introduction to pharmaceutical microbiology

2. DEFINATION
The word Microbiology was derived from Greek: mīkros ("small")+ bios
("life")
It is the branch of science that is concerned primarily with the biology of
microorganisms and their effects on other living organisms.
Microbiology is the study of living organisms of microscopic size which
include Bacteria, Protozoa, Algae, Fungus andViruses.

3. Microbiology may be Pure or Applied:
 Pure microbiology:
• Includes bacteriology, mycology, protozoology, phycology, parasitology,
virology, nematology, cellular microbiology, evolutionary microbiology,
molecular microbiology, etc.
 Applied microbiology:
• Includes medical microbiology, pharmaceutical microbiology,
industrialmicrobiology, food microbiology, soil microbiology, water
microbiology, air microbiology and microbial biotechnology.
When microbiological concepts, processes and techniques are applied to
pharmaceutical operations, the subject is then called ‘pharmaceutical
microbiology’.

5. Pure microbiology
• Bacteriology (Study of Bacteria)
• Mycology(Study of fungi)
• Protozoalogy (Study of Protozoa)
• Phycology/Agrology (Study of Algae)
• Parasitology (Study of Parasites)
• MicrobialTaxonomy(Rank based classification of bacteria)
• Genetics
• Immunlogy

6. Applied Microbiology
• Medical Microbiology – Deal with causative agents
• Pharmaceutical Microbiology- Deal with microorganism which produces
Antiboitics, Enzymes andVaccines.
• Industrial Microbiology- Deals with beverages production and Amino acid
• Microbial Microbiology- It is scientific manipulation at molecular and genetic level.
• Food Microbiology- Deals with interaction of microorganism and food.
• Soil Microbiology- Deals with interaction of Soil microorganism.
• Agricultural Microbiology- Relation with microorganism and crops.
• Aquatic Microbiology – Study of Microorganisms with fresh and marine water
includes water purification etc.
• Air Microbiology – Deals with contamination and spoilage of food through air.
• Epidemiology- Concerned with monitoring, control and spread of disease in
communities.

7. History of Microbiology
• Louis Pasteur is regarded as the father of microbiology whereas Robert Koch is
the father of medical microbiology.
1. Antonie Philips van Leeuwenhoek (24 October 1632 – 26 August 1723)
• He was a Dutch Merchant and keen in nature.
• His hobby was glass grinding and preparation of lenses and this
led to him to assemble 250 simple microscopes.
• He use magnifying glass to study weaves in various types of cloths.
• Observed and measured a large no of microorganism.
• He was the first to observe and accurately describe the shape of human RBC’s and
little agents of disease ‘animalcules’
• He also communicate his work to Royal Society of
London in 1684 in his letter.
• In his experiment he described inhibitory
effect of acetic acid on microorganisms.

8. 2. Louis Pasteur(1822-1895)
• French Organic Chemist
• Considered as father of modern microbiology.
• Foundation to bacteriology.
• He demonstrated that air contains microscopically organized structures, so
he explained it with these experiment.

9. • He performed series of operation to prove M.O. were present in air
• they were not spontaneously produced. So for these he performed Swan
neck flask experiment

10. • Fermentation process: beer/wine not produced without microbes and
wine does not undergo spoilage when kept for few minutes at 50-60°C.
• Bcoz of these arise a new process of preserving wine, fruit juice and milk c/a
Pasteurization.
• He designated life in presence of O2 as aerobic , in absence of O2 anaerobic
and in presence and in absence of oxygen as ‘Facultative anaerobic
• He introduced the method of sterilization by repeated heating with
appropriate intervals, method known as ‘Tyndallisation ’
• In 1880,he isolated bacterium responsible for Chicken Cholera and grew
in pure culture.

11. • Pasteur also introduced a vaccine for a Anthrax and Rabies.
• GermTheory of Disease - He investigated silkworm disease and proved
that disease was due to protozoan parasite and he also proved that bacteria
are the causative agent of some disease.
• Diffrenet places/Diffrenet conc. Of microbes
• Disproved spontaneous generation

12. Robert Koch (1843-1910)
• German physician
• Considered as father of medical microbiology
• First to demonstrate the role of bacteria in causing diseases
• First to isolate Bacillus anthracis
• Discovered Mycobacterium tuberculosis,1882 andVibrio
cholera,1883, awarded a Nobel prize in 1905
• Developed relationship between microorganism and diseases

13. •Koch's postulates:
•The microorganism must be present in every case of
disease
•The microorganisms must be isolated from the
diseased animal and grown in pure culture
•The disease must be reproduced when a pure culture
of the microorganisms is inoculated into a susceptible
host
•The same microorganism must be desolated from the
experiment from the experimentally infected host

14. Scopes and importance
oProduction of antibiotics:
• 2/3rd of antibiotics are produced from microorganisms.
• Many antibiotics are isolated from natural
microorganisms by the process of fermentation.
• Eg: Penicillin from Penicilium species,
Streptomycin from Streptomyces griseus,
Tetracycline from Streptomyces aureofaciens,
• Chloramphenicol from Streptomyces venezuelae.

15. oProduction of enzymes, vaccines and other pharmaceutical products:
• Microbial cells produce intracellular and extracellular enzymes
and these enzymes are important for the success of
pharmaceutical fermentation process.
• Eg: amylase, protease, lipase, invertase, oxidase,
catalase, cortison reductase, etc.
• Bio-surfactants have lot of applications in agriculture,
food industries, industrial cleaning, leather, paper and metal industries,
textiles, cosmetics and the pharmaceutical industry.
• Different types of bio-surfactants are synthezed by a number of microorganisms
• eg: Acinetobacter speciesa, Bacillus species, Pseudomonas species, Rhodococcus
species,etc.

16. oDiagnosis of diseases and treatment:
• Different tests are used to detect
infectious microorganisms, eg: ELISA,
Widal test,TUBERCULLIN SKINTEST,
oTreatment of industrial waste material:
• Most industrial processes produce
waste water, salts, organic matter and spent
media and these are toxic.
• Many microbial species are used for decomposition
of such waste materials and organic
components, eg: Actinomycetes, fungi, protozoa, etc.

17. oPlant growth promotion:
• Many microbial cells present in soil, play an important role
in soil fertility, herbicidal resistance, insect resistance,
change in protein/oil content and enhanced quality of plant
products, eg: Rhizobium species, Rhodospirillum species,
Azotobacter species, Agrobacterium rhizogenes,
Pseudomonas species, Acetobacter species.
• Azotobacter chroococcum, living nitrogen fixing
bacterium capable of synthezing and secreting
plant growth promoting substances like
thiamine, riboflavin, IAA, gibberellin,etc

18. oSterile product preparation:
• Ph. Microbiology plays a major role in preparation of
sterile products.
• Deals with sterile rooms, aseptic techniques, detection
of microbes by sampling and sterility testing of
different sterile preparations.
oSterilization:
• Any processes/methods that eliminates, removes, kills
or deactivates all forms of life and other biological
agents(bacteria, fungi, viruses, etc) called sterilization.
• Moist heat sterilization, dry heat sterilization,
membrane filtration, gaseous sterilization and
chemical sterilization are the methods used for killing
microorganisms.

19. oSteroid bio-transformation:
• Steroids are physiologically active compounds of complex structure, eg:
cholesterol, ergosterol, testosterone, progesterone, etc.
• Important steroids can be produced by microbiological transformations of
naturally occuring steroids, eg: Steptomyces species, Rhizopus species,
Aspergillus species, Penicillium species.
oIdentification of microorganisms:
• One of the vital function of pharmaceutical microbiology is identification of
microorganisms found in products and the manufacturing environment.
• The microorganisms are isolated and identified by morphological,
biochemical, cultural, microscopic characteristics and genetic studies.

20. oTesting of pharmaceutical:
• Raw material and finished products:
• The Presence of microorganisms like E. coli, Salmonella
species, Pseudomonas aeruginosa and Staphylococcus aureus,
in the raw materials and finished products may deteriorate their
efficacy and potency.
• Test the presence of microbes are described in th IP, USP, EP and
BP.
• Limit on the total number of viable microbes in given
product(TVC) and involved in exclusion of specific pathogens.
• Water act as a vehicle-always tested forTMC.

21. oMicrobiological assays of antibiotics:
• The antibiotic bioassay provides a potency for the over all
biological activity of an antibiotic preparation.
• Agar diffusion(plate assay) and turbidometric assay.
oEvaluation of disinfectant :
• Effective cleaning and disinfection of manufacturing facilities are
crucial to the achievement and maintenance of high quality
standard required for medicine and medical devices.
• Phenol coefficient test, cup plate method, turbidometric method
and kelsey Sykes test.

22. oAntimicrobial Preservative efficacy testing:
• Different types of pharmaceutical products and cosmetics are
protected from attack by microorganisms by using preservatives.
Eg:
• Methyl, ethyl, propyl and butyl Parabens, Sorbic acid.
• The preservatives should be non-toxic, easily available and have
broad spectrum activity.
• Cup-plate method, turbidometric method and total viable
counting techniques are used for determination of anti-microbal
efficacy of preservatives.

23. oEndo-toxin testing:
• Bacterial endotoxin or pyrogens have a number of physiological
effects following intravenous injections such as fever, activation
of cytokine system, endothelial cell damage and intravascular
coagulation.
• The Bacterial Endo-toxinsTest(BET) is most important in
microbiology laboratories involved with quality control of
parenterals and medical devices.
• LALTest, gel-clot method, chromogenic end point, kinetic
turbidometric assay and kinetic chromogenic assay are also used
to detect endotoxins.

24. oTest for support of the sterility assurance system:
•Sterility test mainly applicable in running an aseptic area
and microbiology lab.
•It is mainly performed in laminar air flow cabinets for
parenteral preparations and other sterile
pharmaceuticals to detect presence of microorganism.
•Bioburden testing is used to estimate the number of
microorganism in the product prior to sterilization