This presentation is Useful for B. Pharmacy SEM III Students to study the Topic Fungi According to PCI Syllabus.
It Consist of Morpholoy of Fungi, Cultivation , Replication and Classification of Virud
Morphology, Classification, Cultivation and Reproduction of FungiKrutika Pardeshi
This presentation is Useful for B. Pharmacy SEM III Students to study the Topic Fungi According to PCI Syllabus.
It Consist of Morpholoy of Fungi, Cultivation , Reproduction and Classification of Fungi.
Evaluation of Bactericidal and BacteriostaticRajsingh467604
What are disinfectants?
As per the definition given by WHO ( World health organization ) : a disinfectant is a chemical agent, which destroys or inhibits growth of pathogenic microorganisms in the non-sporing or vegetative state.
Why Evaluation?
Evaluation of disinfectants is used to check the ability or efficacy of any disinfectant against specific microorganisms to establish its effectiveness.
Evaluation tests of bactericide.
1. RIDEAL WALKER TEST
This test is also known as the phenol coefficient test,in which any chemical is compared with phenol for its antimicrobial activity.
The result is shown in the form of phenol coefficient.
▪ If a phenol coefficient of a given test disinfectant is less than 1, it means that disinfectant is less effective than phenol.
▪ If a phenol coefficient of a given test disinfectant is more than 1, it means that disinfectant is more effective than phenol.
Procedure
1.1 Different dilutions of the test disinfectant and phenol are prepared and 5 ml of each dilution is inoculated with 0.5ml of the 24 hour growth culture of the organisms.
1.2 All tubes(Disinfectant + organisms & phenol + organisms) are placed in a water bath ( at 17.5° C)
1.3 Subcultures of each reaction mixture are taken and transferred to 5ml sterile broth at an interval of 2.5 minutes from zero to 10 mintues.
1.4 Broth tubes are incubated at 37° C for 2 to 3 days & examined for the presence or absence of the growth.
1.5 Then the Rideal Walker coefficient is calculated :
2. CHICK MARTIN TEST.
CHICK MARTIN test is performed in the much similar way as the RIDEAL Walker test but with a little variation.
Principle : This test is carried out in the presence of organic matter like 3% human feces or dried yeast.
Procedure
2.1 Serial dilutions of test solution and phenol is prepared in distilled water.
2.2 To this 3% yeast suspension is also added.
2.3 To this solution the S. typhi is added
2.4 After contact time of 30 mins the above mixture is transferred to the freshly prepared 10 ml of broth.
2.5 The test tubes are incubated at 37°C for 48 hours.
2.6 Presence or absence of the growth is calculated.
Evaluation tests of Bacteriostatic.
1. Tube dilution & Agar plate Method
1.1 The chemical agent is incorporated into nutrient broth or agar medium and inoculated with test micro-organisms.
1.2 These tubes are incubated at 30° TO 35°C for 2 to 3 days and then the results in the form of turbidity or colonies are observed.
1.3 The results are recorded and the activity of the given disinfectant is compared.
2. Cup plate method
2.1 Agar is melted and cooled at 45° Celsius.
2.2 Then inoculated with test micro-organisms and poured into a sterile petri plate.
2.3 In the cup plate method, when the inoculated agar has solidified, holes around 8mm in diameter are cut in the medium with a steel cork borer.
2.4 Now the antimicrobial agents are directly placed in the holes.
Preservation of pharmaceutical products using antimicrobial agents. PHARMACEU...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-VPart-3
Preservation of pharmaceutical products using antimicrobial agents.
Introduction. Ideal Properties of Preservatives:
Antimicrobial Chemical Preservatives
Development of a Preservative System.
Factors affecting efficacy of a preservative: 1. Interaction With components of the formulation
2. Properties of the Preservatives:
3) Effect of Containers.
4) Type of microbes:
5) Influence of pH:
Challenge Test: Efficacy Test of Preservative : Medium used, Choice of test organism:
Preparation of the inoculum:
Procedure:
Interpretation of Results:
Evaluation of Bactericidal and Bacteriostatic (Disinfectant). PHARMACEUTICAL ...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-III Part-5 Evaluation of Bactericidal and Bacteriostatic (Disinfectant). The common methods used for evaluation of a disinfectant are as follows,
Tube Dilution Method.
Agar Plate Method.
Filter Paper & Cup Plate Method.
Ditch-Plate Method.
Phenol Coefficient Method.
The official phenol coefficient tests include,
Rideal-Walker Test (RW Test).
Chick-Martin Test.
United States FDA Test for Phenol Coefficient. (FDA Test)
The US Association of Official Agricultural Chemists Test (FDA Test)
A. Rideal-Walker Test:
Kelsey Sykes Method
Animal Cell Culture: Growth of animal cells in culture. PHARMACEUTICAL MICROB...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-VPart-4
Animal Cell Culture: Growth of animal cells in culture.
Introduction: Histroy, The culture media used for animal cell culture are classified as,
Natural, Artificial, Synthesized
Natural Culture Media:
a. Blood Plasma:
b. Blood Serum:
c. Tissue Extracts:
Artificial Media
Some common examples of artificial media are,
Minimal Essential Medium (MEM),
CMRL 1066,
RPMI 1640.
Synthetic media re classified as,
Serum Containing Media.
Serum Free Media.
a. Serum Containing Media:
b. Serum Free Media:
Physicochemical Parameters needed for growth animal cell culture:
General procedure for cell Culture.
Isolation of the tissue:
Disaggregation of the Tissue:
Mechanical disaggregation
b. Enzymatic Disaggregation
. Trypsin based disaggregation or trypsinization:
Warm trypsinization:
Cold trypsinization:
Drawbacks of trypsin disaggregation:
B. Collagenase based disaggregation:
C. Chelating Agents:
3. Seeding of Culture:
Types of spoilage, factors affecting the microbial spoilage of pharmaceutical...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-V Part-1
Types of spoilage, factors affecting the microbial spoilage of pharmaceutical products, source and type of contaminants. Introduction: Defintion Types of Microbial Spoilage:
1. Infection induced due to contaminated pharmaceutical products: Table no. 1.1 Common pathogens spoiling pharmaceutical products:
2. Physicochemical spoilage –
i) Viable growth ii) Gas production
iii) Colouration / Decolouration
iv) Odour formation
v) Taste change
3. Physical Spoilage:
Cracking of emulsion:
Odor changes
4. Biological spoilage:
Microbial Toxins
Microbial Metabolites
5. Chemical spoilage: Table 1.2 Susceptibility of pharmaceutical ingredients to microbial contamination
Factors affecting microbial spoilage
Size of contaminant inoculum
Nutritional factors
Moisture content
pH
Storage temperature
Redox potential
Packaging design
Sources and Types Of Contamination:
Personnel,
Poor facility design,
Incoming ventilation air,
Machinery and other equipment for production,
Raw material and semi-finished material,
Packaging material,
Utilities,
Different media used in the production process as well as for cleaning and Cleanroom clothing.
Morphology, Classification, Cultivation and Reproduction of FungiKrutika Pardeshi
This presentation is Useful for B. Pharmacy SEM III Students to study the Topic Fungi According to PCI Syllabus.
It Consist of Morpholoy of Fungi, Cultivation , Reproduction and Classification of Fungi.
Evaluation of Bactericidal and BacteriostaticRajsingh467604
What are disinfectants?
As per the definition given by WHO ( World health organization ) : a disinfectant is a chemical agent, which destroys or inhibits growth of pathogenic microorganisms in the non-sporing or vegetative state.
Why Evaluation?
Evaluation of disinfectants is used to check the ability or efficacy of any disinfectant against specific microorganisms to establish its effectiveness.
Evaluation tests of bactericide.
1. RIDEAL WALKER TEST
This test is also known as the phenol coefficient test,in which any chemical is compared with phenol for its antimicrobial activity.
The result is shown in the form of phenol coefficient.
▪ If a phenol coefficient of a given test disinfectant is less than 1, it means that disinfectant is less effective than phenol.
▪ If a phenol coefficient of a given test disinfectant is more than 1, it means that disinfectant is more effective than phenol.
Procedure
1.1 Different dilutions of the test disinfectant and phenol are prepared and 5 ml of each dilution is inoculated with 0.5ml of the 24 hour growth culture of the organisms.
1.2 All tubes(Disinfectant + organisms & phenol + organisms) are placed in a water bath ( at 17.5° C)
1.3 Subcultures of each reaction mixture are taken and transferred to 5ml sterile broth at an interval of 2.5 minutes from zero to 10 mintues.
1.4 Broth tubes are incubated at 37° C for 2 to 3 days & examined for the presence or absence of the growth.
1.5 Then the Rideal Walker coefficient is calculated :
2. CHICK MARTIN TEST.
CHICK MARTIN test is performed in the much similar way as the RIDEAL Walker test but with a little variation.
Principle : This test is carried out in the presence of organic matter like 3% human feces or dried yeast.
Procedure
2.1 Serial dilutions of test solution and phenol is prepared in distilled water.
2.2 To this 3% yeast suspension is also added.
2.3 To this solution the S. typhi is added
2.4 After contact time of 30 mins the above mixture is transferred to the freshly prepared 10 ml of broth.
2.5 The test tubes are incubated at 37°C for 48 hours.
2.6 Presence or absence of the growth is calculated.
Evaluation tests of Bacteriostatic.
1. Tube dilution & Agar plate Method
1.1 The chemical agent is incorporated into nutrient broth or agar medium and inoculated with test micro-organisms.
1.2 These tubes are incubated at 30° TO 35°C for 2 to 3 days and then the results in the form of turbidity or colonies are observed.
1.3 The results are recorded and the activity of the given disinfectant is compared.
2. Cup plate method
2.1 Agar is melted and cooled at 45° Celsius.
2.2 Then inoculated with test micro-organisms and poured into a sterile petri plate.
2.3 In the cup plate method, when the inoculated agar has solidified, holes around 8mm in diameter are cut in the medium with a steel cork borer.
2.4 Now the antimicrobial agents are directly placed in the holes.
Preservation of pharmaceutical products using antimicrobial agents. PHARMACEU...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-VPart-3
Preservation of pharmaceutical products using antimicrobial agents.
Introduction. Ideal Properties of Preservatives:
Antimicrobial Chemical Preservatives
Development of a Preservative System.
Factors affecting efficacy of a preservative: 1. Interaction With components of the formulation
2. Properties of the Preservatives:
3) Effect of Containers.
4) Type of microbes:
5) Influence of pH:
Challenge Test: Efficacy Test of Preservative : Medium used, Choice of test organism:
Preparation of the inoculum:
Procedure:
Interpretation of Results:
Evaluation of Bactericidal and Bacteriostatic (Disinfectant). PHARMACEUTICAL ...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-III Part-5 Evaluation of Bactericidal and Bacteriostatic (Disinfectant). The common methods used for evaluation of a disinfectant are as follows,
Tube Dilution Method.
Agar Plate Method.
Filter Paper & Cup Plate Method.
Ditch-Plate Method.
Phenol Coefficient Method.
The official phenol coefficient tests include,
Rideal-Walker Test (RW Test).
Chick-Martin Test.
United States FDA Test for Phenol Coefficient. (FDA Test)
The US Association of Official Agricultural Chemists Test (FDA Test)
A. Rideal-Walker Test:
Kelsey Sykes Method
Animal Cell Culture: Growth of animal cells in culture. PHARMACEUTICAL MICROB...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-VPart-4
Animal Cell Culture: Growth of animal cells in culture.
Introduction: Histroy, The culture media used for animal cell culture are classified as,
Natural, Artificial, Synthesized
Natural Culture Media:
a. Blood Plasma:
b. Blood Serum:
c. Tissue Extracts:
Artificial Media
Some common examples of artificial media are,
Minimal Essential Medium (MEM),
CMRL 1066,
RPMI 1640.
Synthetic media re classified as,
Serum Containing Media.
Serum Free Media.
a. Serum Containing Media:
b. Serum Free Media:
Physicochemical Parameters needed for growth animal cell culture:
General procedure for cell Culture.
Isolation of the tissue:
Disaggregation of the Tissue:
Mechanical disaggregation
b. Enzymatic Disaggregation
. Trypsin based disaggregation or trypsinization:
Warm trypsinization:
Cold trypsinization:
Drawbacks of trypsin disaggregation:
B. Collagenase based disaggregation:
C. Chelating Agents:
3. Seeding of Culture:
Types of spoilage, factors affecting the microbial spoilage of pharmaceutical...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-V Part-1
Types of spoilage, factors affecting the microbial spoilage of pharmaceutical products, source and type of contaminants. Introduction: Defintion Types of Microbial Spoilage:
1. Infection induced due to contaminated pharmaceutical products: Table no. 1.1 Common pathogens spoiling pharmaceutical products:
2. Physicochemical spoilage –
i) Viable growth ii) Gas production
iii) Colouration / Decolouration
iv) Odour formation
v) Taste change
3. Physical Spoilage:
Cracking of emulsion:
Odor changes
4. Biological spoilage:
Microbial Toxins
Microbial Metabolites
5. Chemical spoilage: Table 1.2 Susceptibility of pharmaceutical ingredients to microbial contamination
Factors affecting microbial spoilage
Size of contaminant inoculum
Nutritional factors
Moisture content
pH
Storage temperature
Redox potential
Packaging design
Sources and Types Of Contamination:
Personnel,
Poor facility design,
Incoming ventilation air,
Machinery and other equipment for production,
Raw material and semi-finished material,
Packaging material,
Utilities,
Different media used in the production process as well as for cleaning and Cleanroom clothing.
VIRUS PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-IIIPart-2Study of morphology, ...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-IIIPart-2Study of morphology, classification, reproduction/replication and cultivation of Virus. Introduction, Def General characteristics of Viruses: small size characteristic shapes, obligate intracellular parasites no built-in metabolic machinery no ribosomes
only one type of nucleic acid
do not grow in size. Morphology of Virus: Helical, Polyhedral (Icosahedral) Viral Envelop, Complex virus, Classification of virus. Viral Replication LIFE CYCLE OF BACTIRIOPHAGES Lytic cycle: Attachment, Penetration, Biosynthesis, Maturation and Release of progeny Phage Particles. The Lysogenic Cycle, Cultivation of virus : Animal inoculation, Embryonated eggs or chick embryo method and Tissue culture or cell culture: Organ cultures Explant culture and Cell culture. Types of cell culture
1.Primary cell culture: 2. Diploid cell culture (Semi-continuous cell lines):3. Heteroploid cultures (Continuous cell lines):
MULTIPLICATION OF HUMAN VIRUS:1. Attachment of Viral Particles 2. Penetration 3. Uncoating 4. Replication Of Viral Nucleic Acids And Translation Of The Genome 5) Maturation Or Assembly Of Virions. ) 6. Release Of Virions Into The Surrounding Environment
Assessment of microbial contamination and spoilage. PHARMACEUTICAL MICROBIOLO...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-VPart-2
Assessment of microbial contamination and spoilage.
Assessment of microbial contamination and spoilage
1. Physical and chemical changes:
2. Assessment of viable microorganisms in non-sterile products:
3. Sterility test:
4. Estimation of pyrogens:
Microbial Limit Tests:
Total Aerobic Microbial Count:
Membrane Filtration.
Plate Count Methods.
Pour Plate Method.
Surface spread Method.
Most Probable Number(MPN)
Biochemical tests are based on reactions that takes place in various living rganisms. In microbiology these are useful for identification of various microorganisms like identification and differentiation of various bacterial species. IMViC test is a group of test that are used to differentiate between Escheritia and Enterobacter species.
Evaluation of the efficiency of sterilization methods.Sterility indicatorsMs. Pooja Bhandare
Evaluation of the efficiency of sterilization methods.Sterility indicators
Sterility criteria: Bioburden ,Sensitivity of microorganisms
Death rate or Survivor curve,D- Value or Decimal reduction time,Z- value or Thermal reduction time, f- value, Q10 Value or Temperature Coefficient, Inactivation Factor:
STERILITY INDICATORS : Physical Indicators, Chemical Indicators
Biological Indicators
1. Physical Indicators: i) Moist heat Indicator ii) Dry heat iii) Radio sterilization iv) Gaseous methods v) Filtration 2.CHEMICAL INDICATORS : I) Browne’s tubes II) WITTNESS TUBES IV) Royce Sachet V) Chemical Dosimeter 3.BIOLOGICAL INDICATORS
Pharmaceutical Microbiology Unit-2
Identification of Bacteria using staining techniques(Simple, Gram’s & Acid fast staining) and Biochemical Test (IMViC).1. INDOLE TEST 2. METHYL RED (MR) TEST 3. VOGES-PROSKAUR (VP) TEST 4. CITRATE UTILIZATION TEST
Microbiological Assay of Vitamin & Amino acid Assessment of a New Antibiotic...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T) Unit-IV Part-3
Microbiological Assay of Vitamin & Amino acid Assessment of a New Antibiotic: Introduction:
Principle
Microbiological Assay of Cynocobalamin (Vitamin B12):
Tritrimetric Method.
Turbidimetric Method.
Preparation of Standard Cynocobalmine stock solution:
Preparation of Basal Medium Stock Solution:
Test Solution of the material to be assayed Preparation of inoculum: Procedure of Titrimetric method: Turbidimetric Method: Microbiological assay of Amino acids. Assessment of a New Antibiotic.
Introduction:
MIC of an antibiotic is tested either by one of the following ways,
Liquid Dilution Method.
Solid Dilution Method
Microbiological assay-Principles and methods of different microbiological assay.someshwar mankar
Principles and methods of different microbiological assay. Methods for standardization of
antibiotics, vitamins and amino acids. Assessment of a new antibiotic.
Disinfection, Definition, classification,Mode of action, factors affecting & ...someshwar mankar
Disinfection, Definition, classification,Mode of action, factors affecting & Evaluation of disinfectant as per bacteriostatic & Bacteriocidal action
Department of Pharmaceutics,PRCOP,Loni
Two general methods are used for microbiological assays
Method A: Cylinder plate method or cup plate method.
Method B: Tube assay method or titrimetric method.
Nutritional requirements of bacteria and nutrient media (2) copyvinaya warad
To understand nutritional requirements of bacteria
To study nutritional classification of bacteria
To study constituents of nutrient media
To understand types of nutrient media.
To understand uses of different nutrient media
Classification and mode of action of disinfectants PHARMACEUTICAL MICROBIOLOG...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-III Classification and mode of action of disinfectants. DISINFECTANT
Definition: Ideal properties of disinfectants: CLASSIFICATION OF DISINFECTANTS: Based on consistency 1. Liquid (E.g., Alcohols, Phenols) 2.Gaseous (Formaldehyde vapor, Ethylene oxide). Based on spectrum of activity 1. High level disinfectant
2. Intermediate level disinfectant
3. Low level disinfectant .Based on mechanism of action: 1.Action on membrane2.Denaturation of cellular proteins 3.Damage to nucleic acids 4.Oxidation of essential sulfhydryl groups of enzymes 5.Alkylation of amino-, carboxyl- and hydroxyl group. MODE OF ACTION AND APPICATION OF DISINFECTANT
Acid and alkalies
Halogens
Heavy metals
Phenols and its derivatives
Alcohol
Aldehydes
Dyes:
Quaternary ammonium compounds
Detergents and soaps.
Principles and methods of different microbiological assay, methods for standa...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-IV Part-2 Principles and methods of different microbiological assay, methods for standardization of antibiotics.
Introduction: Principles Advantages of Microbial Assay: Disadvantages of Microbial Assay: MICROBIOLOGICAL ASSAY OF ANIBIOTICS PRINCIPLE Media used for antibiotics assay Standard Preparation. Buffer Solutions Preparation of the Sample Solution: Test Organisms Preparation of inoculum: Methods of preparation of test organism suspension: Assay Methods: Method A: Cup-plate or Cylinder Plate Method.
Method B: Turbidimetric or Tube assay Method
VIRUS PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-IIIPart-2Study of morphology, ...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-IIIPart-2Study of morphology, classification, reproduction/replication and cultivation of Virus. Introduction, Def General characteristics of Viruses: small size characteristic shapes, obligate intracellular parasites no built-in metabolic machinery no ribosomes
only one type of nucleic acid
do not grow in size. Morphology of Virus: Helical, Polyhedral (Icosahedral) Viral Envelop, Complex virus, Classification of virus. Viral Replication LIFE CYCLE OF BACTIRIOPHAGES Lytic cycle: Attachment, Penetration, Biosynthesis, Maturation and Release of progeny Phage Particles. The Lysogenic Cycle, Cultivation of virus : Animal inoculation, Embryonated eggs or chick embryo method and Tissue culture or cell culture: Organ cultures Explant culture and Cell culture. Types of cell culture
1.Primary cell culture: 2. Diploid cell culture (Semi-continuous cell lines):3. Heteroploid cultures (Continuous cell lines):
MULTIPLICATION OF HUMAN VIRUS:1. Attachment of Viral Particles 2. Penetration 3. Uncoating 4. Replication Of Viral Nucleic Acids And Translation Of The Genome 5) Maturation Or Assembly Of Virions. ) 6. Release Of Virions Into The Surrounding Environment
Assessment of microbial contamination and spoilage. PHARMACEUTICAL MICROBIOLO...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-VPart-2
Assessment of microbial contamination and spoilage.
Assessment of microbial contamination and spoilage
1. Physical and chemical changes:
2. Assessment of viable microorganisms in non-sterile products:
3. Sterility test:
4. Estimation of pyrogens:
Microbial Limit Tests:
Total Aerobic Microbial Count:
Membrane Filtration.
Plate Count Methods.
Pour Plate Method.
Surface spread Method.
Most Probable Number(MPN)
Biochemical tests are based on reactions that takes place in various living rganisms. In microbiology these are useful for identification of various microorganisms like identification and differentiation of various bacterial species. IMViC test is a group of test that are used to differentiate between Escheritia and Enterobacter species.
Evaluation of the efficiency of sterilization methods.Sterility indicatorsMs. Pooja Bhandare
Evaluation of the efficiency of sterilization methods.Sterility indicators
Sterility criteria: Bioburden ,Sensitivity of microorganisms
Death rate or Survivor curve,D- Value or Decimal reduction time,Z- value or Thermal reduction time, f- value, Q10 Value or Temperature Coefficient, Inactivation Factor:
STERILITY INDICATORS : Physical Indicators, Chemical Indicators
Biological Indicators
1. Physical Indicators: i) Moist heat Indicator ii) Dry heat iii) Radio sterilization iv) Gaseous methods v) Filtration 2.CHEMICAL INDICATORS : I) Browne’s tubes II) WITTNESS TUBES IV) Royce Sachet V) Chemical Dosimeter 3.BIOLOGICAL INDICATORS
Pharmaceutical Microbiology Unit-2
Identification of Bacteria using staining techniques(Simple, Gram’s & Acid fast staining) and Biochemical Test (IMViC).1. INDOLE TEST 2. METHYL RED (MR) TEST 3. VOGES-PROSKAUR (VP) TEST 4. CITRATE UTILIZATION TEST
Microbiological Assay of Vitamin & Amino acid Assessment of a New Antibiotic...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T) Unit-IV Part-3
Microbiological Assay of Vitamin & Amino acid Assessment of a New Antibiotic: Introduction:
Principle
Microbiological Assay of Cynocobalamin (Vitamin B12):
Tritrimetric Method.
Turbidimetric Method.
Preparation of Standard Cynocobalmine stock solution:
Preparation of Basal Medium Stock Solution:
Test Solution of the material to be assayed Preparation of inoculum: Procedure of Titrimetric method: Turbidimetric Method: Microbiological assay of Amino acids. Assessment of a New Antibiotic.
Introduction:
MIC of an antibiotic is tested either by one of the following ways,
Liquid Dilution Method.
Solid Dilution Method
Microbiological assay-Principles and methods of different microbiological assay.someshwar mankar
Principles and methods of different microbiological assay. Methods for standardization of
antibiotics, vitamins and amino acids. Assessment of a new antibiotic.
Disinfection, Definition, classification,Mode of action, factors affecting & ...someshwar mankar
Disinfection, Definition, classification,Mode of action, factors affecting & Evaluation of disinfectant as per bacteriostatic & Bacteriocidal action
Department of Pharmaceutics,PRCOP,Loni
Two general methods are used for microbiological assays
Method A: Cylinder plate method or cup plate method.
Method B: Tube assay method or titrimetric method.
Nutritional requirements of bacteria and nutrient media (2) copyvinaya warad
To understand nutritional requirements of bacteria
To study nutritional classification of bacteria
To study constituents of nutrient media
To understand types of nutrient media.
To understand uses of different nutrient media
Classification and mode of action of disinfectants PHARMACEUTICAL MICROBIOLOG...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-III Classification and mode of action of disinfectants. DISINFECTANT
Definition: Ideal properties of disinfectants: CLASSIFICATION OF DISINFECTANTS: Based on consistency 1. Liquid (E.g., Alcohols, Phenols) 2.Gaseous (Formaldehyde vapor, Ethylene oxide). Based on spectrum of activity 1. High level disinfectant
2. Intermediate level disinfectant
3. Low level disinfectant .Based on mechanism of action: 1.Action on membrane2.Denaturation of cellular proteins 3.Damage to nucleic acids 4.Oxidation of essential sulfhydryl groups of enzymes 5.Alkylation of amino-, carboxyl- and hydroxyl group. MODE OF ACTION AND APPICATION OF DISINFECTANT
Acid and alkalies
Halogens
Heavy metals
Phenols and its derivatives
Alcohol
Aldehydes
Dyes:
Quaternary ammonium compounds
Detergents and soaps.
Principles and methods of different microbiological assay, methods for standa...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-IV Part-2 Principles and methods of different microbiological assay, methods for standardization of antibiotics.
Introduction: Principles Advantages of Microbial Assay: Disadvantages of Microbial Assay: MICROBIOLOGICAL ASSAY OF ANIBIOTICS PRINCIPLE Media used for antibiotics assay Standard Preparation. Buffer Solutions Preparation of the Sample Solution: Test Organisms Preparation of inoculum: Methods of preparation of test organism suspension: Assay Methods: Method A: Cup-plate or Cylinder Plate Method.
Method B: Turbidimetric or Tube assay Method
General Characters and Classification of Viruses. Includes ICTV classification and Baltimore classification of viruses. A brief explanation of the Viral structure and Lifecycle.
micro teaching on communication m.sc nursing.pdfAnurag Sharma
Microteaching is a unique model of practice teaching. It is a viable instrument for the. desired change in the teaching behavior or the behavior potential which, in specified types of real. classroom situations, tends to facilitate the achievement of specified types of objectives.
Title: Sense of Smell
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journeygreendigital
Tom Selleck, an enduring figure in Hollywood. has captivated audiences for decades with his rugged charm, iconic moustache. and memorable roles in television and film. From his breakout role as Thomas Magnum in Magnum P.I. to his current portrayal of Frank Reagan in Blue Bloods. Selleck's career has spanned over 50 years. But beyond his professional achievements. fans have often been curious about Tom Selleck Health. especially as he has aged in the public eye.
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Introduction
Many have been interested in Tom Selleck health. not only because of his enduring presence on screen but also because of the challenges. and lifestyle choices he has faced and made over the years. This article delves into the various aspects of Tom Selleck health. exploring his fitness regimen, diet, mental health. and the challenges he has encountered as he ages. We'll look at how he maintains his well-being. the health issues he has faced, and his approach to ageing .
Early Life and Career
Childhood and Athletic Beginnings
Tom Selleck was born on January 29, 1945, in Detroit, Michigan, and grew up in Sherman Oaks, California. From an early age, he was involved in sports, particularly basketball. which played a significant role in his physical development. His athletic pursuits continued into college. where he attended the University of Southern California (USC) on a basketball scholarship. This early involvement in sports laid a strong foundation for his physical health and disciplined lifestyle.
Transition to Acting
Selleck's transition from an athlete to an actor came with its physical demands. His first significant role in "Magnum P.I." required him to perform various stunts and maintain a fit appearance. This role, which he played from 1980 to 1988. necessitated a rigorous fitness routine to meet the show's demands. setting the stage for his long-term commitment to health and wellness.
Fitness Regimen
Workout Routine
Tom Selleck health and fitness regimen has evolved. adapting to his changing roles and age. During his "Magnum, P.I." days. Selleck's workouts were intense and focused on building and maintaining muscle mass. His routine included weightlifting, cardiovascular exercises. and specific training for the stunts he performed on the show.
Selleck adjusted his fitness routine as he aged to suit his body's needs. Today, his workouts focus on maintaining flexibility, strength, and cardiovascular health. He incorporates low-impact exercises such as swimming, walking, and light weightlifting. This balanced approach helps him stay fit without putting undue strain on his joints and muscles.
Importance of Flexibility and Mobility
In recent years, Selleck has emphasized the importance of flexibility and mobility in his fitness regimen. Understanding the natural decline in muscle mass and joint flexibility with age. he includes stretching and yoga in his routine. These practices help prevent injuries, improve posture, and maintain mobilit
Flu Vaccine Alert in Bangalore Karnatakaaddon Scans
As flu season approaches, health officials in Bangalore, Karnataka, are urging residents to get their flu vaccinations. The seasonal flu, while common, can lead to severe health complications, particularly for vulnerable populations such as young children, the elderly, and those with underlying health conditions.
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This year, the flu season is expected to coincide with a potential increase in other respiratory illnesses. The Karnataka Health Department has launched an awareness campaign highlighting the significance of flu vaccinations. They have set up multiple vaccination centers across Bangalore, making it convenient for residents to receive their shots.
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Report Back from SGO 2024: What’s the Latest in Cervical Cancer?bkling
Are you curious about what’s new in cervical cancer research or unsure what the findings mean? Join Dr. Emily Ko, a gynecologic oncologist at Penn Medicine, to learn about the latest updates from the Society of Gynecologic Oncology (SGO) 2024 Annual Meeting on Women’s Cancer. Dr. Ko will discuss what the research presented at the conference means for you and answer your questions about the new developments.
Couples presenting to the infertility clinic- Do they really have infertility...Sujoy Dasgupta
Dr Sujoy Dasgupta presented the study on "Couples presenting to the infertility clinic- Do they really have infertility? – The unexplored stories of non-consummation" in the 13th Congress of the Asia Pacific Initiative on Reproduction (ASPIRE 2024) at Manila on 24 May, 2024.
Anti ulcer drugs and their Advance pharmacology ||
Anti-ulcer drugs are medications used to prevent and treat ulcers in the stomach and upper part of the small intestine (duodenal ulcers). These ulcers are often caused by an imbalance between stomach acid and the mucosal lining, which protects the stomach lining.
||Scope: Overview of various classes of anti-ulcer drugs, their mechanisms of action, indications, side effects, and clinical considerations.
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3. Viruses:
An infective agent that typically consists of a nucleic acid
molecule in a protein coat, is too small to be seen by light
microscopy, and is able to multiply only within the living
cells of a host.
4. Introduction to viruses
Viruses do not have cells that divide; new viruses are
assembled in the infected host cell
But unlike still simpler infectious agents, viruses contain
genes, which gives them the ability to mutate and evolve.
Evolved from plasmids: pieces of DNA that can move between
cells
while others may have evolved from bacteria.
Over 5,000 species of viruses have been discovered.
5. Introduction to viruses
A virus consists of two or three parts:
genes, made from either DNA or RNA,
long molecules that carry genetic information
protein coat that protects the genes; and in some viruses, an
envelope of fat
Viruses vary in shape from the simple
helical and icosahedral to more
complex structures.
Viruses range in size from 20 to 300 nanometres; it would
take 30,000 to 750,000 of them, side by side, to stretch to 1
centimeter.
6. Viruses spread in many ways. Just as many viruses are
very specific as to which host species or tissue they
attack,
Plant viruses are often spread from plant to plant by
insects and other organisms, known as vectors.
Some viruses of animals, including humans, are spread
by exposure to infected bodily fluids
Viruses such as influenza are spread through the air by
droplets of moisture when people cough or sneeze.
Viruses such as norovirus are transmitted by the
faecal–oral route, which involves the
contamination of hands, food and water.
Spreading , Vectors:
7. The human immunodeficiency virus, HIV, is transmitted by
bodily fluids transferred during sex.
Dengue virus, are spread by blood-sucking insects.
Rotavirus is often spread by direct contact with infected
children.
Antibiotics have no effect on viruses,
but antiviral drugs have been developed to treat life-
threatening
infections. Vaccines that produce lifelong
immunity can prevent some viral infections.
8. Characteristics
Obligate intracellular parasites of bacteria, protozoa, fungi, algae, plants, and
animals.
Ultramicroscopic size, ranging from 20 nm up to 450 nm
(diameter).
Not cellular in nature; structure is very compact and economical.
Do not independently fulfill the characteristics of life.
Inactive macromolecules outside the host cell and active only
inside host cells.
Basic structure consists of protein shell (capsid) surrounding nucleic acid core.
• Nucleic acid can be either DNA or RNA but notboth
9. • Nucleic acid can be double-stranded DNA, single- stranded DNA single-
stranded RNA, ordouble-stranded RNA.
• Molecules on virus surface impart high specificity for
• attachment to host cell.
• Multiply by taking control of host cell’s genetic material and regulating the
synthesis and assembly of new viruses.
• Lack enzymes for most metabolic processes.
• Lack machinery for synthesizing proteins.
• Most RNA viruses multiply in & are released fromthe cytoplasm.
• Viral infections range from very mild to life threatening.
11. • Viruses have no nucleus, no organelles, no cytoplasm or
cell membrane—Non-cellular
12. Size of virus ?
• Smallest infectious agents
• Most are so small, they can only be seen with an electron microscope
• Proviruses- around 20 nm in diameter
• Mimi viruses- up to 450 nm in length
• Special stains and an electron microscope
• Negative staining outlines the shape
• Positive staining shows internal details
13.
14. Classification
• Viruses are classified on the basis of habitat (host).which is
trivial system beside this Viruses are classified on following
criteria.
• Structure
• Chemical composition
• Similarities in genetic makeup
• International Committee on the Taxonomy of Viruses, which
includes
• 3 orders
• 63 families “-viridae”
• 263 genera “-virus”
15. Types of Classification:
• 3 Types of systems were proposed to classify the viruses:
• Baltimore Classification.
• Classical System Classification.
• Genetic Classification.
16. 7 groups were made.
Its principles are fundamental to an understanding of virus
classification and genome replication.
The Baltimore classification has + RNA as its central point.
I: dsDNA viruses (e.g. Adenoviruses, Herpesviruses, Poxviruses)
II: ssDNA viruses (+ strand or "sense") DNA (e.g. Parvoviruses)
III: dsRNA viruses (e.g. Reoviruses)
IV: (+)ssRNA viruses (+ strand or sense) RNA
(e.g. Picornaviruses, Togaviruses)
V: (−)ssRNA viruses (− strand or antisense) RNA
(e.g. Orthomyxoviruses, Rhabdoviruses)
VI: ssRNA viruses (+ strand or sense) RNA with DNA
intermediate in life-cycle (e.g. Retroviruses)
VII: dsDNA viruses (e.g. Heptadnaviruses)
Baltimore Classification:
17. Classification on basis of
host
• Animal viruses:
• Viruses of animal host
• Rabies, Polio, Mumps, Chicken pox, Small pox, and
Influenza.
• Plant Viruses:
• viruses which show their live characteristics when
attached to plants.
• Tobacco mosaic virus (TMV), Banana streak virus,
Carrot thin leaf virus
• Bacterial Virus: Bacteriophages ( T1, T2, T3, andT4.)
18.
19. Classification on
Genetics basis
• According to genetic consequences viruses are classified as.
DNA Viruses and RNAViruses
• Genes may be linear or circular
• The smallest have only 4 genes and largest have several
hundred.
• DNAViruses
• DNA Viruses are the viruses which consist of DNA genome .
They complete their activities by transcription and most of
them attack on organisms of similar genome.
• RNAViruses
• RNA Viruses are the viruses which consist of RNAgenome.
They complete their activities by reverse transcription.
20. Classification on
structural basis
• With relevant to morphology of viral structure viruses are
organized as Enveloped and Nonenveloped viruses.
• However they are also arranges subclasses of DNAand
RNAviruses
21. Structure of virus
• Questions Relating to Structure
• Is it rigid?
• How big is it?
• Is it flexible?
• Structure Must Serve Virus
• It should provide protection for genome
• It should allow virus to move from one host to next
• It should allow for attachment of virus on to new host
24. Capsids:
Viruses consists of nucleic acid core surrounded by a protein
called capsid.
Capsid is composed of large number of capsomer which is
made up of polypeptide molecules.
The capsid with the enclosed nucleic acid is known as
nucleocapsid.
Fully formed virus that is able to establish an infection in a host
cell is known as virion.
25.
26. Functions of Capsid
It protects the viral genome from physical
destruction and enzymatic inactivation by
nucleases in biological material.
It provides the binding site which enable the virus to
attach to specific site on the host cell.
It facilitates the assembly and packaging of viral
genetic information.
It serves as a vehicle of transmission from host to another.
It is antigenic and specific for each viruses
It provides the structural symmetry to the virus
particle.
27. Nucleic Acids:
• Genome:
• the sum total of the genetic information carried by an organism
• Number of viral genes compared with a cell- quite small
• They only have the genes necessary to invade host cells and redirect
their activity
30. RNA Viruses:
• Mostly single-stranded
• Positive-sense RNA: genomes that are ready for immediate
translation into proteins
• Negative-sense RNA: genomes have to be converted into
the proper form to be made into protein
32. Enveloped Virus
The envelop or outer covering of virus
containing lipid is derived from the
plasma membrane of the host cell
during the release by budding from the
cell surface.
The envelop is glycoprotein in nature.
Enveloped viruses are susceptible to the
action of lipid solvent such as ether,
chloroform and detergent.
Eg. Herpes virus, Hepatitis B virus,
HIV virus
Non Enveloped Virus
Viruses which does not have
outer covering.
Naked viruses are more likely
to be resistant to lipid solvents
like ether, chloroform and
detergent.
Viral Envelop
33.
34. In mature virus particle, the glycoproteins often appear as
projecting spikes on the outer surface of the envelop
which are known as peplomers.
A virus may have more than one type of
peplomers. E.g the influenza virus carries two
types of peplomers, the hemagglutinin and
neuraminidase
Peplomers
35. Functions of Peplomers
It helps for attachment of virus to the host cell receptors
to initiate the entrance of the virion into the cell.
It attach to receptors on red blood cells, causing
these cell to agglutinate.
It has enzymatic activity like neuraminidase which cleave
neuraminic acid from host cell glycoproteins.
It has antigenic properties.
36.
37. Structural Symmetries:
Icosahedral Symmetry
An icosahedral (icosa, meaning 20 in greek) is a polgon with 12
vertices or corners and 20 facets or sides.
Each facet is in the shape of an equilateral triangle.
Pentagonal capsomers at the vertices
(pentons) and hexagonal capsomers
making up the facets (hexons)
Eg. Adeno viruses
38. Helical Symmetry
The nucleic acid and capsomers are wound together in
the form of helix or spiral.
Eg. Influenza virus, parainfluenz virus, rabies
virus.
Structural Symmetries:
39. Complex Symmetry
Viruses which don not show either icoshedral or helical
symmetry
due to complexity of their
structure are referred to have
complex symmetry.
Eg. Pox viruses
Structural Symmetries:
40.
41. The genomic information necessary for viral replication is contained in the
viral nucleic acid but lacking biosynthetic enzymes.
The virus depend on the synthetic machinery of the host cell for replication.
Viral multiplication proceeds as following manner.
• Adsorption,
• Penetration,
• Uncoating,
• Synthesis,
• Assembly and Release
• Adsorption.
Viral Replication
42. Adsorption/Attachment
• Virus encounters susceptible host cells
• Adsorbs specifically to receptor sites on the cell
membrane
• Because of the exact fit required, viruses have a limited
host range
43. Penetration
• Flexible cell membrane of the host is penetrated by the
whole virus or its nucleic acid
• Endocytosis: entire virus engulfed by the cell and
enclosed in a vacuole or vesicle
• The viral envelope can also directly fuse with the host
cell membrane
44. Uncoating
• Enzymes in the vacuole dissolve the envelope and capsid
• The virus is now uncoated
45.
46. Synthesis
• Free viral nucleic acid exerts control over the host’s
synthetic and metabolic machinery
• DNA viruses- enter host cell’s nucleus where theyare
replicated and assembled
• DNA enters the nucleus and is transcribed into RNA
• The RNA becomes a message for synthesizing viral
proteins (translation)
• New DNA is synthesized using host nucleotides
• RNA viruses- replicated and assembled in thecytoplasm
48. Release
• Nonenveloped and complex viruses are released when the
cell lyses or ruptures
• Enveloped viruses are liberated by budding or exocytosis
• Anywhere from 3,000 to 100,000 virions may be
released, depending on the virus
• Entire length of cycle- anywhere from 8 to 36 hours
51. BACTERIOPHAGE
-Bactria Eater
- Virus that infect the bacteria
-T4 Bacteriophage is one of the phage from
total number of Bacteriophage of E. Coli
- Total Number – T1-T7
-Which replicate only by Lytic Cycle.
52. VIRULANT
VIRUS
VIRAL
NUCLEIC ACID
REPLICATE BY
RUPTURING
HOST CELL
VIRAL DNA
REPLICATE
SEPRATELY
WITHOUT
FUSING WITH
HOST DNA
NON
VIRULANT
VIRUS /
TEMPERATE
VIRUS
VIRAL
NUCLEIC ACID
REPLICATE BY
FUSION WITH
HOST CELL
VIRAL DNA
FUSED AND
CONVERTED
TO PROPHAGE
LYTIC CYCLE
LYSOGENIC CYCLE
53. CULTIVATION OF VIRUS
PURPOSE OF VIRUS CULTIVATION
• To isolate and identify viruses in clinical samples.
• To prepare viruses for vaccine production.
• To do research on viral structure, replication , genetics
and effects on host cells.
54. METHODS FOR CULTIVATION OF VIRUSES
Inoculation of Virus into
Animal
Inoculation of Virus into
Embryonated Eggs.
Tissue culture
55. Inoculation of Virus in Animals
1. Viruses which are not cultivated in embryonated egg and tissue culture
are cultivated in laboratory animals. e.g: mice, guinea pig , hamester ,
rabbits and primates are used.
2. The selected animals should be healthy and free from any communicable
diseases.
3. Suckling mice (less than 48 hours old) are most commonly used.
4. Route for Inoculation-
Intracerebral.
Subcutaneous.
Intraperitoneal.
Intranasal.
56. It is administered into the cerebrum. It means
when a diseased blood vessel within the brain
bursts allowing blood to leak inside the brain.
A subcutaneous injection is an injection in
which a needle is inserted just under the skin.
Intraperitoneal injection is given to peritoneal
cavity.
It lying within or administered by way of the
nasal structure.
57. ADVANTAGES
• Production of antibodies can be identified.
• Diagnosis, pathogenesis and clinical symptoms
are determined.
• Primary isolation of certain viruses.
• Mice provide a reliable model for studying viral
replication.
• Used for the study of immune responses,
epidemiology and oncogenesis
DISADVANTAGES
• Expensive and difficulties to maintained animals.
• Difficulty in choosing of animals for particular
virus.
• Some human viruses cannot be grown in animals
or can be grown but do not cause diseases.
• Mice do not provide models for vaccine
development.
58. Inoculation of Virus in Egg Embryo
1. The process of cultivation of viruses in embryonated eggs depend
upon the type of egg being used.
2. Egg provide a suitable means for :
-The primary isolation and identification of viruses.
-The production of vaccines.
-The maintained of stock culture.
59. After incubation , the egg is broken and virus is
isolated from tissue of egg.
For inoculation , eggs are first prepared for
cultivation , the sheel surface are first prepared for
cultivation , the shell surface is first disinfected with
iodine and penetrated with a small steriledrill.
Viruses are inoculated into chick embryo of 7-12
days old.
60.
61. Virus growth and multiplication in the egg embryo is
indicated by the death of the embryo , by embryo cell
damage , or by the formation of typical pocks or
lesions on the egg membrane.
Viruses can be cultivated in various pats of egg like :
1. Chorioallantoic membrane (CAM)
2. Allantoic cavity
3. Amniotic sac
4. Yolk sac
62. 1.Chorioallantoic Membrane (CAM)
• Inoculation is mainly for growing provirus.
• After incubation visible lesions called pocks are observed ,
which is grey white area in transparent CAM.
• Herpes simplex virus isalso grown.
• Single virus gives single pocks.
• Thismethod is suitable for plaque studies.
63. 2. Allantoic Cavity
• Inoculation is mainly done for production of vaccine of
influenza virus , yellow fever , rabies.
• Most of avaian viruses can be isolated using this method.
• Allantoic inoculation is a quick and easy method that yields
large amounts (8-15ml) of virus-infected egg fluids.
64. 3.Amniotic Sac
• Inoculation is mainly done for primary isolation of
influenza virus and the mumps virus.
• Growth and replication of virus in egg embryo can be
detected by haemagglutination assay.
• The virus is introduced directly into the amniotic fluid that
bathes the developing embryo.
65. 4. Yolk Sac
• Inoculation is done for some bacteria and some viruses.
• Thismethod is simple for cultivation and multiplication of virus.
66. ADVANTAGES
• Widely used method for the isolation of virus and growth.
• Cost effective and maintenance is much easier.
• The embryonated eggs are readily available.
• They are free from contaminating bacteria and many
latent viruses.
• Ideal substrate for the viral growth and replication.
• less labor is needed.
• Widely used method to grow virus for some vaccine
production.
• Defense mechanisms are not involved in embryonated
eggs.
DISADVANTAGES
• The site of inoculation for varies with different virus . That
is , each virus have different sites for growth and
replication.
67. Inoculation of Virus Using Tissue Culture
There are three types of tissue culture:
- Organ culture.
- Explant culture.
- Cell culture.
68. 1. Small bits of organs can be maintained in vitro for days
and weeks, preserving their original architecture and
function.
2. Organs culture are useful for the isolation of some
viruses which appear to be highly specialised parasite of
certain organs.
3. For example, the tracheal ring organ culture is employed
for the isolation of coronavirus, a respiratory pathogen.
Inoculation of Virus Using Organ Culture
69. 1. Fragments of minced tissue can be grown as ‘explants’
embedded in plasma clots.
2. They may also be cultivated in suspension.
3. This method is now seldom employed in virology.
4. Adenoid tissue explant culture were used for the
isolation of adenoviruses.
Inoculation of Virus Using Explant Culture
70. This is the type of culture routinely employed for growing
viruses
Inoculation of Virus Using Cell Culture
Cell
Culture
Primary
Cell Culture
Diploid Cell
Culture
Continuous
Cell Lines
71. - Tissue are dissociated into the component cell by the
action of proteolytic enzyme.
-The cells are washed , counted and suspended in a
growth medium.
-Such media will enable most cell types to multiply with
a division time of 24-48 hrs.
- The cell suspension is dispensed in bottles, tubes or
petridishes
- The cell adhere to the glass surface and on
incubation, divide to form a confluent monolayer sheet
of cells covering the surface within about a week.
72.
73. 1. These are normal cells obtained from fresh organs of
animals or human being and cultured.
2. They are capable of only limited growth in culture and
cannot be maintained in serial culture.e.g. monkey
kidney cell culture.human embryonic kidney. chick
embryo cell culture.
3. They are commonly employed for primary isolation of
viruses and in preparation of vaccine.
Primary Cell Culture
74. 1. These are cells of single type, contain the same number
of chromosome as the parent cells and are diploid.
2. The diploid cell strains can be subculture for limited
number of times.
3. After about 50 serial passage they undergo senescence.
4. They are also employed for the production of viral
vaccine. Eg. human embryonic lung cell strain WI-38
Diploid Cell Culture
75. 1. Animal cells capable of indefinite growth are called continuous cell lines or cell
lines.
2. These are the cells of a single type , usually derived from cancer cells , that are
capable of continuous serial cultivation indefinitely.
3. Standard cell lines derived from human cancers , such as HeLa , HEp – 2 and
KB cell lines have been used in laboratories throughout the world for many
years.
4. These cell linesmay be maintained by serial subcultivation or stored in the cold (
5. -70⁰C ) for use when necessary.
6. Some cell lines are now permitted to be used for vaccine manufacture, for
example: Vero cells for rabies vaccine.
Continuous Cell Line
76. Advantages
• Relative ease, broad
spectrum, cheaper and
sensitivity
Disadvantages
• The process requires
trained technicians with
experience in working on a
full time basis.
• Tissue or serum for
analysis is sent to
central laboratories to
identify virus.
• State health laboratory and
hospital Laboratory do not
isolate and identify virus
for clinical work.