“INHIBITORY POTENTIAL OF FLOWER EXTRACTS ON MRSA FROM CELL PHONES”

8/10/2019 INHIBITORY POTENTIAL OF FLOWER EXTRACTS ON MRSA FROM CELL PHONES

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Department of Microbiology Page 1

INHIBITORY POTENTIAL OF FLOWER EXTRACTS ON

MRSA FROM CELL PHONES

INTRODUCTION:

Staphylococcus aureus is a Gram positive, coagulase positive coccus in the family

Staphylococcaceae. Staphylococcus aureusis aGram-positivecocci bacterium that is a

member of theFirmicutes,and is frequently found in the human respiratory tract and on the

skin. It is positive for catalase and nitrate reduction. Although S. aureusis not always

pathogenic, it is a common cause of skin infections (e.g. boils), respiratory disease

(e.g.sinusitis), andfood poisoning. Disease-associated strains often promote infections by

producing potent proteintoxins,and expressing cell-surface proteins thatbind and inactivate

antibodies.The emergence ofantibiotic-resistant forms of pathogenic S. aureus(e.g.MRSA)

is a worldwide problem in clinical medicine.

Staphylococcus was first identified in 1880 inAberdeen,United Kingdom, by the surgeon

SirAlexander Ogston inpus from a surgical abscess in a knee joint. This name was later

appended to Staphylococcus aureusby Rosenbach who was credited by the official system of

nomenclature at the time. It is estimated that 20% of the human population are long-term

carriers of S. aureuswhich can be found as part of the normalskin flora and in anterior nares

of the nasal passages. S. aureusis the most common species of staphylococcus to

causeStaphinfections and is a successful pathogen due to a combination of nasal carriage

and bacterial immuno-evasive strategies.S. aureuscan cause a range of illnesses, from minor

skin infections, such aspimples, impetigo, boils (furuncles), cellulitis, folliculitis,

carbuncles, scalded skin syndrome, and abscesses, to life-threatening diseases such

aspneumonia,meningitis,osteomyelitis,endocarditis,toxicshocksyndrome (TSS),bacteremia

andsepsis.Its incidence ranges from skin, soft tissue, respiratory, bone, joint, endovascular

towound infections. It is still one of the five most common causes ofnosocomial
http://en.wikipedia.org/wiki/Gram-positivehttp://en.wikipedia.org/wiki/Coccushttp://en.wikipedia.org/wiki/Bacteriumhttp://en.wikipedia.org/wiki/Firmicuteshttp://en.wikipedia.org/wiki/Pathogenhttp://en.wikipedia.org/wiki/Boilshttp://en.wikipedia.org/wiki/Sinusitishttp://en.wikipedia.org/wiki/Food_poisoninghttp://en.wikipedia.org/wiki/Exotoxinhttp://en.wikipedia.org/wiki/Protein_Ahttp://en.wikipedia.org/wiki/Protein_Ahttp://en.wikipedia.org/wiki/Antibiotic-resistanthttp://en.wikipedia.org/wiki/Methicillin-resistant_Staphylococcus_aureushttp://en.wikipedia.org/wiki/Aberdeenhttp://en.wikipedia.org/wiki/United_Kingdomhttp://en.wikipedia.org/wiki/Alexander_Ogstonhttp://en.wikipedia.org/wiki/Pushttp://en.wikipedia.org/wiki/Skin_florahttp://en.wikipedia.org/wiki/Staph_infectionhttp://en.wikipedia.org/wiki/Staph_infectionhttp://en.wikipedia.org/wiki/Staph_infectionhttp://en.wikipedia.org/wiki/Infectionhttp://en.wikipedia.org/wiki/Pimplehttp://en.wikipedia.org/wiki/Impetigohttp://en.wikipedia.org/wiki/Boilhttp://en.wikipedia.org/wiki/Cellulitishttp://en.wikipedia.org/wiki/Carbunclehttp://en.wikipedia.org/wiki/Scalded_skin_syndromehttp://en.wikipedia.org/wiki/Abscesshttp://en.wikipedia.org/wiki/Pneumoniahttp://en.wikipedia.org/wiki/Meningitishttp://en.wikipedia.org/wiki/Osteomyelitishttp://en.wikipedia.org/wiki/Endocarditishttp://en.wikipedia.org/wiki/Toxic_shock_syndromehttp://en.wikipedia.org/wiki/Bacteremiahttp://en.wikipedia.org/wiki/Sepsishttp://en.wikipedia.org/wiki/Wound_infectionhttp://en.wikipedia.org/wiki/Nosocomial_infectionhttp://en.wikipedia.org/wiki/Nosocomial_infectionhttp://en.wikipedia.org/wiki/Wound_infectionhttp://en.wikipedia.org/wiki/Sepsishttp://en.wikipedia.org/wiki/Bacteremiahttp://en.wikipedia.org/wiki/Toxic_shock_syndromehttp://en.wikipedia.org/wiki/Endocarditishttp://en.wikipedia.org/wiki/Osteomyelitishttp://en.wikipedia.org/wiki/Meningitishttp://en.wikipedia.org/wiki/Pneumoniahttp://en.wikipedia.org/wiki/Abscesshttp://en.wikipedia.org/wiki/Scalded_skin_syndromehttp://en.wikipedia.org/wiki/Carbunclehttp://en.wikipedia.org/wiki/Cellulitishttp://en.wikipedia.org/wiki/Boilhttp://en.wikipedia.org/wiki/Impetigohttp://en.wikipedia.org/wiki/Pimplehttp://en.wikipedia.org/wiki/Infectionhttp://en.wikipedia.org/wiki/Staph_infectionhttp://en.wikipedia.org/wiki/Skin_florahttp://en.wikipedia.org/wiki/Pushttp://en.wikipedia.org/wiki/Alexander_Ogstonhttp://en.wikipedia.org/wiki/United_Kingdomhttp://en.wikipedia.org/wiki/Aberdeenhttp://en.wikipedia.org/wiki/Methicillin-resistant_Staphylococcus_aureushttp://en.wikipedia.org/wiki/Antibiotic-resistanthttp://en.wikipedia.org/wiki/Protein_Ahttp://en.wikipedia.org/wiki/Protein_Ahttp://en.wikipedia.org/wiki/Exotoxinhttp://en.wikipedia.org/wiki/Food_poisoninghttp://en.wikipedia.org/wiki/Sinusitishttp://en.wikipedia.org/wiki/Boilshttp://en.wikipedia.org/wiki/Pathogenhttp://en.wikipedia.org/wiki/Firmicuteshttp://en.wikipedia.org/wiki/Bacteriumhttp://en.wikipedia.org/wiki/Coccushttp://en.wikipedia.org/wiki/Gram-positive


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infections and is often the cause of postsurgical wound infections. Each year, some 500,000

patients in United States' hospitals contract a staphylococcal infection.

Role in Disease:

S. aureusis responsible for many infections but it may also occur as acommensal. The

presence of S. aureusdoes not always indicate infection. S. aureuscan survive from hours to

weeks, or even months, on dry environmental surfaces, depending on strain.

S. aureuscan infect tissues when the skin or mucosal barriers have been breached. This can

lead to many different types of infections includingfuruncles andcarbuncles (a collection of

furuncles).

S. aureus infections can spread through contact with pus from an infected wound, skin-to-

skin contact with an infected person by producinghyaluronidase that destroys tissues, and

contact with objects such as towels, sheets, clothing, or athletic equipment used by an

infected person. Deeply penetrating S. aureusinfections can be severe. Prosthetic joints put a

person at particular risk ofseptic arthritis, and staphylococcalendocarditis (infection of the

heart valves) andpneumonia. Strains of S. aureuscan hostphages, such as -PVL

(producesPanton-Valentine leukocidin), that increase virulence.

Antibiotic Resistance in Staphylococcus aureus:

In the 1940s, penicillin was introduced for the treatment of infection; as early as 1942, strains

of S. aureusresistant to penicillin had been detected in hospitals. Within 2 decades, 80% of

both hospital- and community-acquired S. aureusisolates was penicillin resistant. The

introduction of methicillin in 1961 was rapidly followed by reports of methicillin resistance

in S. aureus.Today, MRSA strains are found worldwide, and most are multidrug resistant.

Recently even the Vancomycin resistant Staphylococci were recovered from Hospitals, and

Cell phones.
http://en.wikipedia.org/wiki/Nosocomial_infectionhttp://en.wikipedia.org/wiki/Commensalhttp://en.wikipedia.org/wiki/Furunclehttp://en.wikipedia.org/wiki/Carbunclehttp://en.wikipedia.org/wiki/Hyaluronidasehttp://en.wikipedia.org/wiki/Septic_arthritishttp://en.wikipedia.org/wiki/Endocarditishttp://en.wikipedia.org/wiki/Pneumoniahttp://en.wikipedia.org/wiki/Phagehttp://en.wikipedia.org/wiki/Phagehttp://en.wikipedia.org/wiki/Panton-Valentine_leukocidinhttp://en.wikipedia.org/wiki/Panton-Valentine_leukocidinhttp://en.wikipedia.org/wiki/Phagehttp://en.wikipedia.org/wiki/Pneumoniahttp://en.wikipedia.org/wiki/Endocarditishttp://en.wikipedia.org/wiki/Septic_arthritishttp://en.wikipedia.org/wiki/Hyaluronidasehttp://en.wikipedia.org/wiki/Carbunclehttp://en.wikipedia.org/wiki/Furunclehttp://en.wikipedia.org/wiki/Commensalhttp://en.wikipedia.org/wiki/Nosocomial_infection


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Staphylococcus aureus is an opportunistic pathogen often carried asymptomatically on the

human body. Methicillin-resistant S. aureus (MRSA) strains have acquired a gene that makes

them resistant to all beta-lactam antibiotics. Hospital-associated strains of this organism are

serious nosocomial pathogens that have become resistant to most common antibiotics, and

treatment can be challenging. Community-associated MRSA strains occur in people who

have not been hospitalized or recently had invasive procedures. They first appeared in high-

risk populations (e.g., intravenous drug users, people with chronic illnesses), but are now

found even in healthy children. Until recently, community-associated strains were susceptible

to many antibiotics other than beta-lactams; however, resistance seems to be increasing, and

multiple antibiotic resistant strains have started to emerge. Human-adapted MRSA can be

transmitted to animals in close contact, which can sometimes act as carriers and re-infect

people. However, the recent magnitude and trend of these infections have not been reported.

We used national hospitalization and resistance data to estimate the annual number of

hospitalizations and deaths associated with S. aureus and MRSA from 1999 through 2005.

During this period, the estimated number of S. aureusrelated hospitalizations increased 62%,

from294,570 to 477,927, and the estimated number of MRSA related hospitalizations more

than doubled, from 127,036to 278,203. Our findings suggest that S. aureus and MRSA

should be considered a national priority for disease control. Although, the clinical

significance of methicillin resistance has been questioned in the past, there is now widespread

acknowledgement of the pathogenicity of MRSA. It has emerged as a significant cause of

both nosocomial and community-acquired infections. Recent report of strains of MRSA

isolated from children in the community has led to speculation that the epidemiology of S.

aureus is changing. the magnitude of the effect and trend in the incidence and associated

mortality rates n of infections related to S. aureus and MRSA over a 7-year period, from 1999

through 2005, paying particular attention to the overall S. aureus infection level and the trend


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of typical community-associated infections. Evidence on the magnitude and trend of the

problem on a national level informs rational, evidence-based decisions about how to allocate

resources and adjust healthcare policy to address this issue. Infection trends are useful to

clinicians, hospital administrators, insurers, and policymakers who make decisions regarding

control measures, especially infection control measures to contain the spread of nosocomial

and community-associated pathogens.

Methicillin-resistant S. aureus strains have acquired the mec A gene, which is carried on a

large mobile genetic element called the staphylococcal chromosomal cassette mec (SCCmec).

This gene codes for a penicillin binding protein, PBP2a, which interferes with the effects of

beta lactam antibiotics (e.g. penicillins and cephalosporins) on cell walls. It confers virtually

complete resistance to all beta-lactam antibiotics including the semi-synthetic penicillins.

Intervention studies have been published to explore alternative antimicrobial agents to control

and prevent diseases due to multidrug resistant S. aureus. Although other practices have been

explored such as bacterial interference therapy and phage therapy medicinal plants have also

been considered by some researchers since they are frequently used in popular medicine as

remedies for many infectious diseases. The aim of the present study was to determine the

inhibitory effect of different flower extracts on the growth of multiple resistan ts of S.

Aureus.


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Flower details:

Campanulais one of severalgenera in thefamilyCampanulaceae with the common

name bellflower. It takes both its common and its scientific name from its bell-

shapedflowers campanulaisLatin for "little bell". The genus includes over 500species and

severalsubspecies,distributed across the temperate and subtropical regions of theNorthern

Hemisphere,with the highest diversity in theMediterranean region east to theCaucasus.The

range also extends into mountains in tropical regions ofAsia andAfrica. The species

includeannual,biennial andperennialplants, and vary in habit from dwarf arctic and alpine

species under 5 cm high, to largetemperategrassland andwoodland species growing to 2

meters (6 ft 7 in) tall.

Theleaves are alternate and often vary in shape on a single plant, with larger, broader leaves

at the base of the stem and smaller, narrower leaves higher up; the leaf margin may be either

entire or serrated (sometimes both on the same plant). Many species contain white latex in the

leaves and stems.

The flowers are produced inpanicles (sometimes solitary), and have a five-lobedcorolla,

typically large (25 cm or more long), mostly blue to purple, sometimes white or pink. Below

the corolla, 5 leaf-likesepals form the calyx. Some species have a small additional leaf-like

growth termed an

"appendage" between each sepal, and the presence or absence, relative size,

and attitude of the appendage is often used to distinguish between closely related species.

Thefruit is acapsule containing numerous smallseeds.Campanulaspecies are used as food

plants by the larvae of someLepidoptera species includingCommon Pug (recorded on

Harebell),Dot Moth,Ingrailed Clay (recorded on Harebell),Lime-speck Pug andMouse

Moth.
http://en.wikipedia.org/wiki/Genushttp://en.wikipedia.org/wiki/Family_(biology)http://en.wikipedia.org/wiki/Campanulaceaehttp://en.wikipedia.org/wiki/Flowerhttp://en.wikipedia.org/wiki/Latinhttp://en.wikipedia.org/wiki/Specieshttp://en.wikipedia.org/wiki/Subspecieshttp://en.wikipedia.org/wiki/Northern_Hemispherehttp://en.wikipedia.org/wiki/Northern_Hemispherehttp://en.wikipedia.org/wiki/Mediterraneanhttp://en.wikipedia.org/wiki/Caucasushttp://en.wikipedia.org/wiki/Asiahttp://en.wikipedia.org/wiki/Africahttp://en.wikipedia.org/wiki/Annual_planthttp://en.wikipedia.org/wiki/Biennial_planthttp://en.wikipedia.org/wiki/Perennial_planthttp://en.wikipedia.org/wiki/Temperatenesshttp://en.wikipedia.org/wiki/Grasslandhttp://en.wikipedia.org/wiki/Woodlandhttp://en.wikipedia.org/wiki/Leafhttp://en.wikipedia.org/wiki/Latexhttp://en.wikipedia.org/wiki/Panicleshttp://en.wikipedia.org/wiki/Corolla_(flower)http://en.wikipedia.org/wiki/Sepalshttp://en.wikipedia.org/wiki/Fruithttp://en.wikipedia.org/wiki/Capsule_(fruit)http://en.wikipedia.org/wiki/Seedhttp://en.wikipedia.org/wiki/Larvahttp://en.wikipedia.org/wiki/Lepidopterahttp://en.wikipedia.org/wiki/Common_Pughttp://en.wikipedia.org/wiki/Dot_Mothhttp://en.wikipedia.org/wiki/Ingrailed_Clayhttp://en.wikipedia.org/wiki/Lime-speck_Pughttp://en.wikipedia.org/wiki/Mouse_Mothhttp://en.wikipedia.org/wiki/Mouse_Mothhttp://en.wikipedia.org/wiki/Mouse_Mothhttp://en.wikipedia.org/wiki/Mouse_Mothhttp://en.wikipedia.org/wiki/Lime-speck_Pughttp://en.wikipedia.org/wiki/Ingrailed_Clayhttp://en.wikipedia.org/wiki/Dot_Mothhttp://en.wikipedia.org/wiki/Common_Pughttp://en.wikipedia.org/wiki/Lepidopterahttp://en.wikipedia.org/wiki/Larvahttp://en.wikipedia.org/wiki/Seedhttp://en.wikipedia.org/wiki/Capsule_(fruit)http://en.wikipedia.org/wiki/Fruithttp://en.wikipedia.org/wiki/Sepalshttp://en.wikipedia.org/wiki/Corolla_(flower)http://en.wikipedia.org/wiki/Panicleshttp://en.wikipedia.org/wiki/Latexhttp://en.wikipedia.org/wiki/Leafhttp://en.wikipedia.org/wiki/Woodlandhttp://en.wikipedia.org/wiki/Grasslandhttp://en.wikipedia.org/wiki/Temperatenesshttp://en.wikipedia.org/wiki/Perennial_planthttp://en.wikipedia.org/wiki/Biennial_planthttp://en.wikipedia.org/wiki/Annual_planthttp://en.wikipedia.org/wiki/Africahttp://en.wikipedia.org/wiki/Asiahttp://en.wikipedia.org/wiki/Caucasushttp://en.wikipedia.org/wiki/Mediterraneanhttp://en.wikipedia.org/wiki/Northern_Hemispherehttp://en.wikipedia.org/wiki/Northern_Hemispherehttp://en.wikipedia.org/wiki/Subspecieshttp://en.wikipedia.org/wiki/Specieshttp://en.wikipedia.org/wiki/Latinhttp://en.wikipedia.org/wiki/Flowerhttp://en.wikipedia.org/wiki/Campanulaceaehttp://en.wikipedia.org/wiki/Family_(biology)http://en.wikipedia.org/wiki/Genus


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Daturais agenus of nine species of poisonousvespertineflowering plantsbelonging to the

familySolanaceae.They are known as angel's trumpets, sometimes sharing that name with

the closely related genusBrugmansia, and commonly as daturas. They are also sometimes

called moonflowers, one of several plant species to be so. Its precise and natural distribution

is uncertain, owing to its extensivecultivation andnaturalization throughout the temperate

and tropical regions of the globe. Its distribution within theAmericas andNorth Africa,

however, is most likely restricted to theUnited States and Mexico in North America,

andTunisia in Africa, where the highest species diversity occurs.

All species ofDaturaare poisonous, especially their seeds and flowers.

SomeSouth Americanplants formerly thought of asDaturaare now treated as belonging to

the distinct genusBrugmansia (Brugmansiadiffers fromDaturain that it is woody,

makingshrubs or smalltrees, and it has pendulous flowers, rather than erect ones). Other

related genera includeHyoscyamusandAtropa.

REVIEW OF LITERATURE:

Staphylococci are Gram-positive bacteria, with diameters of 0.5 1.5 m and characterized

by individual cocci, which divide in more than one plane to form grape-like clusters. To date,

there are 32 species and eight sub-species in the genus Staphylococcus, many of which

preferentially colonies the human body (Kloos and Bannerman, 1994),

However Staphylococcus aureus and Staphylococcus epidermidis are the two most

characterized and studied strains. The staphylococci are non-motile, non-spore forming

facultative anaerobes that grow by aerobic respiration or by fermentation. Most species have

a relative complex nutritional requirement, however; in general they require an organic

source of nitrogen, supplied by 5 to 12 essential amino acids, e.g. arginine, valine, and B

vitamins, including thiamine and nicotinamide (Kloos and Schleifer, 1986; Wilkinson, 1997).
http://en.wikipedia.org/wiki/Genushttp://en.wikipedia.org/wiki/Vespertine_(biology)http://en.wikipedia.org/wiki/Flowering_planthttp://en.wikipedia.org/wiki/Solanaceaehttp://en.wikipedia.org/wiki/Brugmansiahttp://en.wikipedia.org/wiki/Brugmansiahttp://en.wikipedia.org/wiki/Brugmansiahttp://en.wikipedia.org/wiki/Plant_cultivationhttp://en.wikipedia.org/wiki/Introduced_specieshttp://en.wikipedia.org/wiki/Americashttp://en.wikipedia.org/wiki/North_Africahttp://en.wikipedia.org/wiki/United_Stateshttp://en.wikipedia.org/wiki/Mexicohttp://en.wikipedia.org/wiki/Tunisiahttp://en.wikipedia.org/wiki/South_Americahttp://en.wikipedia.org/wiki/Brugmansiahttp://en.wikipedia.org/wiki/Brugmansiahttp://en.wikipedia.org/wiki/Brugmansiahttp://en.wikipedia.org/wiki/Shrubhttp://en.wikipedia.org/wiki/Treehttp://en.wikipedia.org/wiki/Henbanehttp://en.wikipedia.org/wiki/Henbanehttp://en.wikipedia.org/wiki/Henbanehttp://en.wikipedia.org/wiki/Atropahttp://en.wikipedia.org/wiki/Atropahttp://en.wikipedia.org/wiki/Atropahttp://en.wikipedia.org/wiki/Atropahttp://en.wikipedia.org/wiki/Henbanehttp://en.wikipedia.org/wiki/Treehttp://en.wikipedia.org/wiki/Shrubhttp://en.wikipedia.org/wiki/Brugmansiahttp://en.wikipedia.org/wiki/South_Americahttp://en.wikipedia.org/wiki/Tunisiahttp://en.wikipedia.org/wiki/Mexicohttp://en.wikipedia.org/wiki/United_Stateshttp://en.wikipedia.org/wiki/North_Africahttp://en.wikipedia.org/wiki/Americashttp://en.wikipedia.org/wiki/Introduced_specieshttp://en.wikipedia.org/wiki/Plant_cultivationhttp://en.wikipedia.org/wiki/Brugmansiahttp://en.wikipedia.org/wiki/Solanaceaehttp://en.wikipedia.org/wiki/Flowering_planthttp://en.wikipedia.org/wiki/Vespertine_(biology)http://en.wikipedia.org/wiki/Genus


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Members of this genus are catalase-positive and oxidase-negative, distinguishing them from

the genus streptococci, which are catalase-negative, and have a different cell wall

composition to staphylococci (Wilkinson, 1997).

Staphylococci are tolerant to high concentrations of salt (Wilkinson, 1997) and show

resistance to heat (Kloos and Lambe 1991). Pathogenic staphylococci are commonly

identified by their ability to produce coagulase, and thus clot blood (Kloos and Musselwhite,

1975). This distinguishes the coagulase positive strains, S. aureus (a human pathogen), and S.

intermedius and S. hyicus (two animal pathogens), from the other staphylococcal species such

as S. epidermidis, that are coagulase-negative (CoNS).

Staphylococcus aureus is an opportunistic pathogen often carried asymptomatically on the

human body. Methicillin-resistant S. aureus (MRSA) strains have acquired a gene that makes

them resistant to all beta-lactam antibiotics. Infections caused by Staphylococcus aureus

pose serious threat in health care institutions. (Panlilio et al. 1992; and NNIS 2001, 2004). It

is one of the most widely spread and virulent nosocomial pathogen and is usually resistant to

multiple antibiotics making infections difficult to treat (Cooper et al. 2004). It appears to add

to the total burden of Staphylococcus infections in the hospitals, rather than replacing

sensitive S. aureus, and is associated with sharp risk in mortality attributable to

Staphylococcal infection (Crowcroft & Catchpole 2002).

Staphylococcus aureus strains continue to be a major problem in many healthcare institutions

especially with emergence of Methicillin resistant Staphylococcus aureus (MRSA) and now

account for more than 50% of S. aureus recovered from patients in intensive care units and

about 40% of S. aureus isolated from non intensive care unit (Boyce 2003).

Although, the clinical significance of methicillin resistance has been questioned in the past,

there is now widespread acknowledgement of the pathogenicity of MRSA. It has emerged as

a significant cause of both nosocomial and community-acquired infections. Recent report of


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strains of MRSA isolated from children in the community has led to speculation that the

epidemiology of S. aureus is changing (CDC 1999; and Boyce 1998).

Traditionally, MRSA infections have been acquired almost exclusively in hospitals, long-

term care facilities or similar institutional settings (Thompson et al. 1982). Health-care

associated infection commonly caused by MRSA includes surgical site infections, bacteremia

and endocarditic, pneumonia, soft-tissue infections and urinary tract infections. However, the

emergence of community-associated MRSA (CA-MRSA) infections is of major concern to

both public health officials and clinicians.

The first report of CA-MRSA infection occurred among Australian aboriginals and Native

Americans in Canada in the early 1990s (Boyce 2003). The earliest reported cases of CA-

MRSA infection in the United States occurred in children with little or no recognized contact

with the hospitals or other health care institutions (Herold et al. 1998). Coagulase negative

Staphylococci (CNS) belong to the group of opportunistic pathogens since they are found as

normal flora of the skin and mucus membranes in different part of the body (Einsenstein and

Schaechter 1994). For this reason, CNS are often reported without further specification,

assuming that they are contaminating clinical samples but are not involved in the primary

infection.

However, there is mounting evidence that these bacteria may be responsible for primary

infections as a result of increased use of medical in dwelling plastic devices and

compromised or immune depressed patients (Jarvis andMartone1994; and Kloos and

Bannerman 1994). Methicillin resistance among CNS is particularly important due to cross

resistance to virtually al B-lactam agents and other antimicrobial classes. As a result,

therapeutic approaches are restricted to glycopetide and new antimicrobial agents as

Linezolid (Woods et al. 2002). Therefore, an accurate analysis of resistance between clinical


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and community strains may allow the provision of better antimicrobial therapy. Besides, the

importance for patient care the detection also has implications for the validity of antibiotic

resistance surveillance. Hence the purpose of this study is to isolate Staphylococcus species

from both clinical and community based samples and to determine the antibiogram of the

isolates against some selected commercial antibiotics.

Methicillin resistant S. aureus (MRSA) is a bacterium that has developed resistance to most

antibiotics such as methicillin, gentamycin, fucidic acid and clindamycin that are commonly

used for Staphylococcus infections, unfortunately, leading to failure of treatment (Shai et al.,

2004). The two major strains of MRSA are known to be hospital-acquired (HA) MRSA and

community-acquired (CA) MRSA. HA-MRSA includes cases in which the patient has had a

current or recent hospitalization receives dialysis, or resides in a long-term care facility.

During the period 1970 to 2010, strains of S. aureus resistant to multiple antibiotics including

methicillin were increasingly responsible for outbreaks of nosocomial infections in countries

around the world, for example, Saudi Arabia (Madani et al., 2001), Argentina (Reyes et al.,

2009), South Africa (Shittu et al., 2009), Italy (Soavi et al., 2010) and the United States

(Boyce, 1990). In many instances, these outbreaks were associated with individual wards,

neonatal, intensive care and burns units (Liu et al., 2011). Furthermore, increasing incidence

of CA-MRSA has been a growing public health concern (Mandell et al., 2005; Ma et al.,

2007) and has emerged as the predominant cause of skin infections in the USA (Stevens et

al., 2010).

Only few intervention studies have been published to explore alternative antimicrobial agents

to control and prevent diseases due to multidrug resistant S. aureus. Although other practices

have been explored such as bacterial interference therapy (Maibach and Aly, 1981) and phage

therapy (Jikia et al., 2005), medicinal plants have also been considered by some researchers

since they are frequently used in popular medicine as remedies for many infectious diseases


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(Geyid et al., 2005; Mohana et al., 2008; Rahim et al., 2010). The aim of the present study

was to determine the inhibitory effect of different plant extracts on the growth resistant

strains of S. aureus.

OBJECTIVES:

Isolation of Staphylococcifrom Cell Phones.

Characterization Staphylococcus aureus.

Kirby Bauer Antibiotic Sensitivity Test.

Inhibitory Effect of Flower Extracts on Drug Resistant S. aureus.

MATERIALS AND METHODS:

3.1 ISOLATION AND CHARACTERIZATION OF STAPHYLOCOCCI

A. SAMPLING:

Collection of microbial samples from cell phones was carried out by swabbing the cell phone surfaces

on back and front by swabs immersed in sterile saline tubes (Duramz, et al., 2000). Same procedure

was followed for all the samples collected from different population groups .Samplings were

processed immediately after collection for isolation of bacterial pathogens (Kapdi, et al.2008).b.

B. ISOLATION:

The swabbed samples were serially diluted up to 10-9 dilution and plated by pour plate technique on

selective media like Nutrient agar media. The colonies were picked from these selective media by

noting down their colony characters and subjected for characterization studies by culturing them on

slants of respective selective media (Kolpin ,et al.,200 observed under microscope (Sepehri, et al,

2009).


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C. CHARACTERIZATION:

BIOCHEMICAL TESTS:

KOH solubility test:

3% KOH solution was prepared and loop full of test culture was mixed with 3% KOH solution then

observed for the thread like appearance to identify Gram positive and Gram negative bacteria ( Aneja,

1996).

Starch hydrolysis: All the isolated organisms were inoculated on starch agar medium and

after incubation the clear zone was observed for the hydrolysis of starch (Siddiqui, et al,

1999).

Gelatin hydrolysis: Gelatin liquifaction was observed in all the test isolates when the

inoculated tubes were kept in at 4c to observe gelatin hydrolysis (Siddiqui, et al, 1999).

Sugar fermentation:All the test organisms were inoculated on 9ml of phenol red sucrose,

lactose; dextrose broth with 1ml broth was filled with Durhams tube. After incubation the

tubes were observed for acid and gas production (Arora, et al, 2009).

Catalase test:all the isolates were subjected for the catalase activity when the test organisms

were treated with 3% hydrogen peroxide by noticing the effervescence (Akinyemi, et al,

2009).

IMViC Test:

1)Indole production test:the test organism was inoculated on 1% tryptone broth after incubation

added with one ml of Kovacksreagent for indole production.

2) Methyl red test: Test organism was inoculated on methyl red broth. After incubation by adding

Five drops of methyl red indicator colour change was observed (Akinyemi, et al, 2009).

3) Voges-Proskaurs test: test organism were inoculated on V-P broth after incubation by adding

12 drops of VP-1 reagent and 3 drops of VP-2 reagent to observe the colour change from yellow to

ruby pink..(Akinyemi, et al, 2009). 4). Citrate utilization test: the test isolates were inoculated on

simmon citrate agar medium after incubation to observe the colour change from green to blue.


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5) Haemolysis test:All the test isolates were inoculated on blood agar medium after incubation

alpha, beta and gamma haemolysis was observed (Akinyemi, et al, 2009).

6) Mannitol fermentation: All the test organisms were inoculated on mannitol salt agar medium

after incubation observe the colour change from red to yellow was observed (Anderson, 2006).

3.2 ANTIBIOTIC ASSAY:

A. SCREENING:

Kirby-Bauer antibiotic sensitivity assay was followed for the selected isolates based on their

characterization studies. Different antibiotics like Methicillin, Vancomycin, Ampicillin and

Penicillin (at 10 micro gram concentration) (Hirematsuet al.,2008).Since all these were found to be

effective antimicrobial antibiotics (Fereen, et al.,2008).Broth cultures of test bacterial isolates were

swabbed on solidified nutrient agar media with sterile swabs (Anderson,2006). The antibiotic discs

were placed at equidistance on the agar surface and plates were incubated at 370C for 24 hours, the

sensitivity or resistance was passed by measuring the zone of inhibition in millimetre (Boyce, 2008).

3.3 ANIBACTERIAL POTENTIAL OF FLOWER EXTRACT

AQEOUS EXTRACT PREPARATION:

Fresh flowers, leaves of datura (dark lavender and light lavender), bell flower and fresh fruit of bell

flower were collected from the surrounding of Shivamogga, Holehonnur and Arasalu. The fresh

samples were washed with water and air dried. About 5 grams of samples were weighed and crushed

with the pestle and mortar respectively. The extract was mixed with 10ml of sterile water respectively

for sterile water extract. Well in agar method was followed to screen the antibacterial activity of

aqueous extracts.

SOLVENT EXTRACT PREPARATION:

Fresh flowers, leaves of datura (dark lavender and light lavender),bell flower and fresh fruit of bell

flower were collected from the surrounding of Shimogga, Holehonnur ,Arasalu. The fresh samples


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were washed with water and air dried. About five grams of samples were weighed and crushed with

the pestle and mortar respectively. The extract was mixed with 10ml of different solvents like

chloroform, petroleum ether, methanol, ethyl acetate respectively for solvent extract.

WELL IN AGAR METHOD:

Sterilized Nutrient media was prepared and poured into sterile Petri plates.

Allow the plates were allowed for solidification.

The plates were labeled with the name of the bacterial culture Streptococcus aureus

which was swabbed on each plate and the type of compound was also labeled

respectively.

After solidification, swab was done on the surface of NA media with the selected

bacterial cultures.

After swabbing the well was digged using sterilized cork borer.

100-200L of the 10% solvent extract was pipette out using micropipette and added

into the well which was digged.

The different solvent extract was added to each well.

The plates were kept for incubation at 37 degree C for 24 hours.

After incubation the zone of inhibition was observed in those plates, and was

measured by millimeter ruler. (Snehayadavet al, 2011).

RESULTS AND DISCUSSION:

Present study revealed the potential of natural products, like flower extracts on Methicillin

resistant S.aureus isolates from Cell Phones. Earlier, antimicrobial activity on natural

products was reported by Alamshel et.al (2009). Antimicrobial activity of datura and


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bellflower solvent extract was tested against S.aureus by WELL IN AGAR method. The

results indicated that the sterile water extract of datura leaf and other solvent extract show

better antimicrobial activity. In vitroantibacterial assay of flower extracts exhibited that the

solvent extracts contain bioactive compounds that can inhibit the growth of S.aureus.

When datura flower with dark lavender colour extract was mixed with sterile water was

transferred to a media which was swabbed with S.aureus organisms shown that zone of

inhibition of microorganism was found to be 26mm.When the Datura flower with light

lavender colour was tested to the above mentioned solvent, the zone of inhibition of

microorganism was found to be 30mm. The same light lavender colour flower was tested to

the solvents like ethyl acetate and methanol, the zone of inhibition of micro organism was

found to be 18mm and 26mm respectively. Where as in petroleum ether solvent, the flower

does not show any zone of inhibition of micro organism rather the microbial growth was very

abundantly that we could found any zone of inhibition. In our experiment, we can see that the

zone of inhibition was more in the solvent sterile water that is 30mm and zone of inhibition is

in ethyl acetate is 18mm with less growth . So the solvent extract, sterile water extract having

more antimicrobial potential and it can be used as drug (Table no2)

When Datura leaf extract treated with solvents was transferred to media we can see that the

zone of inhibition of micro organism is more in methanol 22mm, sterile water 18mm , ethyl

acetate 10mm and in chloroform and petroleum ether zone of inhibition is least (Table no3 ).

When bell flower fruit extract treated with solvents was transferred to media which was

swabbed with S.aureusshows that the zone of inhibition of micro organism in sterile water is

15mm and methanol is 22mm. Whereas zone of inhibition least in chloroform, petroleum

ether and ethyl acetate (Table no4 ).


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In this zone of inhibition of micro organism was more in sterile water 20mm , and chloroform

22mm where less growth of micro organism was found ( Table no 5).

Table no 1: Kirby Bauer Antibiotic Sensitivity Test:

Sl

No

Bacterial

isolates

Zone of inhibition in mm

Penicillin Amphicillin Methicilin Vancomycin

01 S.aureus - - - 17

02 S.aureus - - - 18

03 S.aureus - - - 18

04 S.aureus


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Table no 3: INHIBITORY EFFECT OF DATURA LEAF EXTRACTS ONMRSA

Sl No Solvent Extract Zone of inhibition in mm

control Test

1 Sterile water 16 18

2 Ethyl acetate 10 10

3 Chloroform - -

4 Methanol - 22

5 Petroleum ether - -

Table no 4: INHIBITORY EFFECT OF BELL FLOWER FRUIT EXTRACT ON MRSA

Staphylococcus aureus:


control Test

1 Sterile water 11 15

2 Methanol 22

Table no 5: INHIBITORY EFFECT OF BELL FLOWER LEAF EXTRACT ON MRSA

Staphylococcus aureus:


control Test

1 Sterile water - 20

2 Ethyl acetate 28 32

3 Chloroform - 22

4 Methanol 14 -


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SUMMARY:

The present project work concentrated on isolation and characterisation of Staphylococcus aureus

isolates from cell phones. These isolates when tested for antibiotic sensitivity test by Kirby Bauer disc

diffusion method, few isolates exhibited resistance to the tested standard antibiotics like Penicillin,

Ampicillin, Methicillin and sensitive to Vancomycin.

Methicillin Resistant S.aureus isolates were subjected for further studies. Inhibitory effect of natural

products like flower extract of Datura and Bellflower were tested on these isolates by well in agar

method. 5% concentration of petroleum ether, ethyl acetate, chloroform and methanol extract were

capable of inhibiting MRSA to different extends. Among the solvent extract tested sterile water and

ethyl acetate were best in extracting the active fractions from flower and fruit extracts of Datura and

Bellflowers .


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CONCLUSION:

This study probably suggest the possibility of concurrent use of these plant extracts in

solvents like sterile water , ethyl acetate, methanol, chloroform in combination in

treating infection caused by S.aureus.

This study on antimicrobial potential of Datura and bellflower extract revealed that

the sterile water extract was more potent than other solvent extract.

Therefore our results revealed the importance of Datura and Bellflower of solvent

extracts when compared with antibiotics to control the S.aureus.


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NATURAL PRODUCTS

Bell flower Datura flower Datura flower

Bell flower fruit Datura flower Datura flower


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PHOTOS

Staphylococcus aureusfrom Cell phones:

Antibiotic sensitivity test:


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INHIBITORY EFFECT OF NATURAL PRODUCTS:

Documents

“INHIBITORY POTENTIAL OF FLOWER EXTRACTS ON MRSA FROM CELL PHONES”