The Effect of Age on Isolated Bacteria from Cancer Patients

7/30/2019 The Effect of Age on Isolated Bacteria from Cancer Patients

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Mashooma Khalifa

BIO 415N

Spring 2011


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Recently in the Indian Journal of Cancer the

article, The effect of age on the bacteria

isolated and the antibiotic-sensitivity pattern in

infections among cancerpatients was published

in December of 2010.

Focus of this article is how age can have an

effect on the types of bacteria present in cancer

patients as well as its antibiotic treatment


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As one ages, the cells of the individual start to loseits ability and the individual begins to respond less

frequently to stress while an increase in homeostatic

imbalances occurs.

The body becomes susceptible to cancer as well as

to vast amounts of infectious diseases


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As one ages, do the types of bacteria differ from agegroups?

Does the effect of antibiotics increase or decrease as

one ages, especially if diagnosed with cancer?

Can the effectiveness of the antibiotics be

controlled?


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*Patients with cancer are predisposed to infections,and on many occasions, no focus of infection can be

established

*Pathologists enforce the treatment ofempirical

antibiotic therapy in these patients

*This therapy involves prescribing certain

medications even though an accurate diagnosis

cannot be made [trial & error]


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*It can be hypothesized, that the effectiveness of the

antibiotics may change over time & as age progresses,

which would result in changes in the antibiotic

sensitivity (Kumar et all. 2010).

*That is why, the main purpose of this experiment is to

test such a hypothesis on seriously ill patients, such ascancer patients


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*Laboratory experiments were conducted at Tata Memorial

Hospital in Mumbai, India during the fall of 2010.

*The experiment had to be conducted secretly so that the

neither the patients nor their family & friends be affected in

any physical, emotional or mental way


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Bacterial samples were selected from patients with a

constant history of infections, were selected for this

experiment

A total of 935 isolates were collected of patients

admitted under the Medical Oncology service

Antibiotic sensitivity & microbiological profile of

infections were divided according to three age

groups: 12-19 years, 20-49 years & >50 years.


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Five pathogens were isolated/used:

Pseudomonas species

Enterococcus species

Staphylococcus aureus

Escheria coli

Klebsiella species

Figure 1 Figure 2

Figure 3 Figure 4

Figure 5
http://upload.wikimedia.org/wikipedia/commons/8/85/Pseudomonas_aeruginosa_01.jpghttp://en.wikipedia.org/wiki/File:Enterococcus_histological_pneumonia_01.pnghttp://en.wikipedia.org/wiki/File:Staphylococcus_aureus_VISA_2.jpghttp://en.wikipedia.org/wiki/File:EscherichiaColi_NIAID.jpghttp://en.wikipedia.org/wiki/File:EscherichiaColi_NIAID.jpghttp://en.wikipedia.org/wiki/File:Staphylococcus_aureus_VISA_2.jpghttp://en.wikipedia.org/wiki/File:Enterococcus_histological_pneumonia_01.pnghttp://upload.wikimedia.org/wikipedia/commons/8/85/Pseudomonas_aeruginosa_01.jpg


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The pathogens were easily identifiable via the

Analytical Profile Index or API test

It is a rapid biochemical test system that

allows to identify a limited number of Gram-

Negative Enterobacteriacea


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All test chambers are rehydrated by inoculation with a saline

suspension of a pure culture of the bacterial strain

After incubation in a humidity chamber for 18-24 hours at 37C,the color reactions are read. The results of the test reactions are

converted to a seven-digit code. The code can then be looked upin the database book or software

Common substrates tested on the API is:

CITCitrate for the reaction: citrate utilization UREUrea for the reaction: urea hydrolysis

INDTryptophan for the reaction: indole production

SACSucrose for the reaction: fermentation/oxidation

OXOxidase for the reaction: Oxidation

H2SNa Thiosulfate for the reaction: H2S Production


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Inoculation of chambers with the

saline bacterial culture

Code is interpreted by only writing the

+ reactions that occurred, for

example in set of boxes, two out of

four reactions were positivethus the#2 is written in the box below

Sample of the API Test, the roman numerals on top indicate the 7 set of

boxes in which 7 digits will be recorded

Figure 6

Figure 7

Figure 8


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The antibiotic-sensitivity pattern wasidentifiable via the Kirby-Bauer AntimicrobialSusceptibility Test or the KB Test

This test was done following standardprotocol, such as the CLSI standard (Clinical

& Laboratory Standards Institute)

The samples obtained were then mapped onspecial microbiological software


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An agar plate is inoculated with the bacterial culture

Filter paper is impregnated with an antibiotic that is

then placed on the plate using an Automatic Disk

Dispenser

The antibiotic would diffuse from the disk onto the

agar plate

Based on the characteristics of the bacteria, around

each disk there will be a zone of inhibition

(diameter) that is measured to determine the

resistance and susceptibility values


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Example:

The antibiotic Penicillin Staphylococci in the presence of

Penicillin, has a resistance value of 28mm and a

susceptible value of 29mm

For a pathogen to be resistant to Penicillin, it must have a

diameter of < 28mm

Sample of a KB Test, in which disks of Erthromycin [E-2],

Novobiocin [NA-30], Penicillin [P-10], Polymyxin B [PB-

300], Vancomycin [VA-30] and Streptomycin [S-10] were

impregnated on a bacterial culture ofEscheria coli.

Figure 9


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The results determined that there was no significant

change in the distribution of Gram-Positive & Gram-

Negative bacteria with age

HOWEVER,

There was an increase in the occurrence of

Enterobacteriacea group with increasing age

There was a decrease in infections caused by

nonlactose fermenters with increasing age

Most antibiotics decreased in activity with increasing

age


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Gram-Negative bacteria were the predominant

group of bacteria66.96% of the isolates

An increase in infections caused byE.coli &

Klebsiella with increasing age was found

There was a decrease in infections caused by the

Pseudomonas species with increasing age


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Site of Infections:51.76% at the blood stream

15.08% at the respiratory tract

12.62% at the GI tract

11.55% at the skin & soft tissue

3.32% at the urinary tract

5.67% at other sitesTable 1 Table 2

Percentage of occurrence of bacterial pathogens

among the three age groups

Organism:% by age group (years)

12 19 20 49 > 50

Pseudomonas sp (245) 30.01 26.01 23.87Enterococcus sp (109) 12.72 11.75 10.70S. aureus sp (107) 12.14 10.40 13.17E. coli (106) 8.67 11.37 13.17Klebsiella sp (99) 7.51 11.37 11.11


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Extended-spectrum -lactamase (ESBL) are enzymes whichhydrolyze extended-spectrum cephalosporin's with anoxyimino side chainESBLs are enzymes made by some germs. Germs can cause you to

get sick and antibiotics are used to treat or kill these germs. When

germs have ESBLs, many antibiotics do not work.

ESBL production increased from 10.52% in patients 12-19years to 19.31% in patients 20-49 years to 24.88% in patients50 years or older

The production of ESBL was statistically significant wheneach age group was compared with each other resulting witha P value of < 0.01


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The most active antibiotic agent against Gram-Positive

bacteria was Linezolid with no resistance, but it wasuniversally sensitive to the Enterococcal species

Vancomycin resistance (83%) was common among

Enterococcal speciesbut there was a decrease infrequency as age increased

Vancomycin, Linezolid and Teicoplanin were most active

again Staphylococcal species with no resistance, irrelevant

with ageDecreasing trend of activity for meropenam was

statistically significant against Pseudomonas species (73.3-

41.2%) and of aminoglycosides forAcinetobacterspecies

(61.1-17.4%) as age increased


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An evident trend of decreasing antibiotic

resistance against pathogens with increasing age

The reasons could be greater exposure to

antibiotics during life, higher ESBL production

and the need for more invasive procedures than

Empirical therapy

Only improving efficacy of polymyxin &colistin again Pseudomonas species (40-91.7%)

was observed


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Article

Kumar P, Medhekar A, Ghadyalpatil N.S, Noronha V, Biswas S, Kurkure P, Nair R, Kelkar R, Banavali S.D. 2010. The

effect of age on the bacteria isolated and the antibiotic-sensitivity pattern in infections among cancer patients. Indian

Journal of Cancer [Internet] [cited 2011 Mar 14]; 47(4):391-396. Available from:

http://corvette.salemstate.edu:2561/ehost/detail?hid=8&sid=cfba2cdc-a31e-4f75-baad-

5d212b89487f%40sessionmgr10&vid=3&bdata=JkF1dGhUeXBlPWNvb2tpZSxpcCxjcGlkJmN1c3RpZD1zc2Mmc2l0Z

T1laG9zdC1saXZlJnNjb3BlPXNpdGU%3d#db=bxh&AN=BACD201100046666

Images

Figure 1: http://phil.cdc.gov/PHIL_Images/20050113/c6f15fe678d24b739134735e2296e3f7/6687_lores.jpg

Figure 2: http://phil.cdc.gov/phil_images/20030114/16/PHIL_2899_lores.jpg

Figure 3: http://phil.cdc.gov/PHIL_Images/11157/11157_lores.jpg

Figure 4: http://www.niaid.nih.gov/SiteCollectionImages/topics/biodefenserelated/e_coli.jpg

Figure 5: http://www.flickr.com/photos/ajc1/3787968951/ Figure 6: http://www.biotech.ug.edu.pl/odl/biochem/picbiochem/api2.jpg

Figure 7: http://www.biotech.ug.edu.pl/odl/biochem/picbiochem/api4.jpg

Figure 8: http://www.biotech.ug.edu.pl/odl/biochem/picbiochem/apiE20.JPG

Figure 9: Microbiology Department. Salem State University. 11/3/2010.

Tables

Table 1: http://www.indianjcancer.com/articles/2010/47/4/images/IndianJournalofCancer_2010_47_4_391_73574_b1.jpg

Table 2: http://www.indianjcancer.com/text.asp?2010/47/4/391/73574

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The Effect of Age on Isolated Bacteria from Cancer Patients