Magazine Issue4

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Microbiology World Mar Apr 2014 ISSN 2350 - 8774

www.microbiologyworld.com www.facebook.com/MicrobiologyWorld

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http://www.microbiologyworld.com/http://www.facebook.com/MicrobiologyWorldhttp://www.facebook.com/MicrobiologyWorldhttp://www.microbiologyworld.com/


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Chief Editor

Mr. Sagar AryalMedical Microbiology (M.Sc), Nepal(Founder)

Reviewers

Mr. Samir AgaDepartment of Immunological Diseases

Medical Technologist, Iraq

Mr. Saumyadip Sarkar

ELSEVIER Student Ambassador South Asia, Reed Elsevier (UK)Ph.D Scholar (Human Genetics), India

EditorsDr. Sao Bang

Hanoi Medical University

Dean of Microbiology Department (Provincial Hospital)Microbiology Specialist, Vietnam

Mr. Tankeshwar AcharyaLecturer: Patan Academy of Health Sciences (PAHS)Medical Microbiology, Nepal

Mr. Avishekh Gautam

Ph.D Scholar, Hallym University, South Korea

Mr. Manish ThapaliyaLecturer: St. Xaviers College

Food Microbiology, Nepal

Mr. Sunil PandeyMedical Microbiology, Nepal


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Table of Content

Page No.

Who are Microbiologist? 4-7

DNA Microarray Technique 8-10

Animal Vaccination 11-15

Microbiology in Pharmaceuticals Industry 16-19

Need of Advocacy and awareness about

cancer in Nepal 20-21

How to Microorganism contribute

to body odor? 22-23

Embryonic Stem cell research 24-28

Staphylococcus aureus And MRSA 29-32


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Who are Microbiologist?

Microbiologists are biological scientists who study organisms so small that, generally, they canonly be seen with a microscope. These microorganisms include bacteria, algae, yeasts, fungi,protozoa, viruses, and other microscopic forms of life. Microbiologists isolate and make culturesof microorganisms, identify their characteristics, and observe their reactions to chemicals and

other kinds of stimuli. They also study how microorganisms develop and reproduce as well astheir distribution in nature.

Many microbiologists work for universities, where they teach and do research. Others work atmedical centers or in private industry. Some work for government agencies. Although their jobs

have different aspects and responsibilities, most microbiologists do some research or laboratorywork. They use special equipment to study microorganisms including light microscopes, electron

microscopes, centrifuges, glass tubes, slides, and computers. They are often assisted bybiologica l technicians.

Microbiology is a broad field that includes the study of viruses as well as microscopic organismsfound in all kingdoms of life: plants, animals, protists, fungi, and bacteria. Some microbiologists

specialize in one type of microorganism. For example, bacteriologists concentrate on bacteriaand virologists study viruses.Microbiologists work in several areas. Many do basic research to increase knowledge about the

life processes common to microbes. Their work helps to answer basic questions such as thosepertaining to the use of food and oxygen in cells. Other microbiologists are employed in

medicine. Medical microbiologists study the relationship between microorganisms and disease.

They isolate and identify disease-producing organisms and study their distribution. They alsostudy the ways that the organisms enter the bodies of humans and animals, establish themselves,

and cause disease. Immunologists, for example, study the body's defensive responses tomicroorganisms.

Microbiologists are also employed in the related field of public health. They work to combatproblems such as outbreaks of epidemics, food poisoning, and the pollution of air and water. For

example, public health microbiologists test blood samples sent in by physicians to see whetherpatients have a communicable disease. They also test drinking water, milk supplies, and other

substances that can affect the health of the general public.

Other fields in which microbiologists work include agriculture, marine microbiology, andindustry. Agricultural microbiologists study the microorganisms found in soil and their effects onplant growth. Marine microbiologists seek ways to control the growth of harmful bacteria in

oceans and rivers. Industrial microbiologists work in a variety of industries, including foodprocessing, chemicals, and drugs. They may work to control the activities of microorganisms insuch processes as the tanning of leather and the fermentation of wine.


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Working Conditions

Working conditions for microbiologists vary. Most spend at least part of their time in clean,well-lighted laboratories. Some microbiologists have to collect samples of soil, seawater, and

other substances that contain microorganisms. Some microbiologists spend part of their time inclassrooms and offices. The workweek for many microbiologists in medical centers and privateindustry is generally forty hours. Those who work in universities and other research centers may

have more flexible hours, but their workweeks generally total more than forty hours. Someovertime or shift work may be necessary when a project must be completed or when an

experiment must be monitored around the clock. Microbiologists usually spend some timereading and studying to keep up with the newest findings of other scientists.

Microbiologists must take precautions to prevent specimens from being contaminated and tokeep harmful microorganisms from reproducing uncontrollably. They should have skill in

scientific experimentation and mathematics and be willing to do the precise, detailed workrequired in microbiology. Microbiologists should be able to work either independently or as partof a team. They must be able to keep careful records and to communicate their ideas and findings

to others.

Description, Areas and Scope of Medical Microbiologist

Medical microbiology course is branch within the field of medicine which focuses on microorganisms of medical interest which include the bacteria, viruses, fungi and parasites which are

of medical importance and are capable of causing infectious diseases in human beings.Microbiologists can rather be called as detectives who investigate in the microscopic world - aworld that holds much wonder and mystery, the world which much people cannot see.

Medical Microbiology Course description includes the study of microbial pathogenesis andepidemiology and is related to the study of disease pathology and immunology. Microbiologists

study micro organisms which can cause disease in people, looking into life cycles of suchorganisms, on how they cause infection, how they spread, and cause disease, the means to treat

the diseases and irradiate the disease causing microbes.

This field of microbiology is constantly engaged with identifying new micro organisms,

monitoring changes in rapidly mutating species and dealing with the ongoing challenges inmedicine, ranging from the development of resistance to antibiotics in bacteria to contamination

of water supplies with protozoans. They work in all ways to contribute to mankind and improvehumans and environmental quality of life and living.


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Scope and career prospectus of microbiologist

TEACHING :If you are interested in teaching as a profession and if you are interested to pursue your masters

degree, and you are basically a science student, then microbiology is the best suited to pursue amasters in. Teaching is a noble profession, and the returns after completing your study istremendous. And again the scope is vast as it is the basic subject of study in all medical colleges

as well as Para medical colleges like dental science, physiotherapy, pharmaceutical science,nursing science, radiography, lab technology etc. It is the most highly paid among the teaching

profession in science, even at the initial stage as a beginner. As well you get an opportunity towork among the dignified doctors and amongst the highly esteemed medical professionals.Academic microbiologists are employed by universities, schools and teaching hospitals.

RESEARCH SCIENTIST:

Research is a never ending field and thus one can take up research. As a research scientist amicrobiologist can work in universities, institutions, industries, hospitals, governmentorganizations and carry out research in laboratories. Many work as associate or assistant

scientists or researchers doing the routine work of conducting experiments, others are seniorscientists or project managers who lead experiments, supervise lab workers, interpret data and

develop new theories and experiments.

CLINICAL ASSOCIATE:

One can get into clinical research as clinical coordinator or clinical trials administrator and lateron generally qualify to move on to a full clinical research associate position.

CLINICAL MICROBIOLOGIST:Clinical microbiologist assist the treating team of doctors with the investigations, diagnosis and

treatment of disease. That analyze and interpret data related to patient samples. They work forthe betterment of the society by engaging in a wider range for complex work in laboratories

especially against incurable, life threatening diseases like cancer, AIDS etc. They adviseclinicians on the use of tests, diagnosis of disease and planning and progress of the treatment.They work to combat problems such as outbreaks of epidemics, food poisoning, pollution of air

and water, for eg., they check blood samples sent in by physicians to check for anycommunicable diseases.

TOXICOLOGIST:Microbiologists are employed as toxicologists in investigatory laboratories. They plan and carry

out laboratory and field studies to identify, monitor and evaluate the impact of toxic materialsand radiations on human and animal health, and on the health and current status of the

environment, as well as the impact of future technologies.


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FORENSIC SCIENTISTS:Microbiologists with appropriate skills are also considered for posts in forensic science.

BIOMEDICAL SCIENTISTS:

Following basic training, most biomedical scientists specialize in one aspect of medicallaboratory science. The main areas are microbiology, clinical chemistry, transfusion science,hematology, histopathology, cytology, immunology and virology.

BIOSTATISTICIAN:

Biostatisticians are statisticians who work in the health related fields. They design researchstudies and collect and analyze data on problems such as how disease progress, how safe is thetreatment, or the impact of certain risk factors associated with medical conditions.

Microbiologists have enough and more jobs as quality assurance specialists in thepharmaceutical companies, food processing industry, diary and milk products, beverages, hotels,biotechnology companies, agriculture, forestry etc.

Microbiology in Nepal

Nepal is a least developed and land locked country. Ecologically it is divided in to Mountainregion, Hilly region and Terai region. Majority of population living in remote and village area

and these areas are lacking many facilities and there is unplanned urbanization in developed partresulting in lack of health care facilities, places and water supply. Most of them have poor

economic status and have little or no knowledge about proper sanitation so that infectious can beprevented. About 70% of all health problems are attributed by infectious diseases in nepal. Manychildren are dying from easily preventable and treatable diseases such as diarrhea and/or

dysentery, and acute respiratory infections. So, Knowledge of microbiology is important inNepal .

- Sunil PandeyNobel College, Kathmandu, Nepal


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DNA Microarray Technique

It is DNA homology analysis for detecting polymorphisms and mutations in both prokaryoticand eukaryotic genomic DNA.

A microarray is a compact device which contains a large number of well defined immobilizedcapture molecules in the form of spots assembled in a known pattern on a solid support, usually

glass slides, nylon membranes or silicon chips. These molecules can be PCR products, proteins,antibodies, oligos.

PRINCIPLE

The basic principle of microarray is based on the hybridization of complimentary probe and

target cells. Each DNA spot contains a small amount of a specific DNA sequence in picomoles(1012 moles), which are known as probes.These can be a short section of a gene or other DNAelement that are used to hybridize a cDNA or cRNA (also called anti-sense RNA) sample (calledtarget) under high-stringency conditions. The complementary nucleic acid sequences specifically

pair with each other by making hydrogen bonds between complementary nucleotide base pairs.Probe-target hybridization is usually detected and quantified by detection of fluorophore, silver,

or chemo-iluminescence-labeled targets to determine relative abundance of nucleic acidsequences in the target.

Thus by using an array containing many DNA samples, scientists can determine the expressionlevel of thousands of genes in a cell by measuring the amount of mRNA bound to each spot on

the array.

HISTORY

Microarray technology is evolved from Southern Blotting, in Southern blotting technique DNA

fragments are attached to a substrate and then they are probed with a known DNA sequencesegment.

In 1987 the arrayed DNA were used for the identification of genes whose expression is regulatedby interferon. These early gene arrays where developed by spotting cDNA on filter papers with

the help of automated pin spotting device.

In 1995 quantitative monitoring of gene expression with a complimentary DNA microarray was

done.


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In 1996 for the first time microarrays were commercialized by an American companyAFFYMETRIX.

In 1997 a complete eukaryotic genome of a yeast Saccharomyces cerevisiae was published on an

microarray.

From 2000-2003 the use of microarray in clinics was introduced for the detection of cancerous

cells.

In 2004 the whole human genome was published on a microarray.

TYPES OF MICROARRAYS

DNA microarrays such as cDNA microarray, oligonucleotide microarrays and SNP microarrays .

MMchips, for surveillance of microRNA populations.Peptide microarrays, for detailed analysis or optimization of protein protein interactions.Tissue microarray.

Chemical compound microarray.Cellular microarray (also called transfection microarrays)

Antibody microarrays.Carbohydrate microarrays (glycoarrays)Phenotype microarrays.

BASIC STEPS INVOLVED IN MICROARRAY TECHNIQUE1. Obtain DNA microarray chip

2. Extraction of RNA from the samples3. Cy3 and Cy5 labeling of the RNA4. Production of cDNA by RT-PCR

5. Hybridization with cDNA probes to DNA array6. Detection of hybridization signal

7. DNA microarray data analysis


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APPLICATIONS OF MICROARRAY

DNA microarray are used to examine gene expression patterns under diseases such as cancer.Tumor profiling using DNA microarrays allows the analysis of development and progression of

such complex diseases.

DNA microarrays are used for virus detection from blood and other samples thus it is used forpathogen detection.

DNA microarrays more recently have been used to indentify inheritable markers, and there usedas genotyping tool.

Used for single nucleotide polymorphism (SNP) detection.

ADVANTAGES OF MICROARRAYS

Microarrays are better than other profiling techniques because:Easier to use

Can analyze thousand of genes or markers at a timeGenerate large amount of data in little timeDo not require large scale sequencing

Allow the quantization of thousand of genes from many samplesCompare two different population like:

wild type vs wild, normal tissues vs cancerous tissue, study specific chromosomal regions.

- Muhammad Hamza Sana (D.V.M)+923436247467

[email protected]

Institute of Microbiology, University of Agriculture, Pakistan.


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Animal Vaccination: low cost insurance against

disease

Introduction

It is always good to prevent rather than to cure a disease and vaccination is the most effectiveway of preventing diseases. Beginning with the variolation the method of prophylaxis reached a

new era with the advent of cell culture techniques in 1950s which led to the development ofmany live attenuated and inactivated virus vaccines. More recently, the field of vaccinology haswitnessed the introduction of novel new generation vaccines produced through various form of

recombinant DNA and related techniques which offer significant improvements and potential

advantages in terms of both their safety and their efficacy. Its all because of vaccination that wewere able to eradicated smallpox and rinderpest globally. Like people, animals also needvaccines. The importance of vaccination is when you are aware of the vaccines of your animaland their schedule and more importantly getting your animal vaccinated regularly at prescribed

time interval. This article provides information to animal owners and pet lovers on routinevaccination programs to prevent common diseases.

When to vaccinate

Animals should be vaccinated for a disease before they will encounter the microorganismscausing it. It takes from 10 to 21 days after vaccination to mount a protective immune response.

The exact length of time depends on the animal, its age and health status, the vaccine itself, andwhether it has been vaccinated before. We have to emphasize the fact that no single schedule is

suitable for every situation. An effective vaccination schedule depends on the type of operation,maternal antibody levels, disease challenge on the farm, age, level of hygiene and many otherfactors. It is best to consult a veterinary specialist to advice a schedule based on your specific

circumstances. Here are the vaccination schedule charts for different animals mentioned belowwhich can serve as a guideline only.

Vaccination of Cattle, Buffalo, Sheep and Goats

A herd health management plan is vital to profitable production. Some producers, however, donot vaccinate until they experience a loss. The investment in disease prevention is less than the

cost of disease treatment. Dont wait until a disease outbreak occurs. For animals to reach theirperformance potential, they must be healthy. Many animal health problems can be controlledwith good management, proper nutrition and vaccination against infectious diseases.


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Here is the table for the vaccination schedule for cattle, buffalo,

sheep and goat:

Diseases Animal PrimaryVaccination

Re-Vaccination

F.M.D. All cloven footedanimals

4 months Twice in a year

H.S. Cattle, Buffalo,Sheep,Goat

6 months Annually before monsoon(twice a year in endemic areas)

Anthrax All species ofAnimals

6 months Once Annually (In Affectedarea only)

B.Q. Cattle,Buffalo,Sheep,Goat 6 months Annually before monsoon

Brucellosis Female cattle &buffalo Calf

4-8 monthsOnly

Only once i.e. at 4-8 months ofage in females in problemherds

Theileriosis Cattle above 2 months Annually

Enterotoxaemia Sheep,Goat 4 months for kid

or lamb (If damis vaccinated).

At the age of 1stweek for kid orlamb (If dam is

not vaccinated)

Before monsoon (preferably in

May).Booster vaccination after 15

days of first vaccination

P.P.R. Sheep,Goat 3 months for kidor lamb

Once in three years

Sheep Pox,

Goat Pox,C.C.P.P.

Sheep,Goat At the age of 3

month & abovefor lamb or kid

Once Annually (December

month)

Rabies All species of

Animals

0, 3,7,14,28 & 90 days post

exposure only


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Vaccination of Dog

There are three major vaccines used in dogs. The vaccination schedule is given below:

Diseases PrimaryVaccination

SecondaryVaccination

Re-Vaccination

Canine Distemper, Canine Hepatitis

(CanineAdenovirus 2), Corona ViralEnteritis, Canine Parainfluenza, Parvo

Virus Infection, Leptospirosis(L.canicola, L.icterohaemorrhagiae)

6-8 weeks of age 2-3 weeks later

upto 16 weeks ofage

Annual

Rabies3 months of age After 3 months

Annual

Bordatella After 1 year Annual

Vaccination of Cat

Diseases Primary Vaccination Secondary Vaccination Re-Vaccination

Panleukopenia,Rhinotracheitis,

Calicivirus

6 weeks of age andrepeat every 3-4 weeks until

16 weeks of age

1 year after completionof initial series

Every 3 years

Felineleukemia

8 weeks of age and repeat in3-4 weeks

1 year after completionof initial series

Annual

Rabies Single dose as early as 8

weeks of age, depending on

the product.

2 doses, 12 months apart Annually or every

3 years,

depending onvaccine used

Bordetella 8 weeks, then 2-4 weeks later 2 doses, 2-4 weeks apart Annually


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Vaccination of Horse

Vaccinate your horse against all diseases he may be exposed to, including at home or if youtravel with your horse. The chart below is a guideline of the vaccines suggested for some generalages and classes of horses.

Diseases Primary Vaccination Re-Vaccination

Tetanus toxoid

From non-vaccinated mare:First dose: 3-4 months

Second dose: 4-5 monthsFrom vaccinated mare:First dose: 6 months

Second dose: 7 monthsThird dose: 8-9 months

Annually

Anthrax 6 months Once Annually (In Affected area only)

Rabies 3 months or above Annually.Post exposure : 3,7,14,28 & 90 days

Some important points to remember:

Practice good management.1. Before any vaccination deworming should be compulsory to get better results.2. Read and follow vaccine labels carefully.3. Always follow the manufacturers recommendations for dosage, method of

administration, number of times given and proper storage.4. Vaccinate to prevent diseases which have a high risk of occurring, not diseases with a

low risk.

5. Herd-specific vaccination programs should always be developed in consultation with aveterinarian.

6. The best way to stay on schedule with vaccinations for your dog or cat is to follow therecommendations of a veterinarian you trust.


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Reference and Readings

1. Greene, C.E., Schultz, R.D. and Ford, R.B. 2001.Canine vaccination. Vet Clin North AmSmall Anim Pract.31:473-492.

2. Scherk, M.A., Ford, R.B., Gaskell, R.M., Hartmann, K., Hurley, K.F., Lappin, M.R.,Levy, J.K., Little, S.E., Nordone, S.K. and Sparkes, A.H. 2013.AAFP Feline Vaccination

Advisory Panel Report. Journal of Feline Medicine and Surgery.15(9):7853. Gaskell, R.M., Gettinby, G., Graham, S.J. and Skilton, D.2002. Veterinary Products

Committee working group report on feline and canine vaccination. Vet Rec.150:126-134.

4. Callan, R.J. 2001. Fundamental considerat ions in developing vaccination protocols.AABP Proceedings.34:14-22.

5. Hunsaker, B.D. and Tripp, S.P. 2007. Vaccine field efficacy: A review of field efficacyreported for vaccine antigens used in beef cattle and dairy practice,1996 to present. AAPBProceedings.40:26-32.

6. Perino, L.J., and Hunsaker, B.D. 1997. A review of bovine respiratory disease vaccinefield efficacy. Bovine Practitioner.31(1):59-66.

7. Radostits, O.M. 2001. Herd Health: Food Animal Production Medicine, 3rd ed.WBSaunders, Philadelphia.

8. Singh, S.N. 2011. Foot And Mouth Disease Control Strategies Globalframe Work.International Journal of Life Science and Pharma Research.288:9-42.

9. Giggins T., Soorej, K. 2010.Current Indian Veterinary Index. VETads Publications,Kerala, pp.375-396.

http://www.naro.go.ug. National Agriculture Research Systemhttp://www.ndri.res.in. National Dairy research Institutehttp://www.cirg.res.in. Central Institute for Research on Goats.

http://www.indg.in. India development gateway.

Vinod Kumar Singh1*, Vipin Kumar Upadhayay1, Sakshi

Bhadouriya1, Vikas Gupta1, Utkarsh Kumar Tripathi2, Yogender

Singh3, Jobin Jose Kattor1, Dash Prakash M.1

1. Indian Veterinary Research Institute, Izatnagar, Bareilly (U.P.)

2. National Dairy Research Institute, Karnal (Haryana)

3. Nanaji Deshmukh Pashu Chikitsa Vigyan Vishvavidyalaya,

Jabalpur (M.P.)
http://www.microbiologyworld.com/http://www.facebook.com/MicrobiologyWorldhttp://www.naro.go.ug/http://www.naro.go.ug/http://www.ndri.res.in/http://www.ndri.res.in/http://www.cirg.res.in/http://www.cirg.res.in/http://www.indg.in/http://www.indg.in/http://www.indg.in/http://www.cirg.res.in/http://www.ndri.res.in/http://www.naro.go.ug/http://www.facebook.com/MicrobiologyWorldhttp://www.microbiologyworld.com/


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Microbiology in Pharmaceuticals Industry

Microbiology is one of the important disciplines of science applied in Pharmaceutical industry inorder to ensure the sterility of the products such as injections, eye drops, ear drops and infusions.These products must be free from all kinds of microbes particularly bacteria and fungi. To ensure

this, a number of procedures are applied which are mentioned below:

1.Sterilization Protocol for working Area/RoomThe purpose of this practice is to minimize contamination during filling of the injection vials andampoules or performing a sterility test to get accurate results.The requirements for this procedure are antiseptic such as 3% chlorhexadine solution, 70%

Isopropyl alcohol (IPA), distilled water and 0.5% Phenol.

As a first step, all types of wastes are removed from the area followed by cleaning with avacuum cleaner. Then the whole area is washed with 3% chlorhexadine and autoclaved water.Doors and windows are cleaned with wet cloth. The laminar flow hood (LFH), filling machineand other equipments placed in the area are then cleaned with 70% v/v Isopropyl alcohol (IPA).

The rooms floor, roof and walls are cleaned with 0.5% Phenol.

2.Bio Burden/Bio Impression Test (General)To confirm that the area/lab is sterilized, bio-impression test is carried out. This test can be done

by two methods.

Taking swab from floor and walls

Exposure of media plates to the area to identify the presence of microorganisms

For taking swab samples, touching walls, equipments or any other items present in the sterile

area should be avoided. One must enter the area in a sterile autoclaved uniform in order to avoidcontamination. The tools should be rinsed with alcohol inside the area. Separate swab should beused for each area such as floor, walls, ceiling and other indoor installations. The swab should

carefully be brought into the LFH. The media plates should be streaked with the swabs and put inthe incubator on 35C for 48 hours. The results should be recorded through colony counter.


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The plates with sterile media (Generally Nutrient agar) should be placed uncovered in the areafor 3-4 hours.

PlateNo.

Filling Area UnderLaminar

Acceptance Criteria

Bacteria Molds

01 Conveyer Belt


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4.Sterility Test of the Product by Membrane Filtration MethodThe purpose of this test is to check the sterility of the vials and ampoules of injections, eye dropsand Ear drops. Chemicals required for this test is Peptone water (Buffer), Fluid Thioglycolate

Media (FTM), Tryptone Soya Broth (TSB), Isopropyl alcohol (IPA).

The test is done in a sterilized microbiology laboratory with a laminar flow hood. Two tubes of

both TSB and FTM are taken to the lab along with a sterilized filtration assembly and amembrane filter with 0.2 m pore size. The filtration assembly is unwrapped inside the LFH and

is connected with the suction pump. The sample ampoules/vials are opened and a small quantityof it is put into the 100 ml peptone water and then into the cup of filtration assembly. Byswitching on the suction pump, the entire liquid pass through the membrane filter and moves to

the bottle attached to the suction pump. The cup is removed from the assembly and filter is cutthrough a sterilized scissors into two pieces. The half of filter paper is placed in one bottle of

FTM and other half in the other bottle of TSB. FTM is incubated 32C 2.5C and TSB is at22C 2.5C for 14 days. The result is dependent on the turbidity of the media in bottles.Anaerobic bacteria grow in the bottom and aerobic on the surface. FTM is used for Bacteria and

TSB is used for fugal culture.

5.Pyrogen/Endotoxine Detection TestPyrogen is the substance which causes fever. The purpose of this test is to find endotoxin ininjection vials and ampoules. For this, a technique known as Limulus Amebocyte Lysate (LAL)

test is performed in the laboratory.

In the procedure of this test 1mg/ml dilution of the sample is prepared and put in a test tube

marked as Test and the two other test tubes with Blank and Standard marks. Depyrogenatedwater (0.05ml) is added to each tube with the help of sterilized pipette. A 1:10 dilution is madeof the test tube marked as Test. Then 0.05 ml of lysate is added to each tube. The tube is then

incubated at 37C for 30 minutes. After incubation the tubes are immediately transferred tochilled water. A 0.250 ml of diazo coupling reagents 8,9,10 (available with the kit) is transferred

to the reaction. The intensity of color formed as a result of the reaction between Endotoxin andLysate is seen. The absorbance of sample test and standard is measured by spectrophotometerwith a wave length of 545nm. If 0.4ml of 8M acetic acid is used instead of Diazo Coupling

Reagent, then the absorbance is measured by 405nm wave length. The following formula is usedfor calculation.

EU/ml =. .( )

(


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6.Air SamplingThis technique is used for the identification of microorganisms in the air of sterilized area. Forthis purpose a machine named as air sampler is used. A sterilized media plate of nutrient agar isplaced inside it. The machine sucks the air and throws it on the plate. It can be run for 5 to10

minutes. The plate is then incubated at 35 C for 48 hours to see the presence of any microbes.

References

1. Handbook of Microbiological Quality Control in Pharmaceuticals and Medical 11 NewFitter Lane, London EC4P 4EE: Taylor & Francis; 2000.

2. Williams RL. United States Pharmacopeia; Microbiological & Biological Tests andAssays 1, 2000 ed2000. p. 1809-23.

- Abid JanInstitute of Basic Medical Sciences (IBMS),

Khyber Medical University, Peshawar, Pakistan


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Need of Advocacy and awareness about cancer in

NepalMany people in Nepal are ignorant about cancer. This is because unlike infectious disease,chronic diseases like cancer comes under least priority area of governmental health policy inNepal. Very few national programs are conducted to educate and aware people about cancer.

Education program are conducted sporadically and only few people have access to such program.In the recent time, Healthy Nepal Foundation (HNF), an organization dedicated to increase

awareness about cancer has done some remarkable work to increase understanding of cancer toNepalese population. Since 2012, HNF has developed several education modules to educatestudents and public about common cancer such as cervical, breast, oral and lung cancer in Nepal.

With the help of volunteers from different colleges, this organization conducts program at

colleges and sometime in the community.

Volunteers is the main working pillars of HFN. With the help of volunteers, we are able toconduct education programs at around twenty schools to date and our main goal is to educate

student and the public about cancer and its prevention, said Kiran Sapkota, co -founder of theorganization. We are dedicated to educate and inform public about risks of cancer and promote

healthy life style to reduce cancer burden in the society, Sapkota said.

Members of healthy Nepal foundation have conducted several education program at secondary

schools, higher secondary colleges and universities around the nation. HN College CoordinatorMr. Sunil Pandey, who is also a student of Medical Microbiology at Nobel College, said we

have got very good response and appreciation from both teachers and guardians from respectiveschools where we conduct cancer education program. Mr. Pandey was so passionate to assistHNF that he allocate his tight time schedule out of his college to conduct programs around the

nation. Mr. Pandey contact schools, recruit volunteers, communicate with teacher and conducteducation awareness program at different schools around the country. With the help of experts

we were able to develop education material for only few cancers and we are still developingadditional cancer programs in the future said Pandey.

Healthy Nepal Foundation is operated by physicians, public health officials, and community andschool volunteers. Till date it has conducted programs in Parwat, Myagdi, Baglung, Pokhara,

Kathmandu, Chitwan and several other districts. Our main aim is to make a disease free healthy

society said Dr. Kalyan Sapkota, another co-founder of the organization. Dr. Sapkota and histeam have conducted cancer screening programs at several communities in Myagdi, Parwat and

Baglung in the past.


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Currently Healthy Nepal has concentrated more of its activities on educating students and publicabout cervical and breast cancer. These two cancers are the significant cause of mortality and

morbidity in women in Nepal said Dr. Sapkota. With the available funding, Healthy NepalFoundation had organized free Pap smear screening camp at different places in Nepal. A pap

test is the best way to detect cell changes that may be early sign of precancerous of the cervix.Dr. Sapkota added that, Breast cancer affects one in eight women during their lives. No oneknows why some women get breast cancer, but there are a number of risk factors. If we reduce

these risk factors, we can minimize the number of cancer patients in Nepal, said Dr. Sapkota.

Sharing his experience while conducting cervical cancer awareness program, Mr. Sunil Pandeysaid, we were so shocked to know that many students do not know much about common cancer,and many have misconception about it. After we conduct program, we do posttest and found that

our program really changed how students perceive about cancer. Their knowledge remarkably

changes just after an hour or two education session, added Mr. Pandey. Many women do notknow about the screening test available to detect cancer at early age. This include Pap smearscreening and self-breast examination in Nepal.

Mr. Sunil Pandey said there are several opportunities available for youth to be involved in cancereducation projects. He is recruiting many volunteers to initiate oral and lung cancer awareness in

the near future.

For more information or to get involved, email [email protected], or

visit the organizations Facebook page athttps://www.facebook.com/healthynepal

- Sunil PandeyMedical Microbiology

Nobel College, Kathmandu, Nepal
http://www.microbiologyworld.com/http://www.facebook.com/MicrobiologyWorldhttps://www.facebook.com/healthynepalhttps://www.facebook.com/healthynepalhttps://www.facebook.com/healthynepalhttps://www.facebook.com/healthynepalhttp://www.facebook.com/MicrobiologyWorldhttp://www.microbiologyworld.com/


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How to Microorganism contribute to body odor?

More than 100 trillion bacteria live in the human body, according to the results of the first DNA-sequence of human microbiome. Humans are an important ecosystem for a wide variety of

microorganisms. The human body is a diverse environment in and on which specific niches areformed. The average adult carries 10 times more microbial cells (10^14) than human cells

(average about 10^3). The adult human is covered with approximately 2m^2 of skin. It has beenconsidered that this surface area supports 10^12 bacteria.

A body odor can be an unpleasant smell generated from the skin. It is as a result of bacteria thatlives on the skin which break down sweat into acid. The bacteria break chemical bond of protein

in the sweat and convert it to acid. In this case we can say that microorganisms and the body

exhibit a type of relationship called Ammensalism (ammensalism is a type of relationship inwhich the product of one organism has a negative effect on the other organism).

Most skin bacteria are found on superficial cells, colonizing dead cells or closely associated with

the oil and sweat glands. Secreations from the sweet glands provide the water, amino acids, urea,electrolytes and specific fatty acids that serve as nutrients primary for S. epidermidis and aerobiccorynebacteria. The oil glands secrete complex lipids that may be partially degraded by the

enzymes from certain gram-positive bacteria (e.g, Propionibacterium acne). These bacteriausually are harmless; however, they are associated with the skin disease acne vulgaris. They can

change the lipids secreted by the oil glands to unsaturated fatty acid such as oleic acid that havestrong antimicrobial activity against gram negative bacteria and some fungi. Some of these fatty

acids are volatile and may have a strong odor.

Sweat interacting with microbes can produce body odor. Examples of microbes that interact with

sweat are Propionobacteria, Staphylococci, Minococci, Corneforms and Pityrosporum. Thereare two types of sweat; Ecrine and apocrine, which are secreated by different types of glands.Bacteria only interact with apocrine sweat, which is fattier and richer and usually occurs in areas

with body hair such as groin and armpits.

The acids produced depends on the species of bacteria;

*Propionic acid (propanoic acid) is commonly found in sweat-propionibacteria break amino

acids down into propionoic acid. Propionicbacteria live in the ducts of sebaceous glands.

*Isovaleric acid (3-methylbutanoic acid) is as a result of the action of bacteria staphyloccoccusepidermidis, which are present in several strong cheese type.


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TREATMENT/PREVENTION

Antibacterial soap and frequent washing can keep bacteria down to a manageable level. Avoidwearing thick clothe as thick clothes absorb sweat and allow the multiplication of bacteria

instead wear clothe that allows sweat to evaporate and this prevent bacteria from breaking downthe sweat. Deodorants that contain antibacterial substances that act selectively against gram-positive bacteria can be used to reduce production of volatile unsaturated fatty acids and body

odor.

- Ayplux


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Embryonic Stem cell research - an ethical dilemma,current scenario & how we can overcome this

Now-a-days we talk a lot about Stem cells. Undoubtedly human stem cells have opened new era

of medical research and applications by empowering to treat diseases like Parkinson's disease,Arthritis, Type I diabetes, Burn victims, cardiovascular diseases and many more. These also givethe opportunity to unlock the secret of life, how life starts, how a fertilized egg get transformed

by cell division and cell differentiation into the diverse range of specialized cells that make a fullgrown baby by following the human developmental pathway. Because diseases like cancer or

conditions as birth defects are thought to occur due to problems in the differentiation process, amore complete understanding of the genetic and molecular controls of these processes of thedevelopment that happens in normal cells will help scientists treat the developmental errors that

can occur and suggest new strategies for therapy. For all of these the key is Embryonic Stemcells, and here the debate beginsan ethical dilemma.

Different countries have chosen different regulatory measures in embryonic stem cell research.Mentioning embryonic stem cells in the pub still divides opinion on the topic. But what exactly

are the ethical arguments and why are they so tricky to resolve?

Research on Embryonic stem cell poses a moral and ethical conflict between two views. It forcesus to choose between one of these two moral principles:

The duty to prevent or alleviate suffering of thousands of human beings The duty to respect and save the value of human life

In this case of embryonic stem cell research, it's virtually not possible to take care of each ethicalprinciples. As embryonic stem cells are derived from the inner cell mass of a blastocyst, an early-

stage pre implantation embryo, the embryo has got to be destroyed first. This implies thedestruction of a possible human life. However embryonic stem cell study could cause the

invention of recent medical treatments and therapies that may save lifetime of many humanbeings. Therefore during this state of affairs, which of these ethical principle ought to have thehigher hand? The solution lies on our tendency to view the embryo. Does it have the status of a

person at all?

1. The embryo has full ethical value from fertilization onwards: An embryo can beviewed as a person or a potential person while it is still an embryo. Because differentpeople gives different definition to be a person, the criteria for personhood is not

clear yet.


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Arguments supporting this view: Development of a zygote into full grown baby is a continuousprocess. A human embryo is nothing but a human being in the embryonic stage. Though theembryo does not have the characteristics of a person right now, it will become a person once and

that is why should be given the minimum respect and dignity of a person.

Arguments against this view: An early embryo that has not yet implanted into the uterus cannever have the psychological, emotional or physical properties that we associate with being aperson. It therefore does not have any interests to be protected and we can use it for the benefit of

patients (who ARE persons).

2. There is a cut-off point at 14 days after fertilization:Some people argue that a human embryo deserves special protection from around day 14 after

fertilization because:

Arguments supporting this view: After 14 days the embryo can no longer split to form twins.Before this point, the embryo could still be split to become two or more babies, or it might fail todevelop at all.

Argument against this view: Before day 14, the embryo has no central nervous system and

therefore no senses. If we can take organs from patients who have been declared brain dead anduse them for transplants, then we can also use hundred-cell embryos that have no nervoussystem.

3. The embryo has increasing status as it develops: Arguments supporting this view: If a life is lost, we tend to feel differently about it dependingon the stage of the lost life. A fertilized egg before implantation in the uterus could be granted a

lesser degree of respect than a human fetus or a born baby. More than half of all fertilized eggsare lost due to natural causes. If the natural process involves such loss, then using some embryos

in stem cell research should not worry us either.

Argument against this view: If we judge the moral status of the embryo from its age, then we

are making arbitrary decisions about who is human. For example, even if we say formation of thenervous system marks the start of personhood, we still would not say a patient who has lost nerve

cells in a stroke has become less human. If we are not sure whether a fertilized egg should be

considered a human being, then we should not destroy it. A hunter does not shoot if he is notsure whether his target is a deer or a man.


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4. The embryo has no moral status at all:An embryo is organic material with a status no different from other body parts.

Arguments supporting this view: Fertilized human eggs are just parts of other peoples bodies

until they have developed enough to survive independently. The only respect due to blastocystsis the respect that should be shown to other peoples property. If we destroy a blastocyst beforeimplantation into the uterus we do not harm it because it has no beliefs, desires, expectations,

aims or purposes to be harmed.

Argument against this view: By taking embryonic stem cells out of an early embryo, weprevent the embryo from developing in its normal way. This means it is prevented frombecoming what it was programmed to become a human being.

Attitude of Different Countries towards this Issue:

1.USA:In March 2009, President Obama issued an executive order intended to bolster human embryonicstem (hES) cell research in the United States. Obama's move overturns an order signed by

President Bush in 2001 that barred the National Institutes of Health from funding research onembryonic stem cells beyond using 60 cell lines that existed at that time. Obama also signed a

presidential memorandum establishing greater independence for federal science policies andprograms. "In recent years, when it comes to stem cell research, rather than furthering discovery,

our government has forced what I believe is a false choice between sound science and moralvalues," President Obama said at the White House.( Ref:3)

2.England:Britain, who many consider to be the leader in stem cell research, opened the worlds first stemcell bank in May of 2004 which currently stores and distributes these cells. This bank serves as arepository for all human stem cell lines produced under conditions of Good Manufacturing

Practice. At the opening of the UK stem cell bank Lord Warner, Minister of Health, addressedmany of the issues surrounding embryonic stem cell research and did an excellent job of

representing the views of the British government and the majority of the British people. He

recognized that there are concerns over what this kind of controversial research could lead to inthe future, but that the benefits outweigh the risks. He also made assurances that there are safety

measures in place to prevent embryonic stem cell research from leading to human reproductivecloning and other scientific technologies that are globally rejected and seen as being morally

unacceptable.


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3.Sweden:The Swedish government, much like that of Great Britain, recognizes the impact that stem cellresearch will have in the next century not only on medicine, but in the way people think about

disease and injury. Sweden has therefore been another forerunner in breakthroughs made in thepast decade in developing stem cell research. As of 2001, 24 of the 64 embryonic stem cell linesavailable globally were originated in Sweden (at both the both the Karolinska Institute and

Gteborg University).

4.Germany:The use of embryos for research is heavily restricted in Germany under the Embryo Protection

Act (Embryonenschutzgesetz) 1991, which makes the derivation of embryonic stem cell lines acriminal offence. The embryo is also protected under the German Constitution (Grundgesetz).

The Basic Law states that human dignity is inviolable and that everyone has the right to lifeand inviolability of his person. Nonetheless, it also states the freedom to pursue science and

research. The derivation of embryonic stem cells is banned but embryonic stem cell lines can beimported specifically for research if the line was generated before a defined cut-off date.

5. Elsewhere, Japan, India, Iran, Israel, South Korea, and China are supportive, Australia ispartially supportive (exempting reproductive cloning yet allowing research on embryonic stem

cells that are derived from the process of IVF); however New Zealand, most of Africa (exceptingSouth Africa) and most of South America (excepting Brazil) are restrictive.

How to solve this critical issue:

The possible options for resolving the conflict is to search for the replacement of hES andhopefully there are options. Life is a journey of gaining some new attributes whereas losing some

old ones. For long time scientists have studied the human development pathway from a singleunspecialized cell to a full grown specialized baby. They have seen that in this peculiar processas the zygote divides and differentiate some classes of genes are switched off and some are

switched on. There are very much sophisticated information and special routes for every cell tobecome specialized from unspecialized state. In the specialized adult cells, the earlier genes

(when it was a stem cell) are switched off forever. Then scientists thought if somehow they can

again turn on those genes in an adult cell, they can reverse the route to make a stem cell from anadult cell. Induced pluripotent stem cells (also known as iPS cells or iPSCs) are a type of

pluripotent stem cell that can be generated directly from adult cells. The iPSC technology waspioneered by Shinya Yamanakas lab in Kyoto, Japan, who showed in 2006 that the introduction

of four specific genes could convert adult cells to pluripotent stem cells. He was awarded the


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2012 Nobel Prize along with Sir John Gurdon "for the discovery that mature cells can bereprogrammed to become pluripotent." Pluripotent stem cells hold great promise in the field ofregenerative medicine. Because they can propagate indefinitely, as well as give rise to every

other cell type in the body (such as neurons, heart, pancreatic, and liver cells), they represent asingle source of cells that could be used to replace those lost to damage or disease. Since iPSCs

can be derived directly from adult tissues, they not only bypass the need for embryos, but can bemade in a patient-matched manner, which means that each individual could have their ownpluripotent stem cell line. These unlimited supplies of autologous cells could be used to generate

transplants without the risk of immune rejection. While the iPSC technology has not yetadvanced to a stage where therapeutic transplants have been deemed safe, iPSCs are readily

being used in personalized drug discovery efforts and understanding the patient-specific basis ofdisease.

No need to say it is very difficult to come to a conclusion on the ethical dilemma of stem cell

research. But, as iPS cells have the potential to develop into a human embryonic stem cell, thedebate over stem cell research is becoming increasingly irrelevant.

References:

1. Nature Cell Biology (http://www.nature.com/ncb/journal/v12/n7/full/ncb0710-627.html )

2. EuroStem Cell : Embryonic stem cell research: an ethical dilemma , 23 Mar 2011 (http://www.eurostemcell.org/factsheet/embyronic-stem-cell-research-ethical-d ilemma )3. CNNPolitics.com March 9, 2009 -- Updated 1643 GMT (0043 HKT) Obama overturns

Bush policy on stem cells (http://edition.cnn.com/2009/POLITICS/03/09/obama.stem.cells/ )4. Stem Cell Research : International Policies

(http://www.mtholyoke.edu/~amhunsak/europe.htm )5. Regulation of stem cell research in Germany Kate Doherty(http://www.eurostemcell.org/regulations/regulat ion-stem-cell-research-germany )

6. Wikipedia: Induced pluripotent stem cell(http://en.wikipedia.org/wiki/Induced_pluripotent_stem_cell )

- Bharat Manna

B.Tech Student, Department of Biotechnology,Bengal College of Engineering & Technology

West Bengal, India.


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Staphylococcus aureusAnd MRSA

Staphylococci are spherical gram-positive cocci arranged in irregular grapelike clusters. Allstaphylococci produce catalase, whereas no streptococci do (catalase degrades H2O2 into O2 andH2O). Catalase is an important virulence factor because H2O2 is microbicidal and its

degradation limits the ability of neutrophils to kill. Although S. aureus is not always pathogenic,it is a common cause of skin infections (e.g. boils), respiratory disease (e.g. sinusitis), and food

poisoning. Disease-associated strains often promote infections by producing potent proteintoxins, and expressing cell-surface proteins that bind 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 in Aberdeen, United Kingdom, by the surgeon Sir

Alexander Ogston in pus from a surgical abscess in a knee joint. This name was later appendedto Staphylococcus aureus by 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 carriersof S. aureus which can be found as part of the normal skin flora and in anterior nares of the nasal

passages. S. aureus is the most common species of staphylococcus to cause Staph infections andis a successful pathogen due to a combination of nasal carriage and bacterial immuno-evasivestrategies. S. aureus can cause a range of illnesses, from minor skin infections, such as pimples,

impetigo, boils (furuncles), cellulitis folliculitis, carbuncles, scalded skin syndrome, andabscesses, to life-threatening diseases such as pneumonia, meningitis, osteomyelitis,

endocarditis, toxic shock syndrome (TSS), bacteremia, and sepsis. Its incidence ranges fromskin, soft tissue, respiratory, bone, joint, endovascular to wound infections. It is still one of the

five most common causes of nosocomial infections and is often the cause of postsurgical woundinfections. Each year, some 500,000 patients in American hospitals contract a staphylococcalinfection.

Methicillin-Resistant Staphylococcus aureus (MRSA)

Staphylococcus aureus (S. aureus) is the leading cause of gram positive bacterial infections andproduces a wide spectrum of diseases, ranging from minor skin infections to fatal necrotizing

pneumonia. Although S. aureus infections were historically treatable with common antibiotics,emergence of drug-resistant organisms is now a major concern. Methicillin-resistant

Staphylococcus aureus (MRSA) was endemic in hospitals by the late 1960s, but it appearedrapidly and unexpectedly in communities in the 1990s and is now prevalent worldwide.

S. aureus is notorious for its ability to become resistant to antibiotics. Infections that are caused

by antibiotic-resistant strains often occur in epidemic waves that are initiated by one or a few


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successful clones. MRSA features prominently in these epidemics. Historically associated withhospitals and other health care settings, MRSA has now emerged as a widespread cause ofcommunity infections. Community or community-associated MRSA (CA-MRSA) can spread

rapidly among healthy individuals. Outbreaks of CA-MRSA infections have been reportedworldwide.

The frequency of MRSA infections continues to grow in hospital-associated settings, and morerecently, in community settings globally. The increase in the incidence of infections due to S.

aureus is partially a consequence of advances in patient care and also of the pathogen's ability toadapt to a changing environment. Infection due to S. aureus imposes a high and increasing

burden on health care resources. A growing concern is the emergence of MRSA infections inpatients with no apparent risk factors. The growing problem in the Indian scenario is that MRSAprevalence has increased from 12% in 1992 to 80.83% in 1999. MRSA in tonsils may serve as a

potential source for the spread of these organisms to other body sites as well to other individuals.

MRSA is prevalent in many hospitals and often reflects the difficulties in hospitals and the healthservice generally, in terms of the control and prevention of healthcare-associated infection.Multidrug-resistant bacteria, such as MRSA, are endemic in healthcare settings in the United

States and many other countries of the world. Nosocomial transmission of MRSA serves as asource of hospital outbreaks, and recent reports of vancomycin resistant S. aureus strains in the

United States emphasize the need for better control of MRSA and other resistant bacteria withinhealthcare settings.

Pathogenesis:

Staph.aureus present in the nose of 30% of healthy people and may be found on the skin. Itcauses infection most commonly at sites of lowered host resistance, such as damaged skin (e.g.,surgical site infection) or mucous membranes (e.g. ventila tor-associated pneumonia).

Staphylococcal Toxins:

Enterotoxins:

Enterotoxins, types A-E,G, H, I & J are commonly produced by up to 65% of strains of Staph.aureus, sometimes singly & sometimes in combination. These toxic proteins withstand exposure

to 100c for several minutes. When ingested as performed toxins in contaminated food,microgram amounts of toxins, within a minute, induce the symptoms of staphylococcal food

poisoning: nausea, vomiting, & diarrhea. However, enterotoxins, which are super antigens,probably also, play an important role in other serious staphylococcal infections, e.g., bloodstreaminfections, especially accompanied by septic shock.


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Toxic Shock Syndrome Toxin (TSST-1):

This was discovered in early 1980s as result of epidemiological and microbiological

investigations in USA of toxic shock syndrome, a multi system disease caused by staphylococcalTSST-1 or enterotoxins or both. A link was established with the use of highly absorbent tampons

in menstruating women, although non-menstrual cases are now as common. The absence ofcirculating antibodies of TSST-1 is a factor in the pathogenesis of this syndrome. TSST-1 andthe enterotoxins are now recognized as super antigens, that is, they are potent activators, of T

lymphocytes resulting in the liberation of cytokines such as tumor necrosis factor, and they bindwith high affinity to mononuclear cells. These characteristics partly explain the florid &

multisystem nature of the clinical conditions associated with these toxins.

Epidermolytic Toxin:

Two kinds of Epidermolytic toxins (types A & B) are commonly produced by strains that causeblistering diseases. These toxins induce intraepidermal blisters at the granular cell layer. Suchblisters of pemphigus neonatarum. The most dramatic manifestation of Epidermolytic toxin isscalded skin syndrome in small children, where the toxin spreads automatically in individuals

who lack neutralizing antitoxin.

Panton-Valentine Leukocidin (PVL):

This toxin was recognized some decades back but its potential contribution to clinical

manifestations and outcome have been increasingly described in the context of community-acquired MRSA (CA-MRSA). As the name suggests PVL can adversely affect cells, resulting in

leucopenia, but animals studies do not suggest high virulence. Nonetheless, epidemiological datain many countries reveal an association between necrotizing pneumonia and some complicatedskin and soft tissue infections caused by PVL-positive strains of CA-MRSA.

Laboratory Diagnosis:

One or more of the following specimens should be collected to confirm diagnosis;

Pus from abscesses, wounds, burns etc. is much preferred swabs. Sputum from patients with pneumonia, bronchoscopic lavage, is increasingly used in

critically ill patients.

Faeces or vomit from patients with suspected food poisoning, or the remains ofimplicated foods.

Blood from patients with suspected BSI such as septic shock, osteomyelitis, orendocarditis.

Mid-stream urine from patients with suspected cystitis or pyelonephritis.


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Anterior nasal and perineal swabs (moistened in saline or sterile water) from suspectedcarriers, nasal swabs should be rubbed in turn over the anterior walls of both nostrils.

The characteristic clusters of Gram-positive cocci can be often be demonstrated by microscopy,and the organisms cultured readily on blood agar and the most other media within 24 hrs. or less.

The tube or slide coagulase test is performed to distinguish Staph. aureus from coagulasenegative species and AST with ceofoxitin to confirm MRSA using standard methods.

Commercially, available molecular methods using PCR have been developed to reduce the timeto the detection of MRSA from 48-72 h with culture to less than 12 h facilitate earlierpreventative measures. However, the results from trails with PCR to date mixed and do not

clearly indicate that this more expensive approach can assist in reducing MRSA rates in acutehospitals.

- Anil BhujelBsc Microbiology, Pokhara Bigyan Thata Prabidhi Campus

Nayabazzar-9, Pokhara. Nepal

Email: [email protected]

Website: www.microbiollogy.blogspot.com


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You can also send your articles to

[email protected]

or

[email protected]

Selected ones will be published in

our next issue of May-June 2014.

Thanks,

M icrobiology World Team

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