This document provides an overview of different types of culture media used for growing microorganisms. It discusses the history and definitions of culture media. Media are classified based on consistency (liquid, semi-solid, solid) and method of growth detection (conventional vs automated). The document describes various types of media including simple/basal media, enriched media, enrichment media, selective media, differential media, transport media, and media for specific microorganisms. It provides examples of commonly used media and their purposes. The preparation, sterilization, testing and storage of media are also outlined.
The minimum inhibitory concentration (MIC) is the lowest concentration of an antimicrobial agent that inhibits visible growth of a microorganism. MIC testing determines if a microbe is susceptible, resistant, or has intermediate resistance to an antibiotic. MIC is measured through agar or broth dilution methods by incubating microbes in solutions with increasing concentrations of an antibiotic and observing growth. The MIC provides guidance on effective antibiotic treatment strategies.
Biochemical tests for characterization of bacteriaDr. Pavan Kundur
The document describes various biochemical tests used to characterize bacteria, including the IMViC tests, oxidase test, gelatin hydrolysis test, and starch hydrolysis test. The IMViC tests include indole, methyl red, Voges-Proskauer, and citrate tests used to identify members of the Enterobacteriaceae family. The oxidase test detects the enzyme cytochrome c oxidase to distinguish between oxidase-positive and -negative bacteria. The gelatin hydrolysis test identifies bacteria that can produce the protease gelatinase to liquefy gelatin. The starch hydrolysis test detects bacteria that produce the amylase enzyme to break down starch.
Coagulase positive
Staphylococcus aureus
Human pathogen
Coagulase negative
Staphylococcus epidermidis
Normal flora, disease under special circumstances
Staphylococcus saprophyticus
UTI in young females
Aerobic
Facultative anaerobic
Catalase positive
Non motile
Non sporulating
Pigment – white, golden yellow
This document discusses the IMViC biochemical tests used to identify bacteria. The IMViC tests include Indole production, Methyl red, Voges-Proskauer, and Citrate utilization tests. It focuses on explaining the Indole production test in detail. This test determines if a bacteria can produce the aromatic compound indole from tryptophan. Bacteria are grown in tryptophan broth, and Kovac's or Ehrlich's reagent is added to detect the presence of indole, indicated by a pink color ring formation. The document provides the principles, requirements, procedures, and interpretation of the Indole production biochemical test used in microbial identification.
Methyl Red (MR) and Voges-Proskauer (VP) Test principle, Method, Interpretation & QC #MR & VP
Mallu Medicos Lounge
As the channel name suggests, our channel will be a perfect lounge for the malayali medicos..we wil be covering videos which will be like lecture classes related to the subjects biochemistry and microbiology in which we are specialised.. It will be a better learning experience for the students especially for those who are not able to understand and follow the normal classes in college..we assure the students that you will get a basic idea regarding the topic and extra reading can be done from the reference textbooks..
Our Partner Channel
Health & Voyage channel link - https://youtu.be/nzKqRVjlwc0
Qualification
Maneesha M Joseph
MSc MLT (Microbiology)
Assistant Professor
Baby memorial college of allied Health science
Kozhikode
#Methyl Red Test
#Voges-Proskauer Test
#MRVP Procedure
#MRVP Tests
#MR-VP Tests
#VP (Voges Proskaeur) Test
#MR/VP Test
#Methyl red (MR) and Voges-Proskauer (VP)
#Methyl red test in Microbiology
#Medical
#Microbiology
#MR/VP Test malayalam lecturer
#Mallu Medicos Lounge
#MalluMedicosLounge
#MLT
#Channel introduction
#HealthAndVoyage
This document discusses media used for growing fungi. It describes potato dextrose agar used for growing isolated fungal strains and observing morphology. The ingredients for potato dextrose agar are listed as 1000ml water, 4g sliced and washed potatoes, 20g dextrose, 20g agar powder, with a final pH of 5.6±0.2 at 25°C. Corn meal agar and dichloran 18% glycerol agar are also described as media used to identify yeasts.
Sabouraud dextrose agar (SDA) is used to isolate and cultivate fungi and yeasts from clinical specimens. It contains nutrients like dextrose and enzymatic digest of casein to support fungal growth, and antibiotics to inhibit bacteria. The document outlines the materials, composition, and procedure to prepare SDA media. Colonies are examined after incubation and typical morphologies can indicate fungal species present. However, SDA may not promote conidiation in some fungi and antimicrobials could inhibit some pathogens.
Actinomycetes are filamentous, Gram-positive bacteria that resemble fungi in morphology. They are commonly found in soil. Actinomyces species can cause cervicofacial, thoracic, abdominal or pelvic infections in humans through trauma or poor dental hygiene. Infections appear as indurated swellings with sinuses draining sulfur granules. Nocardia species are aerobic Actinomycetes that can cause cutaneous or systemic infections, especially in immunocompromised individuals. Actinomycotic mycetoma presents as a localized tumor of the foot or hand with multiple draining sinuses.
The minimum inhibitory concentration (MIC) is the lowest concentration of an antimicrobial agent that inhibits visible growth of a microorganism. MIC testing determines if a microbe is susceptible, resistant, or has intermediate resistance to an antibiotic. MIC is measured through agar or broth dilution methods by incubating microbes in solutions with increasing concentrations of an antibiotic and observing growth. The MIC provides guidance on effective antibiotic treatment strategies.
Biochemical tests for characterization of bacteriaDr. Pavan Kundur
The document describes various biochemical tests used to characterize bacteria, including the IMViC tests, oxidase test, gelatin hydrolysis test, and starch hydrolysis test. The IMViC tests include indole, methyl red, Voges-Proskauer, and citrate tests used to identify members of the Enterobacteriaceae family. The oxidase test detects the enzyme cytochrome c oxidase to distinguish between oxidase-positive and -negative bacteria. The gelatin hydrolysis test identifies bacteria that can produce the protease gelatinase to liquefy gelatin. The starch hydrolysis test detects bacteria that produce the amylase enzyme to break down starch.
Coagulase positive
Staphylococcus aureus
Human pathogen
Coagulase negative
Staphylococcus epidermidis
Normal flora, disease under special circumstances
Staphylococcus saprophyticus
UTI in young females
Aerobic
Facultative anaerobic
Catalase positive
Non motile
Non sporulating
Pigment – white, golden yellow
This document discusses the IMViC biochemical tests used to identify bacteria. The IMViC tests include Indole production, Methyl red, Voges-Proskauer, and Citrate utilization tests. It focuses on explaining the Indole production test in detail. This test determines if a bacteria can produce the aromatic compound indole from tryptophan. Bacteria are grown in tryptophan broth, and Kovac's or Ehrlich's reagent is added to detect the presence of indole, indicated by a pink color ring formation. The document provides the principles, requirements, procedures, and interpretation of the Indole production biochemical test used in microbial identification.
Methyl Red (MR) and Voges-Proskauer (VP) Test principle, Method, Interpretation & QC #MR & VP
Mallu Medicos Lounge
As the channel name suggests, our channel will be a perfect lounge for the malayali medicos..we wil be covering videos which will be like lecture classes related to the subjects biochemistry and microbiology in which we are specialised.. It will be a better learning experience for the students especially for those who are not able to understand and follow the normal classes in college..we assure the students that you will get a basic idea regarding the topic and extra reading can be done from the reference textbooks..
Our Partner Channel
Health & Voyage channel link - https://youtu.be/nzKqRVjlwc0
Qualification
Maneesha M Joseph
MSc MLT (Microbiology)
Assistant Professor
Baby memorial college of allied Health science
Kozhikode
#Methyl Red Test
#Voges-Proskauer Test
#MRVP Procedure
#MRVP Tests
#MR-VP Tests
#VP (Voges Proskaeur) Test
#MR/VP Test
#Methyl red (MR) and Voges-Proskauer (VP)
#Methyl red test in Microbiology
#Medical
#Microbiology
#MR/VP Test malayalam lecturer
#Mallu Medicos Lounge
#MalluMedicosLounge
#MLT
#Channel introduction
#HealthAndVoyage
This document discusses media used for growing fungi. It describes potato dextrose agar used for growing isolated fungal strains and observing morphology. The ingredients for potato dextrose agar are listed as 1000ml water, 4g sliced and washed potatoes, 20g dextrose, 20g agar powder, with a final pH of 5.6±0.2 at 25°C. Corn meal agar and dichloran 18% glycerol agar are also described as media used to identify yeasts.
Sabouraud dextrose agar (SDA) is used to isolate and cultivate fungi and yeasts from clinical specimens. It contains nutrients like dextrose and enzymatic digest of casein to support fungal growth, and antibiotics to inhibit bacteria. The document outlines the materials, composition, and procedure to prepare SDA media. Colonies are examined after incubation and typical morphologies can indicate fungal species present. However, SDA may not promote conidiation in some fungi and antimicrobials could inhibit some pathogens.
Actinomycetes are filamentous, Gram-positive bacteria that resemble fungi in morphology. They are commonly found in soil. Actinomyces species can cause cervicofacial, thoracic, abdominal or pelvic infections in humans through trauma or poor dental hygiene. Infections appear as indurated swellings with sinuses draining sulfur granules. Nocardia species are aerobic Actinomycetes that can cause cutaneous or systemic infections, especially in immunocompromised individuals. Actinomycotic mycetoma presents as a localized tumor of the foot or hand with multiple draining sinuses.
This document provides information on various types of culture media used for growing microorganisms in the laboratory. It discusses liquid, solid and semisolid media, and differentiates between simple, complex, enriched, selective, differential, indicator and transport media. Specific examples of commonly used media are provided for each type, along with their compositions and uses. The document aims to outline the basic requirements, classifications and characteristics of different culture media.
I. This presentation discusses three bacterial identification tests: the catalase test, urease test, and triple sugar iron (TSI) test.
II. It provides details on the principles, procedures, and results of the urease test and TSI test. The urease test identifies Proteus bacteria based on their production of urease enzymes, while the TSI test distinguishes between Enterobacteriaceae based on their sugar fermentation patterns.
III. The presentation is delivered by five group members and discusses the applications and limitations of the TSI test for bacterial identification.
The document discusses methods for identifying bacteria, including phenotypic, immunological, and genetic techniques. Phenotypic methods examine bacterial morphology, staining characteristics, and biochemical reactions. Tests like Gram staining, colony morphology, and catalase can provide initial identification. Further tests of carbohydrate use, enzyme production, and sensitivity to inhibitors allow identification to the species level. Immunological methods detect bacterial antigens, while genetic techniques like PCR and nucleic acid analysis provide accurate identification by examining microbial DNA. Both traditional and molecular methods are used to fully characterize unknown bacteria.
The indole test is used to differentiate bacteria based on their ability to produce indole from the amino acid tryptophan. Bacteria containing the enzyme tryptophanase can break down tryptophan into indole, ammonia, and pyruvic acid. The test involves inoculating tryptophan broth with a bacterial sample and incubating. Kovac's reagent is then added, which will produce a red color in the presence of indole, indicating a positive result. Examples of bacteria that test positive include Klebsiella oxytoca and Proteus species, while Salmonella, Pseudomonas, and Yersinia species typically test negative.
This document discusses the classification, diagnosis, and laboratory identification of fungal infections. It covers topics such as the classification of fungi based on morphology and disease, common oral fungal infections, specimen collection guidelines, and various staining and culture techniques used to identify fungi under the microscope or in culture media. These include wet mount preparations with potassium hydroxide or India ink, differential stains like Gram or Gomori's silver, and fungal culture media like Sabouraud dextrose agar or birdseed agar.
Sugar fermentation tests, Cetrimide agar medium, Hugh Leifson medium Shivam kumar Sriwas
1. The document discusses sugar fermentation, how to test for it using media like phenol red carbohydrate broth, and the interpretation of results.
2. Hugh Leifson medium and Cetrimide agar are described as media used to differentiate bacterial metabolism and isolate Pseudomonas aeruginosa, respectively.
3. Key components, principles, preparation, and expected results are outlined for both Hugh Leifson medium and Cetrimide agar tests.
A simple lecture for the description of the various culture media used for isolation of different bacteria in a pure form for further identification procedures.
Cultivation of bacteria and culture methodsAshfaq Ahmad
Cultivation of bacteria allows for the isolation, growth, and study of microorganisms. There are various culture methods and media that support the growth of bacteria. Liquid broths and solid agar plates can be used with different nutrient formulations to selectively grow specific bacteria. Streaking, lawning, stabbing, and pour plating are common culture techniques used to isolate pure colonies for analysis. Specialized enriched, selective, differential, and transport media help optimize bacterial growth and identification.
This lecture discusses laboratory methods for determining antibiotic susceptibility, including disk diffusion tests, minimum inhibitory concentration (MIC) tests, and minimum bactericidal concentration (MBC) tests. Filter paper disks containing different antibiotic concentrations are applied to bacterial lawns to measure susceptibility. The MIC is the lowest concentration that inhibits growth, while the MBC kills bacteria. These methods help evaluate antibiotic effectiveness and monitor emerging resistance.
1. There are two main types of bacterial counts - total bacterial count and total viable count. Total bacterial count includes both living and dead cells while total viable count only measures living cells.
2. Bacterial enumeration is important for comparing growth under different conditions, and in industries like dairy, food, and water microbiology.
3. Methods of enumeration include direct counting using microscopy or Coulter counter, and indirect counting of viable cells using serial dilution plating or membrane filtration. Other methods determine cell mass through dry weight, nitrogen content, or turbidity measurements.
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.
Bacterial culture involves growing bacterial colonies in artificial media containing necessary nutrients and under suitable conditions. Ideal culture media support satisfactory, rapid, and reproducible bacterial growth while revealing bacterial characteristics. Common media components include water, agar, peptones, and extracts. Media are classified by consistency (liquid, semi-solid, solid), constituents (simple, complex, synthetic), or special properties (enriched, selective, differential). Sterilization methods include autoclaving or hot air oven.
This document discusses clinical microbiology and methods used in the field. It provides definitions of clinical microbiology as the study of bacteria and their relation to medicine. Key methods discussed include culture-based techniques of growing bacteria in controlled conditions to isolate, identify, and determine antibiotic sensitivity. Additional non-culture methods described are use of monoclonal antibodies, enzyme immunoassays, probes, polymerase chain reaction (PCR), and ligase chain reaction.
The document provides instructions for performing several microbiological tests, including sugar fermentation tests, indole production, methyl red, Voges-Proskauer, citrate utilization, nitrate reduction, urease, triple sugar iron, oxidase, catalase, amylase, and lipase. It explains the principles, expected results, and interpretations for each test. The tests can be used to differentiate bacterial species and determine their metabolic abilities.
The CAMP test identifies Streptococcus agalactiae (Group B Strep) and Listeria species by their production of a substance called the CAMP factor. The CAMP factor acts synergistically with Staphylococcus aureus to induce enhanced hemolysis, appearing as an arrowhead shape. Discovered in 1944 by Christie, Atkins, Munch, and Petersen, the CAMP test remains an important method for identifying S. agalactiae and Listeria monocytogenes.
The document provides information on differential leukocyte counting (DLC). It defines DLC as the relative proportion of different leukocytes expressed as a percentage. The main types of white blood cells (WBCs) described are granulocytes (neutrophils, eosinophils, basophils) and agranulocytes/mononuclear cells (lymphocytes, monocytes). Pathologic variations in DLC counts include neutrophilia, neutropenia, lymphocytosis, lymphopenia, monocytosis, eosinophilia, eosinopenia, and basophilia. Causes of each variation are outlined. Automated methods for differential counting are also discussed and compared to manual microscopic methods
The TSI test is used to determine carbohydrate fermentation and hydrogen sulfide production in bacteria. It contains glucose, sucrose, and lactose sugars, along with peptones and phenol red pH indicator. Carbohydrate fermentation produces acid, changing the color from red to yellow, while peptone metabolism makes it more alkaline. Hydrogen sulfide production is shown by black precipitate. Results are interpreted based on color changes in the slant and butt portions, indicating which sugars are fermented and if H2S is produced.
The Ziehl-Neelsen stain is used to detect acid-fast bacteria like mycobacteria. It works by using carbolfuchsin as a primary stain that is retained in acid-fast bacteria even after treatment with an acid decolorizer due to their high lipid and mycolic acid content. Non-acid fast cells are then counterstained blue while acid-fast bacteria appear red. The stain involves heating slides coated with carbolfuchsin, treating with an acid decolorizer like sulfuric acid, and counterstaining with methylene blue to identify acid-fast mycobacteria under the microscope.
culture media
CULTURE – Is term given to microorganisms that are cultivated in the lab for the purpose of studying them.
MEDIUM – Is the term given to the combination of ingredients that will support the growth & cultivation of microorganisms outside their natural habitats.
Necessary Requirements for Growth of Bacteria
Distilled Water
Nitrogen containing compounds
Peptone- Golden granular powder
Complex mixture of partially digested protiens by proteolytic
enzymes pepsin, trysin or papain
Peptones, Proteoses, polypeptides, aminoacids, inorganic salts like phosphates
potassium & magnesium
Accessory growth factors like nicotinic acid & riboflavin
Energy sources
Suitable Ph- 7.2 – 7.4
Solidifying agents:
Gelatin– Protien
Agar— Chief component is Long chain Polysaccharide
Melts at 95°c & solidify only when cooled to about 42°c
1- 2% yields a suitable gel eg. Non-nutritive agar
According to Physical State:
Liquid – Peptone Water, Nutrient Broth
Semisolid – Nutrient Agar Stabs
Solid – Blood Agar
According to Oxygen requirement:
Aerobic Medium
Anaerobic Media
- Definition
- Uses of culture media
- Basic composition of culture media
- Types of culture media
--Based on physical state
----solid medium
----semi solid medium
----liquid medium
--Based on ingredients
----Simple or basal medium.
----Complex medium.
----Synthetic or defined medium.
----Semisynthetic medium.
--Special medium
----Enriched media
----Enrichment media
----Selective media
----Differential media
----Indicator media
----Transport media
----Anaerobic media
-Media preparation
-Culture method
--Streak culture
--Lawn culture
-references
This document provides information on various types of culture media used for growing microorganisms in the laboratory. It discusses liquid, solid and semisolid media, and differentiates between simple, complex, enriched, selective, differential, indicator and transport media. Specific examples of commonly used media are provided for each type, along with their compositions and uses. The document aims to outline the basic requirements, classifications and characteristics of different culture media.
I. This presentation discusses three bacterial identification tests: the catalase test, urease test, and triple sugar iron (TSI) test.
II. It provides details on the principles, procedures, and results of the urease test and TSI test. The urease test identifies Proteus bacteria based on their production of urease enzymes, while the TSI test distinguishes between Enterobacteriaceae based on their sugar fermentation patterns.
III. The presentation is delivered by five group members and discusses the applications and limitations of the TSI test for bacterial identification.
The document discusses methods for identifying bacteria, including phenotypic, immunological, and genetic techniques. Phenotypic methods examine bacterial morphology, staining characteristics, and biochemical reactions. Tests like Gram staining, colony morphology, and catalase can provide initial identification. Further tests of carbohydrate use, enzyme production, and sensitivity to inhibitors allow identification to the species level. Immunological methods detect bacterial antigens, while genetic techniques like PCR and nucleic acid analysis provide accurate identification by examining microbial DNA. Both traditional and molecular methods are used to fully characterize unknown bacteria.
The indole test is used to differentiate bacteria based on their ability to produce indole from the amino acid tryptophan. Bacteria containing the enzyme tryptophanase can break down tryptophan into indole, ammonia, and pyruvic acid. The test involves inoculating tryptophan broth with a bacterial sample and incubating. Kovac's reagent is then added, which will produce a red color in the presence of indole, indicating a positive result. Examples of bacteria that test positive include Klebsiella oxytoca and Proteus species, while Salmonella, Pseudomonas, and Yersinia species typically test negative.
This document discusses the classification, diagnosis, and laboratory identification of fungal infections. It covers topics such as the classification of fungi based on morphology and disease, common oral fungal infections, specimen collection guidelines, and various staining and culture techniques used to identify fungi under the microscope or in culture media. These include wet mount preparations with potassium hydroxide or India ink, differential stains like Gram or Gomori's silver, and fungal culture media like Sabouraud dextrose agar or birdseed agar.
Sugar fermentation tests, Cetrimide agar medium, Hugh Leifson medium Shivam kumar Sriwas
1. The document discusses sugar fermentation, how to test for it using media like phenol red carbohydrate broth, and the interpretation of results.
2. Hugh Leifson medium and Cetrimide agar are described as media used to differentiate bacterial metabolism and isolate Pseudomonas aeruginosa, respectively.
3. Key components, principles, preparation, and expected results are outlined for both Hugh Leifson medium and Cetrimide agar tests.
A simple lecture for the description of the various culture media used for isolation of different bacteria in a pure form for further identification procedures.
Cultivation of bacteria and culture methodsAshfaq Ahmad
Cultivation of bacteria allows for the isolation, growth, and study of microorganisms. There are various culture methods and media that support the growth of bacteria. Liquid broths and solid agar plates can be used with different nutrient formulations to selectively grow specific bacteria. Streaking, lawning, stabbing, and pour plating are common culture techniques used to isolate pure colonies for analysis. Specialized enriched, selective, differential, and transport media help optimize bacterial growth and identification.
This lecture discusses laboratory methods for determining antibiotic susceptibility, including disk diffusion tests, minimum inhibitory concentration (MIC) tests, and minimum bactericidal concentration (MBC) tests. Filter paper disks containing different antibiotic concentrations are applied to bacterial lawns to measure susceptibility. The MIC is the lowest concentration that inhibits growth, while the MBC kills bacteria. These methods help evaluate antibiotic effectiveness and monitor emerging resistance.
1. There are two main types of bacterial counts - total bacterial count and total viable count. Total bacterial count includes both living and dead cells while total viable count only measures living cells.
2. Bacterial enumeration is important for comparing growth under different conditions, and in industries like dairy, food, and water microbiology.
3. Methods of enumeration include direct counting using microscopy or Coulter counter, and indirect counting of viable cells using serial dilution plating or membrane filtration. Other methods determine cell mass through dry weight, nitrogen content, or turbidity measurements.
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.
Bacterial culture involves growing bacterial colonies in artificial media containing necessary nutrients and under suitable conditions. Ideal culture media support satisfactory, rapid, and reproducible bacterial growth while revealing bacterial characteristics. Common media components include water, agar, peptones, and extracts. Media are classified by consistency (liquid, semi-solid, solid), constituents (simple, complex, synthetic), or special properties (enriched, selective, differential). Sterilization methods include autoclaving or hot air oven.
This document discusses clinical microbiology and methods used in the field. It provides definitions of clinical microbiology as the study of bacteria and their relation to medicine. Key methods discussed include culture-based techniques of growing bacteria in controlled conditions to isolate, identify, and determine antibiotic sensitivity. Additional non-culture methods described are use of monoclonal antibodies, enzyme immunoassays, probes, polymerase chain reaction (PCR), and ligase chain reaction.
The document provides instructions for performing several microbiological tests, including sugar fermentation tests, indole production, methyl red, Voges-Proskauer, citrate utilization, nitrate reduction, urease, triple sugar iron, oxidase, catalase, amylase, and lipase. It explains the principles, expected results, and interpretations for each test. The tests can be used to differentiate bacterial species and determine their metabolic abilities.
The CAMP test identifies Streptococcus agalactiae (Group B Strep) and Listeria species by their production of a substance called the CAMP factor. The CAMP factor acts synergistically with Staphylococcus aureus to induce enhanced hemolysis, appearing as an arrowhead shape. Discovered in 1944 by Christie, Atkins, Munch, and Petersen, the CAMP test remains an important method for identifying S. agalactiae and Listeria monocytogenes.
The document provides information on differential leukocyte counting (DLC). It defines DLC as the relative proportion of different leukocytes expressed as a percentage. The main types of white blood cells (WBCs) described are granulocytes (neutrophils, eosinophils, basophils) and agranulocytes/mononuclear cells (lymphocytes, monocytes). Pathologic variations in DLC counts include neutrophilia, neutropenia, lymphocytosis, lymphopenia, monocytosis, eosinophilia, eosinopenia, and basophilia. Causes of each variation are outlined. Automated methods for differential counting are also discussed and compared to manual microscopic methods
The TSI test is used to determine carbohydrate fermentation and hydrogen sulfide production in bacteria. It contains glucose, sucrose, and lactose sugars, along with peptones and phenol red pH indicator. Carbohydrate fermentation produces acid, changing the color from red to yellow, while peptone metabolism makes it more alkaline. Hydrogen sulfide production is shown by black precipitate. Results are interpreted based on color changes in the slant and butt portions, indicating which sugars are fermented and if H2S is produced.
The Ziehl-Neelsen stain is used to detect acid-fast bacteria like mycobacteria. It works by using carbolfuchsin as a primary stain that is retained in acid-fast bacteria even after treatment with an acid decolorizer due to their high lipid and mycolic acid content. Non-acid fast cells are then counterstained blue while acid-fast bacteria appear red. The stain involves heating slides coated with carbolfuchsin, treating with an acid decolorizer like sulfuric acid, and counterstaining with methylene blue to identify acid-fast mycobacteria under the microscope.
culture media
CULTURE – Is term given to microorganisms that are cultivated in the lab for the purpose of studying them.
MEDIUM – Is the term given to the combination of ingredients that will support the growth & cultivation of microorganisms outside their natural habitats.
Necessary Requirements for Growth of Bacteria
Distilled Water
Nitrogen containing compounds
Peptone- Golden granular powder
Complex mixture of partially digested protiens by proteolytic
enzymes pepsin, trysin or papain
Peptones, Proteoses, polypeptides, aminoacids, inorganic salts like phosphates
potassium & magnesium
Accessory growth factors like nicotinic acid & riboflavin
Energy sources
Suitable Ph- 7.2 – 7.4
Solidifying agents:
Gelatin– Protien
Agar— Chief component is Long chain Polysaccharide
Melts at 95°c & solidify only when cooled to about 42°c
1- 2% yields a suitable gel eg. Non-nutritive agar
According to Physical State:
Liquid – Peptone Water, Nutrient Broth
Semisolid – Nutrient Agar Stabs
Solid – Blood Agar
According to Oxygen requirement:
Aerobic Medium
Anaerobic Media
- Definition
- Uses of culture media
- Basic composition of culture media
- Types of culture media
--Based on physical state
----solid medium
----semi solid medium
----liquid medium
--Based on ingredients
----Simple or basal medium.
----Complex medium.
----Synthetic or defined medium.
----Semisynthetic medium.
--Special medium
----Enriched media
----Enrichment media
----Selective media
----Differential media
----Indicator media
----Transport media
----Anaerobic media
-Media preparation
-Culture method
--Streak culture
--Lawn culture
-references
This document summarizes culture methods used to grow bacteria. Culture media is used to isolate bacteria from infected material. Originally, Louis Pasteur used urine or meat broth as liquid media while Robert Koch used cooked cut potato as a solid medium. Agar was later introduced as a gelling agent for solid media. Various culture methods are described including streak, pour plate, and stab cultures. Biochemical tests like oxidase and indole are used to further identify bacteria grown on culture media.
Culture is the term for microorganisms grown in the lab, and medium provides nutrients for their growth. Culture media are used to isolate, study, and identify bacteria. Media are classified by consistency (liquid, solid, semisolid) and oxygen requirements. Different media, like nutrient agar or blood agar, support growth of various microbes. Selective and differential media help isolate specific bacteria. Culture media must provide appropriate nutrients, pH, and be sterilized before use to isolate bacteria and study their properties.
Culture media are used to support the growth of microorganisms. They can be liquid, semisolid, or solid and contain various nutrients like water, electrolytes, peptone, agar, meat extract, and blood or serum. Culture media are classified based on consistency and purpose of use. Common types include basal media, enriched media, selective media, differential media, transport media, and anaerobic and blood culture media. Automated systems now allow continuous monitoring of blood cultures for faster detection of microbial growth compared to conventional methods.
Culture media are used to grow microorganisms outside the body. Agar is commonly used to solidify culture media into a solid or semi-solid state. Culture media contain nutrients to support microbial growth and are classified based on physical state, ingredients, and purpose. Blood agar and chocolate agar contain blood to enrich growth of fastidious bacteria. Selective media like MacConkey agar inhibit some bacteria to isolate target microbes. Transport media preserve delicate microbes during specimen transport for testing.
Culture media are used to support the growth of microorganisms. They can be liquid, solid, or semisolid depending on their consistency. Culture media contain various nutrients like peptone, agar, and electrolytes. They are classified based on their oxygen requirement, nutrition type, and special characteristics. Examples of types of culture media include enriched, selective, differential, anaerobic, and blood culture media. Automated blood culture systems allow for continuous monitoring and faster detection of microbial growth compared to conventional methods.
Culture media are used to support the growth of microorganisms. There are different types of culture media based on consistency (liquid, solid, semisolid) and special characteristics (enriched, selective, differential). Common constituents include water, electrolytes, peptone, agar. Culture media can be classified based on consistency, oxygen requirement, nutrition level and special characteristics. Examples of specialized media include blood agar, chocolate agar, Lowenstein-Jensen medium and Thiosulfate Citrate Bile salts Sucrose agar. Automated systems like BacT/ALERT can rapidly detect microbial growth in blood cultures.
Bacteria must be cultured to study their properties and identify them. Different culture media and methods are used depending on the purpose. Agar is commonly used to make solid media for growing isolated colonies. Media can be simple, complex, or selective depending on nutritional needs. Streaking is used to isolate pure cultures from clinical specimens. Pour plating quantifies viable bacteria while lawns are used for antibiotic testing. Transport media maintain delicate bacteria during specimen travel.
Bacteria must be cultured to study their properties and identify them. Different culture media and methods are used depending on the purpose. Agar is commonly used to make solid media for growing isolated colonies. Media can be simple, complex, or selective depending on ingredients. Streaking is used to isolate bacteria from clinical specimens onto solid media. Pour plating and liquid cultures are used for quantitative analysis and transporting samples respectively. Proper culture technique allows growth and study of bacteria.
1. The document discusses various culture media used for cultivating microorganisms from clinical specimens. It describes the composition and purpose of different types of media including basal media, enriched media, selective media, and transport media.
2. Specific media are described for cultivating gram positive cocci like Staphylococci and Streptococci. Mannitol salt agar, tellurite glycine agar, and DNase test agar are discussed for isolating and identifying Staphylococcus. Todd Hewitt broth and crystal violet blood agar are mentioned for Streptococci.
3. Modified Thayer Martin medium and modified New York City medium are highlighted as selective media used for isolating Neisseria
This document discusses culture media and culture methods used to grow bacteria. It describes the various types of culture media including solid, liquid, and semi-solid media made with ingredients like agar. Different media are used for specific purposes such as enrichment, selection, and indication of bacterial properties. Culture methods like streaking, lawning, stabbing, and pouring are explained for isolating pure cultures of bacteria and studying their characteristics. Liquid cultures are also used for certain applications.
This document provides an overview of culture media and culture methods used in microbiology. It discusses the different types of culture media based on consistency (solid, liquid, semi-solid), ingredients (simple, complex, synthetic), and oxygen requirements (aerobic, anaerobic). Common media like nutrient agar, blood agar and selective media are described. The preparation of culture media and methods like streak plating, pour plating, and stab culturing are explained. The aim is to learn about culture media, demonstrate preparation techniques, and different culturing methods for isolating and growing bacteria.
Culture media are used to grow bacteria for identification and study. They contain nutrients like carbon, nitrogen, water and growth factors. Common ingredients include meat extract and peptone. Media can be solid, liquid, or semi-solid depending on whether agar is added. Special media like selective, differential and enrichment media allow isolation of specific bacteria. Proper preparation, sterilization and storage of media is required for pure cultures.
Culture Techniques and Maintenance of selected culturesMiracleLivinus1
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This document provides information on culture media and methods used for growing and identifying bacteria. It discusses the various types of culture media including solid, liquid, and semi-solid media as well as specialized media like selective, differential and indicator media. Specific media like blood agar, MacConkey agar and triple sugar iron agar are described. Culture methods like streak plating, pour plating, and stab and stroke cultures are outlined. The document also covers biochemical tests performed using culture media, such as oxidase, indole, citrate, and urease tests to identify bacterial properties. Finally, techniques for culturing anaerobic bacteria in an oxygen-free environment are summarized.
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3. Describe the components of a normal electrocardiogram of a typical bipolar lead (limb II)
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Study Resources:
1. Chapter 11, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. Chapter 3, Cardiology Explained, https://www.ncbi.nlm.nih.gov/books/NBK2214/
7. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
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Overall life span (LS) was 1671.7±1721.6 days and cumulative 5YS reached 62.4%, 10 years – 50.4%, 20 years – 44.6%. 94 LCP lived more than 5 years without cancer (LS=2958.6±1723.6 days), 22 – more than 10 years (LS=5571±1841.8 days). 67 LCP died because of LC (LS=471.9±344 days). AT significantly improved 5YS (68% vs. 53.7%) (P=0.028 by log-rank test). Cox modeling displayed that 5YS of LCP significantly depended on: N0-N12, T3-4, blood cell circuit, cell ratio factors (ratio between cancer cells-CC and blood cells subpopulations), LC cell dynamics, recalcification time, heparin tolerance, prothrombin index, protein, AT, procedure type (P=0.000-0.031). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and N0-12 (rank=1), thrombocytes/CC (rank=2), segmented neutrophils/CC (3), eosinophils/CC (4), erythrocytes/CC (5), healthy cells/CC (6), lymphocytes/CC (7), stick neutrophils/CC (8), leucocytes/CC (9), monocytes/CC (10). Correct prediction of 5YS was 100% by neural networks computing (error=0.000; area under ROC curve=1.0).
TEST BANK For Community Health Nursing A Canadian Perspective, 5th Edition by...Donc Test
TEST BANK For Community Health Nursing A Canadian Perspective, 5th Edition by Stamler, Verified Chapters 1 - 33, Complete Newest Version Community Health Nursing A Canadian Perspective, 5th Edition by Stamler, Verified Chapters 1 - 33, Complete Newest Version Community Health Nursing A Canadian Perspective, 5th Edition by Stamler Community Health Nursing A Canadian Perspective, 5th Edition TEST BANK by Stamler Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Pdf Chapters Download Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Pdf Download Stuvia Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Study Guide Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Ebook Download Stuvia Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Questions and Answers Quizlet Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Studocu Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Quizlet Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Stuvia Community Health Nursing A Canadian Perspective, 5th Edition Pdf Chapters Download Community Health Nursing A Canadian Perspective, 5th Edition Pdf Download Course Hero Community Health Nursing A Canadian Perspective, 5th Edition Answers Quizlet Community Health Nursing A Canadian Perspective, 5th Edition Ebook Download Course hero Community Health Nursing A Canadian Perspective, 5th Edition Questions and Answers Community Health Nursing A Canadian Perspective, 5th Edition Studocu Community Health Nursing A Canadian Perspective, 5th Edition Quizlet Community Health Nursing A Canadian Perspective, 5th Edition Stuvia Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Pdf Chapters Download Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Pdf Download Stuvia Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Study Guide Questions and Answers Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Ebook Download Stuvia Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Questions Quizlet Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Studocu Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Quizlet Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Stuvia
Cell Therapy Expansion and Challenges in Autoimmune DiseaseHealth Advances
There is increasing confidence that cell therapies will soon play a role in the treatment of autoimmune disorders, but the extent of this impact remains to be seen. Early readouts on autologous CAR-Ts in lupus are encouraging, but manufacturing and cost limitations are likely to restrict access to highly refractory patients. Allogeneic CAR-Ts have the potential to broaden access to earlier lines of treatment due to their inherent cost benefits, however they will need to demonstrate comparable or improved efficacy to established modalities.
In addition to infrastructure and capacity constraints, CAR-Ts face a very different risk-benefit dynamic in autoimmune compared to oncology, highlighting the need for tolerable therapies with low adverse event risk. CAR-NK and Treg-based therapies are also being developed in certain autoimmune disorders and may demonstrate favorable safety profiles. Several novel non-cell therapies such as bispecific antibodies, nanobodies, and RNAi drugs, may also offer future alternative competitive solutions with variable value propositions.
Widespread adoption of cell therapies will not only require strong efficacy and safety data, but also adapted pricing and access strategies. At oncology-based price points, CAR-Ts are unlikely to achieve broad market access in autoimmune disorders, with eligible patient populations that are potentially orders of magnitude greater than the number of currently addressable cancer patients. Developers have made strides towards reducing cell therapy COGS while improving manufacturing efficiency, but payors will inevitably restrict access until more sustainable pricing is achieved.
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2. Index
Introduction
Definition
History
Classification
Preparation, sterilization and testing of media
Simple media, Enriched media and Enrichment media
Selective and Differential media
Transport media
Biochemical media
Fungal, Parasitic and Viral media
Anaerobic media
Disposal of media
Non culturable microorganisms
3. Introduction
Microorganisms have to be grown (cultured)for them to be
identified.
Cultivation is growing microorganisms in culture by taking
bacteria from infection site by specimen collection and
growing them in artificial environment.
Culture is the most common diagnostic method used for
detection of bacterial infections.
6. A.Based on consistency
Liquid media or broth:no agar
Semi solid media:0.5%agar
Solid media:2%agar
7. B.Based on the method of growth detection
Conventional culture media
- Simple or basal media
- Enriched media
- Enrichment broth
- Selective media
- Differential media
- Transport media
- Anaerobic media
Automated culture media
8. Preparation of culture media
Weigh the dehydrated media in required quantity.
Dissolve in purified/distilled water as per the manufacturer instruction
Heat to boiling to dissolve the medium completely
Check the pH of the media by using narrow range pH paper
Compare the colour of the pH against the chart provided.
9. Sterilization of culture media
• The following methods are used to sterilize culture media:
>Autoclaving
>Steaming at 100⁰C
>Inspissation
>Filtration
10. Dispensing sterile media into petri dishes
Pour plates under sterile conditions within the hood
Lay out the sterile petri dishes on clean surfaces
Name of the medium and date of preparation should be mentioned on the plate.
Media should not be warmer than 60⁰C when poured
Mix the medium gently by rotating the flask,avoid air bubbles
Hold the plug of flask in such a position that it does not get contaminated
Flame sterilize the neck of the flask
Pour 15-20ml of medium into each disc(90-100mm diameter)
11. Storage of culture media
>Stored at dark place
>At 2-8⁰C
Testing of culture media
>Sterility testing-
• Done for routine media to which sheep blood or other
substances are added after autoclaving.
• 5% batch incubated at 35-37⁰C overnight.
• All media to be checked visually before using.
12. Name of the medium Controls
Sheep blood agar Alpha hemolysis -Streptococcus viridans
Beta hemolysis -Streptococcus pyogens
Gamma hemolysis -Enterococcus spp
Chocolate agar Alpha hemolysis -Streptococcus
pneumoniae
Nutrient agar Pigmented strain of Pseudomonas
MacConkey agar LF –Escherichia coli
NLF-Pseudomonas
TCBS(Thiosulfate-citrate-bile
salts-sucrose agar)
Sucrose fermenting-Vibrio cholerae
Wilson Blair H2S producing-Salmonella typhi
H2S non producing-Salmonella para typhi A
Alkaline peptone water Positive control-Vibrio cholerae
Negative control-Escherichia coli
>Performance testing
13. Name of the medium Controls
Selenite F broth Positive control-Salmonella
Negative control-Escherichia coli
Potassium tellurite agar Black coloured colonies-Corynebacterium
diphtheriae
CLED(Cystine-Lactose-
Electrolyte-Deficient agar)
Staphylococcus aureus-yellow colonies
Escherichia coli-yellow colonies
Pseudomonas-green colonies
Triple sugar iron agar Acid/acid , with/without gas-Escherichia coli
Acid/acid,H2S-Proteus
Alkaline/no change-Pseudomonas
Simmon’s citrate medium Positive control-Klebsiella Pneumoniae
Negative control-Escherichia coli
Christensen’s urea medium Positive control-Proteus spp
Negative control-Escherichia coli
Bile esculin agar Positive control-Enterococcus species
Negative control-Streptococcus viridans
Cetrimide agar Positive control-Pseudomonas aeruginosa
Negative control-Escherichia coli
14. Simple/basal media:
They contain minimum ingredients that support the growth of non
fastidious bacteria.
Name of media Components Primary purpose
Peptone water Peptone(1%)NACI,(0.5%)water >Used as growth medium
>Base for carbohydrate
media
>Used for indole test
Nutrient broth Peptone water , meat
extract(1%)
Isolation of non-fastidious
organisms
Nutrient agar Nutrient broth,2%agar Isolation of non-fastidious
organisms
Semi-solid agar Nutrient broth,0.2 to 0.5%agar Determination of bacterial
motility
16. Enriched media:
Substances like blood , serum , egg are added to the simple/basal
medium to support growth of fastidious bacteria.
Enriched media components Primary purpose
Blood agar 5-10%sheep blood added to
molten NA at 45⁰C- 55⁰C
Isolation of Staphylococcus ,
Streptococcus.
Chocolate agar 5-10%sheep blood added to
molten NA at 72⁰C
Hemoglobin , hemin(X factor)
and Co-enzyme nicotinamide
adenine dinucleotide(NAD or V
factor)
Isolation of Streptococcus
pneumonae , Neisseria gonorrhoeae
, Hemophilus spp.
20. Name of the media Components Primary purpose
Serum containing enriched
medium
• Loeffler serum slope
Nutrient broth, glucose,
ox/sheep/horse serum
Isolation of
Corynebacterium
diphtheriae.
Egg containing enriched
medium
• Lowenstein Jensen
medium
• Dorset egg medium
>Potato flour, L-Asparagine ,
Monopotassium phosphate ,
Magnesium citrate ,
Malachite green , glycerol ,
Egg suspension , distilled
water
>Nutrient broth , whole
fresh eggs.
>Isolation of Mycobacterium
tuberculosis and Atypical
mycobacteria .
>Isolation of
Corynebacterium
diphtheriae.
Buffered charcoal yeast
extract agar
Yeast extract, agar, charcoal
and salts supplemented with
L-cysteine , HCL , ferric
pyrophosphate and alpha-
ketoglutarate.
>Enrichment media for
Legionella spp.
>Supports growth of
Nocardia.
21. Loefflers serum slope Lowenstein-Jensen medium Dorset egg agar Buffered charcoal yeast
extract agar
22. Enrichment media:
They are liquid media added with some inhibitory
agents which selectively allow certain organism to
grow and inhibit others.
23. Enrichment media components Primary purpose
Selenite F broth Peptone-base broth ; sodium
selenite toxic for most
Enterobacteriaceae
Isolation of Samonella species
Tetrathionate broth Peptone-base broth;iodine and
potassium iodide,bile salts and
sodium thiosulfate inhibit Gram
positive organisms and
Enterobacteriaceae
Isolation of Salmonella and
Shigella spp.
Alkaline peptone water Peptone ,sodium
chloride,distilled water.
Isolation of Vibrio cholerae
Gram negative broth Peptone-base broth with glucose
and mannitol;sodium citrate and
sodium deoxycholate act as
inhibitory agents
Isolation of Enteric pathogens
26. NAME OF THE
MEDIUM
CONSTITUENTS USE
MacConkey
Agar
Peptone base with lactose
Bile salts (Sodium
taurocholate)
Indicator: Neutral Red
SELECTIVE - Gram
Negative bacilli
DIFFERENTIAL - Lactose
fermentation
CLED
(Cystine lysine
electrolyte deficient
agar)
Peptone base with lactose,
tryptone, L-cystine
Indicator: Bromothymol
blue
DIFFERENTIAL - Lactose
fermentation
(mostly for bacteria from
the urinary tract)
EMB AGAR
(Eosin Methylene
Blue)
Peptone base with lactose
Indicators: Eosin Y and
Methylene blue
DIFFERENTIAL - Lactose
fermentation
(for Enteric bacilli)
28. NAME OF THE
MEDIUM
CONSTITUENTS USE
BEA
(Bile Esculin Agar)
Nutrient agar base with ferric
citrate
Brown colour of the medium is
due to hydrolysis of esculin
Indicator: Sodium
Desoxycholate (Bile salt)
DIFFERENTIAL - Isolation and
presumptive identification of
group D streptococci and
enterococci
POTASSIUM
TELLURITE
AGAR
Sheep blood agar base with
potassium tellurite
SELECTIVE: Corynebacterium
diphtheriae
(Black coloured colonies)
TCBS AGAR
(Thiosulfate citrate
bile sucrose)
Peptone base agar with yeast
extract, bile salts, citrate,
sucrose, ferric citrate and
sodium thiosulfate
Indicator: Bromothymol blue
SELECTIVE: Vibrio spp
(Yellow coloured colonies)
DIFFERENTIAL: Sucrose
fermentation
30. NAME OF THE MEDIUM CONSTITUENTS USE
SS AGAR
(Salmonella Shigella Agar)
Peptone base with lactose, ferric citrate and
sodium citrate
Inhibitors: Brilliant green and bile salts
Indicator: Neutral red
SELECTIVE - Salmonella and some Shigella spp
WILSON BLAIR
AGAR
Peptone agar base with beef extract,
dextrose, disodium phosphate and Bismuth
sulphite
Inhibitor: Brilliant green dye
Indicators: Ferrous sulphate
SELECTIVE – Isolation and preliminary
identification of Salmonella spp from clinical
specimens
DIFFERENTIAL – Dextrose fermentation
DCA
(Deoxycholate citrate agar)
Peptone base agar with lactose, sodium
citrate, sodium thiosulfate, ferric citrate
Inhibitor: Sodium deoxycholate (Inhibits Gram
positive organisms)
Indicator: Neutral Red
DIFFERENTIAL - Salmonella and Shigella spp from
other Gram negative enteric bacilli
XLD AGAR
(Xylose Lysine
Deoxycholate)
Yeast extract agar with lysine, xylose, lactose,
sucrose and ferric ammonium citrate
Inhibitor: Sodium deoxycholate (Inhibits Gram
positive organisms)
Indicator: Phenol Red
DIFFERENTIAL - Salmonella and Shigella spp from
other Gram negative enteric bacilli
HE AGAR
(Hektoen Enteric Agar)
Peptone base agar with bile salts, lactose,
sucrose, salicin and ferric ammonium citrate
Indicators: Bromothymol blue and acid
fuschin
SELECTIVE - Salmonella and some Shigella spp
DIFFERENTIAL - Salmonella and Shigella spp from
other Gram negative enteric bacilli
34. NAME OF THE
MEDIUM
CONSTITUENTS USE
BORDET-
GENGOU AGAR
Potato-glycerol-based medium enriched with 15 to
20% defibrinated blood
Inhibitor: Methicillin (2.5 microgram/mL)
SELECTIVE – Bordetella pertussis and
Bordetella parapertussis
REGAN LOWE
AGAR
Charcoal agar with horse blood, cephalexin and
amphotericin B
ENRICHMENT and SELECTIVE –
Isolation of Bordetella pertussis
THAYER MARTIN
AGAR
Blood agar base enriched with haemoglobin and
supplement B
Inhibitors: Colistin, Nystatin, Vancomycin,
Trimethoprim
SELECTIVE – N. gonorrhoeae and N.
meningitidis
NYC AGAR
Peptone agar base with cornstarch supplemented with
yeast dialysate, 3% hemoglobin and horse plasma
Antibiotic supplements include:
1. Vancomycin (2 microgram/mL)
2. Colistin (5.5 microgram/mL)
3. Amphoterecin B (1.2 microgram/mL)
4. Trimethoprim (3 microgram/mL)
SELECTIVE – Neisseria gonorrhoeae
Also supports growth of Ureaplasma
urealyticum and some Mycoplasma
spp.
37. NAME OF THE
MEDIUM
CONSTITUENTS USE
CAMPY
BLOOD
AGAR
Brucella agar base with sheep blood and the
following:
Vancomycin(10 mg/L)
Trimethoprim(5 mg/L)
Polymyxin B(2500 U/L)
Amphoterecin B(2 mg/L)
Cephalothin(15 mg/L)
SELECTIVE – Campylobacter
spp.
CVA AGAR
(Cefoperazone,
Vancomycin,
Amphoterecin meduim)
Blood supplemented with the 3 drugs to inhibit
growth of most Gram-negative bacteria, Gram-
positive bacteria and yeast, respectively.
SELECTIVE – Campylobacter
spp
SKIRROW AGAR
Peptone and soy protein base agar with lysed horse
blood
Inhibitors:
1. Vancomycin – Gram positive organisms
2. Polymyxin B and Trimethoprim – most Gram
negative organisms
SELECTIVE – Campylobacter
spp
39. NAME OF THE
MEDIUM
CONSTITUENTS USE
BURKHOLDERIA
CEPACIA SELECTIVE
AGAR
Bile salts
Gentamicin
Ticarcillin
Polymyxin B
Peptone
Yeast extract
SELECTIVE - Burkholderia cepacia
(Cystic fibrosis patients)
CNA AGAR
(Columbia Colistin-
Nalidixic acid agar)
Columbia agar base with
10 mg/L colistin
15 mg/L Nalidixic acid
5% sheep blood
SELECTIVE - Isolation of Gram-
positive cocci
CIN AGAR
(Cefsulodin-Irgasan-
Novobiocin agar)
Peptone base with yeast extract, mannitol
and bile salts
Inhibitors: drugs
Indicator: Neutral red and Crystal violet
SELECTIVE - Isolation of Yersinia spp
and Aeromonas spp
DNase agar
Tryptose, Deoxyribonucleic acid, Sodium
Chloride, Agar
DIFFERENTIAL – Deoxyribonuclease
enzyme producing property
Eg. Serratia marcescens
41. NAME OF THE
MEDIUM
CONSTITUENTS USE
MANNITOL SALT
AGAR
Peptone base, mannitol
Inhibitor: 7.5% concentrated salt
Indicator: Phenol red
SELECTIVE DIFFERENTIATION -
Staphylococci
PEA
(Phenylethyl alcohol
agar)
Nutrient agar base
Inhibitor: Phenylmethanol
(inhibits Gram negative
organisms)
SELECTIVE - Isolation of:
Aerobic Gram positive
cocci and bacilli
Anaerobic Gram positive
cocci and negative bacilli
SSA
(Streptococcal selective
agar)
5% sheep blood agar base with
colistin and trimethoprim-
sulfamethoxazole
Indicator: Crystal violet
SELECTIVE - Streptococcus
pyogenes and Streptococcus
agalactiae
45. MILK AGAR: NON-SELECTIVE SOLID MEDIUM
CONSTITUENTS:
Peptone agar base with yeast extract and milk solids
USE:
Identification of organisms that can hydrolyse casein eg. Streptomyces,
Pseudomonas and Actinomadura.
For differentiation of aerobic actinomycetes based on casein proteolysis.
It is used to demonstrate the pigment producing properties of certain
organisms.
46. MUELLER HINTON AGAR
Non-selective, non-differential medium
Constituents: Beef Extract, Acid Hydrolysate of Casein, Starch and Agar.
Uses
1. Routine susceptibility testing of non-fastidious microorganisms by the Kirby-Bauer disk
diffusion technique.
2. It can be used to cultivate Neisseria
3. It is specified in FDA Bacteriological Analytical Manual for food testing, and procedures
commonly performed on aerobic and facultative anaerobic bacteria.
48. NAME OF THE
MEDIUM
CONSTITUENTS USE
CARY-BLAIR
Soft agar semisolid medium with
sodium thioglycolate, disodium
hydrogen phosphate, NaCl and
CaCl2
To transport enteric pathogens
Eg. Shigella, Salmonella, Vibrio
cholerae and E. coli
For detection of Campylobacter spp
from faeces (when transport time <
2 hours)
STUART’S
Soft agar medium with sodium
thioglycolate (mercaptoacetate),
sodium glycerophosphate, CaCl2,
Distilled water and methylene blue
To maintain viability of gonococci on
swabs during their transmission.
Also for Shigella and E. coli
AMIES
Soft agar medium with sodium
thioglycolate (mercaptoacetate),
NaCl, KCl, CaCl2, MgCl2, Na2PO4,
Potassium dihydrogen phosphate,
CHARCOAL (finely powdered),
distilled water
To maintain viability of gonococci on
swabs during their transmission.
Also for Shigella and E. coli
50. NAME OF THE
MEDIUM
CONSTITUENTS USE
PIKE’S
Blood agar containing
1 in 10 lakh Crystal violet
1 in 16000 Sodium azide
Distributed as for stab cultures
in tubes or bottles
Preservation of Streptococcus
pyogenes, pneumococci and
Haemophilus influenzae in
nose and throat swabs
SACH’S
BUFFERED
GLYCEROL
SALINE
Distilled water and glycerol with
NaCl, Disodium hydrogen
phosphate, potassium
dihydrogen phosphate and
phenol red
Preservation and transport
of Salmonella and Shigella
species and Escherichia coli in
facial specimens. It is not
suitable for Vibrio cholera,
Campylobacter species
or Yersinia enterocolitica.
53. FERMENTATION TEST MEDIA
Constituents:
1. Nutrient medium base: Peptone water, serum peptone water and serum agar
2. Carbohydrate or related compound under test (SUGARS):
MONOSACCHARIDES Pentoses (Arabinose/ Xylose/ Rhamnose)
Hexoses (Glucose/ Fructose/ Mannose/ Sorbose/ Galactose)
DISACCHARIDES Sucrose/ inulin/ Dextrin/ Glycogen
POLYHYDRIC
ALCOHOLS
Glycerol/ Erythritol/ Adonitol/ Mannitol/ Dulcitol/ Sorbitol/ Inositol
GLYCOSIDES Salicin/ Coniferin/ Aesculin
ORGANIC ACIDS Tartrate(Dextro/ Laevo/ Meso) / Citrate/ Mucate/ Gluconate/ Malonate
3. Suitable Indicator:
ANDRADE’S INDICATOR (NaOH with acid fuchsin till colour becomes yellow)
BROMOCRESOL PURPLE
PHENOL RED
BROMOTHYMOL BLUE
4. A small inverted tube (DURHAM TUBE) completely filled with liquid and
containing no air bubbles is usually included in each culture tube or bottle to
detect gas.
54. TSI (Triple Sugar Iron): DIFFERENTIAL MEDIUM
CONSTITUENTS: Agar slant of special medium with Phenol Red (pH
sensitive dye), 1% lactose, 1% sucrose, 0.1% glucose, sodium
thiosulfate and ferrous sulfate.
Carbohydrate fermentation is indicated by the production of gas and a
change in the color of the pH indicator from red to yellow.
USES:
•Differentiate members of the Enterobacteriaceae family from other
gram-negative rods
•Differentiation among Enterobacteriaceae on the basis of their sugar
fermentation patterns
55. INTERPRETATION OF TSI:
An alkaline/acid (red slant/yellow butt) reaction: Indicative of dextrose fermentation only
An acid/acid (yellow slant/yellow butt) reaction: Fermentation of dextrose, lactose and/or
sucrose
An alkaline/alkaline (red slant, red butt) reaction: Absence of carbohydrate fermentation
Blackening of the medium: production of hydrogen sulphide
Gas production: Bubbles or cracks (formation of CO2 and H2)
56. OXIDATIVE FERMENTIVE MEDIA
A semi-solid tubed medium containing the carbohydrate along with a pH
indicator.
Constituents: Peptone base agar with NaCl, Dipotassium hydrogen phosphate,
distilled water and bromothymol blue.
Fermenting organisms: Enterobacteriaceae, Aeromonas, Vibrio
Oxidizing organisms: Pseudomonas
Non-fermenting organisms: Alcaligenes faecalis
57. DECAROXYLASE MEDIUM
Constituents: Decarboxylase test medium base (Peptone, meat extract,
glucose, pyridoxal, bromocresol blue, cresol red and distilled water)
The media is divided into four equal parts. One part is tubed without the
addition of any amino acid, and the tube is labeled as ‘Control’. The remaining
three parts are dispensed onto three tubes, to which L-lysine hydrochloride,
L-arginine hydrochloride, and L-ornithine hydrochloride are added separately.
Uses:
•Decarboxylase test is used to differentiate the members of the
Enterobacteriaceae family with closely related physiological characteristics on
the basis of their ability to produce the enzyme decarboxylase
59. NAME OF THE
MEDIUM
CONSTITUENTS USE
MR
(METHYL RED)
Ingredients per liter of
deionized water:
buffered peptone= 7.0 gm
glucose= 5.0 gm
dipotassium phosphate
Positive Reaction: A distinct red color
Examples: E. coli, Yersinia sps, etc.
Negative Reaction: A yellow color
Examples: Enterobacter aerogenes, Klebsiella
pneumoniae, etc
INDOLE
KOVACS
MEDIUM
P-dimethylaminobenzaldehyde
Hydrochloric acid (37%)
Amyl alcohol
Positive: Formation of a pink to red color (“cherry-
red ring”) in the reagent layer on top of the medium
within seconds of adding the reagent.
Examples: Escherichia coli, Haemophilus
influenzae, Klebsiella oxytoca, Proteus sp., Enterococcus
faecalis, and Vibrio sp.
Negative: No color change
Eg most Haemophilus sp.,
most Klebsiella sp., Neisseria sp., Proteus mirabilis, P.
penneri, Pseudomonas sp.,Salmonella sp., Serratia sp., Yer
sinia sp
60.
61. NAME OF THE
MEDIUM
CONSTITUENTS USE
VP
(VOGES-
PROSKAUER)
VP REAGENT A: BARRITT’S
REAGENT A
Alpha naphthol 5%
Absolute alcohol
VP REAGENT B: BARITT’S
REAGENT B
Potassium hydroxide
Deionized water
Positive Reaction: A pink-red color at the
surface
Examples: Viridans group streptococci,
Enterobacter, Klebsiella, Serratia
marcescens, Vibrio eltor.
Negative Reaction: A lack of a pink-red
color
Examples: Streptococcus mitis, Citrobacter
sp., Shigella, Yersinia, Salmonella, Vibrio
parahaemolyticus
62.
63. NAME OF THE MEDIUM CONSTITUENTS USE
SIMMONS’
CITRATE
MEDIUM
Koser’s medium with agar and
Bromothymol blue as the indicator.
Koser’s medium includes:
NaCl, MgSO4, Ammonium dihydrogen
phosphate, potassium dihydrogen
phosphate and sodium citrate
Positive Reaction: Growth with color change
from green to intense blue along the slant.
Examples: Salmonella, Edwardsiella,
Citrobacter, Klebsiella, Enterobacter,
Serratia, Providencia, etc.
Negative Reaction: No growth and No color
change; Slant remains green.
Examples: Escherichia, Shigella,
Morganella, Yersinia etc.
CHRISTENSEN’S
UREASE
MEDIUM
Peptone agar base with NaCl, dipotassium
hydrogen phosphate, Glucose (10%), Urea
(20%)
Indicator: Phenol red
to distinguish urease-positive
Enterobacteriaceae.
can also be used to differentiate between
yeasts : Candida albicans (negative urease)
and Cryptococcus neoformans (rapid
production of urease).
detect the presence of Helicobacter pylori
65. NAME OF THE
MEDIUM
CONSTITUENTS USE
MALONATE
AGAR
Distilled water with yeast extract,
ammonium sulphate,
dipotassium hydrogen phosphate,
potassium dihydrogen phosphate,
NaCl and sodium malonate
Indicator: Bromothyol Blue
Differentiate among Enterobacteriaceae:
Klebsiella pneumoniae is positive (blue at
24 hours), Escherichia coli is negative(cult
ure remains green).
Also used to separate Salmonella arizonae
(positive) from other Salmonella spp(neg
ative).
Differentiate Enterobacteriaceae in food
and dairy products.
CETRIMIDE
AGAR
Distilled water, agar, glycerine,
MgCl2, Potassium sulfate,
pancreatic digest of gelatin,
cetyltrimethylammonium
bromide
selective medium for the isolation
of Pseudomonas aeruginosa
used for determining the ability of an
organism to produce fluorescein and
pyocyanin (Antibiotica)
66.
67. NAME OF THE
MEDIUM
CONSTITUENTS USE
HYDROLYSIS OF
TRIBUTYRIN
Peptone agar base with
water, yeast extract and
Tributyrin
Lipolytic organisms
remove the opacity by
converting the fat to
water soluble butyric
acid