for B.PHARM 3RD SEM

1. PHARMACEUTICAL
MICROBIOLOGY
Subject Code: BP 303T

2. Introduction
• 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. e.g., Bacteria, Protozoa, Algae, Fungus and Viruses.
• 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, industrial
microbiology, 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’.

3. • Objectives of pharmaceutical microbiology to ensure
safety and efficacy of pharmaceutical products.
• It embraces to validation of disinfectants, evaluation of
the efficacy of disinfectants in suspension, on surfaces
and through field trials.
• It offers protocols and techniques associated with the
operation and assurance of clean-room, aseptic-room
and controlled environments for preventing any
possible microbial contamination, and introduces risk
assessment and practical contamination control
strategies.
Louis Pasteur is regarded as the father of microbiology
whereas Robert Koch is the father of medical
microbiology.

4. Histroy
• 1673-1723, Antony Van
Leeuwenhoek (Dutch) described
live microorganisms that he
observed in teeth scrapings and
rain water.
• First person to actually see living
microorganisms.

5. History
• 1822-1895, Louis Pasteur
• A French organic chemist
• Considered as father of modern
microbiology
• Fermentation process: beer/wine not
produced without microbes
• Microbes causes spoilage
• Demonstrated guncotton experiment
• Performed swan neck flask experiment
• Diffrenet places/Diffrenet conc. of
microbes
• Disproved spontaneous generation
• Developed Pasteurisation process(50-
60°C)
• Virulence power
• Developed Rabies/Anthrax

6. History
• 1843-1910, Robert Koch
• 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 and Vibrio
cholera,1883, awarded a Nobel prize in 1905
• Developed relationship b/t microorganism and diseases
 Koch's postulates:
I. The microorganism must be present in every case of
disease
II. The microorganisms must be isolated from the diseased
animal and grown in pure culture
III. The disease must be reproduced when a pure culture of
the microorganisms is inoculated into a susceptible host
IV. The same microorganism must be desolated from the
experiment from the experimentally infected host

8. Prokaryotic vs. Eukaryotic Cell

9. Eukaryotic cell

10. Differentiate between Prokaryotes and Eukaryoties
Sl.No. Prokaryotes Eukaryotes
1. Nuclear membrane is absent Nuclear membrane is present
2. Nucleolus is absent Nucleolus is present
3. Single circular Chromosome More than one Linear chromosome
4. Mitotic division is absent Mitotic division is present
5. Deoxyribonucleoprotein is absent Deoxyribonucleoprotein is present
6. Cytoplasm streaming is absent Cytoplasm streaming is present
7. Pinocytosis is absent Pinocytosis is present
8. Have Pili and fimbriae(for adhesion),
flagella for propulsion
Have cillia and flagella(for movement)
9. Cell size ranges from 0.5-100µm Cell size ranges from 10-150µm
10. Contained only one copy of each
gene(haploid)
Some eukaryotics genome are organised
into operons
11. No histone in chromosome DNA bounded around Histone
12. Asexual reproduction(binary fission) Sexual reproduction(mitosis & meiosis)

11. 13 Mitochondria is absent Mitochondria is present
14. Lysosomes are absent Lysosomes are present
15. Golgi apparatuis is absent Golgi apparatuis is present
16. Endoplasmic reticulum absent Endoplasmic reticulum present
17. Chloroplasts are absent Chloroplasts are may be present
18. Ribosomes is 70S, distributed in the
cytoplasm
80S arrayed one membranes as in
endoplasmic reticulum, 70S in mitochondria
and chloroplasts
19. In Cytoplasmic membranes sterols
absent
Sterols present
20. Cell wall contain peptidoglycan Peptidoglycan absent
21. Locomotor organelles is simple fibril Locomotor organelles is multifibrilled
22. Pseudopodia absent Pseudopodia present
23. Eg: Bacteria, rickettsiae, chlamydiae,
actinomycetes, etc
Eg: Fungi, protozoa, algae, plants, animals

12. Scopes and importance
1. Production 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.

13. 2. Production 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.

14. 3. Diagnosis of diseases and
treatment:
 Different tests are used to detect
infectious microorganisms, eg: ELISA,
Widal test, TUBERCULLIN SKIN TEST,
4. Treatment 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.

16. 5. Plant 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

17. 6. Sterile 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.
7. Sterilization:
 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.

18. 8. Steroid 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.
9. Identification 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.

22. 10. Testing 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 the
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 for TMC.

23. Microbiological 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.
Evaluation 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.

24. Antimicrobial 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.

25. Endo-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-toxins Test(BET) is most
important in microbiology laboratories involved
with quality control of parenterals and medical
devices.
• LAL Test, gel-clot method, chromogenic end point,
kinetic turbidometric assay and kinetic chromogenic
assay are also used to detect endotoxins.

26. Test 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