

ence

of p

oten

tial

path

ogen

s, a

nd p

atie

nts’

oral

cav

ity m

icro

flora

. Num

erou

s st

udie

s em

phas

ize

the

need

for

effe

ctiv

e m

echa

nism

s to

redu

ce th

e m

icro

bial

con

tam

inat

ion

in D

UW

L an

d hi

ghlig

ht th

e ri

sk fo

r cr

oss-

infe

ctio

n in

gen

eral

pra

ctic

e, e

spec

ially

in v

iew

of t

he e

ver-

incr

easi

ng n

umbe

r of

imm

unoc

ompr

omis

ed p

erso

ns w

ho p

rese

nt a

t out

patie

nt d

enta

l clin

ics.

Szym

an-

ska

et a

l. 20

08

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

636

Den

tal o

ffice

env

iron

men

tBi

olog

ical

fact

ors a

re c

onst

antly

pre

sent

dur

ing

dent

al p

roce

dure

s. Th

ey a

re c

ause

s of c

ross

-infe

ctio

ns in

a d

enta

l offi

ce, c

onst

itut-

ing

occu

patio

nal h

azar

ds fo

r den

tal p

erso

nnel

exp

osed

to th

em fo

r lon

g pe

riod

s of t

ime.

The

auth

ors r

evie

w th

e cu

rren

t kno

wle

dge

on b

acte

rial

fact

ors p

rese

nt in

the

envi

ronm

ent o

f a d

enta

l offi

ce a

nd lo

ok a

t the

infe

ctio

us fa

ctor

s spr

ead

via

bloo

d-bo

rne

rout

e,

dire

ct c

onta

ct w

ith a

pat

ient

and

con

tam

inat

ed e

quip

men

t, an

d ae

roso

ls em

itted

from

bot

h th

e pa

tient

’s m

outh

and

the

dent

al u

nit

itsel

f. Th

e au

thor

s des

crib

e im

port

ant s

ourc

es a

nd ro

utes

of i

nfec

tion

spec

ific

for a

den

tal o

ffice

- bi

ofilm

as a

sour

ce o

f den

tal u

nit

wat

er c

onta

min

atio

n, m

icro

bial

con

tam

inat

ion

of w

ater

in d

enta

l uni

ts a

nd m

icro

bial

con

tam

inat

ion

of a

ir in

den

tal o

ffice

s.

Szym

an-

ska

and

Sitk

owsk

a 20

12

Free

-liv

ing

amoe

bae

in d

rink

ing

wat

erT

here

is a

n ex

pand

ing

body

of e

vide

nce

that

free

-liv

ing

amoe

bae

(FLA

) inc

reas

e bo

th th

e nu

mbe

rs a

nd v

irul

ence

of w

ater

-ba

sed,

hum

an-p

atho

geni

c, a

moe

ba-r

esis

ting

mic

roor

gani

sms

(AR

M).

Legi

onel

la sp

p., M

ycob

acte

rium

spp

., an

d ot

her

oppo

r-tu

nist

ic h

uman

pat

hoge

ns a

re k

now

n to

be

both

AR

M a

nd a

lso

the

etio

logi

c ag

ents

of p

oten

tially

fata

l hum

an lu

ng in

fect

ions

. H

owev

er, c

ompa

rativ

ely

little

is k

now

n ab

out t

he F

LA th

at m

ay fa

cilit

ate

AR

M g

row

th in

dri

nkin

g w

ater

. Thi

s re

view

exa

m-

ines

the

avai

labl

e lit

erat

ure

on F

LA in

trea

ted

drin

king

wat

er s

yste

ms;

in to

tal 2

6 st

udie

s fr

om 1

8 di

ffere

nt c

ount

ries

. FLA

w

ere

repo

rted

to b

reak

thro

ugh

the

wat

er tr

eatm

ent b

arri

er a

nd e

nter

dis

trib

utio

n sy

stem

s, in

add

ition

to th

e ex

pect

ed p

ost-

trea

tmen

t sys

tem

ingr

ess.

Onc

e in

the

dist

ribu

tion

syst

em th

ere

is e

vide

nce

of F

LA c

olon

izat

ion

and

regr

owth

esp

ecia

lly in

re

serv

oirs

and

in-p

rem

ise

plum

bing

sto

rage

tank

s. A

t the

poi

nt o

f use

the

aver

age

FLA

det

ectio

n ra

te w

as 4

5% b

ut h

ighl

y va

ri-

able

(n =

16,

sig

ma

= 31

) due

to b

oth

diffe

renc

es in

bot

h as

say

met

hods

and

the

type

of w

ater

sys

tem

s ex

amin

ed. T

his

revi

ew

reve

als

that

FLA

are

con

sist

ently

det

ecte

d in

trea

ted

drin

king

wat

er s

yste

ms

arou

nd th

e w

orld

and

pre

sent

a y

et u

nqua

ntifi

ed

emer

ging

hea

lth r

isk.

How

ever

, mor

e re

sear

ch is

urg

ently

requ

ired

bef

ore

accu

rate

ris

ks a

sses

smen

ts c

an b

e un

dert

aken

to

asse

ss th

e im

pact

s on

hum

an h

ealth

, in

hous

ehol

ds a

nd in

stitu

tions

, due

to e

xpos

ure

to F

LA fa

cilit

ated

pat

hoge

nic

AR

M.

Thom

as

and

Ash

-bo

lt 20

11

Wat

erM

. avi

um c

ompl

exM

ycob

acte

rium

avi

um c

ompl

ex (M

AC

) is

a gr

oup

of o

ppor

tuni

stic

pat

hoge

ns o

f maj

or p

ublic

hea

lth c

once

rn. I

t is

resp

onsi

ble

for

a w

ide

spec

trum

of d

isea

se d

epen

dent

on

subs

peci

es, r

oute

of i

nfec

tion

and

patie

nts

pre-

exis

ting

cond

ition

s. P

rese

ntly

, th

ere

is li

mite

d re

sear

ch o

n th

e in

cide

nce

of M

AC

infe

ctio

n th

at c

onsi

ders

bot

h pu

lmon

ary

and

othe

r cl

inic

al m

anife

stat

ions

. M

AC

has

bee

n is

olat

ed fr

om v

ario

us te

rres

tria

l and

aqu

atic

env

iron

men

ts in

clud

ing

natu

ral w

ater

s, e

ngin

eere

d w

ater

sys

tem

s an

d so

ils. I

dent

ifyin

g th

e sp

ecifi

c en

viro

nmen

tal s

ourc

es re

spon

sibl

e fo

r hu

man

infe

ctio

n is

ess

entia

l in

min

imiz

ing

dise

ase

prev

alen

ce. T

his

pape

r re

view

s cu

rren

t lite

ratu

re a

nd c

ase

stud

ies

rega

rdin

g th

e w

ide

spec

trum

of d

isea

se c

ause

d by

MA

C

and

the

role

of p

otab

le w

ater

in d

isea

se tr

ansm

issi

on. P

otab

le w

ater

was

reco

gniz

ed a

s a

puta

tive

path

way

for

MA

C in

fect

ion.

C

onta

min

ated

pot

able

wat

er s

ourc

es a

ssoc

iate

d w

ith h

uman

infe

ctio

n in

clud

ed w

arm

wat

er d

istr

ibut

ion

syst

ems,

sho

wer

s,

fauc

ets,

hou

seho

ld d

rink

ing

wat

er, s

wim

min

g po

ols

and

hot t

ub s

pas.

MA

C c

an m

aint

ain

long

-ter

m c

onta

min

atio

n of

pot

able

w

ater

sou

rces

thro

ugh

its h

igh

resi

stan

ce to

dis

infe

ctan

ts, a

ssoc

iatio

n w

ith b

iofil

ms

and

intr

acel

lula

r pa

rasi

tism

of f

ree-

livin

g pr

otoz

oa. F

urth

er re

sear

ch is

requ

ired

to in

vest

igat

e th

e ef

ficie

ncy

of w

ater

trea

tmen

t pro

cess

es a

gain

st M

AC

and

into

con

-st

ruct

ion

and

mai

nten

ance

of w

arm

wat

er d

istr

ibut

ion

syst

ems

and

the

role

they

pla

y in

MA

C p

rolif

erat

ion.

Whi

ley

et

al. 2

012

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

637

Tabl

e 3.

Myc

obac

teri

a in

wat

er

Type

of w

ater

Myc

obac

teri

a de

tect

edA

bstr

act e

xcer

ptRe

fere

nce

Dri

nkin

g w

ater

M. s

p.M

. gor

dona

eM

. kan

sasii

M. f

ortu

itum

In th

e cu

rren

t stu

dy, w

e de

tect

ed a

nd q

uant

ified

the

pres

ence

of N

TM

by

mea

ns o

f a r

apid

met

hod

in w

ater

sam

ples

take

n fr

om 5

3 co

olin

g to

wer

s of

an

urba

n ar

ea (B

arce

lona

, Spa

in).

A g

enus

-spe

cific

qua

ntita

tive

PCR

(Q-P

CR)

ass

ay w

ith a

qua

ntifi

-ca

tion

limit

(QL)

of 5

00 c

ells

l-1 w

as u

sed.

56%

(30)

of s

ampl

es w

ere

posi

tive

with

a c

once

ntra

tion

rang

e fr

om 4

.6 ×

103 to

1.

79 ×

106 c

ells

l-1. I

n so

me

case

s (9

/30)

, sam

ples

wer

e po

sitiv

e bu

t with

leve

ls b

elow

the

QL.

The

col

oniz

atio

n ra

te c

onfir

med

th

at c

oolin

g to

wer

s co

uld

be c

onsi

dere

d as

a p

oten

tial r

eser

voir

for

NT

M. T

his

stud

y al

so e

valu

ated

Q-P

CR

as a

use

ful

met

hod

to d

etec

t and

qua

ntify

NT

M in

sam

ples

com

ing

from

env

iron

men

tal s

ourc

es.

Adr

ados

et

al.

2011

Dri

nkin

g w

ater

M. s

p.

M. f

ortu

itum

M. t

okai

ense

M. a

ustr

oafr

ican

umM

. van

bale

nii

M. a

ichi

ense

M. h

eide

lber

gens

e

A to

tal o

f 148

soi

l sam

ples

and

24

wat

er s

ampl

es w

ere

colle

cted

from

var

ious

loca

tions

and

exa

min

ed to

det

erm

ine

the

pres

-en

ce o

f myc

obac

teri

a. T

he d

etec

tion

met

hod

invo

lved

sem

isel

ectiv

e cu

lturi

ng a

nd a

cid-

fast

sta

inin

g, fo

llow

ing

deco

ntam

ina-

tion

of s

ampl

es to

enr

ich

myc

obac

teri

a an

d re

duce

the

num

bers

of o

ther

mic

roor

gani

sms,

or

PCR

with

pri

mer

s sp

ecifi

c fo

r th

e m

ycob

acte

rial

16S

rRN

A g

ene,

usi

ng D

NA

ext

ract

ed d

irec

tly fr

om s

oil a

nd w

ater

sam

ples

. Myc

obac

teri

a w

ere

dete

cted

in

the

maj

ority

of t

he s

ampl

es, a

nd s

ubse

quen

t seq

uenc

e an

alys

is o

f PC

R pr

oduc

ts a

mpl

ified

dir

ectly

from

soi

l DN

A in

dica

ted

that

mos

t of t

he p

rodu

cts

wer

e re

late

d to

kno

wn

envi

ronm

enta

l myc

obac

teri

a. F

or b

oth

met

hods

the

rate

s of

reco

very

wer

e co

nsis

tent

ly h

ighe

r fo

r dr

y se

ason

sam

ples

than

for

wet

sea

son

sam

ples

.

Chi

lima

et a

l. 20

06

Dri

nkin

g w

ater

M. a

. par

atub

ercu

losi

sRe

al-t

ime

quan

titat

ive

PCR

assa

ys fo

r H

elic

obac

ter

pylo

ri, Y

ersi

nia

ente

roco

litic

a, a

nd M

ycob

acte

rium

avi

um s

ubsp

. par

atu-

berc

ulos

is, h

uman

pat

hoge

ns w

ith lo

ng-t

ime

surv

ival

cap

acity

in w

ater

, and

for

the

resi

stan

ce g

enes

erm

B, m

ecA

, bla

SHV

-5,

ampC

, tet

O, a

nd v

anA

wer

e ad

apte

d or

dev

elop

ed fo

r w

ater

sam

ples

diff

erin

g in

pol

luta

nt c

onte

nt. T

he re

sist

ance

gen

es a

nd

path

ogen

con

cent

ratio

ns w

ere

dete

rmin

ed a

t fiv

e or

six

sam

plin

g po

ints

for

each

rech

arge

sys

tem

. In

drin

king

and

irri

gatio

n w

ater

, non

e of

the

path

ogen

s w

ere

dete

cted

.

Bock

el-

man

n et

al

. 200

9

Dri

nkin

g w

ater

M. s

p.M

. per

egri

num

M. n

onch

rom

ogen

icum

M. s

meg

mat

isM

. for

tuitu

mM

. avi

um ss

p. h

omin

issu

isM

. aru

pens

eM

. gor

dona

eM

. chi

tae

We

inve

stig

ated

the

pres

ence

of n

ontu

berc

ulou

s m

ycob

acte

ria

(NT

M) i

n th

ree

Mex

ican

aqu

atic

sys

tem

s to

eva

luat

e th

e pr

eval

ence

with

the

dist

ribu

tion

of N

TM

spe

cies

. Key

phy

sico

chem

ical

par

amet

ers

of th

e w

ater

sam

ples

wer

e de

term

ined

to

find

corr

elat

ions

with

the

spec

ies’

dist

ribu

tions

. We

used

mul

tiloc

us s

eque

nce

anal

ysis

(MLS

A) b

ased

on

hsp6

5, r

poB,

and

16

S rR

NA

frag

men

ts to

det

erm

ine

thei

r ta

xono

mic

aff

iliat

ions

. NT

M w

ere

reco

vere

d fr

om w

ater

dis

trib

utio

n sy

stem

s an

d re

clai

med

wat

er fr

om th

e M

exic

o C

ity M

etro

polit

an A

rea

(MC

MA

). T

he is

olat

ed s

peci

es w

ere

asso

ciat

ed w

ith a

tem

pera

ture

of

21

degr

ees

C a

nd p

H >

7.7

. The

phy

loge

netic

ana

lysi

s sh

owed

that

eig

ht o

f the

14

diffe

rent

NT

M s

trai

ns w

ere

unam

bigu

-ou

sly

clas

sifia

ble:

Myc

obac

teri

um p

ereg

rinu

m, M

. non

chro

mog

enic

um (2

), M

. sm

egm

atis

(2),

M. f

ortu

itum

, M. a

vium

ssp

. ho

min

issu

is, M

. aru

pens

e, M

. gor

dona

e, a

nd M

. chi

tae.

Cas

tillo

-Ro

dal e

t al

. 201

2

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

638

Dri

nkin

g w

ater

M. s

p.M

. aru

pens

eM

. gor

dona

e

In th

is st

udy,

to g

ive

insi

ght i

nto

the

bact

eria

l div

ersi

ty o

f bio

film

s fro

m fu

ll-sc

ale

drin

king

wat

er d

istr

ibut

ion

syst

ems (

DW

DSs

), th

e ba

cter

ial c

omm

unity

com

posi

tions

of b

iofil

ms f

rom

two

urba

n D

WD

Ss (G

uang

zhou

and

Bei

jing,

Chi

na) w

ere

dete

rmin

ed u

sing

a

16S

rRN

A g

ene

libra

ry te

chni

que.

Mea

nwhi

le, t

he o

ccur

renc

e an

d di

vers

ity o

f myc

obac

teri

a w

ere

also

ana

lyze

d by

a M

ycob

acte

-ri

um-s

peci

fic h

sp g

ene

assa

y. Th

e bi

ofilm

s fro

m th

e fu

ll-sc

ale

DW

DSs

hav

e co

mpl

ex b

acte

rial

pop

ulat

ions

. Pro

teob

acte

ria

was

the

com

mon

and

pre

dom

inan

t gro

up in

all

biofi

lm sa

mpl

es, i

n ag

reem

ent w

ith p

revi

ous r

epor

ts. Th

e co

mm

unity

stru

ctur

es o

f bac

teri

a at

the

thre

e si

tes i

n G

uang

zhou

DW

DS

wer

e si

gnifi

cant

ly d

iffer

ent,

desp

ite th

e si

mila

r phy

sico

chem

ical

pro

pert

ies o

f por

tabl

e w

ater

. So

me

abun

dant

and

pec

ulia

r bac

teri

al p

hylo

type

s wer

e no

tew

orth

y, in

clud

ing

Met

hylo

philu

s, M

assil

ia, a

nd P

lano

mic

robi

um, m

em-

bers

of w

hich

are

rare

ly fo

und

in D

WD

Ss a

nd th

eir r

oles

in D

WD

S bi

ofilm

s are

still

unc

lear

. The

dive

rsity

of M

ycob

acte

rium

spec

ies

in b

iofil

m sa

mpl

es w

as ra

ther

low.

Myc

obac

teri

um a

rupe

nse a

nd M

ycob

acte

rium

gord

onae

wer

e th

e pr

imar

y M

ycob

acte

rium

spec

ies

in G

uang

zhou

and

Bei

jing

biofi

lms,

resp

ectiv

ely,

indi

catin

g th

at M

. aru

pens

e m

ay b

e m

ore

resi

stan

t to

chlo

ride

than

M. g

ordo

nae.

Liu

et a

l. 20

12

Dri

nkin

g w

ater

M. a

vium

The

spre

ad o

f opp

ortu

nist

ic p

atho

gens

via

pub

lic w

ater

syst

ems i

s of g

row

ing

conc

ern.

The

purp

ose

of th

is st

udy

was

to id

entif

y pa

tter

ns o

f occ

urre

nce

amon

g th

ree

oppo

rtun

istic

pat

hoge

ns (L

egio

nella

pne

umop

hila

, Myc

obac

teri

um a

vium

, and

Pse

udom

onas

ae

rugi

nosa

) rel

ativ

e to

bio

tic a

nd a

biot

ic fa

ctor

s in

two

repr

esen

tativ

e ch

lora

min

ated

dri

nkin

g w

ater

dis

trib

utio

n sy

stem

s usi

ng

cultu

re-in

depe

nden

t met

hods

. Gen

eral

ly, a

hig

h oc

curr

ence

of L

egio

nella

(≥ 6

9.0%

) and

myc

obac

teri

a (1

00%

), lo

wer

occ

urre

nce

of L

. pne

umop

hila

(≤ 2

0%) a

nd M

. avi

um (≤

33.

3%),

and

rare

det

ectio

n of

Pse

udom

onas

aer

ugin

osa

(≤ 1

3.3%

) wer

e ob

serv

ed in

bo

th sy

stem

s acc

ordi

ng to

qua

ntita

tive

PCR

. Als

o, H

artm

anel

la v

erm

iform

is w

as m

ore

prev

alen

t tha

n A

cant

ham

oeba

, bot

h of

w

hich

are

kno

wn

host

s for

opp

ortu

nist

ic p

atho

gen

ampl

ifica

tion,

the

latt

er it

self

cont

aini

ng p

atho

geni

c m

embe

rs. Th

ree-

min

ute

flush

ing

serv

ed to

dis

tingu

ish

dist

ribu

tion

syst

em w

ater

from

plu

mbi

ng in

bui

ldin

gs (i

.e.,

prem

ise

plum

bing

wat

er) a

nd re

sulte

d in

redu

ced

num

bers

of c

opie

s of L

egio

nella

, myc

obac

teri

a, H

. ver

mifo

rmis

, and

16S

rRN

A g

enes

(P <

0.0

5) w

hile

yie

ldin

g di

stin

ct

term

inal

rest

rict

ion

frag

men

t pol

ymor

phis

m (T

-RFL

P) p

rofil

es o

f 16S

rRN

A g

enes

. With

in c

erta

in su

bgro

ups o

f sam

ples

, som

e po

sitiv

e co

rrel

atio

ns, i

nclu

ding

cor

rela

tions

of n

umbe

rs o

f myc

obac

teri

a an

d to

tal b

acte

ria

(16S

rRN

A g

enes

), H

. ver

mifo

rmis

and

to

tal b

acte

ria,

myc

obac

teri

a an

d H

. ver

mifo

rmis

, and

Leg

ione

lla a

nd H

. ver

mifo

rmis

, wer

e no

ted,

em

phas

izin

g po

tent

ial m

icro

-bi

al e

colo

gica

l rel

atio

nshi

ps. O

vera

ll, th

e re

sults

pro

vide

insi

ght i

nto

fact

ors t

hat m

ay a

id in

con

trol

ling

oppo

rtun

istic

pat

hoge

n pr

olife

ratio

n in

real

-wor

ld w

ater

syst

ems.

Wan

g et

al

. 201

2

Dri

nkin

g w

ater

M. s

p.A

met

agen

ome-

base

d ap

proa

ch w

as u

sed

to a

sses

s the

taxo

nom

ic a

ffilia

tion

and

func

tion

pote

ntia

l of m

icro

bial

pop

ulat

ions

in

free

-chl

orin

e-tr

eate

d (C

HL)

and

mon

ochl

oram

ine-

trea

ted

(CH

M) d

rink

ing

wat

er (D

W).

In a

ll, 3

62,6

40 (a

vera

ging

544

bp)

and

15

5,59

3 (a

vera

ging

554

bp)

pyr

oseq

uenc

ing

read

s wer

e an

alyz

ed fo

r the

CH

L an

d C

HM

sam

ples

, res

pect

ivel

y. M

ost a

nnot

ated

pr

otei

ns w

ere

foun

d to

be

of b

acte

rial

ori

gin,

alth

ough

euk

aryo

tic, a

rcha

eal,

and

vira

l pro

tein

s wer

e al

so id

entifi

ed. D

iffer

ence

s in

com

mun

ity st

ruct

ure

and

func

tion

wer

e no

ted.

Mos

t not

ably

, Leg

ione

lla-li

ke g

enes

wer

e m

ore

abun

dant

in th

e C

HL

sam

ples

w

hile

myc

obac

teri

al g

enes

wer

e m

ore

abun

dant

in C

HM

sam

ples

. Gen

es a

ssoc

iate

d w

ith m

ultip

le d

isin

fect

ant m

echa

nism

s wer

e id

entifi

ed in

bot

h co

mm

uniti

es. M

oreo

ver,

sequ

ence

s lin

ked

to v

irul

ence

fact

ors,

such

as a

ntib

iotic

resi

stan

ce m

echa

nism

s, w

ere

obse

rved

in b

oth

mic

robi

al c

omm

uniti

es. Th

is st

udy

prov

ides

new

insi

ghts

into

the

gene

tic n

etw

ork

and

pote

ntia

l bio

logi

cal

proc

esse

s ass

ocia

ted

with

the

mol

ecul

ar m

icro

bial

eco

logy

of D

W m

icro

bial

com

mun

ities

.

Gom

ez-

Alv

arez

et

al. 2

012

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

639

Dri

nkin

g w

ater

and

ice

M. s

p.M

. por

cinu

m

We

iden

tified

M. p

orci

num

from

24

patie

nts a

t a G

alve

ston

uni

vers

ity h

ospi

tal (

Uni

vers

ity o

f Tex

as M

edic

al B

ranc

h) o

ver a

5-y

ear

peri

od. M

. por

cinu

m w

as c

onsi

dere

d a

path

ogen

in 1

1 (4

6%) o

f 24

infe

cted

pat

ient

s, in

clud

ing

4 pa

tient

s with

com

mun

ity-a

cqui

red

dise

ase.

Ret

rosp

ectiv

e pa

tient

dat

a w

ere

colle

cted

, and

wat

er sa

mpl

es w

ere

cultu

red.

Mol

ecul

ar a

naly

sis o

f wat

er is

olat

es, c

lust

ered

cl

inic

al is

olat

es, a

nd 1

5 un

rela

ted

cont

rol s

trai

ns o

f M. p

orci

num

was

per

form

ed. A

mon

g sa

mpl

es o

f hos

pita

l ice

and

tap

wat

er, 6

3%

wer

e po

sitiv

e fo

r RG

M, 5

0% o

f whi

ch w

ere

M. p

orci

num

. Am

ong

sam

ples

of w

ater

from

the

city

of G

alve

ston

, fou

r of fi

ve h

ouse

-ho

lds (

80%

) wer

e po

sitiv

e fo

r M. p

orci

num

. By

puls

ed-fi

eld

gel e

lect

roph

ores

is (P

FGE)

, 8 o

f 10

envi

ronm

enta

l M. p

orci

num

wer

e de

term

ined

to b

elon

g to

two

clos

ely

rela

ted

clon

es.

Brow

n-El

liott

et

al. 2

011

Dri

nkin

g w

ater

M. s

p.M

. muc

ogen

icum

M. r

hode

siae

M. p

ereg

rinu

mM

. for

tuitu

m

This

stud

y ex

amin

ed th

e fr

eque

ncy

of o

ccur

renc

e of

non

-tub

ercu

lous

myc

obac

teri

a (N

TM

) in

pota

ble

wat

er sa

mpl

es fr

om a

mai

n tr

aum

a ho

spita

l in

Mex

ico

City

. Six

ty-n

ine

pota

ble

wat

er sa

mpl

es w

ere

colle

cted

, 23

from

eac

h so

urce

: cis

tern

, kitc

hen

tap

and

bath

room

show

ers.

Of t

he 6

9 sa

mpl

es, 3

6 ha

rbou

red

NT

M sp

ecie

s. Tw

enty

-nin

e of

the

36 is

olat

es w

ere

Myc

obac

teri

um m

ucog

eni-

cum

, tw

o M

ycob

acte

rium

rhod

esia

e, o

ne M

ycob

acte

rium

per

egri

num

, one

Myc

obac

teri

um fo

rtui

tum

and

thre

e w

ere

Myc

o-

bact

eriu

m sp

p. H

ospi

tal p

otab

le w

ater

har

bour

ing

NT

M re

pres

ents

a p

oten

tial s

ourc

e fo

r nos

ocom

ial i

nfec

tions

, the

refo

re w

e su

gges

t tha

t hos

pita

l pot

able

wat

er m

icro

biol

ogic

al g

uide

lines

shou

ld in

clud

e te

stin

g fo

r NT

M sp

ecie

s.

Fern

an-

dez-

Ren-

don

et a

l. 20

12

Dri

nkin

g w

ater

M. s

p.M

. len

tiflav

umM

. tus

ciae

M. g

ordo

nae

Dri

nkin

g w

ater

dis

trib

utio

n sy

stem

s wer

e an

alyz

ed fo

r via

ble

coun

ts o

f myc

obac

teri

a by

sam

plin

g w

ater

from

wat

erw

orks

and

in

diffe

rent

par

ts o

f the

syst

ems.

In a

dditi

on, l

oose

dep

osits

col

lect

ed d

urin

g m

echa

nica

l cle

anin

g of

the

mai

n pi

pelin

es w

ere

sim

i-la

rly

anal

yzed

. The

stud

y co

vere

d 16

syst

ems a

t eig

ht lo

calit

ies i

n Fi

nlan

d. In

an

expe

rim

enta

l stu

dy, m

ycob

acte

rial

col

oniz

atio

n of

bio

film

s on

poly

viny

l chl

orid

e tu

bes i

n a

syst

em w

as st

udie

d. Th

e is

olat

ion

freq

uenc

y of

myc

obac

teri

a in

crea

sed

from

35%

at

the

wat

erw

orks

to 8

0% in

the

syst

em, a

nd th

e nu

mbe

r of m

ycob

acte

ria

in th

e po

sitiv

e sa

mpl

es in

crea

sed

from

15

to

140

CFU

/lite

r, re

spec

tivel

y. M

ycob

acte

ria

wer

e is

olat

ed fr

om a

ll 11

dep

osits

with

an

accu

mul

atio

n tim

e of

tens

of y

ears

and

from

al

l 4 d

epos

its w

hich

had

acc

umul

ated

dur

ing

a 1-

year

follo

w-u

p tim

e. Th

e nu

mbe

rs o

f myc

obac

teri

a w

ere

high

in b

oth

old

and

youn

g de

posi

ts (m

edia

ns, 1

.8 ×

105 a

nd 3

.9 ×

105 C

FU/g

[dry

wei

ght]

, res

pect

ivel

y). B

oth

wat

er a

nd d

epos

it sa

mpl

es y

ield

ed th

e hi

ghes

t num

bers

of m

ycob

acte

ria

in th

e sy

stem

s usi

ng su

rfac

e w

ater

and

app

lyin

g oz

onat

ion

as a

n in

term

edia

te tr

eatm

ent o

r po

sttr

eatm

ent.

The

num

ber a

nd g

row

th o

f myc

obac

teri

a in

syst

em w

ater

s cor

rela

ted

stro

ngly

with

the

conc

entr

atio

n of

ass

imi-

labl

e or

gani

c ca

rbon

in th

e w

ater

leav

ing

the

wat

erw

orks

. The

dens

ities

of m

ycob

acte

ria

in th

e de

velo

ping

bio

film

s wer

e hi

ghes

t at

the

dist

al si

tes o

f the

syst

ems.

Ove

r 90%

of t

he m

ycob

acte

ria

isol

ated

from

wat

er a

nd d

epos

its b

elon

ged

to M

ycob

acte

rium

le

ntifl

avum

, M. t

usci

ae, M

. gor

dona

e, a

nd a

pre

viou

sly

uncl

assi

fied

grou

p of

myc

obac

teri

a. O

ur re

sults

indi

cate

that

dri

nkin

g w

ater

syst

ems m

ay b

e a

sour

ce fo

r rec

ently

dis

cove

red

new

myc

obac

teri

al sp

ecie

s.

Torv

inen

et

al.

2010

Dri

nkin

g w

ater

dis

trib

u-ti

on s

yste

mM

. gor

dona

eM

. kan

sasii

M. i

ntra

cellu

lare

M. c

helo

nae

Myc

obac

teri

a ha

ve e

mer

ged

as a

maj

or c

ause

of o

ppor

tuni

stic

infe

ctio

ns. U

ntil

the

pres

ent,

only

a fe

w st

udie

s hav

e ch

arac

teri

zed

myc

obac

teri

a pr

esen

t in

the

wat

er d

istr

ibut

ion

syst

em o

f urb

an a

reas

. In

this

stud

y, w

e ch

arac

teri

ze th

ese

mic

roor

gani

sms i

n th

e Li

sbon

wat

er d

istr

ibut

ion

syst

em. O

ur re

sults

indi

cate

a h

igh

rate

of p

ositi

vitie

s (90

.5%

) with

mai

nly

sapr

ophy

tic m

ycob

acte

ria.

A

roun

d 63

% o

f the

se re

sults

bel

ong

to st

rain

s of M

ycob

acte

rium

gor

dona

e in

dica

ting

a ge

nera

lized

pro

lifer

atio

n of

this

spec

ies i

n th

e Li

sbon

wat

er d

istr

ibut

ion

syst

em. A

tota

l of 2

1.05

% o

f the

isol

ates

are

from

M. k

ansa

sii, M

. int

race

llula

re, a

nd M

. che

lona

e.

Sant

os e

t al

. 200

5

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

640

Dri

nkin

g w

ater

...T

he n

atur

al re

serv

oirs

of t

hese

non

-pri

mar

y pa

thog

enic

myc

obac

teri

a in

clud

e aq

uatic

and

terr

estr

ial e

nvir

onm

ents

. Und

er

cert

ain

circ

umst

ance

s, e

.g.,

skin

lesi

ons,

pul

mon

ary

or im

mun

e dy

sfun

ctio

ns a

nd c

hron

ic d

isea

ses,

thes

e en

viro

nmen

tal

myc

obac

teri

a (E

M) m

ay c

ause

dis

ease

. EM

suc

h as

M. a

vium

, M. k

ansa

sii,

and

M. x

enop

i hav

e fr

eque

ntly

bee

n is

olat

ed fr

om

drin

king

wat

er a

nd h

ospi

tal w

ater

dis

trib

utio

n sy

stem

s. B

iofil

m fo

rmat

ion,

am

oeba

-ass

ocia

ted

lifes

tyle

, and

resi

stan

ce to

ch

lori

ne h

ave

been

reco

gniz

ed a

s im

port

ant f

acto

rs th

at c

ontr

ibut

e to

the

surv

ival

, col

oniz

atio

n an

d pe

rsis

tenc

e of

EM

in

wat

er d

istr

ibut

ion

syst

ems.

Alth

ough

the

pres

ence

of E

M in

tap

wat

er h

as b

een

linke

d to

nos

ocom

ial i

nfec

tions

and

pse

udo-

infe

ctio

ns, i

t rem

ains

unc

lear

if th

ese

EM p

rovi

de a

hea

lth r

isk

for

imm

unoc

ompr

omis

ed p

eopl

e, in

par

ticul

ar A

IDS

patie

nts.

In

this

rega

rd, c

ontr

ol s

trat

egie

s ba

sed

on m

aint

enan

ce o

f an

effe

ctiv

e di

sinf

ecta

nt re

sidu

al a

nd lo

w c

once

ntra

tion

of n

utri

-en

ts h

ave

been

pro

pose

d to

kee

p EM

num

bers

to a

min

imum

in w

ater

dis

trib

utio

n sy

stem

s.

Vaer

e-w

ijck

et

al. 2

005

Dri

nkin

g w

ater

Free

-liv

ing

amoe

bae

have

bee

n de

tect

ed in

a la

rge

num

ber

of m

an-m

ade

wat

er s

yste

ms,

incl

udin

g dr

inki

ng w

ater

dis

trib

u-tio

n sy

stem

s. S

ome

of th

ese

amoe

bae

can

host

am

oeba

e-re

sist

ing

bact

eria

, and

thus

act

pot

entia

lly a

s re

serv

oirs

and

veh

icle

s fo

r a

num

ber

of p

atho

gens

. The

obj

ectiv

es o

f thi

s st

udy

wer

e to

cha

ract

eriz

e th

e am

oeba

e an

d am

oeba

e-re

sist

ing

bact

eria

pr

esen

t in

diffe

rent

raw

wat

ers

used

for

drin

king

wat

er p

rodu

ctio

n, a

nd to

ass

ess

the

effic

ienc

y of

diff

eren

t tre

atm

ents

app

lied

for

drin

king

wat

er p

rodu

ctio

n in

rem

ovin

g or

inac

tivat

ing

thes

e am

oeba

e. T

he p

relim

inar

y re

sults

of t

his

stud

y co

nfir

m th

e pr

esen

ce o

f am

oeba

e an

d am

oeba

e-re

sist

ing

bact

eria

in r

aw w

ater

s us

ed fo

r dr

inki

ng w

ater

pro

duct

ion.

Due

to th

eir

capa

city

to

enc

yst,

mos

t of t

hese

am

oeba

e ar

e ex

trem

ely

resi

stan

t to

disi

nfec

tion

proc

esse

s. In

thes

e co

nditi

ons,

pre

vent

ing

the

dis-

sem

inat

ion

of th

ese

mic

ro-o

rgan

ism

s th

roug

h dr

inki

ng w

ater

will

mai

nly

requ

ire

thei

r ph

ysic

al re

mov

al b

y cl

arifi

catio

n an

d fil

trat

ion

proc

esse

s. T

he p

artic

ular

haz

ard

that

am

oeba

e-re

sist

ing

bact

eria

repr

esen

t in

drin

king

wat

er p

rodu

ctio

n sh

ould

be

take

n in

to a

ccou

nt in

any

ris

k as

sess

men

t con

duct

ed in

the

fram

ewor

k of

a w

ater

saf

ety

plan

, and

con

trol

str

ateg

ies

base

d on

ph

ysic

al re

mov

al r

athe

r th

an d

isin

fect

ion

shou

ld b

e ad

opte

d w

here

nec

essa

ry.

Lore

t et

al. 2

008

Dri

nkin

g an

d su

rfac

e w

ater

M. g

ordo

nae

M. l

entifl

avum

M. a

vium

com

plex

To in

vest

igat

e th

e oc

curr

ence

and

spe

cies

div

ersi

ty o

f myc

obac

teri

a in

wat

ers,

sur

face

wat

er s

ampl

es w

ere

colle

cted

mon

thly

fr

om th

e H

an R

iver

and

tap

wat

er s

ampl

es a

t the

term

inal

site

s of

the

dist

ribu

tion

syst

em. M

ycob

acte

ria

in e

ach

wat

er s

ampl

e w

ere

isol

ated

by

deco

ntam

inat

ion

usin

g ce

tylp

yrid

iniu

m c

hlor

ide

(CPC

) and

cul

tivat

ion

on M

iddl

ebro

ok 7

H10

aga

r, an

d th

en

iden

tifie

d by

pol

ymer

ase

chai

n re

actio

n-re

stri

ctio

n fr

agm

ent l

engt

h po

lym

orph

ism

ana

lysi

s (P

RA

) and

seq

uenc

ing

of th

e 65

-kD

a he

at-s

hock

pro

tein

gen

e (h

sp65

gen

e). M

ycob

acte

ria

wer

e de

tect

ed in

59%

of t

he s

urfa

ce w

ater

sam

ples

and

26%

of

the

tap

wat

er s

ampl

es. O

ver

half

of th

e 15

8 is

olat

es c

ould

not

be

iden

tifie

d by

hsp

65 P

RA

and

gen

e se

quen

cing

, and

sev

eral

id

entif

icat

ion

disc

repa

ncie

s w

ere

obse

rved

bet

wee

n th

e tw

o m

etho

ds. T

he m

ost f

requ

ently

isol

ated

spe

cies

was

Myc

obac

te-

rium

gor

dona

e in

sur

face

wat

er a

nd M

. len

tifla

vum

in ta

p w

ater

. M. a

vium

com

plex

(MA

C),

the

mos

t im

port

ant p

atho

gen

amon

g en

viro

nmen

tal m

ycob

acte

ria,

was

det

ecte

d in

the

surf

ace

wat

er s

ampl

es b

ut n

ot fo

und

in th

e ta

p w

ater

sam

ples

. The

re

sult

dem

onst

rate

d th

at w

ater

is a

n im

port

ant e

nvir

onm

enta

l sou

rce

of m

ycob

acte

ria

and

the

com

bine

d ap

plic

atio

n of

hsp

65

PRA

and

seq

uenc

ing

was

mor

e re

liabl

e th

an h

sp65

PR

A a

lone

to a

ccur

atel

y id

entif

y m

ycob

acte

ria

pres

ent i

n w

ater

.

Lee

et a

l. 20

08

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

641

Show

er h

eads

M. a

vium

A

lthou

gh o

ppor

tuni

stic

pat

hoge

ns c

omm

only

are

cul

ture

d fr

om sh

ower

faci

litie

s, th

ere

is li

ttle

kno

wle

dge

of e

ither

thei

r pr

eval

ence

or t

he n

atur

e of

oth

er m

icro

orga

nism

s tha

t may

be

deliv

ered

dur

ing

show

er u

sage

. To

dete

rmin

e th

e co

mpo

sitio

n of

sh

ower

head

bio

film

s and

wat

ers,

we

anal

yzed

rRN

A g

ene

sequ

ence

s fro

m 4

5 sh

ower

head

site

s aro

und

the

Uni

ted

Stat

es. W

e fin

d th

at v

aria

ble

and

com

plex

, but

spec

ific,

mic

robi

al a

ssem

blag

es o

ccur

insi

de sh

ower

head

s. P

artic

ular

ly st

riki

ng w

as th

e fin

ding

th

at se

quen

ces r

epre

sent

ativ

e of

non

-tub

ercu

lous

myc

obac

teri

a (N

TM

) and

oth

er o

ppor

tuni

stic

hum

an p

atho

gens

are

enr

iche

d to

hig

h le

vels

in m

any

show

erhe

ad b

iofil

ms,

>10

0-fo

ld a

bove

bac

kgro

und

wat

er c

onte

nts.

We

conc

lude

that

show

erhe

ads m

ay

pres

ent a

sign

ifica

nt p

oten

tial e

xpos

ure

to a

eros

oliz

ed m

icro

bes,

incl

udin

g do

cum

ente

d op

port

unis

tic p

atho

gens

.

Feaz

el e

t al

. 200

9

Dri

nkin

g w

ater

M

. sp.

M

. gor

dona

e M

. gas

tri/

M. k

ansa

siiM

. for

tuitu

m

M. s

imia

eM

. scr

oful

aceu

mM

. szu

igai

This

pap

er p

rese

nts t

he fi

ndin

g of

the

poss

ible

cau

se o

f the

hig

h fa

lse-

posi

tive

rate

in a

cid-

fast

stai

ning

in h

isto

logi

cal e

xam

ina-

tions

. Usi

ng a

cid-

fast

stai

ning

, cul

ture

, and

PC

R, a

cid-

fast

bac

illi w

ere

dete

cted

in 8

3.7%

of 4

9 ho

spita

l tap

wat

er sa

mpl

es a

nd

nont

uber

culo

us m

ycob

acte

ria

(NT

M) w

ere

dete

cted

in 2

0.4%

of t

he sa

me

49 sa

mpl

es. Th

e 10

NT

M is

olat

es w

ere

also

iden

tified

to

the

spec

ies l

evel

usi

ng P

CR-

rest

rict

ion

frag

men

t len

gth

poly

mor

phis

m. O

ur fi

ndin

gs in

dica

te th

at N

TM

in h

ospi

tal t

ap w

ater

ar

e th

e po

ssib

le c

ause

of f

alse

pos

itive

s in

acid

-fas

t sta

inin

g an

d of

nos

ocom

ial i

nfec

tion

in im

mun

ocom

prom

ised

pat

ient

s.

Cha

ng e

t al

. 200

2

Dri

nkin

g w

ater

M. s

p.M

. per

egri

num

M. c

helo

nae

M. a

bsce

ssus

M. g

ordo

nae

Rece

ntly

the

pres

ence

of N

TM

in p

ublic

dri

nkin

g w

ater

and

hos

pita

l wat

er d

istr

ibut

ion

syst

ems h

as b

een

repo

rted

. Thei

r abi

lity

to fo

rm b

iofil

ms a

nd th

eir r

esis

tanc

e to

chl

orin

e bo

th c

ontr

ibut

e to

thei

r sur

viva

l and

col

oniz

atio

n in

wat

er d

istr

ibut

ion

syst

ems.

Her

e w

e an

alyz

ed th

irty

-tw

o ho

spita

l tap

wat

er sa

mpl

es th

at w

ere

colle

cted

from

diff

eren

t loc

atio

ns in

thre

e ho

spita

ls so

as t

o ev

alua

te th

e pr

eval

ence

of N

TM

spec

ies.

The

wat

er sa

mpl

es w

ere

conc

entr

ated

by

mem

bran

e fil

trat

ion

and

then

elu

ted

with

st

erili

zed

wat

er fo

llow

ing

soni

catio

n. T

wo-

step

dir

ect P

CR

targ

etin

g th

e rp

oB g

ene,

rest

rict

ion

frag

men

t len

gth

poly

mor

phis

m

(RFL

P) u

sing

the

Msp

I res

tric

tion

enzy

me,

and

sequ

ence

ana

lysi

s wer

e pe

rfor

med

for i

dent

ifica

tion

of N

TM

to th

e sp

ecie

s le

vel.

The

sequ

ence

s of e

ach

PCR

prod

uct w

ere

anal

yzed

usi

ng B

LAST

N. S

even

sam

ples

(7/3

2, 2

1.9%

) wer

e po

sitiv

e fo

r NT

M

as d

eter

min

ed b

y ne

sted

-PC

R. Th

e PC

R-RF

LP re

sults

indi

cate

d fiv

e di

ffere

nt p

atte

rns a

mon

g th

e se

ven

posi

tive

PCR

sam

ples

. Th

e w

ater

-bor

n N

TM

wer

e id

entifi

ed, i

nclu

ding

M. p

ereg

rinu

m, M

. che

lona

e (2

cas

es),

M. a

bsce

ssus

, M. g

ordo

nae

(2 c

ases

), an

d M

ycob

acte

rium

sp. J

LS. Th

e di

rect

two-

step

PC

R-RF

LP m

etho

d ta

rget

ing

the

rpoB

gen

e w

as e

ffect

ive

for t

he d

etec

tion

and

the

diffe

rent

iatio

n of

NT

M sp

ecie

s fro

m h

ospi

tal t

ap w

ater

.

Shin

et a

l. 20

08

Dri

nkin

g w

ater

M. s

p.M

. xen

opi

M. c

helo

nae

M. m

ucog

enic

umM

. flav

esce

nsM

. fre

deri

ksbe

rgen

seM

. gor

dona

eM

. mor

ioka

ense

M. v

acca

e

Aim

of t

he st

udy

was

to e

stab

lish

high

ly se

nsiti

ve a

nd sp

ecifi

c te

chni

ques

to d

etec

t and

iden

tify

NT

M in

hos

pita

l dri

nkin

g w

ater

. A

 Myc

obac

teri

um g

enus

-spe

cific

16S

rDN

A-b

ased

real

-tim

e Li

ghtC

ycle

r PC

R as

say

with

inte

rnal

inhi

bitio

n co

ntro

l and

a M

. xen

-op

i-sp

ecifi

c PC

R w

ere

deve

lope

d.N

inet

y-th

ree

wat

er sa

mpl

es fr

om 5

3 ta

ps fr

om 4

hos

pita

ls w

ere

inve

stig

ated

. NT

M w

ere

cultu

red

from

21

of 4

9 (4

3%) c

old

and

32 o

f 44

(73%

) war

m w

ater

sam

ples

. M. c

helo

nae,

M. fl

aves

cens

, M. f

rede

riks

berg

ense

, M. g

ordo

nae,

M. m

orio

kaen

se, M

. muc

ogen

icum

, M. v

acca

e, an

d M

. xen

opi w

ere

iden

tified

with

mol

ecul

ar m

etho

ds. A

ll 93

wat

er sa

mpl

es w

ere

posi

tive

in th

e ge

nus-

spec

ific

PCR.

M. x

enop

i DN

A w

as d

etec

ted

in 4

0 of

44

(91%

) war

m a

nd 3

3 of

49

(67%

) col

d w

ater

sam

ples

in

clud

ing

45 o

f 65

(69%

) M. x

enop

i cul

ture

-neg

ativ

e sa

mpl

es.

Hus

sein

et

al.

2009

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

642

Dri

nkin

g w

ater

bio

film

sM

. avi

umM

ycob

acte

rium

avi

um is

a p

oten

tial p

atho

gen

occu

rrin

g in

dri

nkin

g w

ater

sys

tem

s. It

is a

slo

wly

gro

win

g ba

cter

ium

pro

duc-

ing

a th

ick

cell

wal

l con

tain

ing

myc

olic

aci

ds, a

nd it

is k

now

n to

resi

st c

hlor

ine

bett

er th

an m

any

othe

r m

icro

bes.

Sev

eral

st

udie

s ha

ve s

how

n th

at p

atho

geni

c ba

cter

ia s

urvi

ve b

ette

r in

bio

film

s th

an in

wat

er. B

y us

ing

Prop

ella

bio

film

reac

tors

, we

stud

ied

how

fact

ors

gene

rally

influ

enci

ng th

e gr

owth

of b

iofil

ms

(flo

w r

ate,

pho

spho

rus

conc

entr

atio

n, a

nd te

mpe

ratu

re)

influ

ence

the

surv

ival

of M

. avi

um in

dri

nkin

g w

ater

bio

film

s. T

he g

row

th o

f bio

film

s w

as fo

llow

ed b

y cu

lture

and

DA

PI

(4’,6

’-dia

mid

ino-

2-ph

enyl

indo

le) s

tain

ing,

and

con

cent

ratio

ns o

f M. a

vium

wer

e de

term

ined

by

cultu

re a

nd fl

uore

scen

ce in

si

tu h

ybri

diza

tion

met

hods

. The

spi

ked

M. a

vium

sur

vive

d in

bio

film

s fo

r th

e 4-

wee

k st

udy

peri

od w

ithou

t a d

ram

atic

dec

line

in c

once

ntra

tion.

The

add

ition

of p

hosp

horu

s (1

0 µg

/lite

r) in

crea

sed

the

num

ber

of h

eter

otro

phic

bac

teri

a in

bio

film

s bu

t de

crea

sed

the

cultu

rabi

lity

of M

. avi

um. T

he re

ason

for

this

resu

lt is

pro

babl

y th

at p

hosp

horu

s in

crea

sed

com

petit

ion

with

ot

her

mic

robe

s. A

n in

crea

se in

flow

vel

ocity

had

no

effe

ct o

n th

e su

rviv

al o

f M. a

vium

, alth

ough

it in

crea

sed

the

grow

th o

f bi

ofilm

s. A

hig

her

tem

pera

ture

(20

degr

ees

C v

ersu

s 7

degr

ees

C) i

ncre

ased

bot

h th

e nu

mbe

r of

het

erot

roph

ic b

acte

ria

and

the

surv

ival

of M

. avi

um in

bio

film

s. In

con

clus

ion,

the

resu

lts s

how

that

in te

rms

of a

ffect

ing

the

surv

ival

of s

low

ly g

row

ing

M. a

vium

in b

iofil

ms,

tem

pera

ture

is a

mor

e im

port

ant f

acto

r th

an th

e av

aila

bilit

y of

nut

rien

ts li

ke p

hosp

horu

s.

Torv

inen

et

al.

2007

Dri

nkin

g w

ater

M. a

. par

atub

ercu

losi

sA

“clu

ster

” of p

atie

nts

refe

rs to

the

geog

raph

ic p

roxi

mity

of u

nrel

ated

pat

ient

s w

ith th

e sa

me

dise

ase

and

sugg

ests

a c

omm

on

envi

ronm

enta

l cau

se fo

r th

at d

isea

se. C

lust

ers

of p

atie

nts

with

Cro

hn’s

dise

ase

have

bee

n lin

ked

to th

e pr

esen

ce o

f an

infe

c-tio

us m

icro

orga

nism

in u

npas

teur

ized

milk

and

che

ese,

unt

reat

ed w

ater

sup

plie

d by

wel

ls o

r sp

ring

s, a

nim

al m

anur

e us

ed

as fe

rtili

zer

for

fam

ily v

eget

able

gar

dens

, and

bod

ies

of w

ater

con

tam

inat

ed b

y ag

ricu

ltura

l run

off.

Myc

obac

teri

um a

vium

su

bspe

cies

par

atub

ercu

losi

s (M

AP)

is th

e su

spec

ted

caus

e of

Cro

hn’s

dise

ase.

MA

P ca

uses

a d

isea

se in

dai

ry c

ows

and

othe

r an

imal

s th

at is

sim

ilar

to C

rohn

’s di

seas

e, c

alle

d Jo

hne’s

(’Yo

-kne

es’)

dise

ase

or p

arat

uber

culo

sis.

Dai

ry c

ows

with

John

e’s d

is-

ease

sec

rete

MA

P in

to th

eir

milk

and

exc

rete

MA

P in

to th

eir

fece

s. M

AP

is p

rese

nt in

unt

reat

ed w

ater

suc

h as

wel

l wat

er, i

n bo

dies

of w

ater

con

tam

inat

ed b

y ag

ricu

ltura

l run

off,

and

in u

npas

teur

ized

milk

and

che

ese.

The

“tre

atm

ent”

of “

tap”

wat

er to

m

ake

it “d

rink

able

” or

“pot

able

” by

the

proc

esse

s of

sed

imen

tatio

n, fi

ltrat

ion

and

chlo

rina

tion

has

little

to n

o ef

fect

on

MA

P.

MA

P is

so

resi

stan

t to

chlo

rine

dis

infe

ctio

n th

at s

uch

disi

nfec

tion

actu

ally

sel

ects

for

its g

row

th. O

ther

sub

spec

ies

of M

yco-

bact

eriu

m a

vium

gro

w in

bio

film

s pr

esen

t on

tap

wat

er p

ipes

. Des

pite

the

docu

men

ted

pres

ence

of M

AP

in ta

p w

ater

and

its

pro

babl

e gr

owth

on

tap

wat

er p

ipes

, clu

ster

s of

Cro

hn’s

dise

ase

have

not

pre

viou

sly

been

des

crib

ed in

rela

tions

hip

to ta

p w

ater

pip

es s

uppl

ying

pat

ient

s’ ho

mes

. Thi

s re

port

des

crib

es th

ree

unre

late

d in

divi

dual

s w

ho li

ved

on th

e sa

me

bloc

k al

ong

a st

reet

in a

mid

wes

tern

Am

eric

an c

ity a

nd d

evel

oped

Cro

hn’s

dise

ase

with

in fo

ur y

ears

of e

ach

othe

r in

the

1960

’ s. A

com

mon

ta

p w

ater

pip

e su

pplie

d th

eir

hom

es. T

his

is th

e fir

st re

port

ed c

lust

er o

f Cro

hn’s

dise

ase

poss

ibly

link

ed to

fully

trea

ted

drin

k-in

g w

ater

, and

is c

onsi

sten

t with

pre

viou

sly

repo

rted

clu

ster

s of

Cro

hn’s

dise

ase

linke

d to

an

infe

ctio

us m

icro

orga

nism

in

wat

er.

Pier

ce

2009

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

643

Dri

nkin

g w

ater

M. c

helo

nae

M. k

ansa

siiM

. for

tuitu

m

Non

-tub

ercu

lous

myc

obac

teri

a (N

TM

) fou

nd fr

eque

ntly

in ta

p w

ater

and

env

iron

men

t cau

se im

port

ant o

ppor

tuni

stic

infe

c-tio

ns in

imm

unoc

ompr

omis

ed p

atie

nts.

The

aim

of t

his

stud

y w

as to

isol

ate

and

iden

tify

non-

tube

rcul

ous

myc

obac

teri

a in

soi

l, ra

w m

ilk a

nd w

ater

dis

trib

utio

n sy

stem

sam

ples

in M

ersi

n (a

pro

vinc

e lo

cate

d at

Med

iterr

anea

n re

gion

of T

urke

y).

A to

tal o

f 101

wat

er, 1

24 s

oil a

nd 4

0 m

ilk s

ampl

es c

olle

cted

from

the

cent

ral p

art a

nd s

ubur

ban

part

s of

Mer

sin

duri

ng

Nov

embe

r 20

03–M

ay 2

004

peri

od w

ere

incl

uded

in th

e st

udy.

Wat

er s

ampl

es w

ere

colle

cted

from

29

diffe

rent

wat

er d

istr

ibu-

tion

syst

ems;

soi

l sam

ples

from

diff

eren

t par

ks a

nd g

arde

ns a

nd m

ilk s

ampl

es fr

om r

aw m

ilks

sold

at d

iffer

ent d

istr

icts

. Aft

er

the

sam

ples

wer

e pr

oces

sed

by h

omog

eniz

atio

n an

d de

cont

amin

atio

n, a

cid-

fast

sta

inin

g an

d cu

lture

into

Low

enst

ein-

Jens

en

med

ium

wer

e pe

rfor

med

. Aci

d-fa

st b

acill

i iso

late

d fr

om c

ultu

re m

ediu

m w

ere

iden

tifie

d by

usi

ng c

onve

ntio

nal m

etho

ds,

poly

mer

ase

chai

n re

actio

n (P

CR)

-RFL

P (R

estr

ictio

n Fr

agm

ent L

engt

h Po

lym

orph

ism

) and

INN

O-L

IPA

Myc

obac

teri

a m

eth-

ods.

NT

M w

ere

iden

tifie

d fr

om 4

.9%

(5/1

01) o

f wat

er s

ampl

es a

nd 0

.8%

(1/1

24) o

f soi

l sam

ples

by

cultu

re a

nd P

CR

. No

NT

M

wer

e de

tect

ed in

the

raw

milk

sam

ples

. Thr

ee o

f the

NT

M s

trai

ns is

olat

ed fr

om w

ater

sam

ples

wer

e de

fined

as

Myc

obac

te-

rium

che

lona

e ty

pe Il

l and

two

as M

ycob

acte

rium

kan

sasi

i typ

e II

. One

NT

M s

trai

n is

olat

ed fr

om s

oil w

as d

efin

ed a

s M

yco-

bact

eriu

m fo

rtui

tum

. It w

as o

f not

e th

at tw

o of

the

five

NT

M p

ositi

ve w

ater

sam

ples

wer

e ta

p w

ater

sam

ples

col

lect

ed fr

om

hosp

itals

. It w

as c

oncl

uded

that

NT

M c

olon

izat

ion/

cont

amin

atio

n of

wat

er a

nd e

nvir

onm

ent i

n th

e ho

spita

ls w

as a

pot

entia

l ri

sk fa

ctor

in te

rms

of n

osoc

omia

l inf

ectio

ns. T

hus

surv

eilla

nce

cultu

res

of th

e w

ater

sys

tem

s an

d th

e m

edic

al d

evic

es in

the

hosp

ital a

re n

eces

sary

to fi

x th

e so

urce

of N

TM

, to

iden

tify

and

type

the

stra

ins

and

to e

stab

lish

effe

ctiv

e co

ntro

l mea

sure

s su

ch a

s st

erili

zatio

n, d

isin

fect

ion,

mai

nten

ance

and

mod

erni

zatio

n of

wat

er s

yste

ms.

Caf

ri e

t al

. 201

0

Dri

nkin

g w

ater

Non

tube

rcul

ous

myc

obac

teri

a (N

TM

) are

env

iron

men

tal o

ppor

tuni

stic

pat

hoge

ns o

f hum

ans

and

anim

als.

The

y ar

e fo

und

in

a w

ide

vari

ety

of h

abita

ts to

whi

ch h

uman

s ar

e ex

pose

d, in

clud

ing

drin

king

wat

er d

istr

ibut

ion

syst

ems

and

hous

ehol

d w

ater

an

d pl

umbi

ng. I

n th

at re

gard

, the

y ar

e di

stin

ct fr

om th

eir

oblig

ate

path

ogen

ic re

lativ

es, t

he m

embe

rs o

f the

Myc

obac

teri

um

tube

rcul

osis

com

plex

. Ow

ing

to th

e pr

esen

ce o

f NT

M in

the

hum

an e

nvir

onm

ent,

hum

an a

ctiv

ities

hav

e ha

d di

rect

impa

cts

on th

eir

ecol

ogy

and

ther

eby

thei

r ep

idem

iolo

gy. N

TM

are

olig

otro

phic

, abl

e to

gro

w a

t low

org

anic

mat

ter

conc

entr

atio

ns

and

over

a w

ide

rang

e of

tem

pera

ture

s, a

nd e

ven

at lo

w o

xyge

n co

ncen

trat

ions

. Thu

s, N

TM

are

nor

mal

inha

bita

nts

of n

atu-

ral w

ater

s an

d dr

inki

ng w

ater

s. D

isco

very

of t

he p

rese

nce

of N

TM

-pol

lute

d so

ils is

not

sur

pris

ing

in li

ght o

f the

abi

lity

of

NT

M to

deg

rade

a v

arie

ty o

f hyd

roca

rbon

pol

luta

nts.

A m

ajor

hum

an a

ctiv

ity s

elec

ting

for

the

grow

th a

nd p

redo

min

ance

of

myc

obac

teri

a in

hab

itats

is d

isin

fect

ion.

In c

ompa

riso

n to

oth

er b

acte

ria,

NT

M a

re d

isin

fect

ant,

heav

y m

etal

and

ant

ibio

tic

resi

stan

t. T

here

fore

, the

use

of a

ny a

ntim

icro

bial

age

nt s

elec

ts fo

r m

ycob

acte

ria.

Use

of d

isin

fect

ant i

n dr

inki

ng w

ater

trea

t-m

ent s

elec

ts fo

r m

ycob

acte

ria

that

can

gro

w a

nd c

ome

to p

rolif

erat

e in

dri

nkin

g w

ater

dis

trib

utio

n sy

stem

s in

the

abse

nce

of

disi

nfec

tant

-sen

sitiv

e co

mpe

ting

mic

roor

gani

sms.

NT

M s

elec

tion

may

als

o oc

cur

as a

con

sequ

ence

of a

ntib

iotic

s in

dri

nkin

g w

ater

sou

rces

.

Falk

in-

ham

201

0

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

644

Hou

seho

ld w

ater

To d

eter

min

e w

heth

er p

lum

bing

cou

ld b

e a

sour

ce o

f non

tube

rcul

ous

myc

obac

teri

a (N

TM

) inf

ectio

n, d

urin

g 20

07–2

009

I iso

late

d N

TM

from

sam

ples

from

hou

seho

ld w

ater

sys

tem

s of

NT

M p

atie

nts.

Sam

ples

from

22/

37 (5

9%) h

ouse

hold

s an

d 10

9/39

4 (2

8%) t

otal

sam

ples

yie

lded

NT

M. S

even

teen

(46%

) of t

he 3

7 ho

useh

olds

yie

lded

≥ 1

Myc

obac

teri

um s

pp. i

sola

te o

f th

e sa

me

spec

ies

as th

at fo

und

in th

e pa

tient

; in

7 of

thos

e ho

useh

olds

, the

pat

ient

isol

ate

and

1 pl

umbi

ng is

olat

e ex

hibi

ted

the

sam

e re

petit

ive

sequ

ence

-bas

ed P

CR

DN

A fi

nger

prin

t. H

ouse

hold

s w

ith w

ater

hea

ter

tem

pera

ture

s ≤1

25 d

egre

es C

(≤

 50

degr

ees

C) w

ere

sign

ifica

ntly

mor

e lik

ely

to h

arbo

r N

TM

com

pare

d w

ith h

ouse

hold

s w

ith h

ot w

ater

tem

pera

ture

s ≥

130

degr

ees

F (≥

55

degr

ees

C) (

p =

0.01

07).

Alth

ough

hou

seho

lds

with

wat

er fr

om p

ublic

or

priv

ate

wat

er s

yste

ms

serv

ing

mul

tiple

hou

seho

lds

wer

e m

ore

likel

y to

hav

e N

TM

(19/

27, 7

0%) c

ompa

red

with

hou

seho

lds

with

a w

ell p

rovi

ding

wat

er to

on

ly 1

 hou

seho

ld (5

/12,

42%

), th

at d

iffer

ence

was

not

sig

nific

ant (

p =

0.15

32).

Falk

in-

ham

, III

20

11

Dri

nkin

g w

ater

M. a

vium

com

plex

Myc

obac

teri

um a

vium

com

plex

(MA

C) i

s a

grou

p of

opp

ortu

nist

ic p

atho

gens

of m

ajor

pub

lic h

ealth

con

cern

. It i

s re

spon

sibl

e fo

r a

wid

e sp

ectr

um o

f dis

ease

dep

ende

nt o

n su

bspe

cies

, rou

te o

f inf

ectio

n an

d pa

tient

s pr

e-ex

istin

g co

nditi

ons.

Pre

sent

ly,

ther

e is

lim

ited

rese

arch

on

the

inci

denc

e of

MA

C in

fect

ion

that

con

side

rs b

oth

pulm

onar

y an

d ot

her

clin

ical

man

ifest

atio

ns.

MA

C h

as b

een

isol

ated

from

var

ious

terr

estr

ial a

nd a

quat

ic e

nvir

onm

ents

incl

udin

g na

tura

l wat

ers,

eng

inee

red

wat

er s

yste

ms

and

soils

. Ide

ntify

ing

the

spec

ific

envi

ronm

enta

l sou

rces

resp

onsi

ble

for

hum

an in

fect

ion

is e

ssen

tial i

n m

inim

izin

g di

seas

e pr

eval

ence

. Thi

s pa

per

revi

ews

curr

ent l

itera

ture

and

cas

e st

udie

s re

gard

ing

the

wid

e sp

ectr

um o

f dis

ease

cau

sed

by M

AC

an

d th

e ro

le o

f pot

able

wat

er in

dis

ease

tran

smis

sion

. Pot

able

wat

er w

as re

cogn

ized

as

a pu

tativ

e pa

thw

ay fo

r M

AC

infe

ctio

n.

Con

tam

inat

ed p

otab

le w

ater

sou

rces

ass

ocia

ted

with

hum

an in

fect

ion

incl

uded

war

m w

ater

dis

trib

utio

n sy

stem

s, s

how

ers,

fa

ucet

s, h

ouse

hold

dri

nkin

g w

ater

, sw

imm

ing

pool

s an

d ho

t tub

spa

s. M

AC

can

mai

ntai

n lo

ng-t

erm

con

tam

inat

ion

of p

otab

le

wat

er s

ourc

es th

roug

h its

hig

h re

sist

ance

to d

isin

fect

ants

, ass

ocia

tion

with

bio

film

s an

d in

trac

ellu

lar

para

sitis

m o

f fre

e-liv

ing

prot

ozoa

. Fur

ther

rese

arch

is re

quir

ed to

inve

stig

ate

the

effic

ienc

y of

wat

er tr

eatm

ent p

roce

sses

aga

inst

MA

C a

nd in

to c

on-

stru

ctio

n an

d m

aint

enan

ce o

f war

m w

ater

dis

trib

utio

n sy

stem

s an

d th

e ro

le th

ey p

lay

in M

AC

pro

lifer

atio

n.

Whi

ley

et

al. 2

012

Dri

nkin

g w

ater

M. a

. par

atub

ercu

losi

sM

ycob

acte

rium

avi

um s

ubsp

. par

atub

ercu

losi

s (M

ap) i

s th

e ca

use

of Jo

hne’s

dis

ease

, a c

hron

ic in

fect

ion

of th

e gu

t, in

rum

i-na

nt a

nim

als

that

pro

vide

milk

and

/or

mea

t for

hum

an c

onsu

mpt

ion.

Map

als

o m

ay b

e in

volv

ed in

Cro

hn’s

dise

ase

and

type

I d

iabe

tes

in h

uman

s. A

lthou

gh th

e ro

le o

f Map

in h

uman

dis

ease

s ha

s no

t bee

n es

tabl

ishe

d, m

inim

izin

g th

e ex

posu

re o

f hu

man

s to

the

orga

nism

is c

onsi

dere

d de

sira

ble

as a

pre

caut

iona

ry m

easu

re. I

nfec

ted

anim

als

can

shed

Map

in fe

ces

and

milk

, an

d th

e or

gani

sm c

an b

ecom

e di

ssem

inat

ed in

tiss

ues

rem

ote

from

the

gut a

nd it

s as

soci

ated

lym

ph n

odes

. The

pre

senc

e of

at

leas

t som

e M

ap in

raw

milk

and

mea

t and

in n

atur

al w

ater

s is

like

ly, b

ut th

e nu

mbe

rs o

f Map

in th

ose

food

s an

d w

ater

s sh

ould

be

redu

ced

thro

ugh

cook

ing

or p

urifi

catio

n. T

he a

vaila

ble

info

rmat

ion

rela

ting

to M

ap in

milk

and

dai

ry p

rodu

cts,

mea

ts, a

nd

drin

king

wat

er is

revi

ewed

her

e fo

r as

sess

men

t of t

he r

isks

of e

xpos

ure

to M

ap fr

om c

onsu

mpt

ion

of s

uch

food

s an

d w

ater

.

Gill

et a

l. 20

11

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

645

Dri

nkin

g w

ater

In th

is st

udy,

we

enla

rged

our

pre

viou

s inv

estig

atio

n fo

cusi

ng o

n th

e bi

odiv

ersi

ty o

f chl

amyd

iae

and

amoe

bae

in a

dri

nkin

g w

ater

tr

eatm

ent p

lant

, by

the

incl

usio

n of

two

addi

tiona

l pla

nts a

nd b

y se

arch

ing

also

for t

he p

rese

nce

of le

gion

ella

e an

d m

ycob

acte

ria.

A

utoc

htho

nous

am

oeba

e w

ere

reco

vere

d on

to n

on-n

utri

tive

agar

, ide

ntifi

ed b

y 18

S rR

NA

gen

e se

quen

cing

, and

scre

ened

for t

he

pres

ence

of b

acte

rial

end

osym

bion

ts. B

acte

ria

wer

e al

so se

arch

ed fo

r by

Acan

tham

oeba

co-

cultu

re. F

rom

a to

tal o

f 125

sam

ples

, w

e re

cove

red

38 a

moe

bae,

am

ong

whi

ch si

x ha

rbou

red

endo

sym

bion

ts (t

hree

chl

amyd

iae

and

thre

e le

gion

ella

e). I

n ad

ditio

n, w

e re

cove

red

by a

moe

bal c

o-cu

lture

11

chla

myd

iae,

36

legi

onel

lae

(no

L. p

neum

ophi

la),

and

24 m

ycob

acte

ria

(all

rapi

d-gr

ower

s). T

wo

plan

ts p

rese

nted

a si

mila

r per

cent

age

of sa

mpl

es p

ositi

ve fo

r chl

amyd

iae

(11%

), m

ycob

acte

ria

(20%

) and

am

oeba

e (2

7%),

whe

reas

in

the

third

pla

nt th

e nu

mbe

r of r

ecov

ered

bac

teri

a w

as a

lmos

t tw

ice

high

er. E

ach

plan

t exh

ibite

d a

rela

tivel

y hi

gh sp

ecifi

c m

icro

-bi

ota.

Am

oeba

e w

ere

mai

nly

repr

esen

ted

by v

ario

us N

aegl

eria

spec

ies,

Acan

tham

oeba

spec

ies a

nd H

artm

anne

lla v

erm

iform

is.

Para

chla

myd

iace

ae w

ere

the

mos

t abu

ndan

t chl

amyd

iae

(8 st

rain

s in

tota

l), a

nd in

this

stud

y w

e re

cove

red

a ne

w g

enus

-leve

l st

rain

, alo

ng w

ith n

ew c

hlam

ydia

e pr

evio

usly

repo

rted

. Sim

ilarly

, abo

ut 6

6% o

f the

reco

vere

d le

gion

ella

e an

d 47

% o

f the

isol

ated

m

ycob

acte

ria

coul

d re

pres

ent n

ew sp

ecie

s. O

ur w

ork

high

light

ed a

hig

h sp

ecie

s div

ersi

ty a

mon

g le

gion

ella

e an

d m

ycob

acte

ria,

do

min

ated

by

puta

tive

new

spec

ies,

and

it co

nfirm

ed th

e pr

esen

ce o

f chl

amyd

iae

in th

ese

artifi

cial

wat

er sy

stem

s.

Cor

saro

et

al.

2010

Dri

nkin

g w

ater

Dat

a on

the

occu

rren

ce o

f non

-tub

ercu

lous

myc

obac

teri

a (N

TM

), in

par

alle

l with

thos

e ob

tain

ed fo

r bac

teri

al in

dica

tors

and

am

oeba

e, a

re p

rese

nted

with

the

aim

to c

olle

ct in

form

atio

n on

the

spre

ad o

f NT

M in

dri

nkin

g w

ater

dis

trib

utio

n sy

stem

s in

Ital

y. Sa

mpl

es w

ere

colle

cted

from

taps

of h

ospi

tals

and

hou

seho

lds i

n C

entr

al a

nd S

outh

ern

Ital

y. Th

e co

ncen

trat

ion

valu

es o

btai

ned

for t

he m

ore

trad

ition

al m

icro

bial

par

amet

ers c

ompl

ied

with

the

man

dato

ry re

quire

men

ts fo

r dri

nkin

g w

ater

. Con

vers

ely,

mod

erat

e-to

-hig

h m

icro

bial

load

s (til

l 300

CFU

/L) w

ere

obse

rved

for t

he N

TM

. Pos

itive

sam

ples

wer

e ob

tain

ed fr

om 6

2% o

f the

in

vest

igat

ed w

ater

sam

ples

. Ana

logo

us re

sults

wer

e ob

serv

ed fo

r am

oeba

e sh

owin

g a

high

er p

erce

ntag

e of

pos

itive

sam

ples

(76%

). In

term

s of p

ublic

hea

lth, t

he p

rese

nce

of m

ycob

acte

ria

in w

ater

dis

trib

utio

n sy

stem

s may

repr

esen

t a p

oten

tial r

isk

espe

cial

ly fo

r vu

lner

able

peo

ple

such

as c

hild

ren,

the

elde

rly o

r im

mun

ocom

prom

ised

indi

vidu

als.

Bria

n-ce

sco

et

al. 2

010

Dri

nkin

g w

ater

Legi

onel

la a

nd M

ycob

acte

rium

can

pro

lifer

ate

with

in fr

ee-li

ving

am

oeba

e (F

LA) w

here

they

are

pro

tect

ed fr

om d

isin

fect

ants

at

conc

entr

atio

ns th

at c

an k

ill b

acte

ria

but n

ot p

roto

zoa.

Des

pite

effe

ctiv

e tr

eatm

ent o

f dri

nkin

g w

ater

, mic

robe

s can

ent

er w

ater

ut

ility

dis

trib

utio

n sy

stem

s (D

S) a

nd h

ence

the

plum

bing

with

in b

uild

ing

prem

ises

. Add

ition

ally

, bio

film

form

atio

n m

ay a

ccou

nt

for t

he p

ersi

sten

ce o

f mic

robe

s in

the

DS.

In th

e pr

esen

t stu

dy a

dom

estic

wat

er ta

p in

nor

th-c

entr

al U

nite

d St

ates

(USA

) was

sa

mpl

ed in

Mar

ch a

nd S

epte

mbe

r 200

7 an

d an

alys

ed fo

r FLA

, Leg

ione

lla a

nd M

ycob

acte

rium

. Ide

ntifi

catio

n of

org

anis

ms w

as

dete

rmin

ed b

y gr

owth

on

spec

ific

cultu

re m

edia

, lig

ht a

nd e

lect

ron

mic

rosc

opy,

and

ampl

ifica

tion

of D

NA

pro

bes s

peci

fic fo

r ea

ch o

rgan

ism

. In

both

the

spri

ng a

nd fa

ll sa

mpl

es, a

moe

bae,

Leg

ione

lla a

nd M

ycob

acte

rium

wer

e de

tect

ed. H

owev

er, A

can-

tham

oeba

was

pro

min

ent i

n th

e sp

ring

sam

ple

whe

reas

Vah

lkam

pfia

and

Nae

gler

ia w

ere

the

amoe

bae

dete

cted

in th

e au

tum

n.

Bact

eria

l pro

lifer

atio

n in

labo

rato

ry c

ultu

res w

as n

otic

eabl

y en

hanc

ed in

the

pres

ence

of a

moe

bae

and

biofi

lms r

apid

ly fo

rmed

in

mix

ed a

moe

bae

and

bact

eria

cul

ture

s. It

is h

ypot

hesi

zed

that

tem

pera

ture

affe

cted

the

dyna

mic

s of F

LA sp

ecie

s pop

ulat

ion

stru

ctur

e w

ithin

the

DS

and

that

pat

hoge

nic

bact

eria

that

pro

lifer

ate

with

in F

LA, w

hich

are

them

selv

es o

ppor

tuni

stic

pat

hoge

ns,

pose

dua

l pub

lic h

ealth

risk

s.

Mar

-ci

ano-

Cab

ral e

t al

. 201

0

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

646

Am

oeba

mic

robi

al p

atho

gens

Free

-livi

ng a

moe

bae

that

bel

ong

to th

e ge

nus A

cant

ham

oeba

are

wid

espr

ead

in th

e en

viro

nmen

t, in

clud

ing

wat

er. Th

ey a

re

resp

onsi

ble

for h

uman

infe

ctio

ns a

nd c

an h

ost p

atho

geni

c m

icro

orga

nism

s. U

nder

unf

avor

able

con

ditio

ns, t

hey

form

cys

ts w

ith

high

leve

ls o

f res

ista

nce

to d

isin

fect

ion

met

hods

, thu

s pot

entia

lly re

pres

entin

g a

thre

at to

pub

lic h

ealth

. In

the

pres

ent s

tudy

we

eval

uate

d th

e effi

caci

es o

f var

ious

bio

cide

s aga

inst

trop

hozo

ites a

nd c

ysts

of s

ever

al A

cant

ham

oeba

stra

ins.

We

dem

onst

rate

d th

at d

isin

fect

ant e

ffica

cy v

arie

d de

pend

ing

on th

e st

rain

s tes

ted,

with

env

iron

men

tal s

trai

ns d

emon

stra

ting

grea

ter r

esis

tanc

e th

an c

olle

ctio

n st

rain

s. T

roph

ozoi

tes w

ere

inac

tivat

ed b

y al

l tre

atm

ents

exc

ept t

hose

usi

ng g

luta

rald

ehyd

e as

an

activ

e co

m-

poun

d: fo

r the

se tr

eatm

ents

, we

obse

rved

resi

stan

ce e

ven

afte

r 30

min

exp

osur

e. C

ysts

resi

sted

man

y tr

eatm

ents

, inc

ludi

ng

cert

ain

cond

ition

s with

glu

tara

ldeh

yde

and

othe

r bio

cide

s. M

oist

hea

t at 5

5 de

gree

s C w

as n

ot e

ffici

ent a

gain

st c

ysts

, whe

reas

ex

posu

re a

t 65

degr

ees C

was

. Sev

eral

che

mic

al fo

rmul

atio

ns c

onta

inin

g pe

race

tic a

cid,

hyd

roge

n pe

roxi

de, o

r ort

ho-p

htha

lald

e-hy

de p

rese

nted

gre

ater

effi

cacy

than

glu

tara

ldeh

yde,

as d

id e

than

ol a

nd so

dium

hyp

ochl

orite

; how

ever

, som

e of

thes

e tr

eatm

ents

re

quir

ed re

lativ

ely

long

incu

batio

n tim

es to

ach

ieve

cys

t ina

ctiv

atio

n. A

moe

bal c

ysts

can

be

high

ly re

sist

ant t

o so

me

high

-leve

l di

sinf

ecta

nts,

whi

ch h

as im

plic

atio

ns fo

r clin

ical

pra

ctic

e. Th

ese

resu

lts h

ighl

ight

the

need

to c

onsi

der t

he e

ffect

ive

disi

nfec

tion

of p

roto

zoa

in th

eir v

eget

ativ

e an

d re

sist

ant f

orm

s due

to th

eir i

ntri

nsic

resi

stan

ce. Th

is is

impo

rtan

t not

onl

y to

pre

vent

the

tran

smis

sion

of p

roto

zoa

them

selv

es b

ut a

lso

due

to th

e ri

sks a

ssoc

iate

d w

ith a

rang

e of

mic

robi

al p

atho

gens

that

are

foun

d to

be

asso

ciat

ed in

trac

ellu

larl

y w

ith th

ese

mic

roor

gani

sms.

Cou

lon

et

al. 2

010

Dri

nkin

g w

ater

Ther

e is

an

expa

ndin

g bo

dy o

f evi

denc

e th

at fr

ee-li

ving

am

oeba

e (F

LA) i

ncre

ase

both

the

num

bers

and

vir

ulen

ce o

f wat

er-b

ased

, hu

man

-pat

hoge

nic,

am

oeba

-res

istin

g m

icro

orga

nism

s (A

RM).

Legi

onel

la sp

p., M

ycob

acte

rium

spp.

, and

oth

er o

ppor

tuni

stic

hu

man

pat

hoge

ns a

re k

now

n to

be

both

ARM

and

als

o th

e et

iolo

gic

agen

ts o

f pot

entia

lly fa

tal h

uman

lung

infe

ctio

ns. H

owev

er,

com

para

tivel

y lit

tle is

kno

wn

abou

t the

FLA

that

may

faci

litat

e A

RM g

row

th in

dri

nkin

g w

ater

. This

revi

ew e

xam

ines

the

avai

labl

e lit

erat

ure

on F

LA in

trea

ted

drin

king

wat

er sy

stem

s; in

tota

l 26

stud

ies f

rom

18

diffe

rent

cou

ntri

es. F

LA w

ere

repo

rted

to b

reak

-th

roug

h th

e w

ater

trea

tmen

t bar

rier

and

ent

er d

istr

ibut

ion

syst

ems,

in a

dditi

on to

the

expe

cted

pos

t-tr

eatm

ent s

yste

m in

gres

s. O

nce

in th

e di

stri

butio

n sy

stem

ther

e is

evi

denc

e of

FLA

col

oniz

atio

n an

d re

grow

th e

spec

ially

in re

serv

oirs

and

in-p

rem

ise

plum

b-in

g st

orag

e ta

nks.

At t

he p

oint

of u

se th

e av

erag

e FL

A d

etec

tion

rate

was

45%

but

hig

hly

vari

able

(n =

16,

sigm

a =

31) d

ue to

bot

h di

ffere

nces

in b

oth

assa

y m

etho

ds a

nd th

e ty

pe o

f wat

er sy

stem

s exa

min

ed. Th

is re

view

reve

als t

hat F

LA a

re c

onsi

sten

tly d

etec

ted

in tr

eate

d dr

inki

ng w

ater

syst

ems a

roun

d th

e w

orld

and

pre

sent

a y

et u

nqua

ntifi

ed e

mer

ging

hea

lth ri

sk. H

owev

er, m

ore

rese

arch

is

urge

ntly

requ

ired

befo

re a

ccur

ate

risk

s ass

essm

ents

can

be

unde

rtak

en to

ass

ess t

he im

pact

s on

hum

an h

ealth

, in

hous

ehol

ds a

nd

inst

itutio

ns, d

ue to

exp

osur

e to

FLA

faci

litat

ed p

atho

geni

c A

RM.

Thom

as

and

Ash

-bo

lt 20

11

Dri

nkin

g w

ater

Cul

ture

-bas

ed m

etho

ds fo

r fec

al in

dica

tor m

icro

orga

nism

s are

the

stan

dard

pro

toco

l to

asse

ss p

oten

tial h

ealth

risk

from

dri

nk-

ing

wat

er sy

stem

s. H

owev

er, t

hese

trad

ition

al fe

cal i

ndic

ator

s are

inap

prop

riat

e su

rrog

ates

for d

isin

fect

ion-

resi

stan

t fec

al p

atho

-ge

ns a

nd th

e in

dige

nous

pat

hoge

ns th

at g

row

in d

rink

ing

wat

er sy

stem

s. Th

ere

is n

ow a

rang

e of

mol

ecul

ar-b

ased

met

hods

, suc

h as

qua

ntita

tive

PCR

, whi

ch a

llow

det

ectio

n of

a v

arie

ty o

f pat

hoge

ns a

nd a

ltern

ativ

e in

dica

tors

. Hen

ce, i

n ad

ditio

n to

targ

etin

g to

tal E

sche

rich

ia c

oli (

i.e.,

dead

and

aliv

e) fo

r the

det

ectio

n of

feca

l pol

lutio

n, v

ario

us a

moe

bae

may

be

suita

ble

to in

dica

te th

e po

tent

ial p

rese

nce

of p

atho

geni

c am

oeba

-res

istin

g m

icro

orga

nism

s, su

ch a

s Leg

ione

llae.

Ther

efor

e, m

onito

ring

am

oeba

leve

ls

by q

uant

itativ

e PC

R co

uld

be a

use

ful t

ool f

or d

irec

tly a

nd in

dire

ctly

eva

luat

ing

heal

th ri

sk a

nd c

ould

als

o be

a c

ompl

emen

tary

ap

proa

ch to

cur

rent

mic

robi

al q

ualit

y co

ntro

l str

ateg

ies f

or d

rink

ing

wat

er sy

stem

s.

Cod

ony

et

al. 2

012

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

647

Dri

nkin

g w

ater

M. m

ucog

enic

umO

bjec

tive

to b

ette

r und

erst

and

the

mec

hani

sm o

f chl

orin

e re

sist

ance

of m

ycob

acte

ria

and

eval

uate

the

effici

ency

of v

ario

us

disi

nfec

tion

proc

esse

s. M

etho

ds In

activ

atio

n ex

peri

men

ts o

f one

stra

in M

ycob

acte

ria

muc

ogen

icum

, iso

late

d fr

om a

dri

nkin

g w

ater

dis

trib

utio

n sy

stem

in S

outh

Chi

na w

ere

cond

ucte

d w

ith v

ario

us c

hlor

ine

disi

nfec

tant

s. In

activ

atio

n effi

cien

cy a

nd d

is-

infe

ctan

t res

idua

l, as

wel

l as t

he fo

rmat

ion

of o

rgan

ic c

hlor

amin

es, w

ere

mea

sure

d du

ring

the

expe

rim

ents

. Res

ults

: This

stra

in

of M

. muc

ogen

icum

show

ed h

igh

resi

stan

ce to

chl

orin

e. Th

e C

T v

alue

s of 9

9.9%

inac

tivat

ion

by fr

ee c

hlor

ine,

mon

ochl

oram

ine

and

chlo

rine

dio

xide

wer

e de

tect

ed a

s 29.

6 ±

1.46

, 170

± 6

.16,

and

10.

9 ±

1.55

min

.(mg/

L) re

spec

tivel

y, in

dica

ting

that

chl

orin

e di

oxid

e ex

hibi

ted

sign

ifica

ntly

hig

her e

ffici

ency

than

free

chl

orin

e an

d m

onoc

hlor

amin

e. It

was

als

o fo

und

that

M. m

ucog

enic

um

reac

ted

with

chl

orin

e di

sinf

ecta

nts m

ore

slow

ly th

an S

. aur

eus,

but c

onsu

med

mor

e ch

lori

ne d

isin

fect

ants

dur

ing

long

er ti

me

of

cont

act.

Lipi

d an

alys

is o

f the

cel

l con

stru

ctio

n re

veal

ed th

at 9

5.7%

of c

ell m

embr

ane

lipid

of M

. muc

ogen

icum

was

com

pose

d of

sa

tura

ted

long

cha

in fa

tty

acid

s. S

atur

ated

fatt

y ac

ids w

ere

rega

rded

as m

ore

stab

le a

nd m

ore

hydr

ophi

lic w

hich

ena

bled

the

cell

mem

bran

e to

pre

vent

the

diffu

sion

of c

hlor

ine.

Con

clus

ion

It w

as c

oncl

uded

that

diff

eren

t com

posi

tions

of c

ell m

embr

ane

mig

ht

endo

w M

. muc

ogen

icum

with

a h

ighe

r chl

orin

e re

sist

ance

.

Che

n et

al

. 201

2

Ther

apy

pool

wat

erM

. sp.

...w

e co

nduc

ted

a m

ultis

easo

n su

rvey

of m

icro

orga

nism

s pr

esen

t in

this

ther

apy

pool

wat

er, i

n bi

ofilm

s as

soci

ated

with

the

pool

con

tain

men

t wal

ls, a

nd in

air

imm

edia

tely

abo

ve th

e po

ol. T

he s

urve

y us

ed c

ultu

re, m

icro

scop

y, a

nd c

ultu

re-i

ndep

end-

ent m

olec

ular

phy

loge

netic

ana

lyse

s. A

lthou

gh o

utfit

ted

with

a s

tate

-of-

the

art U

V-p

erox

ide

disi

nfec

tion

syst

em, t

he n

umbe

rs

of b

acte

ria

in th

e th

erap

y po

ol w

ater

wer

e re

lativ

ely

high

com

pare

d w

ith th

e po

tabl

e w

ater

use

d to

fill

the

pool

. Reg

ardl

ess

of

the

sour

ce, d

irec

t mic

rosc

opic

cou

nts

of m

icro

bes

wer

e ro

utin

ely

1,00

0 tim

es g

reat

er th

an c

onve

ntio

nal p

late

cou

nts.

Ana

ly-

sis

of c

lone

libr

arie

s of

sm

all s

ubun

it rR

NA

gen

es fr

om e

nvir

onm

enta

l DN

A p

rovi

ded

phyl

ogen

etic

div

ersi

ty e

stim

ates

of t

he

mic

roor

gani

sms

colle

cted

in a

nd a

bove

the

pool

. A s

urve

y of

>1,

300

rRN

A g

enes

yie

lded

a to

tal o

f 628

uni

que

sequ

ence

s, th

e m

ost c

omm

on o

f whi

ch w

as n

earl

y id

entic

al to

that

of M

. avi

um s

trai

ns. T

he h

igh

prop

ortio

n of

clo

nes

with

diff

eren

t Myc

o-ba

cter

ium

spp

. rRN

A g

enes

sug

gest

ed th

at s

uch

orga

nism

s co

mpr

ised

a s

igni

fican

t fra

ctio

n of

mic

robe

s in

the

pool

wat

er (t

o >3

0%) a

nd p

refe

rent

ially

par

titio

n in

to a

eros

ols

(to

>80%

) rel

ativ

e to

oth

er w

ater

born

e ba

cter

ia p

rese

nt.

Ang

enen

t et

al.

2005

Hot

tub

aero

sol

M. s

p.H

uman

act

iviti

es a

ssoc

iate

d w

ith a

eros

ol-g

ener

atin

g ho

t wat

er so

urce

s are

incr

easi

ngly

pop

ular

. Rec

ently

, a h

yper

sens

itivi

ty

pneu

mon

itis (

HP)

-like

gra

nulo

mat

ous l

ung

dise

ase,

with

non

-tub

ercu

lous

myc

obac

teri

a fr

om e

xpos

ure

to h

ot w

ater

aer

osol

s fr

om h

ot tu

bs/s

pas,

show

ers,

and

indo

or sw

imm

ing

pool

s, ha

s bee

n de

scri

bed

in im

mun

ocom

pete

nt in

divi

dual

s (al

so c

alle

d “h

ot

tub

lung

”). O

ur o

bjec

tive

in th

is st

udy

was

to e

xam

ine

four

add

ition

al c

ases

of h

ot tu

b lu

ng a

nd c

ompa

re th

ese

case

s with

oth

ers

repo

rted

in th

e En

glis

h pr

int l

itera

ture

on

this

dis

ease

. We

retr

ospe

ctiv

ely

revi

ewed

all

case

s (n

= 4)

of p

resu

mpt

ivel

y di

agno

sed

hot t

ub lu

ng in

imm

unoc

ompe

tent

indi

vidu

als a

t the

var

ious

phy

sici

an p

ract

ices

in S

prin

gfiel

d, Il

linoi

s, du

ring

200

1–20

05. I

n ad

ditio

n, w

e se

arch

ed M

EDLI

NE

for c

ases

of h

ot tu

b lu

ng d

escr

ibed

in th

e lit

erat

ure.

We

sum

mar

ized

the

clin

ical

pre

sent

atio

n an

d in

vest

igat

ions

of f

our p

resu

mpt

ive

case

s and

revi

ewed

pre

viou

sly re

port

ed c

ases

of h

ot tu

b lu

ng. Th

ere

is a

deb

ate

in th

e lit

erat

ure

whe

ther

hot

tub

lung

is a

n H

P or

a d

irec

t inf

ectio

n of

the

lung

by

nont

uber

culo

us m

ycob

acte

ria.

Pri

mar

y pr

even

tion

of th

is d

isea

se re

lies o

n ve

ntila

tion

and

good

use

pra

ctic

es. S

econ

dary

pre

vent

ion

of th

is d

isea

se re

quire

s edu

catio

n of

bot

h th

e ge

nera

l pub

lic a

nd c

linic

ians

to a

llow

for t

he e

arly

dia

gnos

is o

f thi

s dis

ease

.

Sood

et

al. 2

007

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

648

Hot

tubs

M. a

vium

To a

sses

s th

e cu

rren

t spe

ctru

m o

f cau

ses

and

clin

ical

feat

ures

ass

ocia

ted

with

hyp

erse

nsiti

vity

pne

umon

itis (

HP)

. We

stud

ied

cons

ecut

ive

patie

nts

with

HP

diag

nose

d at

the

May

o C

linic

in R

oche

ster

, Min

n, fr

om Ja

nuar

y 1,

199

7, th

roug

h D

ecem

ber 

31,

2002

. Dia

gnos

tic c

rite

ria

for

HP

incl

uded

the

follo

win

g: (1

) pre

senc

e of

resp

irat

ory

sym

ptom

s, (2

) rad

iolo

gic

evid

ence

of

diff

use

lung

dis

ease

, (3)

kno

wn

expo

sure

or

a po

sitiv

e se

rolo

gic

test

resu

lt to

an

inci

ting

antig

en, a

nd (4

) no

othe

r id

entif

i-ab

le c

ause

for

the

lung

dis

ease

. If t

here

was

no

iden

tifia

ble

inci

ting

antig

en, I

of t

he fo

llow

ing

2 cr

iteri

a w

as re

quir

ed: (

1) lu

ng

biop

sy s

peci

men

that

dem

onst

rate

d fe

atur

es o

f HP

or (2

) bro

ncho

alve

olar

lava

ge ly

mph

ocyt

osis

and

hig

h-re

solu

tion

com

-pu

ted

tom

ogra

phic

evi

denc

e of

gro

und-

glas

s op

aciti

es o

r ce

ntri

lobu

lar

nodu

les

bila

tera

lly. T

he m

eant

SO

age

of t

he 8

5 st

udy

patie

nts

was

53

± 14

yea

rs; 5

3 pa

tient

s (6

2%) w

ere

wom

en. O

nly

2 pa

tient

s (2

%) w

ere

curr

ent s

mok

ers.

Chr

onic

(A m

onth

s)

resp

irat

ory

sym

ptom

s w

ere

pres

ent i

n 66

pat

ient

s (7

8%).

His

topa

thol

ogic

con

firm

atio

n w

as o

btai

ned

in 6

4 pa

tient

s (7

5%).

The

cau

se w

as id

entif

ied

in 6

4 pa

tient

s (7

5%),

and

the

mos

t com

mon

cau

ses

wer

e av

ian

anti-

gens

(34%

) and

Myc

obac

teri

um

aviu

m c

ompl

ex in

hot

tub

wat

er (2

1%).

Farm

er’s

lung

dis

ease

acc

ount

ed fo

r 3.

1% o

f cas

es, a

nd a

n ad

ditio

nal 9

% w

ere

rela

ted

to h

ouse

hold

mol

d ex

posu

re. T

he in

citin

g an

tigen

was

not

iden

tifia

ble

in 2

5% o

f pat

ient

s. M

ost p

atie

nts

with

HP

seen

at t

his

tert

iary

car

e re

ferr

al c

ente

r in

the

Mid

wes

t reg

ion

of th

e U

nite

d St

ates

had

chr

onic

HIP

, and

the

mos

t com

mon

cau

ses w

ere

expo

sure

to b

irds

and

exp

osur

e to

hot

tubs

.

Han

ak e

t al

. 200

7

Hot

tubs

M. a

vium

The

obje

ctiv

e of

our

stud

y w

as to

des

crib

e th

e C

T fe

atur

es o

f “ho

t tub

lung

” cau

sed

by e

xpos

ure

to M

ycob

acte

rium

avi

um c

om-

plex

(MA

C) o

rgan

ism

s in

cont

amin

ated

wat

er C

hart

revi

ew w

as p

erfo

rmed

to id

entif

y al

l pat

ient

s with

a h

isto

logi

c di

agno

sis

of g

ranu

lom

atou

s pne

umon

itis a

nd p

ositi

ve c

ultu

res f

or M

AC

bet

wee

n Ja

nuar

y 1,

199

5, a

nd Ju

ly 1

, 200

4. In

divi

dual

s ide

ntifi

ed

who

als

o ha

d a

hot t

ub w

ere

incl

uded

in th

e st

udy.

Twel

ve p

atie

nts,

seve

n fe

mal

es a

nd fi

ve m

ales

with

an

aver

age

age

of 5

0 ye

ars

(ran

ge, 1

3–66

yea

rs),

who

had

a C

T sc

an w

ere

iden

tified

. The

CT

imag

es w

ere

revi

ewed

by

two

thor

acic

radi

olog

ists

who

as

sess

ed th

e im

ages

for t

he p

rese

nce

of a

ny p

aren

chym

al a

bnor

mal

ities

, inc

ludi

ng n

odul

es, a

reas

of g

roun

d-gl

ass a

tten

uatio

n,

retic

ular

opa

citie

s, a

nd a

ir tr

appi

ng, o

n ex

pira

tory

imag

es. W

hen

nodu

les,

retic

ular

opa

citie

s, a

reas

of g

roun

d-gl

ass a

tten

uatio

n,

or a

com

bina

tion

of th

ese

findi

ngs w

as p

rese

nt, t

he re

view

ers v

isua

lly d

eter

min

ed th

e ex

tent

of i

nvol

vem

ent o

f the

lung

s usi

ng

a sc

ale

of <

10%

, 10–

40%

, or >

40%

. They

als

o re

cord

ed th

e di

stri

butio

n of

the

invo

lvem

ent b

oth

ceph

aloc

auda

l and

tran

saxi

al.

Dec

isio

ns w

ere

reac

hed

by c

onse

nsus

of t

he re

view

ers.

Nod

ules

wer

e pr

esen

t in

10 (8

3%) o

f 12

patie

nts.

In e

ight

(80%

) of

10 p

atie

nts,

the

nodu

les w

ere

diffu

se w

ith a

cen

trilo

bula

r dis

trib

utio

n. In

the

othe

r tw

o, th

e no

dule

s wer

e ra

ndom

ly d

istr

ibut

ed

with

an

uppe

r lun

g pr

edom

inan

ce. I

n fiv

e pa

tient

s the

nod

ules

show

ed a

reas

of g

roun

d-gl

ass a

tten

uatio

n, w

here

as in

the

othe

r fiv

e th

e no

dule

s wer

e so

lid. A

reas

of g

roun

d-gl

ass a

tten

uatio

n w

ere

pres

ent i

n ei

ght (

75%

) of 1

2 pa

tient

s and

wer

e bi

late

ral i

n al

l ca

ses.

The

area

s of g

roun

d-gl

ass a

tten

uatio

n w

ere

diffu

se in

the

ceph

aloc

auda

l pla

ne w

ith a

rand

om d

istr

ibut

ion

in th

e tr

ansa

xial

pl

ane

in se

ven

(88%

) of e

ight

cas

es. I

n th

e re

mai

ning

cas

e, th

e ar

eas o

f gro

und-

glas

s att

enua

tion

had

a lo

wer

lung

pre

dom

inan

ce

with

a ra

ndom

dis

trib

utio

n in

the

tran

saxi

al p

lane

. Exp

irat

ory

imag

es sh

owed

evi

denc

e of

air

trap

ping

in a

ll se

ven

case

s for

w

hich

thes

e im

ages

wer

e av

aila

ble.

In o

ne p

atie

nt, a

ir tr

appi

ng w

as th

e on

ly a

bnor

mal

ity id

entifi

ed. Th

e C

T fi

ndin

gs in

pat

ient

s w

ith h

ot tu

b lu

ng in

clud

e ar

eas o

f gro

und-

glas

s att

enua

tion,

cen

trilo

bula

r nod

ules

, and

air

trap

ping

on

expi

rato

ry im

ages

. Thes

e fin

ding

s are

sim

ilar t

o pr

evio

usly

pub

lishe

d fin

ding

s of s

ubac

ute

hype

rsen

sitiv

ity p

neum

oniti

s. Th

eref

ore,

in c

ases

in w

hich

CT

fin

ding

s sug

gest

hyp

erse

nsiti

vity

pne

umon

itis,

hot

tub

lung

shou

ld a

lso

be a

dia

gnos

tic c

onsi

dera

tion.

Har

tman

et

al.

2007

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

649

Show

er w

ater

and

aer

osol

M. s

p.M

. muc

ogen

icum

M. p

hoca

icum

To q

uant

ify th

e m

icro

bial

load

in sh

ower

wat

er a

nd a

eros

ol sa

mpl

es, w

e us

ed c

ultu

re, m

icro

scop

ic, a

nd q

uant

itativ

e PC

R m

eth-

ods

to in

vest

igat

e fo

ur s

how

er s

talls

in a

ste

m c

ell t

rans

plan

t uni

t at B

arne

s-Je

wis

h H

ospi

tal i

n St

. Lou

is, M

O. W

e al

so te

sted

m

embr

ane-

inte

grat

ed s

how

erhe

ads

as a

pos

sibl

e m

itiga

tion

stra

tegy

. In

addi

tion

to q

uant

ifica

tion,

a 1

6S r

RNA

gen

e se

quen

c-in

g su

rvey

was

use

d to

cha

ract

eriz

e th

e ab

unda

nt b

acte

rial

pop

ulat

ions

with

in s

how

er w

ater

and

aer

osol

s. T

he a

vera

ge to

tal

bact

eria

l cou

nts

wer

e 2.

2 ×

107 c

ells

/lite

r in

sho

wer

wat

er a

nd 3

.4 ×

104 c

ells

/m3 in

sho

wer

aer

osol

, and

thes

e co

unts

wer

e re

duce

d to

6.3

× 1

04 cel

ls/l

iter

(99.

6% e

ffic

ienc

y) a

nd 8

.9 ×

103 c

ells

/m3 (8

2.4%

eff

icie

ncy)

, res

pect

ivel

y, a

fter

mem

bran

e-in

tegr

ated

sho

wer

head

s w

ere

inst

alle

d. P

oten

tially

pat

hoge

nic

orga

nism

s w

ere

foun

d in

bot

h w

ater

and

aer

osol

sam

ples

from

th

e co

nven

tiona

l sho

wer

s. M

ost n

otab

le w

as th

e pr

esen

ce o

f Myc

obac

teri

um m

ucog

enic

um (9

9.5%

iden

tity)

in th

e w

ater

and

Ps

eudo

mon

as a

erug

inos

a (9

9.3%

iden

tity)

in th

e ae

roso

l sam

ples

.

Perk

ins e

t al

. 200

9

Hot

tub

air

and

wat

erM

. sp.

Hot

tub

expo

sure

has

bee

n ca

usal

ly a

ssoc

iate

d w

ith a

ste

roid

-res

pons

ive,

gra

nulo

mat

ous

lung

dis

ease

feat

urin

g no

ntub

ercu

-lo

us m

ycob

acte

rial

(NT

M) g

row

th in

bot

h cl

inic

al a

nd e

nvir

onm

enta

l sam

ples

. Litt

le is

kno

wn

rega

rdin

g pr

eval

ence

of a

nd

risk

fact

ors

for

NT

M-c

onta

min

atio

n an

d as

soci

ated

illn

ess

in th

ese

sett

ings

. In

this

stu

dy, t

he fr

eque

ncy

of N

TM

gro

wth

and

ae

roso

lizat

ion

in 1

8 pu

blic

hot

tubs

and

war

m w

ater

ther

apy

pool

s an

d th

e fa

ctor

s as

soci

ated

with

myc

obac

teri

al g

row

th w

ere

anal

yzed

. Eac

h si

te w

as c

hara

cter

ized

by

wat

er c

hem

istr

y an

alys

is; a

que

stio

nnai

re o

n m

aint

enan

ce, d

isin

fect

ion,

and

wat

er

qual

ity; a

nd a

ir a

nd w

ater

sam

plin

g fo

r qu

antit

ativ

e N

TM

cul

ture

. NT

M w

ere

dete

cted

in a

ir o

r w

ater

from

13/

18 (7

2%) s

ites;

a st

rong

cor

rela

tion

was

foun

d be

twee

n th

e m

axim

um a

ir a

nd w

ater

NT

M c

once

ntra

tions

(rho

0.4

9, p

= 0

.04)

. Use

of h

alog

en

(chl

orin

e or

bro

min

e) d

isin

fect

ion

was

ass

ocia

ted

with

sig

nific

antly

low

er a

ir a

nd w

ater

con

cent

ratio

ns o

f NT

M c

ompa

red

with

dis

infe

ctio

n us

ing

ultr

avio

let l

ight

and

hyd

roge

n pe

roxi

de (p

= 0

.01-

0.04

). H

ighe

r w

ater

turn

over

rat

es w

ere

also

ass

oci-

ated

with

low

er a

ir a

nd w

ater

NT

M c

once

ntra

tions

(p =

0.0

2-0.

03).

The

se fi

ndin

gs s

ugge

st th

at N

TM

are

freq

uent

ly d

etec

t-ab

le in

the

air

and

wat

er o

f spa

s an

d th

erap

y po

ols

and

that

par

ticul

ar m

aint

enan

ce a

nd d

isin

fect

ion

appr

oach

es a

ffect

NT

M

bioa

eros

ol c

once

ntra

tions

in th

ese

sett

ings

.

Gla

zer e

t al

. 200

7

Whi

rlpo

ol s

pa w

ater

M. s

p.A

n ou

tbre

ak o

f occ

upat

iona

l hot

tub

lung

nec

essi

tate

d qu

antit

ativ

e an

alys

is o

f myc

obac

teri

a in

wat

er sa

mpl

es. W

e co

mbi

ned

proc

edur

es fo

r cul

tivat

ion

of m

ycob

acte

ria

in u

rine

and

qua

ntita

tive

anal

yses

of d

ialy

sis w

ater

. Whi

rlpo

ol sp

a w

ater

sam

ples

w

ere

anal

yzed

show

ing

prom

isin

g re

sults

. In

conc

lusi

on, q

uant

itativ

e m

ycob

acte

rial

cul

ture

of w

ater

is p

ossi

ble

by a

dapt

ing

met

hods

rout

inel

y us

ed in

clin

ical

labo

rato

ries

.

Sven

sson

et

al.

2011

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

650

Bot

tled

wat

erM

. avi

umA

dhes

ion

of th

e ba

cter

ia C

ampy

loba

cter

jeju

ni a

nd M

ycob

acte

rium

avi

um o

nto

poly

ethy

lene

tere

phta

late

(PET

), a

poly

mer

wid

ely

used

with

in th

e bo

ttled

wat

er in

dust

ry w

as m

easu

red

in tw

o di

ffere

nt g

roun

dwat

er so

lutio

ns. F

rom

this

, it w

as fo

und

that

whi

lst th

e pe

rcen

tage

cel

l adh

esio

n fo

r a g

iven

stra

in d

id n

ot c

hang

e be

twee

n gr

ound

wat

er ty

pes,

subs

tant

ial v

aria

tion

was

obt

aine

d be

twee

n th

e tw

o ba

cter

ial s

peci

es te

sted

: M. a

vium

(10–

30%

adh

ered

cel

ls) a

nd C

. jej

uni (

1–2%

) and

no

maj

or v

aria

tions

wer

e m

easu

red

as

a fu

nctio

n of

gro

undw

ater

com

posi

tion

for a

giv

en st

rain

. To

expl

ain

this

, the

inte

rfac

ial e

lect

ro-h

ydro

dyna

mic

pro

pert

ies o

f the

ba

cter

ia w

ere

inve

stig

ated

by

mic

roel

ectr

opho

resi

s, w

ith th

e re

sulta

nt d

ata

anal

ysed

on

the

basi

s of e

lect

roki

netic

theo

ry fo

r sof

t bi

ocol

loid

al p

artic

les.

The

resu

lts o

btai

ned

show

ed th

at M

. avi

um c

arri

es a

sign

ifica

nt v

olum

e ch

arge

den

sity

and

that

its p

erip

hera

l la

yer e

xhib

its li

mite

d hy

drod

ynam

ic fl

ow p

erm

eatio

n co

mpa

red

to th

at o

f C. j

ejun

i. It

was

als

o de

mon

stra

ted

that

ster

ic h

indr

ance

to

flow

pen

etra

tion

and

the

degr

ee o

f hyd

roph

obic

ity w

ithin

/of t

he o

uter

bac

teri

al in

terf

ace

are

larg

er fo

r M. a

vium

cel

ls. I

n lin

e w

ith th

is, t

he la

rger

am

ount

of M

. avi

um c

ells

depo

site

d on

to P

ET su

bstr

ates

as c

ompa

red

to th

at o

f C. j

ejun

i can

be

expl

aine

d by

hy

drop

hobi

c at

trac

tion

and

chem

ical

bin

ding

bet

wee

n hy

drop

hobi

c PE

T a

nd o

uter

soft

surf

ace

laye

r of t

he b

acte

ria.

Hyd

roph

obic

-ity

of P

ET w

as a

ddre

ssed

by

com

bini

ng c

onta

ct a

ngle

ana

lyse

s and

forc

e sp

ectr

osco

py u

sing

CH

3-te

rmin

ated

AFM

tip.

Tatc

hou-

Nya

msi

-K

onig

et

al. 2

008

Bot

tled

wat

erM

. avi

umTh

e m

ain

obje

ctiv

e of

our

stud

y w

as to

ass

ess t

he p

ersi

sten

ce o

f Myc

obac

teri

um a

vium

in a

n ol

igot

roph

ic e

nvir

onm

ent s

uch

as

bott

led

grou

ndw

ater

. Filt

ered

gro

undw

ater

sam

ples

wer

e sp

iked

with

was

hed

Myc

o. a

vium

susp

ensi

on a

nd st

ored

in d

ark

and

unde

r sta

tic c

ondi

tions

, at 2

0 de

gree

s C, f

or 3

mon

ths i

n 50

0 m

l PET

bot

tles.

The

loss

of M

yco.

avi

um c

ultiv

abili

ty w

as sl

ow in

w

ater

. On

the

cont

rary

, aft

er a

3-m

onth

stor

age

at 2

0 de

gree

s C, g

row

th o

f att

ache

d ce

lls w

as o

bser

ved

and

cell

adhe

sive

ness

to

the

PET

wal

l inc

reas

ed w

ith ti

me.

It c

ould

pro

babl

y be

bec

ause

of t

he p

rese

nce

of a

n ex

trac

ellu

lar m

atri

x. Th

is st

udy

has s

how

n th

e gr

eat s

tabi

lity

of M

yco.

avi

um in

bul

k w

ater

as w

ell a

s the

ir a

dhes

iven

ess a

nd th

eir g

row

th o

n a

PET

bot

tle w

all i

n an

olig

o-tr

ophi

c en

viro

nmen

t. Sl

owly

gro

win

g m

ycob

acte

ria

are

wel

l ada

pted

to o

ligot

roph

ic e

nvir

onm

ents

such

as g

roun

dwat

er. A

s the

y st

ick

very

wel

l to

surf

aces

, the

y co

uld

be u

sed

for d

eter

min

ing

the

effici

ency

of t

he c

lean

ing

of c

onta

min

ated

surf

aces

.

Tatc

hou-

Nya

msi

-K

onig

et

al. 2

009

Indu

stry

wat

erM

. sp.

Bact

eria

wer

e en

umer

ated

by

conv

entio

nal c

ultu

re m

etho

d an

d flu

ores

cent

vita

l sta

inin

g. A

ctiv

ated

car

bon

trea

tmen

t and

stor

-ag

e in

a ta

nk p

rovi

ded

favo

urab

le e

nvir

onm

ents

for b

acte

rial

gro

wth

. The

bact

eria

l pop

ulat

ion

of th

e w

ater

in b

oth

the

post

-ac

tivat

ed c

arbo

n tr

eatm

ent a

nd th

e ta

nk w

as a

naly

sed

by d

enat

urin

g gr

adie

nt g

el e

lect

roph

ores

is (D

GG

E) w

ith P

CR-

ampl

ified

16

S rD

NA

frag

men

ts in

clud

ing

V6,

-7, a

nd -8

regi

ons.

The

bact

eria

l com

mun

ity st

ruct

ure

in a

ctiv

ated

car

bon

trea

ted

wat

er w

as

stab

le th

roug

hout

the

year

. Sev

eral

kin

ds o

f bac

teri

a su

ch a

s gen

us A

quas

piri

llum

and

Met

hylo

bact

eriu

m w

ere

foun

d in

the

wat

er

afte

r act

ivat

ed c

arbo

n tr

eatm

ent.

The

bact

eria

l com

mun

ity st

ruct

ure

was

cha

nged

and

oth

er b

acte

ria

such

as m

ycob

acte

ria

wer

e de

tect

ed a

fter

stor

age.

Myc

obac

teri

a w

ere

quan

tified

in w

ater

sam

ples

usi

ng re

al-t

ime

PCR

targ

etin

g th

e 16

S rD

NA

gen

e. M

yco-

bact

eria

wer

e al

so d

etec

ted

in ta

p w

ater

and

thei

r num

ber w

as in

crea

sed

103 –1

04 -fol

d hi

gher

aft

er st

orag

e.

Kaw

ai e

t al

. 200

4

Indu

stry

wat

erM

. a. p

arat

uber

culo

sis

Cul

ture

-inde

pend

ent t

echn

ique

s wer

e us

ed fo

r the

det

ectio

n of

pat

hoge

nic

bact

eria

in d

rink

ing

wat

er a

t pot

entia

lly c

ritic

al c

ontr

ol

poin

ts a

long

the

prod

uctio

n lin

es a

t a G

erm

an d

airy

com

pany

and

a S

pani

sh d

ry c

ured

ham

com

pany

. Den

atur

ing

grad

ient

gel

ele

c-tr

opho

resi

s (D

GG

E) w

as u

sed

to d

escr

ibe

bact

eria

l pop

ulat

ion

shift

s ind

icat

ing

biol

ogic

al in

stab

ility

in th

e dr

inki

ng w

ater

sam

ples

. A

utoc

htho

nous

bac

teri

a w

ere

iden

tified

by

sequ

enci

ng th

e ex

cise

d D

GG

E D

NA

ban

ds. M

ore

spec

ifica

lly, r

eal-t

ime

PCR

was

app

lied

to d

etec

t a n

umbe

r of p

atho

geni

c ba

cter

ia, i

.e. L

iste

ria

mon

ocyt

ogen

es, M

ycob

acte

rium

avi

um su

bsp.

par

atub

ercu

losis

, Cam

pylo

- ba

cter

jeju

ni, E

nter

ococ

cus s

pp.,

Salm

onel

la sp

p., E

sche

rich

ia co

li, a

nd P

seud

omon

as a

erug

inos

a.

Vill

arre

al

et a

l. 20

10

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

651

Dri

nkin

g w

ater

Free

-livi

ng a

moe

bae

Free

-livi

ng a

moe

bae

cons

titut

e re

serv

oirs

for m

any

bact

eria

incl

udin

g no

t onl

y w

ell-k

now

n pa

thog

ens b

ut a

lso

emer

ging

pat

ho-

gens

resp

onsi

ble

for r

espi

rato

ry d

isea

ses,

and

con

trib

ute

to th

e pr

otec

tion,

surv

ival

and

dis

sem

inat

ion

of th

ese

bact

eria

in w

ater

sy

stem

s, d

espi

te th

e ap

plic

atio

n of

dis

infe

ctio

n or

ther

mal

trea

tmen

ts. I

n th

is a

rtic

le w

e re

view

the

avai

labl

e in

form

atio

n on

the

pres

ence

of f

ree-

livin

g am

oeba

e an

d am

oeba

e-re

sist

ing

bact

eria

in d

rink

ing

wat

er sy

stem

s, o

n th

e fa

ctor

s tha

t con

trib

ute

to th

eir

pres

ence

in th

e w

ater

and

/or t

he b

iofil

ms,

on

the

poss

ible

con

trol

mea

sure

s and

thei

r effe

ctiv

enes

s, a

nd w

e id

entif

y so

me

gaps

in

curr

ent k

now

ledg

e ne

edin

g fu

rthe

r res

earc

h.

Lore

t and

G

reub

20

10

Wat

er u

sed

for

swim

min

gTh

e fir

st o

f thi

s thr

ee-p

art s

erie

s on

wat

er-r

elat

ed d

erm

atos

es in

volv

ing

the

athl

ete

will

incl

ude

spor

ts o

ccur

ring

with

the

maj

or-

ity o

f tim

e sp

ent i

n th

e w

ater

. Thes

e sp

orts

incl

ude

swim

min

g, d

ivin

g, s

cuba

, sno

rkel

ing

and

wat

er p

olo.

Num

erou

s aut

hors

hav

e de

scri

bed

derm

atol

ogic

con

ditio

ns c

omm

only

see

n in

sw

imm

ers.

This

ser

ies p

rovi

des a

n up

date

d an

d co

mpr

ehen

sive

revi

ew

of th

ese

wat

er d

erm

atos

es. I

n or

der t

o or

gani

ze th

e va

st n

umbe

r of s

kin

cond

ition

s rel

ated

to w

ater

exp

osur

e, w

e di

vide

d th

e sk

in c

ondi

tions

into

gro

upin

gs o

f inf

ectio

us a

nd o

rgan

ism

-rel

ated

der

mat

oses

, irr

itant

and

alle

rgic

der

mat

oses

and

mis

cella

ne-

ous d

erm

atos

es. Th

e va

st m

ajor

ity o

f ski

n co

nditi

ons i

nvol

ving

the

wat

er a

thle

te re

sult

from

che

mic

als a

nd m

icro

bes i

nhab

iting

ea

ch e

nvir

onm

ent.

Whe

n co

nsid

erin

g th

e eff

ects

of s

wim

min

g on

one

’s sk

in, i

t is a

lso

usef

ul to

diff

eren

tiate

bet

wee

n ex

posu

re

to fr

eshw

ater

(lak

es, p

onds

and

sw

imm

ing

pool

s) a

nd e

xpos

ure

to sa

ltwat

er. Th

is re

view

will

ser

ve a

s a g

uide

for d

erm

atol

ogis

ts,

spor

ts m

edic

ine

phys

icia

ns a

nd o

ther

med

ical

pra

ctiti

oner

s in

reco

gniti

on a

nd tr

eatm

ent o

f the

se c

ondi

tions

.

Tlo

ugan

et

al.

2010

a,b,

c

Wat

erin

g tr

ough

sM

. a. p

arat

uber

culo

sis

Myc

obac

teri

um a

vium

subs

p. p

arat

uber

culo

sis (M

ap) i

s the

cau

sativ

e ag

ent o

f Joh

ne’s

dise

ase,

a c

hron

ic e

nter

ic in

fect

ion

that

aff

ects

rum

inan

ts. D

espi

te th

e ub

iqui

tous

occ

urre

nce

of M

ycob

acte

rium

sp. i

n na

ture

and

the

fact

that

John

e’s d

isea

se h

as b

een

repo

rted

wor

ldw

ide,

littl

e re

sear

ch h

as b

een

done

to a

sses

s its

surv

ival

in a

gric

ultu

ral e

nviro

nmen

ts. Th

e go

al o

f thi

s 365

-day

stud

y w

as to

eva

luat

e th

e ab

ility

of M

ap to

per

sist

in m

ixed

-com

mun

ity b

iofil

ms o

n m

ater

ials

com

mon

ly u

sed

to c

onst

ruct

live

stoc

k w

ater

ing

trou

ghs.

Map

was

inoc

ulat

ed in

to 3

2 1

of tr

ough

wat

er c

onta

inin

g ei

ther

con

cret

e, p

last

ic, g

alva

nize

d O

F st

ainl

ess s

teel

tr

ough

mat

eria

ls. Th

e co

ncen

trat

ion

of M

ap w

as d

eter

min

ed b

y us

ing

quan

titat

ive,

real

-tim

e PC

R to

targ

et th

e IS

900

sequ

ence

in

DN

A e

xtra

cts.

Hig

h co

ncen

trat

ions

of M

ap w

ere

dete

cted

on

all t

roug

h m

ater

ials

afte

r 3 d

ays (

arou

nd 1

× 1

05 cel

ls/cm

2 ). Ba

sed

on

the

best

-fit s

lope

s, th

e tim

e re

quire

d fo

r a 9

9% re

duct

ion

(t 99) i

n bi

ofilm

-ass

ocia

ted

Map

cel

ls w

as 1

44 a

nd 1

15 d

ays f

or p

last

ic a

nd

stai

nles

s ste

el tr

ough

mat

eria

ls, r

espe

ctiv

ely.

Map

con

cent

ratio

ns d

id n

ot d

ecre

ase

on c

oncr

ete

and

galv

aniz

ed st

eel t

roug

h m

ater

i-al

s. Th

ese

resu

lts su

gges

t tha

t Map

surv

ives

wel

l in

biofi

lms p

rese

nt o

n liv

esto

ck w

ater

ing

trou

gh m

ater

ials

. To

inhi

bit s

prea

d of

this

or

gani

sm a

nd e

xpos

ure

of su

scep

tible

ani

mal

s to

Map

on

infe

cted

farm

s, be

st m

anag

emen

t pra

ctic

es a

imed

at m

aint

aini

ng b

iofil

m-

free

trou

gh su

rfac

es sh

ould

be

incl

uded

in a

ny Jo

hne’s

con

trol

pla

n.

Coo

k et

al

. 201

0

Nat

ural

wat

er (r

iver

or

lake

)M

. sp.

M. a

vium

Wat

er, s

edim

ent,

and

stem

s and

root

s of c

omm

on re

ed (P

hrag

mite

s aus

tral

is) a

nd g

reat

er re

edm

ace

(Typ

ha la

tifol

ia) w

ere

take

n fr

om 1

5 lo

catio

ns w

ithin

the

reed

bed

plu

s site

s ups

trea

m a

nd d

owns

trea

m. S

ampl

es w

ere

anal

ysed

for m

ycob

acte

ria

usin

g PC

R an

d sp

ecifi

cally

for M

. avi

um u

sing

nes

ted

PCR

. Env

iron

men

tal m

ycob

acte

ria

wer

e fo

und

thro

ugho

ut th

e en

tire

reed

bed

but

M

. avi

um w

as n

ot fo

und

dow

nstr

eam

of t

he fi

rst v

eget

atio

n gr

owth

. The

reed

bed

was

foun

d to

effe

ctiv

ely

rem

ove

M. a

vium

fr

om th

e w

ater

thro

ugh

a co

mbi

natio

n of

sedi

men

tatio

n an

d ad

sorp

tion

onto

veg

etat

ion

stem

s.

Dre

we

et

al. 2

009

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

652

Lake

wat

erM

. sp.

The

act

inob

acte

rial

com

mun

ities

pre

sent

in tw

o Fi

nnis

h la

kes

and

in th

e su

rrou

ndin

g co

nife

r fo

rest

soi

l wer

e in

vest

igat

ed

usin

g D

NA

bas

ed m

etho

ds. T

he d

omin

ant a

ctin

obac

teri

a in

the

soil

wer

e fo

und

to b

elon

g to

gen

us M

ycob

acte

rium

. The

refo

re

spec

ific

prim

ers

wer

e de

sign

ed a

nd te

sted

to s

tudy

the

myc

obac

teri

al c

omm

uniti

es in

bor

eal e

nvir

onm

ent m

ore

clos

ely.

The

de

natu

ring

gra

dien

t gel

ele

ctro

phor

esis

(DG

GE)

and

seq

uenc

ing

anal

ysis

sho

wed

that

the

mic

robi

al p

opul

atio

ns in

lake

s w

ere

diffe

rent

from

thos

e in

the

surr

ound

ing

soil.

Thu

s, e

ach

of th

e en

viro

nmen

ts h

ad th

eir

own

actin

obac

teri

al a

nd m

ycob

acte

rial

po

pula

tions

. The

maj

ority

of t

he o

btai

ned

myc

obac

teri

al s

eque

nces

wer

e cl

osel

y re

late

d to

the

desc

ribe

d sp

ecie

s of

env

iron

-m

enta

l myc

obac

teri

a, s

ome

of w

hich

are

pat

hoge

nic.

Niv

a et

al.

2006

Nat

ural

wat

erM

. a. p

arat

uber

culo

sis

A li

mite

d su

rvey

was

und

erta

ken

in N

orth

ern

Irel

and

to te

st fo

r M

. avi

um s

ubsp

. par

atub

ercu

losi

s in

untr

eate

d w

ater

ent

er-

ing

nine

wat

er tr

eatm

ent w

orks

(WT

Ws)

ove

r a

1-ye

ar p

erio

d. T

hree

det

ectio

n m

etho

ds w

ere

empl

oyed

, viz

., im

mun

omag

-ne

tic s

epar

atio

n-PC

R an

d cu

lture

on

Her

rold

’s eg

g yo

lk m

ediu

m (H

EYM

) and

BA

CT

EC 1

2B m

ediu

m, t

he la

tter

bot

h su

p-pl

emen

ted

with

myc

obac

tins.

Of t

he 1

92 u

ntre

ated

wat

er s

ampl

es te

sted

, 15

(8%

) tes

ted

M. a

vium

sub

sp. p

arat

uber

culo

sis

posi

tive

by o

ne o

r m

ore

of th

e th

ree

dete

ctio

n m

etho

ds.

Wha

n et

al

. 200

5

Riv

er w

ater

and

sed

imen

tM

. sho

ttsii

M. p

seud

osho

tsii

Stri

ped

bass

(Mor

one

saxa

tilis

) in

the

Che

sape

ake

Bay

are

curr

ently

exp

erie

ncin

g a

very

hig

h pr

eval

ence

of m

ycob

acte

rios

is

asso

ciat

ed w

ith n

ewly

des

crib

ed M

ycob

acte

rium

spe

cies

, Myc

obac

teri

um p

seud

osho

ttsi

i and

M. s

hott

sii.

The

eco

logy

of t

hese

m

ycob

acte

ria

outs

ide

the

stri

ped

bass

hos

t is

curr

ently

unk

now

n. In

this

wor

k, w

e de

velo

ped

quan

titat

ive

real

-tim

e PC

R as

says

for

M. p

seud

osho

ttsi

i and

M. s

hott

sii a

nd a

pplie

d th

ese

assa

ys to

DN

A e

xtra

cts

from

Che

sape

ake

Bay

wat

er a

nd s

edi-

men

t sam

ples

, as

wel

l as

to ti

ssue

s fr

om tw

o do

min

ant p

rey

of s

trip

ed b

ass,

Atla

ntic

men

hade

n (B

revo

orti

a ty

rann

us) a

nd b

ay

anch

ovy

(Anc

hoa

mit

chill

i). M

ycob

acte

rium

pse

udos

hott

sii w

as fo

und

to b

e ub

iqui

tous

in w

ater

sam

ples

from

the

mai

n st

em

of th

e C

hesa

peak

e Ba

y an

d w

as a

lso

pres

ent i

n w

ater

and

sed

imen

ts fr

om th

e Ra

ppah

anno

ck R

iver

, Vir

gini

a. M

. pse

udos

hott

sii

was

als

o de

tect

ed in

men

hade

n an

d an

chov

y tis

sues

. In

cont

rast

, M. s

hott

sii w

as n

ot d

etec

ted

in w

ater

, sed

imen

t, or

pre

y fis

h tis

sues

. In

conj

unct

ion

with

its

nonp

igm

ente

d ph

enot

ype,

whi

ch is

freq

uent

ly fo

und

in o

blig

atel

y pa

thog

enic

myc

obac

teri

a of

hu

man

s, th

is p

atte

rn o

f occ

urre

nce

sugg

ests

that

M. s

hott

sii m

ay b

e an

obl

igat

e pa

thog

en o

f str

iped

bas

s.

Gau

thie

r et

al.

2010

Riv

er w

ater

M. a

. par

atub

ercu

losi

sIn

Sou

th W

ales

, Uni

ted

Kin

gdom

, a p

opul

ated

coa

stal

regi

on li

es b

enea

th h

ill p

astu

res

graz

ed b

y liv

esto

ck in

whi

ch M

yco-

bact

eriu

m a

vium

sub

sp. p

arat

uber

culo

sis i

s en

dem

ic. T

he T

aff i

s a

spat

e ri

ver

runn

ing

off t

he h

ills

and

thro

ugh

the

prin

cipa

l ci

ty o

f Car

diff.

We

sam

pled

Taf

f wat

er a

bove

Car

diff

twic

e w

eekl

y fr

om N

ovem

ber

2001

to N

ovem

ber

2002

. M. a

vium

sub

sp.

para

tube

rcul

osis

was

det

ecte

d by

IS90

0 PC

R an

d cu

lture

. Thi

rty-

one

of 9

6 da

ily s

ampl

es (3

2.3%

) wer

e IS

900

PCR

posi

tive,

an

d 12

gre

w M

. avi

um s

ubsp

. par

atub

ercu

losi

s bov

ine

stra

ins.

Pick

up e

t al

. 200

5

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

653

Riv

er w

ater

M. a

. par

atub

ercu

losi

sW

e st

udie

d th

e Ri

ver

Tyw

i in

Sout

h W

ales

, Uni

ted

Kin

gdom

, who

se c

atch

men

t com

pris

es 1

,100

km

2 con

tain

ing

mor

e th

an

a m

illio

n da

iry

and

beef

cat

tle a

nd m

ore

than

1.3

mill

ion

shee

p. T

he R

iver

Tyw

i is

abst

ract

ed fo

r th

e do

mes

tic w

ater

sup

ply.

Betw

een

Aug

ust 2

002

and

Apr

il 20

03, 4

8 of

70

(68.

8%) t

wic

e-w

eekl

y ri

ver

wat

er s

ampl

es te

sted

pos

itive

by

IS90

0 PC

R.

In r

iver

wat

er, t

he o

rgan

ism

s w

ere

asso

ciat

ed w

ith a

sus

pend

ed s

olid

whi

ch w

as d

eple

ted

by th

e w

ater

trea

tmen

t pro

cess

. D

ispo

sal o

f con

tam

inat

ed s

lurr

y ba

ck o

nto

the

land

est

ablis

hed

a cy

cle

of e

nvir

onm

enta

l per

sist

ence

. A c

once

ntra

te fr

om

100 

liter

s of

fini

shed

wat

er te

sted

neg

ativ

e, b

ut 1

of 5

4 do

mes

tic c

old

wat

er ta

nks

test

ed p

ositi

ve, i

ndic

atin

g th

e po

tent

ial

for

thes

e pa

thog

ens

to a

cces

s do

mes

tic o

utle

ts. I

n th

e se

para

te E

nglis

h La

ke D

istr

ict r

egio

n, w

ith h

ills

up to

980

m, t

ests

fo

r M

. avi

um s

ubsp

. par

atub

ercu

losi

s in

the

high

hill

lake

s an

d se

dim

ents

wer

e us

ually

neg

ativ

e, b

ut s

trea

ms

and

sedi

men

ts

beca

me

posi

tive

low

er d

own

the

catc

hmen

t. Se

dim

ents

from

9 o

f 10

maj

or la

kes

rece

ivin

g in

flow

from

thes

e ca

tchm

ents

wer

e po

sitiv

e, w

ith s

edim

ent c

ores

indi

catin

g de

posi

tion

over

at l

east

40

to 5

0 ye

ars.

Tw

o of

12

mon

thly

1-l

iter

sam

ples

of e

fflu

ent

and

a si

ngle

100

-lite

r sa

mpl

e fr

om th

e A

mbl

esid

e se

wag

e tr

eatm

ent w

orks

wer

e po

sitiv

e fo

r M

. avi

um s

ubsp

. par

atub

ercu

lo-

sis.

Sinc

e La

ke A

mbl

esid

e di

scha

rges

into

Lak

e W

inde

rmer

e, w

hich

is a

vaila

ble

for

dom

estic

sup

ply,

ther

e is

a p

oten

tial f

or

thes

e or

gani

sms

to c

ycle

with

in h

uman

pop

ulat

ions

.

Pick

up e

t al

. 200

6

Coa

stal

lago

on w

ater

M

. sp.

Thi

s st

udy

uses

indi

rect

gra

dien

t ana

lysi

s to

illu

stra

te th

e st

rong

rela

tions

hips

that

exi

sts

betw

een

coas

tal w

ater

qua

lity

and

the

abun

danc

e of

Myc

obac

teri

um s

pp. w

ithin

a U

.S. m

id-A

tlant

ic e

mba

ymen

t. M

ycob

acte

rium

spe

cies

abu

ndan

ce a

nd w

ater

qu

ality

con

ditio

ns (b

ased

on

16 p

hysi

cal a

nd c

hem

ical

var

iabl

es) w

ere

exam

ined

sim

ulta

neou

sly

in m

onth

ly s

ampl

es o

btai

ned

at 1

8 M

aryl

and

and

Vir

gini

a co

asta

l bay

sta

tions

from

Aug

ust 2

005

to N

ovem

ber

2006

(n =

212

). A

qua

ntita

tive

mol

ecul

ar

assa

y fo

r M

ycob

acte

rium

spp

. was

eva

luat

ed a

nd a

pplie

d, a

llow

ing

for

rapi

d, d

irec

t enu

mer

atio

n. B

y us

ing

indi

rect

gra

di-

ent a

naly

sis

(env

iron

men

tal p

rinc

ipal

com

pone

nts

anal

ysis

), a

stro

ng li

nkag

e be

twee

n eu

trop

hic

cond

ition

s, c

hara

cter

ized

by

low

dis

solv

ed-o

xyge

n le

vels

and

ele

vate

d nu

trie

nt c

once

ntra

tions

, and

myc

obac

teri

a w

as d

eter

min

ed. M

ore

spec

ifica

lly,

a st

rong

nut

rien

t res

pons

e w

as n

oted

, with

all

nitr

ogen

com

pone

nts

and

turb

idity

mea

sure

men

ts c

orre

latin

g po

sitiv

ely

with

ab

unda

nce

(r v

alue

s of

> 0

.30;

P v

alue

s of

< 0

.001

), w

hile

dis

solv

ed o

xyge

n sh

owed

a s

tron

g ne

gativ

e re

latio

nshi

p (r

= –

0.38

; P

= 0.

01).

Logi

stic

regr

essi

on m

odel

s de

velo

ped

usin

g sa

linity

, dis

solv

ed o

xyge

n, a

nd to

tal n

itrog

en s

how

ed a

hig

h de

gree

of

conc

orda

nce

(83%

). T

hese

resu

lts s

ugge

st th

at c

oast

al re

stor

atio

n an

d m

anag

emen

t str

ateg

ies

desi

gned

to re

duce

eut

roph

ica-

tion

may

als

o re

duce

tota

l myc

obac

teri

a in

coa

stal

wat

ers.

Jaco

bs e

t al

. 200

9

Surf

ace

wat

erM

. ulc

eran

sT

his

stud

y re

port

s th

e fir

st s

ucce

ssfu

l app

licat

ion

of re

al-t

ime

PCR

for

the

dete

ctio

n of

Myc

obac

teri

um u

lcer

ans,

the

caus

ativ

e ag

ent o

f Bur

uli u

lcer

(BU

), in

Gha

na, a

BU

-end

emic

cou

ntry

. Env

iron

men

tal s

ampl

es a

nd o

rgan

s of

sm

all m

amm

als

wer

e an

a-ly

zed.

The

real

-tim

e PC

R as

says

con

firm

ed th

e pr

esen

ce o

f M. u

lcer

ans i

n a

wat

er s

ampl

e co

llect

ed in

a B

U-e

ndem

ic v

illag

e in

th

e A

shan

ti Re

gion

.

Vand

elan

-no

ote

et

al. 2

010

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

654

Tabl

e 4.

Myc

obac

teri

a in

soi

l

Type

of s

oil

Myc

obac

teri

a de

tect

edA

bstr

act e

xcer

pts

Refe

renc

e

Ara

ble

soil

M. s

p.M

. len

tiflav

umM

. hei

delb

erge

nse

M. a

ustr

oafr

ican

um

PCR

prim

ers d

esig

ned

to a

mpl

ify p

art o

f the

myc

obac

teri

al 1

6S rR

NA

gen

e w

ere

appl

ied

to D

NA

ext

ract

ed fr

om c

ultu

red

orga

n-is

ms a

nd so

il. Th

e PC

R pr

oduc

ts fr

om so

il co

ntai

ned

sequ

ence

s with

sim

ilari

ty to

slow

gro

win

g m

ycob

acte

ria

sim

ilar t

o M

yco-

bact

eriu

m le

ntifl

avum

, and

to fa

st g

row

ing

myc

obac

teri

a su

ch a

s the

xen

obio

tic d

egra

ders

PYR

-I a

nd R

JGII

.

Men

dum

et

al.

2000

Bor

eal f

ores

t soi

lM

. sp.

We

appl

ied

a qu

antit

ativ

e sa

ndw

ich

hybr

idiz

atio

n ap

proa

ch fo

r dir

ect d

etec

tion

of m

ycob

acte

rial

16S

rRN

A in

soil

with

out a

nu

clei

c ac

id a

mpl

ifica

tion

step

. The

num

bers

of m

ycob

acte

rial

16S

rRN

A m

olec

ules

foun

d in

the

soil

indi

cate

d th

e pr

esen

ce o

f up

to 1

07 to 1

08 myc

obac

teri

al c

ells

per

gra

m o

f soi

l. Th

ese

num

bers

exc

eed

by fa

ctor

of 1

0 to

100

× th

e pr

evio

us e

stim

ates

of

myc

obac

teri

a in

soil

base

d on

cul

ture

met

hods

. Whe

n re

al-t

ime

PCR

with

myc

obac

teri

a ta

rget

ing

prim

ers w

as u

sed

to e

stim

ate

the

num

ber o

f 16S

rDN

A c

opie

s in

soil,

one

cop

y of

16S

rDN

A w

as d

etec

ted

per 1

04 cop

ies o

f 16S

rRN

A. Th

is is

clo

se to

the

num

ber o

f 16S

rRN

A m

olec

ules

det

ecte

d pe

r cel

l by

the

sam

e m

etho

d in

labo

rato

ry p

ure

cultu

res o

f M. c

hlor

ophe

nolic

um.

Nie

min

en

et a

l. 20

06

Hou

seho

ld s

oil

M. s

p.A

tota

l of 1

48 so

il sa

mpl

es a

nd 2

4 w

ater

sam

ples

wer

e co

llect

ed fr

om v

ario

us lo

catio

ns a

nd e

xam

ined

to d

eter

min

e th

e pr

esen

ce

of m

ycob

acte

ria.

The

dete

ctio

n m

etho

d in

volv

ed se

mis

elec

tive

cultu

ring

and

aci

d-fa

st st

aini

ng, f

ollo

win

g de

cont

amin

atio

n of

sa

mpl

es to

enr

ich

myc

obac

teri

a an

d re

duce

the

num

bers

of o

ther

mic

roor

gani

sms,

or P

CR

with

pri

mer

s spe

cific

for t

he m

yco-

bact

eria

l 16S

rRN

A g

ene,

usi

ng D

NA

ext

ract

ed d

irec

tly fr

om so

il an

d w

ater

sam

ples

. Myc

obac

teri

a w

ere

dete

cted

in th

e m

ajor

ity

of th

e sa

mpl

es, a

nd su

bseq

uent

sequ

ence

ana

lysi

s of P

CR

prod

ucts

am

plifi

ed d

irec

tly fr

om so

il D

NA

indi

cate

d th

at m

ost o

f the

pr

oduc

ts w

ere

rela

ted

to k

now

n en

viro

nmen

tal m

ycob

acte

ria.

For

bot

h m

etho

ds th

e ra

tes o

f rec

over

y w

ere

cons

iste

ntly

hig

her

for d

ry se

ason

sam

ples

than

for w

et se

ason

sam

ples

.

Chi

lima

et a

l. 20

06

Surf

ace

soil

M. u

lcer

ans

This

pap

er d

escr

ibes

the

deve

lopm

ent o

f a T

aqM

an a

ssay

targ

etin

g IS

2404

mul

tiple

xed

with

an

inte

rnal

pos

itive

con

trol

to

mon

itor i

nhib

ition

with

a d

etec

tion

limit

of le

ss th

an 1

gen

ome

equi

vale

nt o

f DN

A. Th

e as

say

impr

oves

the

turn

arou

nd ti

me

for

diag

nosi

s and

repl

aces

con

vent

iona

l gel

-bas

ed P

CR

as th

e ro

utin

e m

etho

d fo

r lab

orat

ory

confi

rmat

ion

of M

. ulc

eran

s inf

ectio

n in

Vic

tori

a, A

ustr

alia

. Fol

low

ing

anal

ysis

of 4

15 c

linic

al sp

ecim

ens,

the

new

test

dem

onst

rate

d 10

0% se

nsiti

vity

and

spec

ifici

ty

com

pare

d w

ith c

ultu

re. A

noth

er m

ultip

lex

TaqM

an a

ssay

targ

etin

g IS

2606

and

the

keto

redu

ctas

e-B

dom

ain

of th

e M

. ulc

eran

s m

ycol

acto

ne p

olyk

etid

e sy

ntha

se g

enes

was

des

igne

d to

aug

men

t the

spec

ifici

ty o

f the

IS24

04 P

CR

for t

he a

naly

sis o

f a v

arie

ty

of e

nvir

onm

enta

l sam

ples

. Ass

ayin

g fo

r the

se th

ree

targ

ets e

nabl

ed th

e de

tect

ion

of M

. ulc

eran

s DN

A in

soil,

sedi

men

t, an

d m

osqu

ito e

xtra

cts c

olle

cted

from

an

area

of e

ndem

icity

for B

urul

i ulc

er in

Vic

tori

a w

ith a

hig

h de

gree

of c

onfid

ence

. Fin

al c

on-

firm

atio

n w

as o

btai

ned

by th

e de

tect

ion

and

sequ

enci

ng o

f var

iabl

e-nu

mbe

r tan

dem

repe

at (V

NT

R) lo

cus 9

, whi

ch m

atch

ed th

e V

NT

R lo

cus 9

sequ

ence

obt

aine

d fr

om th

e cl

inic

al is

olat

es in

this

regi

on. Th

is su

ite o

f new

met

hods

is e

nabl

ing

rapi

d pr

ogre

ss in

th

e un

ders

tand

ing

of th

e ec

olog

y of

this

impo

rtan

t hum

an p

atho

gen.

Fyfe

et a

l. 20

07

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

655

Surf

ace

soil

M. b

ovis

PCR

prim

ers

spec

ific

for

the

Myc

obac

teri

um tu

berc

ulos

is c

ompl

ex w

ere

used

to d

etec

t the

pre

senc

e of

Myc

obac

teri

um

bovi

s BC

G (P

aste

ur) i

n so

il m

icro

cosm

s an

d M

ycob

acte

rium

bov

is in

env

iron

men

tal s

ampl

es ta

ken

from

a fa

rm in

Irel

and

with

a h

isto

ry o

f bov

ine

tube

rcul

osis

. M. b

ovis

gen

es w

ere

dete

cted

in s

oil a

t 4 a

nd 2

1 m

onth

s af

ter

poss

ible

con

tam

inat

ion.

G

ene

leve

ls w

ere

foun

d in

the

rang

e of

1 ×

103 to

3.6

× 1

03 gen

e co

pies

per

g o

f soi

l, de

pend

ing

on th

e sa

mpl

ing

area

. Are

as

arou

nd b

adge

r se

tts

had

the

high

est l

evel

s of

det

ecta

ble

gene

s an

d w

ere

show

n to

hav

e th

e hi

ghes

t lev

els

of g

ene

pers

iste

nce.

M

. bov

is-s

peci

fic 1

6S r

RNA

seq

uenc

es w

ere

dete

cted

, pro

vidi

ng e

vide

nce

of th

e pr

esen

ce o

f via

ble

cells

in Ir

ish

soils

. Stu

dies

of

DN

A tu

rnov

er in

soi

l mic

roco

sms

prov

ed th

at d

ead

cells

of M

. bov

is B

CG

did

not

per

sist

bey

ond

10 d

ays.

Youn

g et

al

. 200

5

Surf

ace

soil

(Ant

arct

ic)

M. s

p.T

he d

iver

sity

of a

ctin

omyc

etes

was

est

imat

ed w

ith tw

o di

ffere

nt s

trat

egie

s th

at u

se P

CR-

dena

turi

ng g

radi

ent g

el e

lect

roph

o-re

sis.

Fir

st, a

PC

R w

as a

pplie

d, u

sing

a g

roup

-spe

cific

pri

mer

that

allo

ws

sele

ctiv

e am

plifi

catio

n of

act

inom

ycet

e se

quen

ces.

Se

cond

, a n

este

d-PC

R ap

proa

ch w

as u

sed

that

allo

ws

the

estim

atio

n of

the

rela

tive

abun

danc

e of

act

inom

ycet

es w

ithin

the

bact

eria

l com

mun

ity. M

olec

ular

iden

tific

atio

n, w

hich

was

bas

ed o

n 16

S rD

NA

seq

uenc

e an

alys

is, r

evea

led

eigh

t gen

era

of

actin

omyc

etes

, Act

inob

acte

rium

, Act

inom

yces

, an

uncu

lture

d A

ctin

omyc

ete,

Str

epto

myc

es, L

eifs

onia

, Fra

nkin

eae,

Rho

doco

ccus

, an

d M

ycob

acte

rium

.

Lear

n-H

an e

t al.

2012

Aqu

ifer

sen

dM

. a. p

arat

uber

culo

sis

To in

vest

igat

e th

e pr

oces

ses

cont

rolli

ng th

e tr

ansp

ort o

f Myc

obac

teri

um a

vium

sub

sp. p

arat

uber

culo

sis (

Map

) thr

ough

aqu

ifer

mat

eria

ls. W

e m

easu

red

two

impo

rtan

t sur

face

cha

ract

eris

tics

know

n to

affe

ct b

acte

rial

att

achm

ent t

o se

dim

ent s

urfa

ces:

surf

ace

char

ge a

nd h

ydro

phob

icity

. We

then

mea

sure

d th

e tr

ansp

ort o

f Map

thro

ugh

labo

rato

ry c

olum

ns p

acke

d w

ith a

quife

r sa

nd w

ith v

aryi

ng io

nic

stre

ngth

sol

utio

ns a

nd s

edim

ent s

urfa

ce c

harg

e. W

e fo

und

that

Map

has

a s

tron

g ne

gativ

e ch

arge

and

is

hig

hly

hydr

opho

bic

and

that

the

tran

spor

t of M

ap th

roug

h po

sitiv

ely

char

ged

Fe-c

oate

d sa

nds

was

redu

ced

com

pare

d w

ith

tran

spor

t thr

ough

neg

ativ

ely

char

ged

clea

n qu

artz

san

d, a

lthou

gh M

ap tr

ansp

ort f

or a

ll tr

eatm

ents

was

low

com

pare

d w

ith

the

tran

spor

t beh

avio

ur re

port

ed in

the

liter

atur

e fo

r ot

her

bact

eria

. Our

resu

lts s

ugge

st th

at th

e po

tent

ial f

or g

roun

dwat

er

cont

amin

atio

n by

Map

is lo

w; h

owev

er, t

he o

rgan

ism

may

rem

ain

boun

d to

the

soil

near

the

surf

ace

whe

re it

can

be

inge

sted

by

gra

zing

ani

mal

s or

be

rele

ased

dur

ing

run

off e

vent

s. T

his

is th

e fir

st s

tudy

look

ing

at th

e su

rfac

e ch

arac

teri

stic

s an

d tr

ansp

ort b

ehav

iour

of M

ap th

roug

h aq

uife

r m

ater

ials

and

ther

efor

e pr

ovid

es im

port

ant i

nfor

mat

ion

for

unde

rsta

ndin

g th

e m

ovem

ent o

f Map

in th

e en

viro

nmen

t.

Bols

ter e

t al

. 200

9

Fore

st s

oil

M. s

p.To

tal m

etag

enom

ic D

NA

was

isol

ated

from

hig

h A

ndea

n fo

rest

soi

l and

sub

ject

ed to

taxo

nom

ical

and

func

tiona

l com

posi

tion

anal

yses

by

mea

ns o

f clo

ne li

brar

y ge

nera

tion

and

sequ

enci

ng .

Mos

t clo

ne s

eque

nces

wer

e cl

assi

fied

as B

acte

ria

belo

ngin

g to

phl

ya A

ctin

obac

teri

a, P

rote

obac

teri

a an

d A

cido

bact

eria

. Am

ong

the

mos

t rep

rese

nted

ord

ers

wer

e A

ctin

omyc

etal

es (3

4%

aver

age)

, Rhi

zobi

ales

, Bur

khol

deri

ales

and

Myx

ococ

cale

s an

d w

ith a

gre

ater

num

ber

of s

eque

nces

in th

e ge

nus

Myc

obac

te-

rium

(7%

ave

rage

), Fr

anki

a, S

trep

tom

yces

and

Bra

dyrh

izob

ium

.

Mon

tana

et

al.

2012

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

656

Aci

dic

fore

st s

oil

M. s

p.M

. sim

iae

M. c

onsp

icuu

mM

. coo

kii

M. h

odle

riM

. aic

hien

seM

. hol

satic

umM

. tus

ciae

M. p

alle

nsM

. gad

ium

The

dive

rsity

of e

nvir

onm

enta

l myc

obac

teri

a w

as st

udie

d in

wat

er-lo

gged

aci

dic

fore

st so

il. M

ycob

acte

ria

wer

e as

sess

ed in

upp

er

and

low

er so

il ho

rizo

ns a

nd su

mm

er a

nd w

inte

r sea

sons

usi

ng T

-RFL

P an

d se

quen

cing

of 1

6S rR

NA

gen

e pr

oduc

ed w

ith M

yco-

bact

eriu

m-s

peci

fic p

rim

ers.

Myc

obac

teri

a di

vers

ity d

iffer

ed b

etw

een

both

the

two

seas

ons a

nd so

il ho

rizo

ns. C

loni

ng re

veal

ed

the

pres

ence

of m

ycob

acte

ria

belo

ngin

g to

thre

e m

ajor

clu

ster

s rec

ogni

zed

with

in th

e ge

nus,

i.e.

fast

-gro

win

g, in

term

edia

te,

and

slow

-gro

win

g sp

ecie

s, w

ith u

npre

cede

nted

abu

ndan

ce a

nd d

iver

sity

of t

he la

tter

. Tw

o no

vel c

lust

ers o

f seq

uenc

es u

nrel

ated

to

the

know

n m

ycob

acte

ria

wer

e id

entifi

ed. Th

is st

udy

rais

es th

e po

ssib

ility

that

fore

st w

etla

nds c

ould

serv

e as

env

iron

men

tal

rese

rvoi

rs fo

r an

unex

plor

ed d

iver

sity

of m

ycob

acte

ria

incl

udin

g th

ose

rela

ted

to p

atho

geni

c sp

ecie

s.

Kop

ecky

et

al.

2011

Gar

den/

Hou

se s

oil

(4''d

eep)

M

. lep

rae

In th

e pr

esen

t stu

dy, w

e ha

ve tr

ied

to d

etec

t via

ble

M. l

epra

e fr

om so

il sa

mpl

es in

end

emic

are

as b

y us

ing

mol

ecul

ar m

etho

ds.

Eigh

ty so

il sa

mpl

es w

ere

colle

cted

from

vill

ages

of t

his a

rea,

DN

A a

nd R

NA

of M

. lep

rae

extr

acte

d an

d id

entifi

ed u

sing

spec

ific

M. l

epra

e pr

imer

s. P

CR

ampl

ifica

tion

was

don

e an

d re

al-t

ime

RT-P

CR

was

use

d to

det

ect v

iabl

e M

. lep

rae.

DN

A ta

rget

ing

the

16S

regi

on o

f M. l

epra

e w

as d

etec

ted

in 3

7.5%

, whe

reas

M. l

epra

e RN

A ta

rget

ing

the

sam

e re

gion

was

det

ecte

d in

35%

of t

hese

sa

mpl

es. O

f the

tota

l 80

sam

ples

, 40

wer

e co

llect

ed fr

om re

side

ntia

l are

as o

f lep

rosy

pat

ient

s whe

reas

40

sam

ples

wer

e fr

om n

o-pa

tient

are

as. F

ifty-

five

perc

ent p

ositi

vity

for 1

6S rR

NA

of M

. lep

rae

was

obs

erve

d fr

om th

e “p

atie

nt” a

rea

in c

ompa

riso

n to

15%

po

sitiv

ity fr

om th

e “n

o-pa

tient

” are

a (p

< 0

.001

). Th

is st

udy

thus

pro

vide

s val

uabl

e in

form

atio

n of

pre

senc

e of

via

ble

M. l

epra

e in

so

il sp

ecim

ens,

whi

ch w

ould

be

of u

se in

inve

stig

atin

g th

e tr

ansm

issi

on d

ynam

ics i

n le

pros

y.

Lava

nia

et

al. 2

008

Hou

seho

ld s

oil

M. l

epra

e...

Evid

ence

sugg

ests

that

hum

idity

may

favo

r sur

viva

l of M

. lep

rae

in th

e en

viro

nmen

t. Se

vera

l rep

orts

show

that

non

-hum

an

sour

ces l

ike

’nat

ural

ly’ i

nfec

ted

arm

adill

os o

r mon

keys

cou

ld a

ct a

s res

ervo

ir fo

r M. l

epra

e. In

anim

ate

obje

cts o

r fom

ites l

ike

artic

les u

sed

by in

fect

ious

pat

ient

s may

theo

retic

ally

spre

ad in

fect

ion.

How

ever

, it i

s onl

y th

roug

h de

taile

d kn

owle

dge

of th

e bi

o-di

vers

ity a

nd e

colo

gy th

at th

e im

port

ance

of t

his m

ode

of tr

ansm

issi

on c

an b

e fu

lly a

sses

sed.

Our

stud

y fo

cuse

s her

e to

dec

iphe

r th

e ro

le o

f env

iron

men

t in

the

tran

smis

sion

of t

he d

isea

se. T

wo

hund

red

and

seve

n so

il sa

mpl

es w

ere

colle

cted

from

a v

illag

e in

en

dem

ic a

rea

whe

re a

ctiv

e ca

ses a

lso

resi

ded

at th

e tim

e of

sam

ple

colle

ctio

n. S

lit sk

in sm

ears

wer

e co

llect

ed fr

om 1

3 m

ultib

acil-

lary

(MB)

lepr

osy

patie

nts a

nd 1

2 ho

useh

old

cont

acts

of t

he p

atie

nts s

uspe

cted

to b

e hi

dden

cas

es. D

NA

and

RN

A o

f M. l

epra

e w

ere

extr

acte

d an

d am

plifi

ed u

sing

M. l

epra

e sp

ecifi

c pr

imer

s. S

even

ty-o

ne so

il sa

mpl

es sh

owed

pre

senc

e of

M. l

epra

e D

NA

w

here

as 1

6S rR

NA

cou

ld b

e de

tect

ed in

twen

ty-e

ight

of t

hese

sam

ples

. Sam

ples

, bot

h fr

om th

e en

viro

nmen

t and

the

patie

nts,

exhi

bite

d th

e sa

me

geno

type

whe

n te

sted

by

sing

le n

ucle

otid

e po

lym

orph

ism

(SN

P) ty

ping

. Gen

otyp

e of

M. l

epra

e fo

und

in th

e so

il an

d th

e pa

tient

s res

idin

g in

the

sam

e ar

ea c

ould

hel

p in

und

erst

andi

ng th

e tr

ansm

issi

on li

nk in

lepr

osy.

Tura

nkar

et

al.

2012

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

657

Poly

cycl

ic a

rom

atic

hy

droc

arbo

ns

– C

onta

min

ated

soi

lM

. sp.

M. t

usci

aeM

. fre

deri

ksbe

rgen

seM

. aus

troa

fric

anum

M. p

etro

leop

hylu

m

To st

udy

the

natu

ral r

ole

and

dive

rsity

of t

he M

ycob

acte

rium

com

mun

ity in

con

tam

inat

ed so

ils, a

cul

ture

-ind

epen

dent

fing

er-

prin

ting

met

hod

base

d on

PC

R co

mbi

ned

with

den

atur

ing

grad

ient

gel

ele

ctro

phor

esis

(DG

GE)

was

dev

elop

ed. N

ew P

CR

prim

-er

s wer

e se

lect

ed w

hich

spec

ifica

lly ta

rget

ed th

e 16

S rR

NA

gen

es o

f fas

t-gr

owin

g m

ycob

acte

ria,

and

sing

le-b

and

DG

GE

profi

les

of a

mpl

icon

s wer

e ob

tain

ed fo

r mos

t Myc

obac

teri

um st

rain

s tes

ted.

Str

ains

bel

ongi

ng to

the

sam

e sp

ecie

s rev

eale

d id

entic

al

DG

GE

finge

rpri

nts,

and

in m

ost c

ases

, but

not

all,

thes

e fin

gerp

rint

s wer

e ty

pica

l for

one

spec

ies,

allo

win

g pa

rtia

l diff

eren

tiatio

n be

twee

n sp

ecie

s in

a M

ycob

acte

rium

com

mun

ity. M

ycob

acte

rium

stra

ins i

nocu

late

d in

soil

wer

e de

tect

ed w

ith a

det

ectio

n lim

it of

106 C

FU/g

of s

oil u

sing

the

new

pri

mer

set a

s suc

h, o

r app

roxi

mat

ely

102 C

FU/g

in a

nes

ted

PCR

appr

oach

com

bini

ng e

ubac

-te

rial

and

the

Myc

obac

teri

um sp

ecifi

c pr

imer

s. U

sing

the

PCR-

DG

GE

met

hod,

diff

eren

t spe

cies

cou

ld b

e in

divi

dual

ly re

cogn

ized

in

a m

ixed

Myc

obac

teri

um c

omm

unity

.

Leys

et a

l. 20

05

Poly

cycl

ic a

rom

atic

hy

droc

arbo

ns

– C

onta

min

ated

soi

lM

. sp.

M. m

onas

cens

eM

. chl

orop

heno

licum

In th

e pr

esen

t stu

dy 1

6S r

RNA

gen

es w

ere

PCR

ampl

ified

usi

ng M

ycob

acte

rium

-spe

cific

pri

mer

s an

d se

para

ted

by te

mpe

ra-

ture

gra

dien

t gel

ele

ctro

phor

esis

(TG

GE)

, and

pro

min

ent b

ands

wer

e se

quen

ced

to c

ompa

re th

e in

dige

nous

Myc

obac

teri

um

com

mun

ity s

truc

ture

s in

four

pai

rs o

f soi

l sam

ples

take

n fr

om h

eavi

ly c

onta

min

ated

and

less

con

tam

inat

ed a

reas

at f

our

dif-

fere

nt s

ites.

Che

ung

and

Kin

kle

2001

Stab

le fl

oor

M. a

. par

atub

ercu

losi

sTh

e ob

ject

ives

of t

he p

rese

nt st

udy

wer

e to

est

imat

e th

e co

rrel

atio

n an

d as

soci

atio

n be

twee

n C

t and

CFU

in fr

esh

and

thaw

ed

pool

ed fe

cal a

nd e

nvir

onm

enta

l sam

ples

. Res

ults

of H

EYM

cul

ture

of 1

,997

poo

led

feca

l sam

ples

from

cow

s in

14 h

erds

, and

80

2 en

viro

nmen

tal s

ampl

es fr

om 1

09 d

airi

es n

atio

nwid

e w

ere

nega

tivel

y (in

vers

ely)

cor

rela

ted

with

thei

r res

pect

ive

real

-tim

e qP

CR

resu

lts. Th

e Sp

earm

an’s

rank

cor

rela

tion

betw

een

Ct a

nd C

FU w

as g

ood

(–0.

66) i

n fr

esh

and

thaw

ed p

oole

d fe

cal s

ampl

es,

and

exce

llent

(–0.

76) a

nd g

ood

(–0.

61) i

n fr

esh

and

thaw

ed e

nvir

onm

enta

l sam

ples

, res

pect

ivel

y. Th

e co

rrel

atio

n va

ried

from

go

od (–

0.53

) to

exce

llent

(–0.

90) d

epen

ding

on

the

num

ber o

f sam

ples

in a

feca

l poo

l. Tr

unca

ted

regr

essi

on m

odel

s ind

icat

ed

a si

gnifi

cant

neg

ativ

e as

soci

atio

n be

twee

n C

t and

CFU

in fe

cal p

ools

and

env

iron

men

tal s

ampl

es. Th

e us

e of

real

-tim

e qP

CR

inst

ead

of H

EYM

can

yie

ld ra

pid,

qua

ntita

tive

estim

ates

of M

AP

load

and

allo

w fo

r inc

orpo

ratio

n of

real

-tim

e qP

CR

resu

lts o

f po

oled

and

env

iron

men

tal s

ampl

es in

test

ing

stra

tegi

es to

iden

tify

dair

y co

w g

roup

s with

the

high

est M

AP

shed

ding

.

Aly

et a

l. 20

10

Stab

le fl

oor

dust

M

. a. p

arat

uber

culo

sis

Envi

ronm

enta

l sam

ples

wer

e co

llect

ed to

inve

stig

ate

the

spat

ial a

nd te

mpo

ral s

prea

d of

Myc

obac

teri

um a

vium

subs

p. p

arat

uber

-cu

losi

s (M

AP)

in a

dai

ry c

attle

bar

n be

fore

and

aft

er th

e in

trod

uctio

n of

two

grou

ps o

f MA

P-sh

eddi

ng a

nim

als.

Sam

ples

col

-le

cted

off

the

floor

of t

he b

arn

refle

cted

the

mom

ent o

f sam

plin

g w

here

as sa

mpl

es c

olle

cted

by

mic

rofib

er w

ipes

at a

min

imal

of

3 m

hei

ght c

onta

ined

the

accu

mul

ated

sett

led

dust

ove

r a 3

-wee

k pe

riod

. Sam

ples

wer

e an

alys

ed b

y IS

900

qPC

R fo

r the

pre

senc

e of

MA

P D

NA

and

by

cultu

re fo

r via

ble

MA

P ba

cter

ia. M

AP

DN

A w

as d

etec

ted

in a

larg

e nu

mbe

r of s

ites b

oth

befo

re a

nd a

fter

in

trod

uctio

n ca

ttle

. MA

P D

NA

was

det

ecte

d in

side

the

barn

in fl

oor a

nd d

ust s

ampl

es fr

om c

ubic

les a

nd sl

atte

d flo

ors a

nd in

set-

tled

dust

sam

ples

loca

ted

abov

e th

e sl

atte

d flo

ors a

nd in

the

vent

ilatio

n ri

dge

open

ing.

Out

side

the

barn

MA

P D

NA

was

det

ecte

d by

PC

R in

sam

ples

refle

ctin

g th

e w

alki

ng p

ath

of th

e fa

rmer

des

pite

hyg

iene

mea

sure

s. N

o vi

able

MA

P w

as d

etec

ted

befo

re th

e in

trod

uctio

n of

shed

der c

attle

. Thre

e w

eeks

late

r via

ble

MA

P w

as fo

und

insi

de th

e ba

rn a

t 7/4

9 lo

catio

ns b

ut n

ot o

utsi

de. F

iftee

n w

eeks

late

r via

ble

MA

P w

as a

lso

dete

cted

in e

nvir

onm

enta

l sam

ples

out

side

the

barn

. In

conc

lusi

on, i

ntro

duct

ion

of M

AP

shed

-di

ng c

attle

lead

to w

ides

prea

d co

ntam

inat

ion

of th

e in

tern

al a

nd e

xter

nal e

nvir

onm

ent o

f a d

airy

bar

n, in

clud

ing

the

pres

ence

of

viab

le M

AP

in se

ttle

d du

st p

artic

les s

ugge

stin

g po

tent

ial t

rans

mis

sion

of M

AP

infe

ctio

n th

roug

h bi

o-ae

roso

ls.

Eise

nber

g et

al.

2009

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

658

Pott

ing

and

gard

en s

oil

M. a

. avi

umM

. a. h

omin

issu

is

In o

rder

to tr

ace

the

sour

ce o

f inf

ectio

n fr

om th

e en

viro

nmen

t, a

met

hod

of D

NA

isol

atio

n fr

om so

il an

d ot

her e

nviro

nmen

tal s

am-

ples

, suc

h as

dus

t, co

bweb

s, an

d co

mpo

st, w

as d

evel

oped

. The

trip

lex

qPC

R ex

amin

atio

n re

veal

ed th

e pr

esen

ce o

f M. a

vium

subs

p.

hom

inis

suis

in a

hig

h pr

opor

tion

of th

e en

viro

nmen

tal s

ampl

es (4

2.8%

in th

e fir

st p

atie

nt’s

hous

e an

d 47

.6%

in th

e se

cond

pat

ient

’s ho

use)

. Bot

h pa

tient

s wer

e al

so e

xpos

ed to

M. a

vium

subs

p. a

vium

, whi

ch w

as p

rese

nt d

ue to

the

bree

ding

of i

nfec

ted

dom

estic

he

ns. Th

e hi

gh in

fect

ious

dos

e of

M. a

vium

subs

p. h

omin

issu

is or

the

incr

ease

d su

scep

tibili

ty o

f hum

ans t

o M

. avi

um su

bsp.

hom

in-

issu

is c

ompa

red

to M

. avi

um su

bsp.

avi

um c

ould

be

the

reas

on w

hy th

e ch

ildre

n w

ere

infe

cted

with

M. a

vium

subs

p. h

omin

issu

is.

Kae

vska

et

al.

2011

Pott

ing

and

gard

en s

oil

M. a

vium

com

plex

Back

grou

nd: M

ycob

acte

rium

avi

um-in

trac

ellu

lare

com

plex

(MA

C) i

s a u

biqu

itous

pat

hoge

n fo

und

in so

il an

d w

ater

. Env

i-ro

nmen

tal e

xpos

ure

is th

e pr

imar

y ro

ute

for M

AC

infe

ctio

n. H

owev

er, s

peci

fic e

nvir

onm

enta

l ris

k fa

ctor

s hav

e be

en p

oorl

y de

term

ined

in im

mun

ocom

pete

nt p

atie

nts w

ith p

ulm

onar

y M

AC

dis

ease

. Met

hods

: A c

ase-

cont

rol s

tudy

was

per

form

ed w

ith

106 

patie

nts w

ith p

ulm

onar

y M

AC

dis

ease

(men

[wom

en],

23 [8

3]; a

ge, 6

4.3

± 9.

2 ye

ars)

and

53

age-

mat

ched

con

trol

pat

ient

s w

ith b

ronc

hiec

tasi

s but

not

pul

mon

ary

MA

C in

fect

ion

(men

[wom

en],

7[46

]; ag

e, 6

3.0

± 11

.0 y

ears

). A

ll pa

rtic

ipan

ts c

ompl

eted

a

stan

dard

ized

que

stio

nnai

re th

at in

clud

ed q

uest

ions

abo

ut m

edic

al h

isto

ry, s

mok

ing

hist

ory,

alco

hol u

sage

, age

at m

enop

ause

, an

d en

viro

nmen

t exp

osur

es. E

nvir

onm

ent e

xpos

ures

incl

uded

soil

expo

sure

from

farm

ing

or g

arde

ning

; wat

er e

xpos

ure

from

ba

thin

g, sh

ower

ing,

hot

tub

use,

dis

hwas

hing

, sw

imm

ing,

and

dri

nkin

g w

ater

; and

pet

exp

osur

e. R

esul

ts: N

o di

ffere

nces

wer

e id

entifi

ed in

the

patie

nt c

hara

cter

istic

s and

und

erly

ing

dise

ases

. Mor

e ca

se p

atie

nts e

xper

ienc

ed h

igh

soil

expo

sure

(≥ 2

per

w

eek)

than

con

trol

pat

ient

s (23

.6%

vs 9

.4%

, P =

.032

); th

is re

mai

ned

sign

ifica

nt a

fter

mul

tivar

iate

ana

lysi

s (O

R, 5

.9; 9

5% C

I,

1.4–

24.7

; P =

.015

). Th

ere

wer

e no

sign

ifica

nt d

iffer

ence

s in

othe

r env

iron

men

tal e

xpos

ures

. Cas

e pa

tient

s with

hig

h so

il ex

po-

sure

wer

e si

gnifi

cant

ly o

lder

than

thos

e w

ith lo

w so

il ex

posu

re (6

7.3

± 7.

3 ye

ars v

s 64.

3 ±

9.5

year

s, P

= .0

37).

Oth

er c

hara

cter

-is

tics,

und

erly

ing

dise

ases

, and

myc

obac

teri

al sp

ecie

s did

not

diff

er b

etw

een

the

two

grou

ps. C

oncl

usio

ns: P

atie

nts w

ith p

ulm

o-na

ry M

AC

dis

ease

had

sign

ifica

ntly

mor

e so

il ex

posu

re th

an n

onin

fect

ed c

ontr

ol p

atie

nts,

whi

ch su

gges

ts th

at e

nvir

onm

enta

l so

il ex

posu

re is

a li

kely

risk

fact

or fo

r the

dev

elop

men

t of p

ulm

onar

y M

AC

dis

ease

.

Mae

kaw

a et

al.

2011

Diff

eren

t soi

l typ

esM

. a. p

arat

uber

culo

sis

Att

achm

ent o

f Myc

obac

teri

um a

vium

subs

p. p

arat

uber

culo

sis to

soil

part

icle

s cou

ld in

crea

se th

eir a

vaila

bilit

y to

farm

ani

mal

s, as

w

ell a

s infl

uenc

e th

e tr

ansp

orta

tion

of M

. avi

um su

bsp.

par

atub

ercu

losis

to w

ater

sour

ces.

To in

vest

igat

e th

e po

ssib

ility

of s

uch

atta

chm

ent,

we

pass

ed a

kno

wn

quan

tity

of M

. avi

um su

bsp.

par

atub

ercu

losis

thro

ugh

chro

mat

ogra

phy

colu

mns

pac

ked

with

cla

y so

il, sa

ndy

soil,

pur

e si

lica,

cla

y-si

lica

mix

ture

, or c

lay-

silic

a co

mpl

exes

and

mea

sure

d th

e or

gani

sms r

ecov

ered

in th

e el

uent

usi

ng

cultu

re o

r qua

ntita

tive

PCR.

Exp

erim

ents

wer

e re

peat

ed u

sing

buff

er a

t a ra

nge

of p

H le

vels

with

pur

e si

lica

to in

vest

igat

e th

e eff

ect

of p

H o

n M

. avi

um su

bsp.

par

atub

ercu

losis

att

achm

ent.

Line

ar m

ixed

-mod

el a

naly

ses w

ere

cond

ucte

d to

com

pare

the

prop

ortio

nal

reco

very

of M

. avi

um su

bsp.

par

atub

ercu

losis

in th

e el

uent

bet

wee

n di

ffere

nt su

bstr

ates

and

pH

leve

ls. O

f the

org

anis

ms a

dded

to

the

colu

mns

, 83

to 1

00%

wer

e es

timat

ed to

be

reta

ined

in th

e co

lum

ns a

fter a

djus

tmen

t for

thos

e re

tain

ed in

em

pty

cont

rol c

olum

ns.

The

prop

ortio

ns re

cove

red

wer

e si

gnifi

cant

ly d

iffer

ent a

cros

s diff

eren

t sub

stra

tes,

with

the

rete

ntio

n be

ing

sign

ifica

ntly

gre

ater

(P

 < 0

.05)

in p

ure

subs

trat

es (s

ilica

and

cla

y-si

lica

com

plex

es) t

han

in so

il su

bstr

ates

(cla

y so

il an

d sa

ndy

soil)

. How

ever

, the

re w

ere

no si

gnifi

cant

diff

eren

ces i

n th

e re

tent

ion

of M

. avi

um su

bsp.

par

atub

ercu

losis

bet

wee

n si

lica

and

clay

-sili

ca c

ompl

exes

or b

etw

een

clay

soil

and

sand

y so

il. Th

e pr

opor

tion

reta

ined

dec

reas

ed w

ith in

crea

sing

pH

in o

ne o

f the

exp

erim

ents

, ind

icat

ing

grea

ter a

dsor

p-tio

n of

M. a

vium

subs

p. p

arat

uber

culo

sis to

soil

part

icle

s at a

n ac

idic

pH

(P <

0.0

5). Th

e re

sults

sugg

est t

hat u

nder

exp

erim

enta

l co

nditi

ons M

. avi

um su

bsp.

par

atub

ercu

losis

ads

orbs

to a

rang

e of

soil

part

icle

s, an

d th

is a

ttac

hmen

t is i

nflue

nced

by

soil

pH.

Dha

nd e

t al

. 200

9b

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

659

Farm

soi

lM

. a. p

arat

uber

culo

sis

Spec

ulat

ion

abou

t the

ass

ocia

tion

of so

il ch

arac

teri

stic

s with

the

expr

essi

on o

f ovi

ne Jo

hne’s

dis

ease

(OJD

) pro

mpt

ed th

is

cros

s-se

ctio

nal s

tudy

. We

enro

lled

92 sh

eep

flock

s in

Aus

tral

ia d

urin

g 20

04–2

005

and

in e

ach

enro

lled

flock

col

lect

ed p

oole

d fa

ecal

sam

ples

from

an

iden

tified

coh

ort (

grou

p of

sam

e ag

e an

d se

x) o

f she

ep a

nd so

il sa

mpl

es fr

om th

e pa

ddoc

ks g

raze

d by

th

is c

ohor

t of s

heep

. Fae

cal p

ools

wer

e cu

lture

d to

cre

ate

thre

e ou

tcom

e va

riab

les:

posi

tive

or n

egat

ive

stat

us o

f fae

cal p

ools

(p

ool O

JD st

atus

, bin

ary)

: the

log

num

ber o

f via

ble

Myc

obac

teri

um a

vium

subs

p. p

arat

uber

culo

sis (

MA

P) o

rgan

ism

s per

gra

m

of fa

eces

(log

poo

l MA

P nu

mbe

r, co

ntin

uous

): an

d th

e pr

eval

ence

of f

aeca

l she

dder

s (co

hort

OJD

pre

vale

nce

leve

l, or

dina

l: lo

w

<2%

, med

ium

2–1

0% a

nd h

igh

>10%

). Se

para

te st

atis

tical

mod

els w

ere

then

dev

elop

ed to

inve

stig

ate

the

asso

ciat

ion

betw

een

soil

char

acte

rist

ics a

nd e

ach

outc

ome

vari

able

. She

ep ra

ised

on

soils

with

a h

ighe

r per

cent

age

of o

rgan

ic c

arbo

n an

d cl

ay h

ad a

hi

gher

OJD

pre

vale

nce

whe

reas

, she

ep g

razi

ng o

n so

ils w

ith a

hig

her c

onte

nt o

f san

d an

d ni

trog

en h

ad a

low

er O

JD p

reva

lenc

e.

Iron

con

tent

of t

he so

il w

as p

ositi

vely

ass

ocia

ted

with

OJD

infe

ctio

n bu

t the

ass

ocia

tion

betw

een

soil

pH a

nd O

JD w

as in

conc

lu-

sive

. Par

ent s

oil t

ype,

the

only

farm

leve

l fac

tor,

was

not

sign

ifica

nt in

any

of t

he fi

nal m

odel

s. S

tudy

resu

lts in

dica

te a

hig

her r

isk

of O

JD in

shee

p ra

ised

on

soils

with

gre

ater

org

anic

mat

ter a

nd c

lay

cont

ent.

We

hypo

thes

ise

that

this

is d

ue to

ads

orpt

ion

of

MA

P to

cla

y an

d th

e co

nseq

uent

rete

ntio

n of

the

bact

eria

in th

e to

psoi

l, th

us m

akin

g th

em a

vaila

ble

in h

ighe

r num

bers

to g

raz-

ing

shee

p.

Dha

nd e

t al

. 200

9a

Past

ure

soil

M. a

. par

atub

ercu

losi

sTh

e ai

m o

f thi

s stu

dy w

as to

dem

onst

rate

the

pers

iste

nce

of M

ycob

acte

rium

avi

um su

bsp.

par

atub

ercu

losi

s (M

AP)

in so

il an

d co

loni

zatio

n of

diff

eren

t pla

nt p

arts

aft

er d

elib

erat

e ex

posu

re to

mou

flon

fece

s nat

ural

ly c

onta

min

ated

with

diff

eren

t am

ount

s of

MA

P. S

ampl

es o

f aer

ial p

arts

of p

lant

s, th

eir r

oots

, and

the

soil

belo

w th

e ro

ots w

ere

colle

cted

aft

er 1

5 w

eeks

and

exa

min

ed

usin

g IS

900

real

-tim

e qu

antit

ativ

e PC

R (q

PCR)

and

cul

tivat

ion.

Alth

ough

the

pres

ence

of v

iabl

e M

AP

cells

was

not

dem

on-

stra

ted,

alm

ost a

ll sa

mpl

es w

ere

foun

d to

be

posi

tive

usin

g qP

CR

. MA

P IS

900

was

not

onl

y fo

und

in th

e up

per g

reen

par

ts, b

ut

also

in th

e ro

ots a

nd so

il sa

mpl

es (f

rom

1.0

0 ×

100 to

6.4

3 ×

103 ).

The

leve

l of s

oil a

nd p

lant

con

tam

inat

ion

was

influ

ence

d m

ainl

y by

moi

stur

e, c

lay

cont

ent,

and

the

dept

h fr

om w

hich

the

sam

ples

wer

e co

llect

ed, r

athe

r tha

n by

the

initi

al c

once

ntra

tion

of M

AP

in th

e fe

ces a

t the

beg

inni

ng o

f the

exp

erim

ent.

Prib

ylov

a et

al.

2011

Peat

M

. sp.

M. a

. hom

inis

suis

M. f

ortu

itum

M. g

ordo

nae

M. c

helo

nae

M. t

erra

eM

. xen

opi

M. fl

aves

cens

M. p

hlei

Exam

inat

ion

of 1

18 sa

mpl

es fr

om v

ario

us ty

pes o

f com

mer

cial

ly a

vaila

ble

peat

(nat

ural

pea

t, pa

cked

pea

t for

hor

ticul

ture

and

sp

ecia

lly p

roce

ssed

pea

t int

ende

d fo

r pig

let f

eedi

ng) s

how

ed th

at P

PM w

ere

pres

ent i

n 84

(71.

1%) s

ampl

es. M

ycob

acte

rium

av

ium

subs

p. h

omin

issu

is (8

2.1%

) was

the

mos

t fre

quen

t myc

obac

teri

al is

olat

e. In

add

ition

, fro

m a

nat

ural

loca

lity

whe

re p

eat

is m

ined

and

stor

ed in

larg

e pi

les f

or u

p to

four

mon

ths,

myc

obac

teri

a w

ere

dete

cted

in p

eat s

ampl

es c

olle

cted

from

the

surf

ace

and

from

up

to 2

5 cm

in d

epth

.

Mat

lova

et

al.

2012

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

660

Floo

r so

il in

phe

asan

t fa

rms

M. s

p.M

. a. h

omin

issu

isM

. che

lona

eM

. for

tuitu

mM

. scr

oful

aceu

mM

. ter

rae

Alth

ough

avi

an m

ycob

acte

rios

is is

not

pre

vale

nt a

mon

g do

mes

tic fo

wl u

sed

for i

nten

sive

hus

band

ry, i

t has

bee

n de

scri

bed

in

both

free

livi

ng b

irds

and

bir

ds in

cap

tivity

, e.g

., zo

olog

ical

gar

dens

and

smal

l fow

l floc

ks. I

n th

is st

udy,

we

exam

ined

305

phe

as-

ants

from

six

flock

s as w

ell a

s 70

othe

r bir

ds b

elon

ging

to 1

4 sp

ecie

s and

97

othe

r ver

tebr

ates

cau

ght i

n a

clos

ed a

rea.

We

also

in

vest

igat

ed th

e pr

eval

ence

of m

ycob

acte

ria

in n

on-v

erte

brat

es (e

arth

wor

ms)

and

soil

in tw

o ph

easa

nt fl

ocks

. Myc

obac

teri

um

aviu

m su

bsp.

avi

um (M

. a. a

vium

) was

isol

ated

in fo

ur fl

ocks

from

17

(5.6

%) p

heas

ants

. In

one

M. a

. avi

um-i

nfec

ted

phea

sant

co

-infe

ctio

n w

ith M

. a. h

omin

issu

is w

as d

iagn

osed

. Gra

nulo

mat

ous i

nflam

mat

ory

lesi

ons w

ere

obse

rved

in li

ver a

nd sp

leen

in

only

four

M. a

. avi

um-in

fect

ed p

heas

ants

ori

gina

ting

from

two

flock

s. F

rom

the

othe

r 38

phea

sant

s oth

er m

ycob

acte

rial

spec

ies

wer

e is

olat

ed, s

uch

as M

. for

tuitu

m, M

. ter

rae,

M. t

rivi

ale,

M. c

helo

nae,

M. s

crof

ulac

eum

, M. s

meg

mat

is, M

. flav

esce

ns, M

. die

rn-

hofe

ri a

nd n

on-id

entifi

able

myc

obac

teri

al sp

ecie

s. In

the

grou

p of

70

bird

s of o

ther

spec

ies,

we

iden

tified

M. a

. avi

um in

two

(2.9

%) g

osha

wks

(Acc

ipite

r gen

tilis

). W

e di

d no

t iso

late

M. a

. avi

um fr

om a

ny o

f the

oth

er 9

7 ve

rteb

rate

s, th

e 39

1 en

viro

nmen

t sa

mpl

es o

r 97

eart

hwor

ms.

Mor

av-

kova

et a

l. 20

11

Floo

r so

il on

cat

tle

farm

M. a

. par

atub

ercu

losi

sTh

e ai

m o

f thi

s stu

dy w

as to

mon

itor t

he p

ersi

sten

ce o

f Myc

obac

teri

um a

vium

subs

p. p

arat

uber

culo

sis i

n en

viro

nmen

tal s

ampl

es

take

n fr

om a

Hol

stei

n fa

rm w

ith a

long

his

tory

of c

linic

al p

arat

uber

culo

sis.

A h

erd

of 6

06 h

ead

was

era

dica

ted,

and

mec

hani

cal

clea

ning

and

dis

infe

ctio

n w

ith c

hlor

amin

e B

with

am

mon

ium

(4%

) was

car

ried

out

on

the

farm

; in

the

surr

ound

ing

area

s (on

th

e fie

ld a

nd fi

eld

mid

den)

lim

e w

as a

pplie

d. E

nvir

onm

enta

l sam

ples

wer

e co

llect

ed b

efor

e an

d ov

er a

per

iod

of 2

4 m

onth

s aft

er

dest

ocki

ng. O

nly

one

sam

ple

out o

f 48

(2%

) exa

min

ed o

n th

e fa

rm (o

rigi

natin

g fr

om a

was

te p

it an

d co

llect

ed b

efor

e de

stoc

king

) w

as p

ositi

ve fo

r M. a

vium

subs

p. p

arat

uber

culo

sis b

y cu

ltiva

tion

on so

lid m

ediu

m (H

erro

ld’s

egg

yolk

med

ium

). Th

e re

sults

usi

ng

real

-tim

e qu

antit

ativ

e PC

R (q

PCR)

show

ed th

at a

tota

l of 8

1% o

f env

iron

men

tal s

ampl

es w

ith a

n av

erag

e m

ean

M. a

vium

subs

p.

para

tube

rcul

osis

cel

l num

ber o

f 3.0

9 ×

103 w

ere

posi

tive

for M

. avi

um su

bsp.

par

atub

ercu

losi

s bef

ore

dest

ocki

ng c

ompa

red

to 4

3% w

ith a

n av

erag

e m

ean

M. a

vium

subs

p. p

arat

uber

culo

sis c

ell n

umbe

r of 5

.86

× 10

2 aft

er 2

4 m

onth

s. M

. avi

um su

bsp.

pa

ratu

berc

ulos

is-p

ositi

ve sa

mpl

es w

ere

dete

cted

in th

e ca

ttle

bar

n as

wel

l as i

n th

e ca

lf ba

rn a

nd su

rrou

ndin

g ar

eas.

M. a

vium

su

bsp.

par

atub

ercu

losi

s was

det

ecte

d fr

om d

iffer

ent m

atri

ces:

floor

and

inst

rum

ent s

crap

ings

, sed

imen

t, or

scra

ping

from

wat

er-

ing

trou

ghs,

was

te p

its, a

nd c

obw

ebs.

M. a

vium

subs

p. p

arat

uber

culo

sis D

NA

was

als

o de

tect

ed in

soil

and

plan

ts c

olle

cted

on

the

field

mid

den

and

the

field

24

mon

ths a

fter

des

tock

ing.

Alth

ough

the

prop

ortio

n of

pos

itive

sam

ples

dec

reas

ed fr

om 6

4% to

23

% o

ver t

ime,

the

num

bers

of M

. avi

um su

bsp.

par

atub

ercu

losi

s cel

ls w

ere

com

para

ble.

Mor

av-

kova

et a

l. 20

12

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

661

Tabl

e 5.

Myc

obac

teri

a in

pla

nts

Type

of p

lant

sM

ycob

acte

ria

dete

cted

Abs

trac

t exc

erpt

sRe

fere

nce

Ree

d M

. avi

umW

ater

, sed

imen

t, an

d st

ems a

nd ro

ots o

f com

mon

reed

(Phr

agm

ites a

ustr

alis

) and

gre

ater

reed

mac

e (T

ypha

latif

olia

) wer

e ta

ken

from

15

loca

tions

with

in th

e re

ed b

ed p

lus s

ites u

pstr

eam

and

dow

nstr

eam

. Sam

ples

wer

e an

alys

ed fo

r myc

obac

teri

a us

ing

PCR

and

spec

ifica

lly fo

r M. a

vium

usi

ng n

este

d PC

R. E

nvir

onm

enta

l myc

obac

teri

a w

ere

foun

d th

roug

hout

the

entir

e re

ed b

ed b

ut

M. a

vium

was

not

foun

d do

wns

trea

m o

f the

firs

t veg

etat

ion

grow

th. Th

e re

ed b

ed w

as fo

und

to e

ffect

ivel

y re

mov

e M

. avi

um

from

the

wat

er th

roug

h a

com

bina

tion

of se

dim

enta

tion

and

adso

rptio

n on

to v

eget

atio

n st

ems.

Dre

we

et

al. 2

009

Sala

d M

. sp.

Food

ass

ocia

ted

indi

geno

us m

icro

bial

com

mun

ities

exe

rt a

ntag

onis

tic e

ffect

s on

path

ogen

s and

may

rout

inel

y de

liver

hea

lth re

le-

vant

mic

roor

gani

sms t

o th

e G

I tra

ct. B

y us

ing

mol

ecul

ar, c

ultu

re in

depe

nden

t met

hods

incl

udin

g PC

R-D

GG

E of

16S

rDN

A-c

od-

ing

regi

ons a

nd re

al-t

ime

PCR

(RT-

PCR)

as w

ell a

s BIO

LOG

met

abol

ic fi

nger

prin

ting,

mic

robi

al c

omm

uniti

es o

n le

ttuc

e w

ere

anal

yzed

in sa

mpl

es fr

om fi

elds

, fro

m su

perm

arke

ts a

nd so

il. A

mpl

ified

16S

rRN

A g

ene

sequ

ence

s (57

.7%

) cou

ld b

e as

sign

ed to

sp

ecie

s pre

viou

sly

repo

rted

as t

ypic

al fo

r the

phy

llosp

here

incl

udin

g Pa

ntoe

a ag

glom

eran

s, Ps

eudo

mon

as fl

aves

cens

, Mor

axel

la

spp.

, and

Myc

obac

teri

um sp

p. 7

1.8%

of t

he se

quen

ces o

btai

ned

repr

esen

ted

so fa

r und

escr

ibed

taxa

.

Zwie

leh-

ner e

t al.

2008

Sila

ge

M. a

. par

atub

ercu

losi

sM

ycob

acte

rium

avi

um su

bsp.

par

atub

ercu

losi

s (M

AP)

is th

e ca

usat

ive

agen

t of p

arat

uber

culo

sis (

John

e’s d

isea

se) i

n ru

min

ants

. Pa

ratu

berc

ulos

is c

an c

ause

seve

re e

cono

mic

loss

es a

nd is

ack

now

ledg

ed a

s one

of t

he m

ost i

mpo

rtan

t dis

ease

s of r

umin

ants

to

day.

Hig

h am

ount

s of M

AP

can

be sh

ed in

the

faec

es o

f inf

ecte

d in

divi

dual

s and

can

surv

ive

for a

long

per

iod

in th

e en

viro

n-m

ent.

In th

e pr

esen

ted

tria

l, ba

led

gras

s sila

ge w

as in

ocul

ated

with

a M

AP-

susp

ensi

on, a

nd th

e vi

abili

ty o

f MA

P w

as st

udie

d ov

er ti

me.

Sam

ples

from

the

bale

s wer

e ta

ken

at in

crea

sing

inte

rval

s and

subs

eque

ntly

test

ed fo

r the

pre

senc

e of

MA

P by

solid

cu

lture

on

Her

rold

’s Eg

g Yo

lk M

edia

(HEY

M),

liqui

d cu

lture

and

real

tim

e Po

lym

eras

e C

hain

Rea

ctio

n (P

CR)

for t

he IS

900

and

F57

frag

men

ts. N

o gr

owth

of M

AP

was

obs

erve

d at

any

tim

e on

solid

or i

n liq

uid

cultu

res,

exc

ept a

t the

tim

e of

inoc

ulat

ion;

PC

R de

tect

ions

wer

e po

sitiv

e in

the

maj

ority

of t

he b

ales

. Fro

m th

e re

sults

of t

he p

rese

nted

stud

y, ba

led

gras

s sila

ge c

an b

e cl

asse

d as

a

min

or ri

sk fo

r the

tran

smis

sion

of M

AP.

Kho

l et a

l. 20

10

Plan

ts g

row

ing

in

cont

amin

ated

soi

lsM

. a. p

arat

uber

culo

sis

The

aim

of t

his s

tudy

was

to d

emon

stra

te th

e pe

rsis

tenc

e of

Myc

obac

teri

um a

vium

subs

p. p

arat

uber

culo

sis (

MA

P) in

soil

and

colo

niza

tion

of d

iffer

ent p

lant

par

ts a

fter

del

iber

ate

expo

sure

to m

ouflo

n fe

ces n

atur

ally

con

tam

inat

ed w

ith d

iffer

ent a

mou

nts

of M

AP.

Sam

ples

of a

eria

l par

ts o

f pla

nts,

thei

r roo

ts, a

nd th

e so

il be

low

the

root

s wer

e co

llect

ed a

fter

15

wee

ks a

nd e

xam

ined

us

ing

IS90

0 re

al-t

ime

quan

titat

ive

PCR

(qPC

R) a

nd c

ultiv

atio

n. A

lthou

gh th

e pr

esen

ce o

f via

ble

MA

P ce

lls w

as n

ot d

emon

-st

rate

d, a

lmos

t all

sam

ples

wer

e fo

und

to b

e po

sitiv

e us

ing

qPC

R. M

AP

IS90

0 w

as n

ot o

nly

foun

d in

the

uppe

r gre

en p

arts

, but

al

so in

the

root

s and

soil

sam

ples

(fro

m 1

.00

× 10

0 to 6

.43

× 10

3 . The

leve

l of s

oil a

nd p

lant

con

tam

inat

ion

was

influ

ence

d m

ainl

y by

moi

stur

e, c

lay

cont

ent,

and

the

dept

h fr

om w

hich

the

sam

ples

wer

e co

llect

ed, r

athe

r tha

n by

the

initi

al c

once

ntra

tion

of M

AP

in th

e fe

ces a

t the

beg

inni

ng o

f the

exp

erim

ent.

Prib

ylov

a et

al.

2011

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

662

Mai

zeM

. phl

eiA

n un

ders

tand

ing

of e

colo

gica

l con

ditio

ns e

ffect

ing

on b

acte

rial

inoc

ulan

ts is

impo

rtan

t whe

n in

trod

ucin

g m

icro

bes f

or in

crea

s-in

g pl

ant g

row

th a

nd p

rodu

ctiv

ity. I

n th

is st

udy

the

influ

ence

of t

wo

diffe

rent

soil

type

s on

the

stim

ulat

ory

effec

t of p

lant

gro

wth

pr

omot

ing

rhiz

obac

teri

a fo

r mai

ze w

as in

vest

igat

ed. Th

e in

vest

igat

ions

wer

e ca

rrie

d ou

t in

pot e

xper

imen

ts w

ith c

alca

reou

s ca

lcis

ol so

il ta

ken

from

Sir

dary

a, U

zbek

ista

n an

d lo

amy

sand

from

Mue

nche

berg

, Ger

man

y. Th

e ba

cter

ia st

rain

s Pse

udom

onas

al

calig

enes

PsA

15, B

acill

us p

olym

yxa

BcP2

6 an

d M

ycob

acte

rium

phl

ei M

bP18

had

a m

uch

bett

er st

imul

ator

y eff

ect o

n pl

ant

grow

th a

nd n

itrog

en (N

), ph

osph

orus

(P) a

nd p

otas

sium

(K) u

ptak

e of

mai

ze in

nut

rien

t defi

cien

t cal

ciso

l soi

l. Th

eir s

timul

ator

y effi

cien

cy re

duce

d in

rela

tivel

y ri

ch lo

amy

sand

soil

whe

re b

acte

rial

inoc

ulan

ts st

imul

ated

onl

y ro

ot g

row

th a

nd N

, K u

ptak

e of

ro

ot. Th

ese

resu

lts su

gges

t tha

t pla

nt g

row

th st

imul

atin

g effi

cien

cy o

f bac

teri

al in

ocul

ants

affe

cted

by

soil

nutr

ition

al c

ondi

tion.

Th

e ba

cter

ial i

nocu

latio

n ha

s a m

uch

bett

er st

imul

ator

y eff

ect o

n pl

ant g

row

th in

nut

rien

t defi

cien

t soi

l tha

n in

nut

rien

t ric

h so

il.

Egam

-be

rdiy

eva

2007

Ric

eM

. bol

letii

In th

is st

udy,

the

effec

ts o

f pla

nt g

enot

ype,

soil

type

and

nut

rien

t use

effi

cien

cy o

n th

e co

mpo

sitio

n of

diff

eren

t bac

teri

al c

om-

mun

ities

ass

ocia

ted

with

rice

root

s wer

e in

vest

igat

ed. Th

us, t

otal

bac

teri

a, A

lpha

- and

Bet

a-pr

oteo

bact

eria

, Pse

udom

onas

and

A

ctin

obac

teri

a w

ere

stud

ied

usin

g PC

R, f

ollo

wed

by

dena

turi

ng g

radi

ent g

el e

lect

roph

ores

is (P

CR-

DG

GE)

. Ric

e ge

noty

pe

dete

rmin

ed, t

o a

larg

e ex

tent

, the

com

posi

tion

of th

e di

ffere

nt b

acte

rial

com

mun

ities

acr

oss c

ultiv

ars.

Sev

eral

cul

tivar

s bel

ong-

ing

to O

ryza

sativ

a ss

p. in

dica

tend

ed to

sele

ct si

mila

r bac

teri

al c

omm

uniti

es, w

here

as th

ose

belo

ngin

g to

subs

peci

es ja

poni

ca

and

arom

atic

a se

lect

ed o

nes w

ith d

iver

gent

com

mun

ity st

ruct

ures

. An

effec

t of s

oil t

ype

was

pro

noun

ced

for t

he A

ctin

obac

teri

a co

mm

uniti

es, w

hile

a sm

all e

ffect

of ’

impr

oved

’ and

’tra

ditio

nal’

plan

ts w

as n

oted

for a

ll co

mm

uniti

es a

naly

zed.

A fe

w d

omin

ant

band

s in

PCR-

DG

GE,

affi

liate

d w

ith R

hizo

bium

radi

obac

ter,

Dic

keya

zea

e, M

ycob

acte

rium

bol

letii

and

with

mem

bers

of t

he

Rhiz

obia

les,

Rho

dosp

irill

acea

e an

d Pa

enib

acill

acea

e, w

ere

spre

ad a

cros

s cul

tivar

s. In

con

tras

t, a

maj

ority

of b

ands

(e.g

. affi

liate

d w

ith E

nter

obac

ter c

loac

ae o

r Bur

khol

deri

a ku

ruri

ensi

s) w

as o

nly

pres

ent i

n pa

rtic

ular

cul

tivar

s or w

as e

rrat

ical

ly d

istr

ibut

ed

amon

g ri

ce re

plic

ates

. Thes

e fin

ding

s sug

gest

ed th

at b

oth

bact

eria

l ada

ptat

ion

and

plan

t gen

otyp

e co

ntri

bute

to th

e sh

apin

g of

th

e dy

nam

ic b

acte

rial

com

mun

ities

ass

ocia

ted

with

root

s of r

ice

plan

ts.

Har

doim

et

al.

2011

Whe

at r

hizo

sphe

re

M. p

hlei

One

of t

he n

atur

al re

serv

oirs

of p

oten

tially

hum

an-p

atho

geni

c ba

cter

ia is

bel

ieve

d to

be

the

rhiz

osph

ere.

The

aim

of t

he p

rese

nt

wor

k w

as to

test

non

tube

rcul

ous m

ycob

acte

rium

Myc

obac

teri

um p

hlei

MbP

18 fo

r its

abi

lity

to c

olon

ize

the

rhiz

osph

ere

of w

heat

an

d to

eva

luat

e its

effe

ct o

n pl

ant g

row

th u

nder

salin

e co

nditi

ons.

In c

ompe

titiv

e w

heat

root

tip

colo

niza

tion

assa

ys, M

. phl

ei

MbP

18 sh

owed

poo

r com

petit

ive

colo

niza

tion

of th

e w

heat

rhiz

osph

ere

com

pare

d to

the

refe

renc

e st

rain

. The

stra

in p

rodu

ced

lipas

e, a

myl

ase,

cel

lula

se, a

nd p

ectin

ase

and

grew

wel

l in

the

pres

ence

of h

igh

salt

(up

to 4

% N

aCl)

and

at h

igh

tem

pera

ture

s (up

to

40

degr

ees C

). It

was

als

o ab

le to

util

ize

a w

ide

rang

e of

car

bohy

drat

es fo

r gro

wth

. The

stra

in p

rodu

ced

indo

le-3

-ace

tic a

cid

and

prov

ed to

be

very

effi

cien

t in

prom

otin

g a

sign

ifica

nt in

crea

se in

the

shoo

t and

root

of w

heat

und

er sa

line

cond

ition

s. In

co

nclu

sion

, the

resu

lts o

f thi

s stu

dy in

dica

te th

at M

. phl

ei M

bP18

has

ben

efici

al e

ffect

s on

plan

t gro

wth

und

er sa

line

cond

ition

s th

roug

h its

abi

lity

to p

rodu

ce d

iffer

ent b

iolo

gica

lly a

ctiv

e co

mpo

unds

such

as c

ell w

all-d

egra

ding

enz

ymes

and

the

phyt

ohor

-m

one

auxi

n. H

owev

er, i

ts c

ompe

titiv

e co

loni

zatio

n ab

ilitie

s in

the

rhiz

osph

ere

are

poor

. In

light

of t

his o

bser

vatio

n, a

ttem

pts

shou

ld b

e m

ade

to m

anag

e th

e rh

izos

pher

e in

ord

er to

pre

vent

col

oniz

atio

n of

the

rhiz

osph

ere

by p

atho

gens

. This

will

hel

p re

mov

e m

ycob

acte

ria

from

hab

itats

whe

re h

uman

s or a

nim

als c

an b

e ex

pose

d.

Egam

-be

rdie

va

2012

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

663

Lett

uce

Tran

sgen

ic le

ttuc

e pl

ants

(Lac

tuca

sativ

a L.

) with

gen

es c

odin

g th

e sy

nthe

sis o

f tub

ercu

lous

ant

igen

s wer

e ob

tain

ed u

sing

the

Agr

obac

teri

um-m

edia

ted

tran

sfor

mat

ion

proc

edur

e. C

otyl

edon

ary

leav

es o

f in

vitr

o le

ttuc

e se

edlin

gs (c

vs. E

rala

sh, S

nezh

inka

, an

d Ru

bino

voe

Kru

zhev

o) w

ere

tran

sfor

med

with

pla

smid

s con

tain

ing

nptI

I, a

sele

ctiv

e ne

omyc

in p

hosp

hotr

ansf

eras

e II

gen

e,

and

ESAT

6, A

g85B

(-T

MD

), an

d ES

AT6:

Ag8

5B (-

TM

D) t

arge

t gen

es. A

PC

R an

alys

is o

f the

gen

ome’s

DN

A c

onfir

med

the

pres

-en

ce o

f bot

h se

lect

ive

and

targ

et g

enes

in a

ll pl

ants

exa

min

ed. A

t the

sam

e tim

e, R

T-PC

R an

alys

is sh

owed

that

, in

the

case

of

stab

le tr

ansc

ript

ion

of th

e np

tII g

ene,

bot

h th

e pr

esen

ce a

nd a

bsen

ce o

f tra

nscr

iptio

n of

the

ESAT

6 ge

ne a

re p

ossi

ble.

Mat

-vi

eiev

a et

al

. 200

9

Chi

cory

cot

yled

ons

Tran

sgen

ic p

lant

s con

tain

ing

eith

er if

n-al

pha

2b g

ene

enco

ding

hum

an le

ukoc

ytic

inte

rfer

on o

r esx

A::f

bpB

ΔT

MD

gen

es e

ncod

-in

g M

ycob

acte

rium

tube

rcul

osis

ant

igen

s ESA

T6

and

Ag8

5B w

ere

rege

nera

ted

from

hai

ry ro

ot c

ultu

res a

fter

tran

sfor

mat

ion

of

chic

ory

coty

ledo

ns (C

icho

rium

inty

bus L

. var

. Fol

iosu

m H

egi)

with

a w

ild-t

ype

A. r

hizo

gene

s A4

stra

in. Th

e di

rect

shoo

t reg

ener

a-tio

n fr

om tr

ansg

enic

root

s with

out c

allu

s for

mat

ion

phas

e on

gro

wth

regu

lato

r-fr

ee n

utri

ent m

ediu

m w

as d

emon

stra

ted.

The

tran

sgen

es tr

ansf

er a

nd tr

ansc

ript

ion

in th

e pl

ants

wer

e co

nfirm

ed b

y th

e re

sults

of R

T-PC

R an

d PC

R an

alys

es.

Mat

-vi

eiev

a et

al

. 201

1

Tube

rcul

osis

tran

smis

-si

on b

y ec

olog

ical

fa

ctor

s

In th

is p

aper

, the

cum

ulat

ive

effec

t of e

colo

gica

l fac

tors

in th

e ha

bita

t on

the

spre

ad o

f tub

ercu

losi

s (T

B) in

hum

an p

opul

atio

n is

m

odel

ed a

nd a

naly

zed.

The

tota

l hum

an p

opul

atio

n is

div

ided

into

two

clas

ses,

susc

eptib

les a

nd in

fect

ives

. It i

s ass

umed

that

T

B is

not

onl

y sp

read

by

dire

ct c

onta

cts w

ith in

fect

ives

in th

e po

pula

tion

but a

lso

indi

rect

ly b

y ba

cter

ia w

hich

are

em

itted

by

infe

ctiv

es in

the

habi

tat.

It is

ass

umed

furt

her t

hat b

acte

ria

surv

ive

due

to c

ondu

cive

eco

logi

cal f

acto

rs su

ch a

s flow

er p

ots,

plan

ts, g

rass

es, h

uman

clo

thes

, etc

. in

the

habi

tat.

The

cum

ulat

ive

dens

ity o

f eco

logi

cal f

acto

rs in

the

habi

tat i

s ass

umed

to b

e go

vern

ed b

y a

popu

latio

n de

nsity

dep

ende

nt lo

gist

ic m

odel

. The

anal

ysis

of t

he m

odel

show

s tha

t as p

aram

eter

s gov

erni

ng th

e co

nduc

ive

ecol

ogic

al fa

ctor

s in

the

habi

tat i

ncre

ase,

the

spre

ad o

f TB

incr

ease

s. Th

e sa

me

resu

lt is

als

o fo

und

with

the

incr

ease

in

the

para

met

er g

over

ning

the

surv

ival

and

acc

umul

atio

n of

bac

teri

a in

the

habi

tat.

It is

furt

her f

ound

that

due

to im

mig

ratio

n of

th

e po

pula

tion

TB

beco

mes

mor

e en

dem

ic. A

num

eric

al st

udy

of th

e m

odel

is a

lso

carr

ied

out t

o su

ppor

t the

ana

lytic

al re

sults

.

Nar

esh

et

al. 2

009

Prot

ozoa

-pla

nt

sym

bios

isTh

e su

rviv

al o

f Sal

mon

ella

ente

rica

was

rece

ntly

show

n to

incr

ease

whe

n th

e ba

cter

ia w

ere

sequ

este

red

in e

xpel

led

food

vac

uole

s (v

esic

les)

of T

etra

hym

ena.

Bec

ause

fres

h pr

oduc

e is

incr

easi

ngly

link

ed to

out

brea

ks o

f ent

eric

illn

ess,

the

pres

ent i

nves

tigat

ion

aim

ed to

det

erm

ine

the

prev

alen

ce o

f pro

tozo

a on

spin

ach

and

lettu

ce a

nd to

exa

min

e th

eir i

nter

actio

ns w

ith S

. ent

eric

a, E

sche

ri-

chia

coli

O15

7:11

7, a

nd L

iste

ria

mon

ocyt

ogen

es. G

lauc

oma

sp.,

Col

poda

stei

nii,

and

Acan

tham

oeba

pal

estin

ensis

wer

e cu

lture

d fr

om

stor

e-bo

ught

spin

ach

and

lettu

ce a

nd u

sed

in o

ur st

udy.

A st

rain

of T

etra

hym

ena

pyri

form

is p

revi

ously

isol

ated

from

spin

ach

and

a so

il-bo

rne

Tetr

ahym

ena

sp. w

ere

also

use

d. W

ashe

d pr

otoz

oa w

ere

allo

wed

to g

raze

on

gree

n flu

ores

cent

pro

tein

-or r

ed fl

uore

scen

t pr

otei

n-la

bele

d en

teri

c pa

thog

ens.

Sign

ifica

nt d

iffer

ence

s in

inte

ract

ions

am

ong

the

vari

ous p

rotis

t-en

teri

c pa

thog

en c

ombi

na-

tions

wer

e ob

serv

ed. V

esic

les w

ere

prod

uced

by

Gla

ucom

a w

ith a

ll of

the

bact

eria

l str

ains

, alth

ough

L. m

onoc

ytog

enes

resu

lted

in th

e sm

alle

st n

umbe

r per

cili

ate.

Ves

icle

pro

duct

ion

was

obs

erve

d al

so d

urin

g gr

azin

g of

Tet

rahy

men

a on

E. c

oli O

157:

117

and

S. en

teri

ca b

ut n

ot d

urin

g gr

azin

g on

L. m

onoc

ytog

enes

, in

vitr

o an

d on

leav

es. A

ll ve

sicl

es c

onta

ined

inta

ct fl

uore

scin

g ba

cter

ia. I

n co

ntra

st, C

. ste

inii

and

the

amoe

ba d

id n

ot p

rodu

ce v

esic

les f

rom

any

of t

he e

nter

ic p

atho

gens

, nor

wer

e pa

thog

ens t

rapp

ed w

ithin

th

eir c

ysts

. Stu

dies

of t

he fa

te o

f E. c

oli O

157:

117

in e

xpel

led

vesi

cles

reve

aled

that

by

4 It

afte

r add

ition

of s

pina

ch e

xtra

ct, t

he b

ac-

teri

a m

ultip

lied

and

esca

ped

the

vesi

cles

. The

pres

ence

of p

roto

zoa

on le

afy

vege

tabl

es a

nd th

eir s

eque

stra

tion

of e

nter

ic b

acte

ria

in

vesi

cles

indi

cate

that

they

may

pla

y an

impo

rtan

t rol

e in

the

ecol

ogy

of h

uman

pat

hoge

ns o

n pr

oduc

e.

Gou

ra-

bath

ini e

t al

. 200

8

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

664

Tabl

e 6.

Myc

obac

teri

a in

air

Type

of s

ampl

es

Myc

obac

teri

a de

tect

edA

bstr

act e

xcer

pts

Refe

renc

e

Ther

apy

pool

aer

osol

an

d ai

rM

. sp.

...w

e co

nduc

ted

a m

ultis

easo

n su

rvey

of m

icro

orga

nism

s pre

sent

in th

is th

erap

y po

ol w

ater

, in

biofi

lms a

ssoc

iate

d w

ith th

e po

ol c

onta

inm

ent w

alls

, and

in a

ir im

med

iate

ly a

bove

the

pool

. The

surv

ey u

sed

cultu

re, m

icro

scop

y, an

d cu

lture

-ind

epen

dent

m

olec

ular

phy

loge

netic

ana

lyse

s. A

lthou

gh o

utfit

ted

with

a st

ate-

of-t

he a

rt U

V-p

erox

ide

disi

nfec

tion

syst

em, t

he n

umbe

rs o

f ba

cter

ia in

the

ther

apy

pool

wat

er w

ere

rela

tivel

y hi

gh c

ompa

red

with

the

pota

ble

wat

er u

sed

to fi

ll th

e po

ol. R

egar

dles

s of t

he

sour

ce, d

irec

t mic

rosc

opic

cou

nts o

f mic

robe

s wer

e ro

utin

ely

appr

oxim

ate

to 1

,000

tim

es g

reat

er th

an c

onve

ntio

nal p

late

cou

nts.

Ana

lysi

s of c

lone

libr

arie

s of s

mal

l sub

unit

rRN

A g

enes

from

env

iron

men

tal D

NA

pro

vide

d ph

ylog

enet

ic d

iver

sity

est

imat

es o

f th

e m

icro

orga

nism

s col

lect

ed in

and

abo

ve th

e po

ol. A

surv

ey o

f >1,

300

rRN

A g

enes

yie

lded

a to

tal o

f 628

uni

que

sequ

ence

s, th

e m

ost c

omm

on o

f whi

ch w

as n

earl

y id

entic

al to

that

of M

. avi

um st

rain

s. Th

e hi

gh p

ropo

rtio

n of

clo

nes w

ith d

iffer

ent M

ycob

ac-

teri

um sp

p. rR

NA

gen

es su

gges

ted

that

such

org

anis

ms c

ompr

ised

a si

gnifi

cant

frac

tion

of m

icro

bes i

n th

e po

ol w

ater

(to

>30%

) an

d pr

efer

entia

lly p

artit

ion

into

aer

osol

s (to

>80

%) r

elat

ive

to o

ther

wat

erbo

rne

bact

eria

pre

sent

.

Ang

enen

t et

al.

2005

Show

er a

eros

olM

. sp.

M. m

ucog

enic

um

To q

uant

ify th

e m

icro

bial

load

in sh

ower

wat

er a

nd a

eros

ol sa

mpl

es, w

e us

ed c

ultu

re, m

icro

scop

ic, a

nd q

uant

itativ

e PC

R m

eth-

ods t

o in

vest

igat

e fo

ur sh

ower

stal

ls in

a st

em c

ell t

rans

plan

t uni

t at B

arne

s-Je

wis

h H

ospi

tal i

n St

. Lou

is, M

O. W

e al

so te

sted

m

embr

ane-

inte

grat

ed sh

ower

head

s as a

pos

sibl

e m

itiga

tion

stra

tegy

. In

addi

tion

to q

uant

ifica

tion,

a 1

6S rR

NA

gen

e se

quen

cing

su

rvey

was

use

d to

cha

ract

eriz

e th

e ab

unda

nt b

acte

rial

pop

ulat

ions

with

in sh

ower

wat

er a

nd a

eros

ols.

The

aver

age

tota

l bac

te-

rial

cou

nts w

ere

2.2

× 10

7 cel

ls/li

ter i

n sh

ower

wat

er a

nd 3

.4 ×

104 c

ells

/m3 in

show

er a

eros

ol, a

nd th

ese

coun

ts w

ere

redu

ced

to

6.3 

× 10

4 cel

ls/li

ter (

99.6

% e

ffici

ency

) and

8.9

× 1

03 cel

ls/m

3 (82.

4% e

ffici

ency

), re

spec

tivel

y, af

ter m

embr

ane-

inte

grat

ed sh

ow-

erhe

ads w

ere

inst

alle

d. P

oten

tially

pat

hoge

nic

orga

nism

s wer

e fo

und

in b

oth

wat

er a

nd a

eros

ol sa

mpl

es fr

om th

e co

nven

tiona

l sh

ower

s. M

ost n

otab

le w

as th

e pr

esen

ce o

f Myc

obac

teri

um m

ucog

enic

um (9

9.5%

iden

tity)

in th

e w

ater

and

Pse

udom

onas

aer

ugi-

nosa

(99.

3% id

entit

y) in

the

aero

sol s

ampl

es.

Perk

ins e

t al

. 200

9

Den

tal u

nit w

ater

lines

ae

roso

lD

enta

l uni

t wat

erlin

es (D

UW

L) su

ppor

t gro

wth

of a

den

se m

icro

bial

pop

ulat

ion

that

incl

udes

pat

hoge

ns a

nd h

yper

sens

itivi

ty-

indu

cing

bac

teri

a, su

ch a

s Leg

ione

lla sp

p. a

nd n

on-t

uber

culo

us m

ycob

acte

ria

(NT

M).

Dyn

amic

den

tal i

nstr

umen

ts c

onne

cted

to

DU

WL

gene

rate

aer

osol

s in

the

wor

k en

viro

nmen

t, w

hich

cou

ld a

llow

wat

erbo

rne

path

ogen

s to

be a

eros

oliz

ed. Th

e us

e of

the

real

-tim

e qu

antit

ativ

e po

lym

eras

e ch

ain

reac

tion

(qPC

R) p

rovi

des a

mor

e ac

cura

te e

stim

atio

n of

exp

osur

e le

vels

com

pare

d w

ith

the

trad

ition

al c

ultu

re a

ppro

ach.

Bio

aero

sol s

ampl

ing

was

per

form

ed 1

3 tim

es in

an

isol

ated

den

tal t

reat

men

t roo

m a

ccor

ding

to

a st

anda

rdiz

ed p

roto

col t

hat i

nclu

ded

four

den

tal p

roph

ylax

is tr

eatm

ents

. Inh

alab

le d

ust s

ampl

es w

ere

take

n at

the

brea

thin

g zo

ne o

f bot

h th

e hy

gien

ist a

nd p

atie

nt a

nd o

utsi

de th

e tr

eatm

ent r

oom

(con

trol

). To

tal b

acte

ria

as w

ell a

s Leg

ione

lla sp

p. a

nd

NT

M w

ere

quan

tified

by

qPC

R in

bio

aero

sol a

nd D

UW

L w

ater

sam

ples

. Den

tal s

taff

and

patie

nts a

re e

xpos

ed to

bac

teri

a ge

ner-

ated

dur

ing

dent

al tr

eatm

ents

(up

to 4

.3 E

+ 0

5 ba

cter

ia p

er m

3 of a

ir).

Beca

use

DU

WL

wat

er st

udie

d w

as w

eakl

y co

ntam

inat

ed

by L

egio

nella

spp.

and

NT

M, t

heir

aer

osol

izat

ion

duri

ng d

enta

l tre

atm

ent w

as n

ot si

gnifi

cant

. As a

resu

lt, in

fect

ious

and

sens

iti-

zatio

n ri

sks a

ssoc

iate

d w

ith le

gion

ella

e an

d N

TM

shou

ld b

e m

inim

al.

Dut

il et

al

. 200

7

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

665

Air

from

pea

t mos

s

proc

essi

ng p

lant

sM

. sp.

M. m

alm

oens

e M

. sm

egm

atis

M

. gra

ceum

M

. boh

emic

umM

. int

erje

ctum

We

eval

uate

d th

e pr

esen

ce o

f myc

obac

teri

a in

air

sam

ples

from

pea

t mos

s pro

cess

ing

plan

ts u

sing

mol

ecul

ar b

iolo

gy a

ppro

ache

s (c

loni

ng-s

eque

ncin

g an

d po

lym

eras

e ch

ain

reac

tion

(PC

R)) a

nd th

e w

orke

rs e

xpos

ure

usin

g im

mun

oglo

bulin

G (I

gG) c

ompl

exes

to

myc

obac

teri

a. In

add

ition

, spe

cies

det

ecte

d in

air

sam

ples

and

in p

eat m

oss w

ere

com

pare

d. T

wo

peat

mos

s pro

cess

ing

plan

ts

wer

e ch

osen

am

ong

14 p

revi

ousl

y st

udie

d. A

tota

l of 4

9 cl

ones

wer

e se

quen

ced.

Rea

l-tim

e PC

R w

as a

lso

perf

orm

ed o

n th

e sa

me

air s

ampl

es to

eva

luat

e th

e ai

rbor

ne c

once

ntra

tion

of m

ycob

acte

ria

and

estim

ate

expo

sure

leve

ls. S

ever

al M

ycob

acte

rium

spec

ies

wer

e pr

esen

t in

the

air s

ampl

es (M

. mal

moe

nse,

M. s

meg

mat

is, M

. gra

ceum

, M. b

ohem

icum

, and

M. i

nter

ject

um).

Myc

obac

teri

um

aviu

m w

as re

cove

red

by c

ultu

re in

pea

t mos

s but

not

in th

e ai

r usi

ng th

e m

olec

ular

app

roac

h. T

otal

air

born

e M

ycob

acte

rium

co

ncen

trat

ion

was

est

imat

ed a

t 8.2

× 1

08 /m3 . W

orke

rs h

ad Ig

G a

gain

st th

e m

ycob

acte

rial

mix

and

M. a

vium

, sug

gest

ing

sign

ifi-

cant

exp

osur

e. Th

e fin

ding

s fro

m a

ir sa

mpl

es, s

uppo

rted

by

IgG

mea

sure

men

ts, d

emon

stra

te th

at p

eat m

oss p

roce

ssin

g pl

ant

wor

kers

are

exp

osed

to m

ycob

acte

ria

in a

dditi

on to

oth

er b

iolo

gica

l age

nts.

Cay

er e

t al

. 200

7

Aer

osol

-gen

erat

ing

acti

viti

esM

. avi

um c

ompl

ex

Ratio

nale

: Myc

obac

teri

um a

vium

com

plex

lung

dis

ease

is a

n in

crea

sing

ly c

omm

on a

nd c

hron

ical

ly d

ebili

tatin

g pr

oble

m. S

ever

al

host

trai

ts h

ave

been

sugg

este

d or

con

firm

ed a

s ris

k fa

ctor

s. P

oten

tial e

nvir

onm

enta

l and

beh

avio

ral r

isk

fact

ors h

ave

also

bee

n pr

opos

ed. F

ew h

ave

been

eva

luat

ed in

com

para

tive

stud

ies.

Obj

ectiv

es: T

o de

term

ine

if ae

roso

l-gen

erat

ing

activ

ities

in th

e ho

me

and

gard

en, f

eatu

res o

f the

hom

e w

ater

supp

ly, o

r sev

eral

pul

mon

ary

and

imm

une-

com

prom

isin

g co

nditi

ons a

re a

ssoc

iate

d w

ith

Myc

obac

teri

um a

vium

com

plex

lung

dis

ease

. Met

hods

: Cas

es w

ere

recr

uite

d fr

om a

cade

mic

med

ical

cen

ters

and

by

info

rmal

re

ferr

als f

rom

non

univ

ersi

ty p

ract

ices

in W

ashi

ngto

n an

d O

rego

n. C

ontr

ol su

bjec

ts w

ere

recr

uite

d by

rand

om-d

igit

dial

ing

and

mat

ched

to c

ases

by

age,

sex,

and

par

tial t

elep

hone

num

ber.

Ass

ocia

tions

wer

e m

easu

red

as o

dds r

atio

s (O

R) e

stim

ated

usi

ng

cond

ition

al lo

gist

ic re

gres

sion

. Mea

sure

men

ts a

nd M

ain

Resu

lts: K

now

n an

d po

tent

ial r

isk

fact

ors w

ere

mea

sure

d by

in-h

ome

inte

rvie

w. F

ifty-

two

mat

ched

pai

rs w

ere

stud

ied.

Six

of 1

2 ex

amin

ed h

ost t

raits

wer

e as

soci

ated

with

dis

ease

, inc

ludi

ng h

isto

ry o

f ch

roni

c ob

stru

ctiv

e pu

lmon

ary

dise

ase

(OR

, 10;

95%

con

fiden

ce in

terv

al [C

I], 1

.2–8

0), p

neum

onia

hos

pita

lizat

ion

(OR

, 3.4

; 95%

C

l, 1.

1–11

), an

d st

eroi

d us

e (O

R, 8

; 95%

CI,

1.6–

41).

In c

ontr

ast,

11 o

f the

14

aero

sol-g

ener

atin

g ac

tiviti

es a

nd a

ll fiv

e fe

atur

es

of h

ome

wat

er su

pply

stud

ied

bore

litt

le o

r no

asso

ciat

ion

with

dis

ease

. Con

clus

ions

: Aer

osol

-gen

erat

ing

activ

ities

seem

not

to

be ri

sk fa

ctor

s for

Myc

obac

teri

um a

vium

com

plex

lung

dis

ease

in H

IV-n

egat

ive

adul

ts, b

ut p

rior

lung

dis

ease

and

imm

une-

sup-

pres

sing

dru

gs se

em to

be

asso

ciat

ed w

ith su

scep

tibili

ty.

Dir

ac e

t al

. 201

2

Aer

osol

infe

ctio

nM

. avi

umIn

a m

ouse

mod

el o

f myc

obac

teri

a-in

duce

d im

mun

opat

holo

gy, w

ild-t

ype

C57

BL/6

(WT

), IL

-18-

knoc

kout

(KO

) and

IFN

-alp

ha

beta

rece

ptor

-KO

mic

e de

velo

ped

circ

umsc

ript

, cen

tral

ly n

ecro

tizin

g gr

anul

omat

ous l

esio

ns in

resp

onse

to a

eros

ol in

fect

ion

with

M. a

vium

, whe

reas

mic

e de

ficie

nt in

the

IFN

-gam

ma

rece

ptor

, STA

T-1

or IR

F-1

did

not e

xhib

it gr

anul

oma

necr

osis

. Com

-pa

rativ

e, m

icro

arra

y-ba

sed

gene

exp

ress

ion

anal

ysis

in th

e lu

ngs o

f inf

ecte

d W

T a

nd IR

F-1-

KO

mic

e id

entifi

ed a

set o

f gen

es

who

se d

iffer

entia

l reg

ulat

ion

was

clo

sely

ass

ocia

ted

with

gra

nulo

ma

necr

osis

, am

ong

them

cat

heps

in K

, cys

tatin

F a

nd m

atri

x m

etal

lopr

otea

se 1

0. F

urth

er m

icro

arra

y-ba

sed

com

pari

son

of g

ene

expr

essi

on in

the

lung

s of i

nfec

ted

WT,

IFN

-gam

ma-

KO

and

IR

F-1-

KO

mic

e re

veal

ed fo

ur d

istin

ct c

lust

ers o

f gen

es w

ith v

aria

ble

depe

nden

ce o

n th

e pr

esen

ce o

f IFN

-gam

ma,

IRF-

1 or

bot

h.

In p

artic

ular

, IRF

-1 a

ppea

red

to b

e di

rect

ly in

volv

ed in

the

diffe

rent

iatio

n of

a ty

pe I

imm

une

resp

onse

to m

ycob

acte

rial

infe

c-tio

n. In

sum

mar

y, IR

F-1,

rath

er th

an b

eing

a m

ere

tran

scri

ptio

n fa

ctor

dow

nstr

eam

of I

FN-g

amm

a, m

ay b

e a

mas

ter r

egul

ator

of

myc

obac

teri

a-in

duce

d im

mun

opat

holo

gy.

Aly

et a

l. 20

09

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

666

Hot

tub

expo

sure

Non

-tub

ercu

lous

m

ycob

acte

ria

Hot

tub

expo

sure

has

bee

n ca

usal

ly a

ssoc

iate

d w

ith a

ster

oid-

resp

onsi

ve, g

ranu

lom

atou

s lun

g di

seas

e fe

atur

ing

non-

tube

rcul

ous

myc

obac

teri

al (N

TM

) gro

wth

in b

oth

clin

ical

and

env

iron

men

tal s

ampl

es. L

ittle

is k

now

n re

gard

ing

prev

alen

ce o

f and

risk

fac-

tors

for N

TM

-con

tam

inat

ion

and

asso

ciat

ed il

lnes

s in

thes

e se

ttin

gs. I

n th

is st

udy,

the

freq

uenc

y of

NT

M g

row

th a

nd a

eros

oliz

a-tio

n in

18

publ

ic h

ot tu

bs a

nd w

arm

wat

er th

erap

y po

ols a

nd th

e fa

ctor

s ass

ocia

ted

with

myc

obac

teri

al g

row

th w

ere

anal

yzed

. Ea

ch si

te w

as c

hara

cter

ized

by

wat

er c

hem

istr

y an

alys

is; a

que

stio

nnai

re o

n m

aint

enan

ce, d

isin

fect

ion,

and

wat

er q

ualit

y; a

nd

air a

nd w

ater

sam

plin

g fo

r qua

ntita

tive

NT

M c

ultu

re. N

TM

wer

e de

tect

ed in

air

or w

ater

from

13/

18 (7

2%) s

ites;

a st

rong

cor

-re

latio

n w

as fo

und

betw

een

the

max

imum

air

and

wat

er N

TM

con

cent

ratio

ns (r

ho 0

.49,

P =

0.0

4). U

se o

f hal

ogen

(chl

orin

e or

br

omin

e) d

isin

fect

ion

was

ass

ocia

ted

with

sign

ifica

ntly

low

er a

ir a

nd w

ater

con

cent

ratio

ns o

f NT

M c

ompa

red

with

dis

infe

ctio

n us

ing

ultr

avio

let l

ight

and

hyd

roge

n pe

roxi

de (P

= 0

.01–

0.04

). H

ighe

r wat

er tu

rnov

er ra

tes w

ere

also

ass

ocia

ted

with

low

er a

ir

and

wat

er N

TM

con

cent

ratio

ns (P

= 0

.02–

0.03

). Th

ese

findi

ngs s

ugge

st th

at N

TM

are

freq

uent

ly d

etec

tabl

e in

the

air a

nd w

ater

of

spas

and

ther

apy

pool

s and

that

par

ticul

ar m

aint

enan

ce a

nd d

isin

fect

ion

appr

oach

es a

ffect

NT

M b

ioae

roso

l con

cent

ratio

ns in

th

ese

sett

ings

.

Gla

zer e

t al

. 200

7

Hot

wat

er a

eros

ols

Non

-tub

ercu

lous

m

ycob

acte

ria

Obj

ectiv

e: H

uman

act

iviti

es a

ssoc

iate

d w

ith a

eros

ol-g

ener

atin

g ho

t wat

er so

urce

s are

incr

easi

ngly

pop

ular

. Rec

ently

, a h

yper

-se

nsiti

vity

pne

umon

itis (

HP)

-like

gra

nulo

mat

ous l

ung

dise

ase,

with

non

-tub

ercu

lous

myc

obac

teri

a fr

om e

xpos

ure

to h

ot w

ater

ae

roso

ls fr

om h

ot tu

bs/s

pas,

show

ers,

and

indo

or sw

imm

ing

pool

s, h

as b

een

desc

ribe

d in

imm

unoc

ompe

tent

indi

vidu

als (

also

ca

lled

“hot

tub

lung

”). O

ur o

bjec

tive

in th

is st

udy

was

to e

xam

ine

four

add

ition

al c

ases

of h

ot tu

b lu

ng a

nd c

ompa

re th

ese

case

s w

ith o

ther

s rep

orte

d in

the

Engl

ish

prin

t lite

ratu

re o

n th

is d

isea

se. D

ata

sour

ces a

nd e

xtra

ctio

n: W

e re

tros

pect

ivel

y re

view

ed

all c

ases

(n =

4) o

f pre

sum

ptiv

ely

diag

nose

d ho

t tub

lung

in im

mun

ocom

pete

nt in

divi

dual

s at t

he v

ario

us p

hysi

cian

pra

ctic

es in

Sp

ring

field

, Illi

nois

, dur

ing

2001

–200

5. In

add

ition

, we

sear

ched

Med

line

for c

ases

of h

ot tu

b lu

ng d

escr

ibed

in th

e lit

erat

ure.

D

ata

synt

hesi

s: W

e su

mm

ariz

ed th

e cl

inic

al p

rese

ntat

ion

and

inve

stig

atio

ns o

f fou

r pre

sum

ptiv

e ca

ses a

nd re

view

ed p

revi

ousl

y re

port

ed c

ases

of h

ot tu

b lu

ng. C

oncl

usio

ns: Th

ere

is a

deb

ate

in th

e lit

erat

ure

whe

ther

hot

tub

lung

is a

n H

P or

a d

irec

t inf

ec-

tion

of th

e lu

ng b

y no

ntub

ercu

lous

myc

obac

teri

a. P

rim

ary

prev

entio

n of

this

dis

ease

relie

s on

vent

ilatio

n an

d go

od u

se p

ract

ices

. Se

cond

ary

prev

entio

n of

this

dis

ease

requ

ires

edu

catio

n of

bot

h th

e ge

nera

l pub

lic a

nd c

linic

ians

to a

llow

for t

he e

arly

dia

gnos

is

of th

is d

isea

se.

Sood

et

al. 2

007

Air

born

e tr

ansm

issi

onM

. bov

isD

espi

te y

ears

of s

tudy

the

prin

cipl

e tr

ansm

issi

on ro

ute

of b

ovin

e tu

berc

ulos

is to

cat

tle re

mai

ns u

nres

olve

d. Th

e di

stri

butio

n of

pa

thol

ogic

al le

sion

s, w

hich

are

con

cent

rate

d in

the

resp

irat

ory

syst

em, a

nd th

e ve

ry lo

w d

ose

of M

ycob

acte

rium

bov

is n

eede

d to

in

itiat

e in

fect

ion

from

a re

spir

ator

y tr

act c

halle

nge

sugg

est t

hat t

he d

isea

se is

spre

ad b

y ai

rbor

ne tr

ansm

issi

on. C

ritic

al to

the

air-

born

e tr

ansm

issi

on o

f a p

atho

geni

c m

icro

orga

nism

is it

s abi

lity

to su

rviv

e th

e st

ress

es in

curr

ed w

hils

t air

born

e. Th

is st

udy

dem

-on

stra

tes t

hat M

. bov

is is

resi

stan

t to

the

stre

sses

impo

sed

imm

edia

tely

aft

er b

ecom

ing

airb

orne

, 94%

surv

ivin

g th

e fir

st 1

0 m

in

afte

r aer

osol

isat

ion.

Onc

e ai

rbor

ne th

e or

gani

sm is

robu

st, i

ts v

iabi

lity

decr

easi

ng w

ith a

hal

f-lif

e of

app

roxi

mat

ely

1.5

hour

s. Th

ese

findi

ngs s

uppo

rt th

e hy

poth

esis

that

air

born

e tr

ansm

issi

on is

the

prin

cipl

e ro

ute

of in

fect

ion

for b

ovin

e tu

berc

ulos

is.

Gan

non

et a

l. 20

07

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

667

Air

M

. bov

isBa

ckgr

ound

: Evi

denc

e ha

s rec

ently

em

erge

d in

dica

ting

that

in a

dditi

on to

larg

e ai

rbor

ne d

ropl

ets,

fine

aer

osol

par

ticle

s can

be

an

impo

rtan

t mod

e of

influ

enza

tran

smis

sion

that

may

hav

e be

en h

ither

to u

nder

estim

ated

. Fur

ther

mor

e, re

cent

per

form

ance

stud

-ie

s eva

luat

ing

airb

orne

infe

ctio

n is

olat

ion

(AII

) roo

ms d

esig

ned

to h

ouse

infe

ctio

us p

atie

nts h

ave

reve

aled

maj

or d

iscr

epan

cies

be

twee

n w

hat i

s pre

scri

bed

and

wha

t is a

ctua

lly m

easu

red.

Met

hods

: We

cond

ucte

d an

exp

erim

enta

l stu

dy to

inve

stig

ate

the

use

of h

igh-

thro

ughp

ut in

-roo

m a

ir d

econ

tam

inat

ion

units

for s

uppl

emen

tal p

rote

ctio

n ag

ains

t air

born

e co

ntam

inat

ion

in a

reas

that

ho

st in

fect

ious

pat

ient

s. Th

e st

udy

incl

uded

bot

h in

trin

sic

perf

orm

ance

test

s of t

he a

ir-de

cont

amin

atio

n un

it ag

ains

t bio

logi

cal

aero

sols

of p

artic

ular

epi

dem

iolo

gic

inte

rest

and

fiel

d te

sts i

n a

hosp

ital A

II ro

om u

nder

diff

eren

t ven

tilat

ion

scen

ario

s. R

esul

ts:

The

unit

test

ed e

ffici

ently

era

dica

ted

airb

orne

H5N

2 in

fluen

za a

nd M

ycob

acte

rium

bov

is (a

4- t

o 5-

log

sing

le-p

ass r

educ

tion)

an

d, w

hen

impl

emen

ted

with

a ro

om e

xtra

ctor

, red

uced

the

peak

con

tam

inat

ion

leve

ls b

y a

fact

or o

f 5, w

ith d

econ

tam

inat

ion

rate

s at l

east

33%

fast

er th

an th

ose

achi

eved

with

the

extr

acto

r alo

ne. C

oncl

usio

n: H

igh-

thro

ughp

ut in

-roo

m a

ir tr

eatm

ent u

nits

ca

n pr

ovid

e su

pple

men

tal c

ontr

ol o

f air

born

e pa

thog

en le

vels

in p

atie

nt is

olat

ion

room

s.

Berg

eron

et

al.

2011

Air

M. p

inni

pedi

iO

bjec

tives

: Ail

outb

reak

of t

uber

culo

sis (

TB)

) in

sea

lions

occ

urre

d re

cent

ly in

a z

oo in

the

Net

herla

nds.

The

dise

ase

was

det

ecte

d in

a c

aptiv

e co

lony

con

sist

ing

of 2

9 an

imal

s kep

t in

an o

pen

air b

asin

with

an

indo

or n

ight

hou

se. A

ppro

xim

atel

y 25

ani

mal

kee

pers

w

ere

in c

lose

con

tact

with

the

anim

als.

Met

hods

: The

sea

lions

wer

e In

vest

igat

ed U

sing

the

tube

rcul

in sk

in te

st (T

ST) w

ith a

vian

an

d bo

vine

pur

ified

pro

tein

der

ivat

ive

(PPD

) and

, in

case

of p

ositi

vity

, nec

rops

ied.

A su

rvey

was

con

duct

ed a

mon

g th

e an

imal

kee

p-er

s Inc

ludi

ng T

STs w

ith M

ycob

acte

rium

tube

rcul

osis

com

plex

PPD

tube

rcul

in, a

che

st X

-ray

and

an

inte

rfer

on-g

amm

a re

leas

e as

say

(IG

RA).

Resu

lts: N

ecro

psy

was

pos

itive

for T

B in

13

of th

e 29

sea

lions

. Thre

e ca

ses o

f pul

mon

ary

invo

lvem

ent w

ere

foun

d. O

nly

one

of th

ese

was

infe

ctio

us a

nd it

was

ther

efor

e re

gard

ed a

s the

sour

ce c

ase.

The

caus

ativ

e M

ycob

acte

rium

was

iden

tified

as

M. p

inni

pedi

i. Si

x of

the

25 a

nim

al k

eepe

rs w

ere

TST

-pos

itive

; in

five

of th

ese,

infe

ctio

n w

as c

onfir

med

by

a po

sitiv

e IG

RA. C

oncl

u-si

on: T

rans

mis

sion

of M

. pin

nipe

dii i

nfec

tion

from

sea

lions

to h

uman

s was

est

ablis

hed

by T

ST. I

GRA

resu

lts la

rgel

y ag

reed

with

the

TST

resu

lts. N

ebul

isat

ion

whe

n cl

eani

ng th

e se

a lio

ns’ e

nclo

sure

was

mos

t lik

ely

the

mai

n ca

use

of tr

ansm

issi

on to

hum

ans.

Kie

rs e

t al

. 200

8

Aer

osol

M. a

. hom

inis

suis

Myc

obac

teri

um a

vium

ssp.

hom

inis

suis

, her

eafte

r ref

erre

d to

as M

. avi

um, f

orm

s bio

film

, a p

rope

rty

that

, in

mic

e, is

ass

ocia

ted

with

lung

infe

ctio

n vi

a ae

roso

l. A

s M. a

vium

mig

ht c

o-in

habi

t the

resp

irato

ry tr

act w

ith o

ther

pat

hoge

ns, t

reat

men

t of t

he c

o-pa

thog

en-a

ssoc

iate

d in

fect

ions

, suc

h as

in b

ronc

hiec

tasi

s, w

ould

exp

ose

M. a

vium

to th

erap

eutic

com

poun

ds th

at m

ay h

ave

thei

r or

igin

in o

ther

org

anis

ms s

hari

ng th

e na

tura

l env

ironm

ents

. Inc

ubat

ion

of M

. avi

um w

ith tw

o co

mpo

unds

pro

duce

d by

env

iron-

men

tal o

rgan

ism

s, st

rept

omyc

in a

nd te

trac

yclin

e, in

vitr

o at

subi

nhib

itory

con

cent

ratio

ns in

crea

sed

biofi

lm fo

rmat

ion

in a

num

ber

of M

. avi

um st

rain

s, al

thou

gh e

xpos

ure

to a

mpi

cilli

n, m

oxifl

oxac

in, r

ifam

pin

and

trim

etho

prim

-sul

pham

etho

xazo

le h

ad n

o eff

ect

on b

iofil

m fo

rmat

ion.

No

sele

ctio

n of

gen

otyp

ical

ly re

sist

ant c

lone

s was

obs

erve

d. A

lthou

gh in

cuba

tion

of b

acte

ria

in th

e pr

esen

ce

of st

rept

omyc

in u

preg

ulat

es th

e ex

pres

sion

of b

iofil

m-a

ssoc

iate

d ge

nes,

the

resp

onse

to th

e an

tibio

tics h

ad n

o as

soci

atio

n w

ith th

e ex

pres

sion

of a

regu

lato

r (Ly

sR) l

inke

d to

the

form

atio

n of

bio

film

in M

. avi

um. B

iofil

ms a

re c

ompo

sed

of p

lank

toni

c an

d se

ssile

ba

cter

ia. W

here

as p

lank

toni

c M

. avi

um is

susc

eptib

le to

cla

rith

rom

ycin

and

eth

ambu

tol (

clin

ical

ly u

sed

antim

icro

bial

s), s

essi

le b

ac-

teri

a ar

e at

leas

t thr

ee-f

old

to fo

ur-f

old

mor

e re

sist

ant t

o an

tibio

tics.

The

sess

ile p

heno

type

, how

ever

, is r

ever

sibl

e, a

nd n

o se

lect

ion

of re

sist

ant c

lone

s was

obs

erve

d. M

ice

infe

cted

thro

ugh

the

airw

ay w

ith b

oth

phen

otyp

es w

ere

infe

cted

with

a si

mila

r num

ber o

f ba

cter

ia, d

emon

stra

ting

no p

heno

type

adv

anta

ge. M

. avi

um b

iofil

m fo

rmat

ion

is e

nhan

ced

by c

omm

only

use

d co

mpo

unds

and

, in

the

sess

ile b

acte

rial

phe

noty

pe, i

s res

ista

nt to

cla

rith

rom

ycin

and

eth

ambu

tol,

in a

reve

rsib

le m

anne

r.

McN

abe

et a

l. 20

11

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

668

Aer

osol

M. t

uber

culo

sis

The

larg

e re

serv

oir o

f hum

an la

tent

tube

rcul

osis

(TB)

con

trib

utes

to th

e gl

obal

succ

ess o

f the

pat

hoge

n, M

ycob

acte

rium

tube

r-cu

losi

s (M

tb).

We

soug

ht to

test

whe

ther

aer

osol

infe

ctio

n of

rabb

its w

ith M

tb H

37Rv

cou

ld m

odel

pau

ciba

cilla

ry h

uman

late

nt

TB.

The

lung

bur

den

of in

fect

ion

peak

ed a

t 5 w

eeks

aft

er a

eros

ol in

fect

ion

follo

wed

by

host

con

tain

men

t of i

nfec

tion

that

was

ac

hiev

ed in

all

rabb

its. O

ne-t

hird

of r

abbi

ts h

ad a

t lea

st o

ne c

aseo

us g

ranu

lom

a w

ith c

ultu

rabl

e ba

cilli

at 3

6 w

eeks

aft

er in

fect

ion

sugg

estin

g pe

rsis

tent

pau

ciba

cilla

ry in

fect

ion.

Cor

ticos

tero

id-in

duce

d im

mun

osup

pres

sion

initi

ated

aft

er d

isea

se c

onta

inm

ent

resu

lted

in re

activ

atio

n of

dis

ease

. Sev

enty

-tw

o pe

rcen

t of r

abbi

ts h

ad c

ultu

rabl

e ba

cilli

in th

e ri

ght u

pper

lung

lobe

hom

ogen

-at

es c

ompa

red

to n

one

of th

e un

trea

ted

cont

rols

. Dis

cont

inua

tion

of d

exam

etha

sone

led

to p

redi

ctab

le ly

mph

oid

reco

very

, with

a

prop

ortio

n of

rabb

its d

evel

opin

g m

ultic

entr

ic la

rge

case

ous g

ranu

lom

a. Th

e de

velo

pmen

t and

seve

rity

of t

he im

mun

e re

con-

stitu

tion

infla

mm

ator

y sy

ndro

me

(IRI

S) w

as d

epen

dent

on

the

antig

en lo

ad a

t the

tim

e of

imm

unos

uppr

essi

on a

nd su

bseq

uent

ba

cilla

ry re

plic

atio

n du

ring

cor

ticos

tero

id-in

duce

d im

mun

osup

pres

sion

. Clin

ical

ly, m

any

aspe

cts w

ere

sim

ilar t

o IR

IS in

seve

rely

im

mun

osup

pres

sed

HIV

-infe

cted

pat

ient

s who

hav

e fu

nctio

nal r

esto

ratio

n of

T c

ells

in re

spon

se to

effe

ctiv

e (h

ighl

y ac

tive)

an

tiret

rovi

ral t

hera

py. Th

is c

ortic

oste

roid

mod

el is

the

only

ani

mal

mod

el o

f the

IRIS

. Fur

ther

stud

y of

the

rabb

it m

odel

of T

B la

tenc

y, re

activ

atio

n an

d IR

IS m

ay b

e im

port

ant i

n un

ders

tand

ing

the

imm

unop

atho

gene

sis o

f the

se p

oorl

y m

odel

led

stat

es a

s w

ell a

s for

impr

oved

dia

gnos

tics f

or sp

ecifi

c st

ages

of d

isea

se.

Man

abe

et a

l. 20

08

Aer

osol

indu

sed

in

fect

ion

M. a

. par

atub

ercu

losi

s

A c

halle

nge

expe

rim

ent w

as p

erfo

rmed

to in

vest

igat

e w

heth

er a

dmin

istr

atio

n of

Myc

obac

teri

um a

vium

subs

p. p

arat

uber

culo

-si

s (M

AP)

via

the

resp

irat

ory

rout

e le

ads t

o M

AP

infe

ctio

n in

cal

ves.

Eig

htee

n ca

lves

from

test

neg

ativ

e da

ms w

ere

rand

omly

al

loca

ted

to fo

ur g

roup

s. S

ix c

alve

s wer

e ch

alle

nged

with

MA

P na

sally

and

six

calv

es w

ere

chal

leng

ed b

y tr

anst

rach

eal i

njec

tion;

th

ree

oral

ly c

halle

nged

cal

ves s

erve

d as

pos

itive

con

trol

s, a

nd th

ree

non-

chal

leng

ed c

alve

s as n

egat

ive

cont

rols

. The

chal

leng

e w

as p

erfo

rmed

as a

nin

e-fo

ld tr

ickl

e do

se, 1

07 CFU

in to

tal.

Bloo

d an

d fa

ecal

sam

ples

wer

e co

llect

ed fr

eque

ntly

. Cal

ves w

ere

euth

aniz

ed th

ree

mon

ths p

ost-

chal

leng

e an

d ex

tens

ivel

y sa

mpl

ed. B

lood

sam

ples

wer

e te

sted

for t

he p

rese

nce

of a

ntib

odie

s and

in

terf

eron

gam

ma

prod

ucin

g ce

lls b

y EL

ISA

. Fae

cal a

nd ti

ssue

sam

ples

wer

e cu

lture

d in

a li

quid

cul

ture

syst

em a

nd th

e pr

esen

ce

of M

AP

was

con

firm

ed b

y IS

900

real

time

PCR

. Fou

rtee

n ou

t of fi

ftee

n ca

lves

had

no

MA

P an

tibod

y re

spon

se. Th

e ne

gativ

e co

n-tr

ols r

emai

ned

nega

tive;

all

posi

tive

cont

rols

bec

ame

infe

cted

. Tw

o na

sally

cha

lleng

ed c

alve

s sho

wed

a P

urifi

ed P

rote

in D

eriv

a-tiv

e A

vian

(PPD

A) s

peci

fic in

terf

eron

gam

ma

resp

onse

. In

all n

asal

ly c

halle

nged

cal

ves,

MA

P po

sitiv

e in

test

inal

sam

ples

wer

e de

tect

ed. I

n th

ree

calv

es o

f the

nas

al g

roup

MA

P po

sitiv

e re

trop

hary

ngea

l lym

ph n

odes

or t

onsi

ls w

ere

dete

cted

. In

all c

alve

s of

the

tran

stra

chea

l gro

up M

AP

posi

tive

inte

stin

al ti

ssue

s wer

e de

tect

ed a

s wel

l and

thre

e ha

d a

MA

P po

sitiv

e tr

ache

obro

nchi

al

lym

ph n

ode.

Thes

e fin

ding

s ind

icat

e th

at in

hala

tion

of M

AP

aero

sols

can

resu

lt in

infe

ctio

n. Th

ese

expe

rim

enta

l res

ults

may

be

rele

vant

for t

rans

mis

sion

und

er fi

eld

cond

ition

s sin

ce v

iabl

e M

AP

has b

een

dete

cted

in d

ust o

n co

mm

erci

al d

airy

farm

s.

Eise

nber

g et

al.

2011

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

669

Des

ert d

ust

M. s

p.A

pre

viou

sly

deve

lope

d re

sequ

enci

ng m

icro

arra

y, “T

ropi

cal a

nd E

mer

ging

Infe

ctio

ns (R

PM-T

EI v

. 1.0

chi

p)”,

desi

gned

to id

entif

y an

d di

scri

min

ate

betw

een

trop

ical

dis

ease

s and

oth

er p

oten

tial b

ioth

reat

age

nts,

thei

r nea

r-ne

ighb

or sp

ecie

s, a

nd/o

r pot

entia

l co

nfou

nder

s, w

as u

sed

to c

hara

cter

ize

the

mic

robe

s pre

sent

in th

e si

lt/cl

ay fr

actio

n of

surf

ace

soils

and

air

born

e du

st c

olle

cted

fr

om th

e M

iddl

e Ea

st. L

ocal

pop

ulat

ions

and

U. S

. mili

tary

per

sonn

el d

eplo

yed

to th

e M

iddl

e Ea

st a

re re

gula

rly

subj

ecte

d to

hig

h le

vels

of a

irbo

rne

dese

rt d

ust c

onta

inin

g a

sign

ifica

nt fr

actio

n of

inha

labl

e pa

rtic

les a

nd so

me

port

ion

requ

ire

clin

ical

aid

. Not

al

l of t

he c

linic

al sy

mpt

oms c

an b

e di

rect

ly a

ttri

bute

d to

the

phys

ical

act

ion

of m

ater

ial i

n th

e hu

man

resp

irat

ory

trac

t. To

bet

ter

unde

rsta

nd th

e po

tent

ial h

ealth

effe

cts o

f the

air

born

e du

st, t

he c

ompo

sitio

n of

the

mic

robi

al c

omm

uniti

es a

ssoc

iate

d w

ith su

r-fa

ce so

il an

d/or

air

born

e du

st (a

ir fi

lter)

sam

ples

from

19

diffe

rent

site

s in

Iraq

and

Kuw

ait w

as id

entifi

ed u

sing

RPM

-TEI

v. 1

.0.

Resu

lts in

dica

ted

that

seve

ral m

icro

orga

nism

s inc

ludi

ng a

cla

ss o

f rap

idly

gro

win

g M

ycob

acte

rium

, Bac

illus

, Bru

cella

, Clo

stri

d-iu

m a

nd C

oxie

lla b

urne

tti,

wer

e pr

esen

t in

the

sam

ples

. The

pres

ence

of t

hese

org

anis

ms i

n th

e su

rfac

e so

ils a

nd th

e in

hala

ble

frac

tion

of a

irbo

rne

dust

ana

lyze

d m

ay p

ose

a hu

man

hea

lth ri

sk a

nd w

arra

nts f

urth

er in

vest

igat

ion.

Bet

ter u

nder

stan

ding

of t

he

fact

ors i

nflue

ncin

g th

e co

mpo

sitio

n of

thes

e m

icro

bial

com

mun

ities

is im

port

ant t

o ad

dres

s que

stio

ns re

late

d to

hum

an h

ealth

an

d is

cri

tical

to a

chie

ving

For

ce H

ealth

Pro

tect

ion

for t

he W

arfig

hter

ope

ratin

g in

the

Mid

dle

East

, Afg

hani

stan

, Nor

th A

fric

a an

d ot

her a

rid

regi

ons.

Lesk

i et

al. 2

010

Dus

tM

. sp.

M. b

arra

ssia

eM

. gilv

umM

. van

bale

nii

Alth

ough

the

link

betw

een

airb

orne

par

ticul

ate

inha

latio

n an

d a

vari

ety

of re

spir

ator

y di

seas

es h

as lo

ng b

een

esta

blis

hed,

litt

le is

kn

own

abou

t the

pat

hoge

nic

role

of t

he m

icro

bial

com

pone

nt o

f the

dus

t. In

this

stud

y, w

e ap

plie

d hi

ghly

mul

tiple

xed

PCR

and

a hi

gh-d

ensi

ty re

sequ

enci

ng m

icro

arra

y (R

PM-T

EI v

ersi

on 1

.0) t

o sc

reen

sam

ples

of fi

ne to

psoi

l par

ticle

s and

air

born

e du

st c

ol-

lect

ed in

19

loca

tions

in Ir

aq a

nd K

uwai

t for

the

pres

ence

of a

bro

ad ra

nge

of h

uman

pat

hoge

ns. Th

e re

sults

indi

cate

d th

e pr

es-

ence

of p

oten

tial h

uman

pat

hoge

ns, i

nclu

ding

Myc

obac

teri

um, B

ruce

lla, C

oxie

lla b

urne

tii, C

lost

ridi

um p

erfr

inge

ns, a

nd B

acill

us.

Lesk

i et

al. 2

011

Met

al w

orki

ng fl

uids

M. i

mm

unog

enum

Hyp

erse

nsiti

vity

pne

umon

itis,

als

o kn

own

as “m

achi

ne o

pera

tor’s

lung

” (M

OL)

, has

bee

n re

late

d to

mic

roor

gani

sms g

row

ing

in

met

alw

orki

ng fl

uids

(MW

Fs),

espe

cial

ly M

ycob

acte

rium

imm

unog

enum

. We

aim

ed to

(i) d

escr

ibe

the

mic

robi

olog

ical

con

tam

ina-

tion

of M

WFs

and

(ii)

look

for c

hem

ical

, phy

sica

l, an

d en

viro

nmen

tal p

aram

eter

s ass

ocia

ted

with

var

iatio

ns in

mic

robi

olog

ical

pr

ofile

s. W

e m

icro

biol

ogic

ally

ana

lyze

d 18

0 M

WF

sam

ples

from

non

auto

mot

ive

plan

ts (e

.g.,

scre

w-m

achi

ning

or m

etal

-cut

ting

plan

ts) i

n th

e Fr

anch

e-C

omte

regi

on in

eas

tern

Fra

nce

and

165

sam

ples

from

thre

e Fr

ench

aut

omot

ive

plan

ts in

whi

ch c

ases

of

MO

L ha

d be

en p

rove

n. O

ur re

sults

reve

aled

two

type

s of m

icro

bial

bio

mes

: the

firs

t was

from

the

nona

utom

otiv

e in

dus-

try,

show

ed p

redo

min

antly

Gra

m-n

egat

ive

rods

(GN

R), a

nd w

as a

ssoc

iate

d w

ith a

low

risk

of M

OL,

and

the

seco

nd c

ame

from

th

e au

tom

otiv

e in

dust

ry th

at w

as a

ffect

ed b

y ca

ses o

f MO

L an

d sh

owed

pre

dom

inan

tly G

ram

-pos

itive

rods

(GPR

). Tr

aces

of

M. i

mm

unog

enum

wer

e sp

orad

ical

ly d

etec

ted

in th

e fir

st ty

pe, w

hile

it w

as h

ighl

y pr

eval

ent i

n th

e au

tom

otiv

e se

ctor

, with

up

to

38%

of s

ampl

es te

stin

g po

sitiv

e. Th

e us

e of

chr

omiu

m, n

icke

l, or

iron

was

ass

ocia

ted

with

gro

wth

of G

ram

-neg

ativ

e ro

ds; c

on-

vers

ely,

grow

th o

f Gra

m-p

ositi

ve ro

ds w

as a

ssoc

iate

d w

ith th

e ab

senc

e of

thes

e m

etal

s. Sy

nthe

tic M

WFs

wer

e m

ore

freq

uent

ly

ster

ile th

an e

mul

sion

s. Ve

geta

ble

oil-b

ased

em

ulsi

ons w

ere

asso

ciat

ed w

ith G

NR

, whi

le m

iner

al o

nes w

ere

asso

ciat

ed w

ith G

PR.

Our

resu

lts su

gges

t tha

t met

al ty

pes a

nd th

e na

ture

of M

WF

play

a p

art i

n M

WF

cont

amin

atio

n, a

nd th

is w

ork

shal

l be

follo

wed

by

furt

her i

n vi

tro

sim

ulat

ion

expe

rim

ents

on

the

kine

tics o

f mic

robi

al p

opul

atio

ns, f

ocus

ing

on th

e ph

enom

ena

of in

hibi

tion

and

syne

rgy.

Mur

at e

t al

. 201

2

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

670

Met

al w

orki

ng fl

uids

M. i

mm

unog

enum

Purp

ose

of re

view

: To

high

light

adv

ance

s in

unde

rsta

ndin

g th

e re

spir

ator

y di

seas

e as

soci

ated

with

met

al m

achi

ning

, a c

omm

on

wor

k pr

oces

s inv

olvi

ng a

ppro

xim

atel

y 1.

2 m

illio

n w

orke

rs in

the

USA

. Rec

ent fi

ndin

gs: R

ecen

t stu

dies

em

phas

ize

that

wor

k-re

late

d as

thm

a an

d hy

pers

ensi

tivity

pne

umon

itis c

ontin

ue to

be

caus

ed b

y ex

posu

re to

met

alw

orki

ng fl

uid.

Iden

tifica

tion

of a

n in

divi

dual

pat

ient

indi

cate

s the

nee

d fo

r fol

low

-up

inve

stig

atio

ns a

t the

wor

k si

te to

pre

vent

add

ition

al d

isea

se a

nd/o

r ide

ntify

ad

ditio

nal e

ffect

ed in

divi

dual

s. Id

entifi

catio

n of

the

caus

al a

gent

for h

yper

sens

itivi

ty p

neum

oniti

s has

cen

tere

d on

mic

robi

al c

on-

tam

inat

ion

of m

etal

wor

king

flui

ds w

ith a

num

ber o

f stu

dies

focu

sing

on

Myc

obac

teri

um im

mun

ogen

um. S

umm

ary:

Bot

h as

thm

a an

d hy

pers

ensi

tivity

pne

umon

itis o

ccur

am

ong

wor

kers

exp

osed

to m

etal

wor

king

flui

d. Th

e in

cide

nce

of th

ese

dise

ases

am

ong

such

wor

kers

is u

nkno

wn.

Out

brea

ks o

f the

se c

ondi

tions

con

tinue

to b

e id

entifi

ed a

mon

g m

etal

mac

hini

sts.

Whe

ther

thes

e ar

e tr

ue o

utbr

eaks

ass

ocia

ted

with

som

e br

eakd

own

in w

orkp

lace

con

trol

s or,

rath

er th

e re

cogn

ition

of o

ngoi

ng e

ndem

ic d

isea

se th

at

is ty

pica

lly m

isdi

agno

sed

as p

neum

onia

or c

omm

on a

dult

onse

t ast

hma,

nee

ds fu

rthe

r eva

luat

ion.

Fur

ther

wor

k to

elu

cida

te th

e sp

ecifi

c ca

usal

age

nt(s

) is n

eces

sary

to a

ffect

effe

ctiv

e w

orkp

lace

con

trol

s. T

reat

ing

an id

entifi

ed in

divi

dual

cas

e as

an

inde

x ca

se

with

a fo

llow

-up

wor

kpla

ce in

vest

igat

ion

will

onl

y be

pos

sibl

e if

prac

ticin

g ph

ysic

ians

inte

ract

with

pub

lic h

ealth

aut

hori

ties t

o re

port

new

ly d

iagn

osed

cas

es.

Rose

n-m

an 2

009

Tabl

e 7.

Dif

fere

nt m

etho

ds fo

r de

tect

ion

and

iden

tifi

cati

on o

f myc

obac

teri

a in

the

envi

ronm

ent

Det

ectio

n m

etho

dM

ycob

acte

ria

dete

cted

Exam

ined

/Pos

itive

sam

ples

Q

uant

ifica

tion

Refe

renc

e

Cul

ture

M. s

p.24

/18

wat

er14

8/76

soil

up to

104 C

FUC

hilim

a et

al.

2006

M. s

p.

49/1

0 w

ater

n-a

Cha

ng e

t al.

2002

M. s

p.49

/21

cold

wat

er44

/32

war

m w

ater

up to

103 C

FU/5

00 m

lH

usse

in e

t al.

2009

M. s

p.50

/8 a

irn-

aC

ayer

et a

l. 20

07M

. sp.

34/2

8 w

ater

n-a

Perk

ins e

t al.

2009

M. s

p.40

/34

wat

ern-

aC

astil

lo-R

odal

et a

l. 20

12M

. sp.

M. p

orci

num

63%

wat

ern-

aBr

own-

Ellio

tt e

t al.

2011

M. s

p.69

/36

wat

ern-

aFe

rnan

dez-

Rend

on e

t al.

2012

M. a

vium

n-a

up to

104 C

FU/m

lSv

enss

on e

t al.

2011

M. a

. par

atub

ercu

losi

s19

2/8

wat

ern-

aW

han

et a

l. 20

05M

. a. p

arat

uber

culo

sis

96/1

2 w

ater

n-a

Pick

up e

t al.

2005

M. s

p.11

8/84

pea

tn-

aM

atlo

va e

t al.

2012

M. s

p.39

1/26

soil

n-a

Mor

avko

va e

t al.

2011

Veterinarni Medicina, 57, 2012 (12): 623–679 Review Article

671

PCR

M. s

p.

49/0

wat

ern-

aC

hang

et a

l. 20

02M

. sp.

24

/13

wat

er14

8/75

soil

n-a

Chi

lima

et a

l. 20

06

IS24

04, I

S260

6/M

. ulc

eran

s35

/2 w

ater

14/

2 de

tritu

s2/

1 ve

geta

tion

n-a

Stin

ear e

t al.

2000

M. b

ovis

/mpb

64, m

pb70

11/4

site

s soi

l10

3 /gYo

ung

et a

l. 20

05

M. l

epra

e/16

S rD

NA

80/3

0 so

iln-

aLa

vani

a et

al.

2008

M. l

epra

e/16

S rD

NA

207/

71 so

iln-

aTu

rank

ar e

t al.

2012

IS90

0/M

. a. p

arat

uber

culo

sis

96/3

1 w

ater

n-a

Pick

up e

t al.

2005

IS90

0/M

. a. p

arat

uber

culo

sis

70/4

8 w

ater

10/9

sedi

men

tn-

aPi

ckup

et a

l. 20

06

IMS-

PCR

IS90

0/M

. a. p

arat

uber

culo

sis

192/

9 w

ater

n-a

Wha

n et

al.

2005

Nes

ted

PCR

rpoB

/ M. s

p.32

/7 w

ater

n-a

Shin

et a

l. 20

08

M. s

p.M

. avi

um45

/25

wat

er45

/5 se

dim

ent

n-a

Dre

we

et a

l. 20

09

Real

tim

e PC

R

IS90

0/M

. a. p

arat

uber

culo

sis

366/

219

dust

n-a

Eise

nber

g et

al.

2011

M. a

. par

atub

ercu

losi

s81

% w

ater

mid

wes

t, 0%

nat

iona

l sur

vey

up to

102 /4

00 m

lBe

umer

et a

l. 20

10M

. a. p

arat

uber

culo

sis

16/0

wat

ern-

aBo

ckel

man

n et

al.

2009

16S

rRN

A/M

. sp.

4/4

bedd

ing

up to

1010

/gPa

kari

nen

et a

l. 20

0716

S rR

NA

/M. s

p.20

/20

dust

n-a

Torv

inen

et a

l. 20

10M

. avi

um

14/1

3 w

ater

32/2

5 bi

ofilm

n-a

Feaz

el e

t al.

2009

M. s

p.53

/30

wat

er10

3 –106 /l

Adr

ados

et a

l. 20

11M

. sp.

21/1

2 w

ater

up to

104 /m

lK

awai

et a

l. 20

0416

S rR

NA

/M. s

p.M

. xen

opi

93/9

3 w

ater

93/7

3 w

ater

n-a

Hus

sein

et a

l. 20

09

IS90

0/M

. a. p

arat

uber

culo

sis

22/0

wat

ern-

aV

illar

real

et a

l. 20

10IS

900,

f57/

M. a

. par

atub

ercu

losi

s19

/10

silla

geup

to 1

07 /gK

hol e

t al.

2010

IS90

0/M

. a. p

arat

uber

culo

sis

19/1

1 so

il57

/43

plan

ts

up to

103 /g

Prib

ylov

a et

al.

2011

IS90

1/M

. a. a

vium

IS12

45/M

. a. h

omin

issu

is28

/10

soil

28/1

3 so

ilup

to 1

07 /gK

aevs

ka e

t al.

2011

ITS/

M. s

p.96

% w

ater

up to

103 /m

lJa

cobs

et a

l. 20

09IS

2404

, IS2

606/

M. u

lcer

ans

1/14

8 w

ater

n-a

Vand

elan

noot

e et

al.

2010

M. l

epra

e/16

S rR

NA

80/2

8 so

ilup

to 1

05 /gLa

vani

a et

al.

2008

M. l

epra

e/16

S rR

NA

207/

28 so

iln-

aTu

rank

ar e

t al.

2012

Review Article Veterinarni Medicina, 57, 2012 (12): 623–679

672

8. Acknowledgements

The critical comments of Professor I. Pavlik, Veterinary Research Institute, Brno, Czech Republic, are greatly appreciated.

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Corresponding Author:

Prof. MVDr. Karel Hruska, CSc., Veterinary Research Institute, Hudcova 70, 621 00 Brno, Czech RepublicTel. +420 533 332 014, E-mail: hruska@vri.cz