ISOLATION AND IDENTIFICATION OF NLF BACTERIA IN VARIOUS SAMPLES.Daisy Saini
IDENTIFICATION AND ISOLATION OF NON-LACTOSE FEREMNTING BACTERIA IN VARIOUS CLINICAL SAMPLES IN A TERTIARY HOSPITAL IN INDIA, INCLUDE BIOCHEMICAL TEST BASE ON THEIR ENZYMATIC ACTIVITY AND GRAPHICAL PRESENTAION OF THEIR DISTRIBUTION ACCORDING TO SEX RATION , AGE GROUP, SAMPLE AND THEIR PROFILE.
This document discusses Staphylococcus aureus and methods for its laboratory diagnosis. It notes that S. aureus is commonly found in the nose and can cause infections, while S. epidermidis lives on the skin and S. saprophyticus in the vagina. Diagnosis involves collecting samples from infections and using gram staining, culturing, and biochemical tests like catalase, coagulase and novobiocin to identify the species. Rapid PCR tests are also now used to diagnose S. aureus.
Practical 40 biochemical test microbiology.pdfssuser668f10
This document outlines various methods used to identify unknown bacterial species, including microscopic examination, culture-based techniques, biochemical tests, serological identification, and molecular methods. Microscopic examination involves staining bacterial smears to determine cell morphology and arrangement. Cultural characteristics like colony morphology, pigmentation, and hemolysis are also examined. Biochemical tests identify metabolic pathways and enzyme production. Serological tests use antibodies to identify species. Molecular methods like PCR and DNA probes allow for rapid and accurate identification compared to older phenotypic techniques. A combination of these methods is usually required to definitively identify an unknown bacterial species.
This document discusses urine specimen collection and processing for microbiological examination. Urine is collected to detect disorders like urinary tract infections. Common microorganisms found in urine include E. coli, Proteus, and Candida species. Mid-stream clean-catch urine is the preferred specimen type. Urine is cultured on agar plates like CLED, blood, and MacConkey to isolate pathogens and test for lactose fermentation. Bacterial colonies are counted and used to interpret urine culture results. Isolated organisms are identified using biochemical tests and antibiotic sensitivity testing.
The document describes several biochemical tests used to identify bacteria, including the catalase test, coagulase test, DNase test, oxidase test, indole test, urease test, citrate test, and triple sugar iron (TSI) test. It provides the purpose, principle, method, and results for each test. It also discusses how to specifically identify Staphylococcus aureus and Streptococcus pyogenes through microscopic examination, culture characteristics, and biochemical properties.
This document summarizes characteristics of the Salmonella genus, including pathogenic species such as Salmonella typhi and Salmonella paratyphia. It describes Salmonella's morphology, optimal growth conditions, virulence factors, mechanisms of pathogenesis, and diseases caused such as gastroenteritis and typhoid fever. The document also outlines methods for laboratory diagnosis of Salmonella including culture-based identification and biochemical testing, as well as treatments including antibiotics and preventive vaccination measures.
Medical Microbiology Laboratory (Enterobacteriaceae - IV)Hussein Al-tameemi
This document discusses Shigella, a genus of bacteria that can cause shigellosis (bacillary dysentery). It belongs to the Enterobacteriaceae family. Shigella are gram-negative, non-spore forming bacilli that are facultative anaerobes. They are identified through their appearance on various culture media like MacConkey's agar and biochemical tests. Common species include S. boydii, S. dysenteriae, S. flexneri, and S. sonnei. Shigella infection is transmitted through the fecal-oral route and causes diarrhea with blood and mucus in stool. Diagnosis involves examining stool samples microscopically and through culture
Biochemical tests for bacterial identificationSuprakash Das
Basic biochemical tests for identification of most common bacteria along with their principles and methods to perform and quality control for UG & PG Students.
ISOLATION AND IDENTIFICATION OF NLF BACTERIA IN VARIOUS SAMPLES.Daisy Saini
IDENTIFICATION AND ISOLATION OF NON-LACTOSE FEREMNTING BACTERIA IN VARIOUS CLINICAL SAMPLES IN A TERTIARY HOSPITAL IN INDIA, INCLUDE BIOCHEMICAL TEST BASE ON THEIR ENZYMATIC ACTIVITY AND GRAPHICAL PRESENTAION OF THEIR DISTRIBUTION ACCORDING TO SEX RATION , AGE GROUP, SAMPLE AND THEIR PROFILE.
This document discusses Staphylococcus aureus and methods for its laboratory diagnosis. It notes that S. aureus is commonly found in the nose and can cause infections, while S. epidermidis lives on the skin and S. saprophyticus in the vagina. Diagnosis involves collecting samples from infections and using gram staining, culturing, and biochemical tests like catalase, coagulase and novobiocin to identify the species. Rapid PCR tests are also now used to diagnose S. aureus.
Practical 40 biochemical test microbiology.pdfssuser668f10
This document outlines various methods used to identify unknown bacterial species, including microscopic examination, culture-based techniques, biochemical tests, serological identification, and molecular methods. Microscopic examination involves staining bacterial smears to determine cell morphology and arrangement. Cultural characteristics like colony morphology, pigmentation, and hemolysis are also examined. Biochemical tests identify metabolic pathways and enzyme production. Serological tests use antibodies to identify species. Molecular methods like PCR and DNA probes allow for rapid and accurate identification compared to older phenotypic techniques. A combination of these methods is usually required to definitively identify an unknown bacterial species.
This document discusses urine specimen collection and processing for microbiological examination. Urine is collected to detect disorders like urinary tract infections. Common microorganisms found in urine include E. coli, Proteus, and Candida species. Mid-stream clean-catch urine is the preferred specimen type. Urine is cultured on agar plates like CLED, blood, and MacConkey to isolate pathogens and test for lactose fermentation. Bacterial colonies are counted and used to interpret urine culture results. Isolated organisms are identified using biochemical tests and antibiotic sensitivity testing.
The document describes several biochemical tests used to identify bacteria, including the catalase test, coagulase test, DNase test, oxidase test, indole test, urease test, citrate test, and triple sugar iron (TSI) test. It provides the purpose, principle, method, and results for each test. It also discusses how to specifically identify Staphylococcus aureus and Streptococcus pyogenes through microscopic examination, culture characteristics, and biochemical properties.
This document summarizes characteristics of the Salmonella genus, including pathogenic species such as Salmonella typhi and Salmonella paratyphia. It describes Salmonella's morphology, optimal growth conditions, virulence factors, mechanisms of pathogenesis, and diseases caused such as gastroenteritis and typhoid fever. The document also outlines methods for laboratory diagnosis of Salmonella including culture-based identification and biochemical testing, as well as treatments including antibiotics and preventive vaccination measures.
Medical Microbiology Laboratory (Enterobacteriaceae - IV)Hussein Al-tameemi
This document discusses Shigella, a genus of bacteria that can cause shigellosis (bacillary dysentery). It belongs to the Enterobacteriaceae family. Shigella are gram-negative, non-spore forming bacilli that are facultative anaerobes. They are identified through their appearance on various culture media like MacConkey's agar and biochemical tests. Common species include S. boydii, S. dysenteriae, S. flexneri, and S. sonnei. Shigella infection is transmitted through the fecal-oral route and causes diarrhea with blood and mucus in stool. Diagnosis involves examining stool samples microscopically and through culture
Biochemical tests for bacterial identificationSuprakash Das
Basic biochemical tests for identification of most common bacteria along with their principles and methods to perform and quality control for UG & PG Students.
Streptococcus and Staphylococcus are common bacterial genera that can cause infections. The document outlines the steps for laboratory diagnosis of infections caused by these bacteria, including specimen collection, culture, staining, and biochemical and antimicrobial testing. Gram staining reveals Gram-positive cocci arranged in clusters for Staphylococcus or chains for Streptococcus. Culture on blood agar shows hemolytic patterns. Biochemical tests help identify pathogenic species like S. aureus and S. pyogenes. Antibiotic susceptibility testing is also important for treatment.
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 document summarizes key information about Staphylococcus, including that it is a genus of gram-positive spherical bacteria that form grape-like clusters. It notes the two main types are coagulase-positive Staphylococcus aureus and coagulase-negative species like S. epidermidis and S. saprophyticus. It provides details on how to differentiate between these types based on laboratory tests like coagulase, catalase, hemolysis, mannitol fermentation and novobiocin sensitivity. Finally, it lists some common diseases that can be caused by Staphylococcus like pyogenic infections, food poisoning and toxic shock syndrome.
This document discusses various methods for enumerating and testing for microorganisms in food. It describes total plate counts, coliform tests, and tests for mesophilic bacteria, staphylococci, Salmonella, Shigella, and other pathogenic bacteria. Specific procedures are outlined, including enrichment, plating, screening, and confirmation steps. A variety of media are used, such as violet red bile agar, Baird-Parker agar, triple sugar iron agar, and lysine iron agar. Colonies are examined for characteristics like color, zone formation, and biochemical reactions to identify microorganisms.
The document discusses laboratory diagnosis of Staphylococcus and Streptococcus bacteria. Key tests include gram stain, catalase test, coagulase test, mannitol salt agar, and novobiocin disc test to differentiate medically important staphylococcal species. For streptococci, hemolytic patterns on blood agar, optochin and bacitracin disc tests are used. Common pathogens include S. aureus, S. epidermidis, S. saprophyticus, S. pyogenes, S. agalactiae. Diseases range from skin infections to bacteremia, with S. aureus a major cause of infections like osteomyelitis, pneumonia and infect
Microbiology of the GIT , Abdallh dwayat.pdf8y4xzv9rqd
1. The document discusses Gram-negative rods related to the enteric tract, including E. coli, Shigella, Vibrio cholerae, and others.
2. It covers the major clinical manifestations of enteric infections like diarrhea, vomiting, and abdominal pain.
3. Laboratory diagnosis of enteric bacteria involves using differential and selective media like MacConkey agar and Eosin methylene blue agar to differentiate lactose fermenters from non-fermenters. Tests like triple sugar iron agar also help identify bacteria.
The document describes experiments performed to identify an unknown bacterial culture. A Gram stain showed the bacteria were Gram-positive cocci arranged in grape-like clusters. Further tests like catalase, oxidase, and growth on various media plates identified the bacteria as Staphylococcus aureus.
L13 medical microbiology laboratory (staphylococcus spp.)Hussein Al-tameemi
This document provides an overview of Staphylococcus bacteria, including taxonomy, characteristics, infections caused, and laboratory identification. It describes the Staphylococcus genus as including over 40 species of gram positive cocci that are an important cause of infections ranging from minor to life-threatening. Key aspects covered include morphology, colonial appearance on different media, biochemical tests for identification of S. aureus and differentiation of coagulase-negative staphylococci.
Staphylococcus and Streptococcus are important gram-positive bacteria. Staphylococcus includes pathogens like S. aureus and S. epidermidis. S. aureus causes skin infections like boils and toxic shock syndrome. S. epidermidis causes UTIs and endocarditis. Streptococcus includes S. pyogenes, S. agalactiae, and S. pneumoniae. S. pyogenes causes strep throat and rheumatic fever. S. agalactiae causes infections in newborns. S. pneumoniae causes pneumonia and ear infections. Identification involves tests like catalase, coagulase, and hemolytic properties on blood agar. Antibiotics like pen
This document summarizes laboratory methods for diagnosing and characterizing Staphylococcus aureus infections. Specimens are collected from infected sites and examined microscopically and through culture-based and biochemical tests to identify S. aureus. Microscopic examination reveals gram-positive cocci arranged in clusters. On culture media, S. aureus forms large, golden-yellow colonies that cause beta-hemolysis. Biochemical tests such as catalase and coagulase are used to distinguish S. aureus from other staphylococci.
Staphylococci and Streptococci organisms.pptvinuthdp
Staphylococci and streptococci are common bacteria that can cause infections in humans. Staphylococcus aureus is a pathogenic staphylococcus that can cause infections on the skin or deeper tissues. It grows aerobically at 37°C and pH 7.4-7.6 on nutrient agar, blood agar, and mannitol salt agar. Virulence factors include toxins and enzymes like hyaluronidase which breaks down hyaluronic acid. Treatment involves antibiotics like cloxacillin or vancomycin for methicillin-resistant strains.
Streptococci are classified by Lancefield grouping based on cell wall carbohydrates and by hemolysis on blood agar
Since Staphylococcus nepalensis were reported for the first time from Nepalese animal specimen, and have been reported from human specimens elsewhere, this bug can be a threat in our part. Protocols must be designed aimed at their identification in our laboratory during microbiological analysis of clinical specimens.
1. The document describes the isolation and biochemical characterization of Staphylococcus aureus from various sweet samples collected from different areas.
2. Methods used include isolating bacteria on nutrient agar plates, morphological and gram staining identification, and biochemical tests to identify S. aureus. Antibiotic sensitivity testing was also performed.
3. Results found S. aureus present in samples, which was identified as gram positive cocci in clusters. Biochemical tests confirmed presence and some isolates were found to be resistant to certain antibiotics like oxacillin and penicillin.
This document discusses the classification and identification of streptococci bacteria. It describes their morphology, culture characteristics, biochemical reactions, Lancefield grouping, and clinical significance. Key streptococci groups discussed include Group A (Streptococcus pyogenes), Group B (Streptococcus agalactiae), Streptococcus pneumoniae, and Enterococcus species. Identification is based on colony appearance on blood agar, Gram staining, and biochemical tests like catalase and bile solubility.
Medical Microbiology Laboratory (Enterobacteriaceae - III)Hussein Al-tameemi
This document provides information about Enterobacteriaceae and Salmonella species. It discusses the taxonomy and classification of Enterobacteriaceae, describing their characteristics. Salmonella is highlighted as an important genus. The document outlines methods for culturing and identifying Salmonella from clinical specimens such as blood and stool. Biochemical tests and serological identification methods like the Widal test are also summarized.
This document discusses three bacterial genera: Citrobacter, Edwardsiella, and Arizonae (Salmonella enterica serovar Arizona).
Citrobacter species are uncommon causes of opportunistic infections and can be found in the normal gut flora. C. freundii has been associated with neonatal meningitis. Edwardsiella tarda is an opportunistic pathogen typically transmitted by ingesting contaminated water, and can cause gastrointestinal or extraintestinal infections. Salmonella Arizonae is a rare human pathogen within the Salmonella family, typically transmitted through contaminated dairy or eggs.
The document discusses various biochemical tests used to identify bacteria based on their reactions. It describes tests like catalase, oxidase, indole, citrate utilization, urea hydrolysis, and triple sugar iron that are used to differentiate bacteria based on their ability to break down substrates or produce certain enzymes. These tests help identify both gram-positive and gram-negative bacteria at the genus or species level based on whether their reactions are positive or negative in the various biochemical tests.
This document provides an overview of common biochemical tests used to differentiate bacterial species, including the catalase test, coagulase test, DNase test, bile solubility test, and litmus milk decolorization test. For each test, the document describes the principle, required materials, procedure, expected results, and positive and negative control organisms. The overall purpose is to teach medical laboratory science students how to perform and interpret the results of various biochemical tests to identify unknown bacterial isolates.
This document discusses urine culture procedures including media used, common microorganisms found in urine infections, specimen collection and culture techniques. It provides details on interpreting culture results and identifying bacteria based on colony morphology on blood, MacConkey and CLED agars. Biochemical tests like SIM, citrate, urease and oxidase are described to differentiate bacteria like E. coli, Klebsiella, Proteus and Pseudomonas. Susceptibility testing is then performed on clinically significant isolates to determine appropriate antibiotic treatment.
Lab diagnosis of diarrhoea involves collecting stool or rectal swab samples and transporting them to the lab within 2 hours using Cary Blair medium. Samples are analyzed through microscopy, culture, biochemical tests and serotyping to identify bacteria like E. coli, Salmonella, Staphylococcus aureus, and Vibrio that can cause diarrhoea based on their characteristics like colony morphology, gram staining, motility, and biochemical reactions. Epidemiological typing through bacteriophage is also done to trace the source of infection for some bacteria.
Streptococcus and Staphylococcus are common bacterial genera that can cause infections. The document outlines the steps for laboratory diagnosis of infections caused by these bacteria, including specimen collection, culture, staining, and biochemical and antimicrobial testing. Gram staining reveals Gram-positive cocci arranged in clusters for Staphylococcus or chains for Streptococcus. Culture on blood agar shows hemolytic patterns. Biochemical tests help identify pathogenic species like S. aureus and S. pyogenes. Antibiotic susceptibility testing is also important for treatment.
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 document summarizes key information about Staphylococcus, including that it is a genus of gram-positive spherical bacteria that form grape-like clusters. It notes the two main types are coagulase-positive Staphylococcus aureus and coagulase-negative species like S. epidermidis and S. saprophyticus. It provides details on how to differentiate between these types based on laboratory tests like coagulase, catalase, hemolysis, mannitol fermentation and novobiocin sensitivity. Finally, it lists some common diseases that can be caused by Staphylococcus like pyogenic infections, food poisoning and toxic shock syndrome.
This document discusses various methods for enumerating and testing for microorganisms in food. It describes total plate counts, coliform tests, and tests for mesophilic bacteria, staphylococci, Salmonella, Shigella, and other pathogenic bacteria. Specific procedures are outlined, including enrichment, plating, screening, and confirmation steps. A variety of media are used, such as violet red bile agar, Baird-Parker agar, triple sugar iron agar, and lysine iron agar. Colonies are examined for characteristics like color, zone formation, and biochemical reactions to identify microorganisms.
The document discusses laboratory diagnosis of Staphylococcus and Streptococcus bacteria. Key tests include gram stain, catalase test, coagulase test, mannitol salt agar, and novobiocin disc test to differentiate medically important staphylococcal species. For streptococci, hemolytic patterns on blood agar, optochin and bacitracin disc tests are used. Common pathogens include S. aureus, S. epidermidis, S. saprophyticus, S. pyogenes, S. agalactiae. Diseases range from skin infections to bacteremia, with S. aureus a major cause of infections like osteomyelitis, pneumonia and infect
Microbiology of the GIT , Abdallh dwayat.pdf8y4xzv9rqd
1. The document discusses Gram-negative rods related to the enteric tract, including E. coli, Shigella, Vibrio cholerae, and others.
2. It covers the major clinical manifestations of enteric infections like diarrhea, vomiting, and abdominal pain.
3. Laboratory diagnosis of enteric bacteria involves using differential and selective media like MacConkey agar and Eosin methylene blue agar to differentiate lactose fermenters from non-fermenters. Tests like triple sugar iron agar also help identify bacteria.
The document describes experiments performed to identify an unknown bacterial culture. A Gram stain showed the bacteria were Gram-positive cocci arranged in grape-like clusters. Further tests like catalase, oxidase, and growth on various media plates identified the bacteria as Staphylococcus aureus.
L13 medical microbiology laboratory (staphylococcus spp.)Hussein Al-tameemi
This document provides an overview of Staphylococcus bacteria, including taxonomy, characteristics, infections caused, and laboratory identification. It describes the Staphylococcus genus as including over 40 species of gram positive cocci that are an important cause of infections ranging from minor to life-threatening. Key aspects covered include morphology, colonial appearance on different media, biochemical tests for identification of S. aureus and differentiation of coagulase-negative staphylococci.
Staphylococcus and Streptococcus are important gram-positive bacteria. Staphylococcus includes pathogens like S. aureus and S. epidermidis. S. aureus causes skin infections like boils and toxic shock syndrome. S. epidermidis causes UTIs and endocarditis. Streptococcus includes S. pyogenes, S. agalactiae, and S. pneumoniae. S. pyogenes causes strep throat and rheumatic fever. S. agalactiae causes infections in newborns. S. pneumoniae causes pneumonia and ear infections. Identification involves tests like catalase, coagulase, and hemolytic properties on blood agar. Antibiotics like pen
This document summarizes laboratory methods for diagnosing and characterizing Staphylococcus aureus infections. Specimens are collected from infected sites and examined microscopically and through culture-based and biochemical tests to identify S. aureus. Microscopic examination reveals gram-positive cocci arranged in clusters. On culture media, S. aureus forms large, golden-yellow colonies that cause beta-hemolysis. Biochemical tests such as catalase and coagulase are used to distinguish S. aureus from other staphylococci.
Staphylococci and Streptococci organisms.pptvinuthdp
Staphylococci and streptococci are common bacteria that can cause infections in humans. Staphylococcus aureus is a pathogenic staphylococcus that can cause infections on the skin or deeper tissues. It grows aerobically at 37°C and pH 7.4-7.6 on nutrient agar, blood agar, and mannitol salt agar. Virulence factors include toxins and enzymes like hyaluronidase which breaks down hyaluronic acid. Treatment involves antibiotics like cloxacillin or vancomycin for methicillin-resistant strains.
Streptococci are classified by Lancefield grouping based on cell wall carbohydrates and by hemolysis on blood agar
Since Staphylococcus nepalensis were reported for the first time from Nepalese animal specimen, and have been reported from human specimens elsewhere, this bug can be a threat in our part. Protocols must be designed aimed at their identification in our laboratory during microbiological analysis of clinical specimens.
1. The document describes the isolation and biochemical characterization of Staphylococcus aureus from various sweet samples collected from different areas.
2. Methods used include isolating bacteria on nutrient agar plates, morphological and gram staining identification, and biochemical tests to identify S. aureus. Antibiotic sensitivity testing was also performed.
3. Results found S. aureus present in samples, which was identified as gram positive cocci in clusters. Biochemical tests confirmed presence and some isolates were found to be resistant to certain antibiotics like oxacillin and penicillin.
This document discusses the classification and identification of streptococci bacteria. It describes their morphology, culture characteristics, biochemical reactions, Lancefield grouping, and clinical significance. Key streptococci groups discussed include Group A (Streptococcus pyogenes), Group B (Streptococcus agalactiae), Streptococcus pneumoniae, and Enterococcus species. Identification is based on colony appearance on blood agar, Gram staining, and biochemical tests like catalase and bile solubility.
Medical Microbiology Laboratory (Enterobacteriaceae - III)Hussein Al-tameemi
This document provides information about Enterobacteriaceae and Salmonella species. It discusses the taxonomy and classification of Enterobacteriaceae, describing their characteristics. Salmonella is highlighted as an important genus. The document outlines methods for culturing and identifying Salmonella from clinical specimens such as blood and stool. Biochemical tests and serological identification methods like the Widal test are also summarized.
This document discusses three bacterial genera: Citrobacter, Edwardsiella, and Arizonae (Salmonella enterica serovar Arizona).
Citrobacter species are uncommon causes of opportunistic infections and can be found in the normal gut flora. C. freundii has been associated with neonatal meningitis. Edwardsiella tarda is an opportunistic pathogen typically transmitted by ingesting contaminated water, and can cause gastrointestinal or extraintestinal infections. Salmonella Arizonae is a rare human pathogen within the Salmonella family, typically transmitted through contaminated dairy or eggs.
The document discusses various biochemical tests used to identify bacteria based on their reactions. It describes tests like catalase, oxidase, indole, citrate utilization, urea hydrolysis, and triple sugar iron that are used to differentiate bacteria based on their ability to break down substrates or produce certain enzymes. These tests help identify both gram-positive and gram-negative bacteria at the genus or species level based on whether their reactions are positive or negative in the various biochemical tests.
This document provides an overview of common biochemical tests used to differentiate bacterial species, including the catalase test, coagulase test, DNase test, bile solubility test, and litmus milk decolorization test. For each test, the document describes the principle, required materials, procedure, expected results, and positive and negative control organisms. The overall purpose is to teach medical laboratory science students how to perform and interpret the results of various biochemical tests to identify unknown bacterial isolates.
This document discusses urine culture procedures including media used, common microorganisms found in urine infections, specimen collection and culture techniques. It provides details on interpreting culture results and identifying bacteria based on colony morphology on blood, MacConkey and CLED agars. Biochemical tests like SIM, citrate, urease and oxidase are described to differentiate bacteria like E. coli, Klebsiella, Proteus and Pseudomonas. Susceptibility testing is then performed on clinically significant isolates to determine appropriate antibiotic treatment.
Lab diagnosis of diarrhoea involves collecting stool or rectal swab samples and transporting them to the lab within 2 hours using Cary Blair medium. Samples are analyzed through microscopy, culture, biochemical tests and serotyping to identify bacteria like E. coli, Salmonella, Staphylococcus aureus, and Vibrio that can cause diarrhoea based on their characteristics like colony morphology, gram staining, motility, and biochemical reactions. Epidemiological typing through bacteriophage is also done to trace the source of infection for some bacteria.
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1. Salmonella, Shigella & Proteus
By SANDEEP KUMAR
Master of Science in Medical Laboratory Science
Delhi Skill and Entrepreneurship University, Dwarka Sec-9, New Delhi
4. Salmonella
Introduction:
1. Infective Dose: High infective dose (102 – 105 Bacilli)
2. Morphology: Salmonella is a rod-shaped bacterium (bacillus). Unlike
other strains of bacilli, salmonella does not produce spores. On
MacConkey agar, salmonella colonies appear colorless and transparent,
sometimes with dark centers.
3. Diseases: Salmonella causes two diseases in humans: Enteric fever
(typhoid) & Gastroenteritis, Doctors refer to both diseases as
“salmonellosis.”
4. Significance: Salmonella is a common foodborne pathogen. It causes
zoonotic infections (transmitted between animals and humans).
5. History: Named after American veterinary surgeon Daniel Elmer Salmon.
5. Salmonella
Morphology:
• Gram Negative Bacilli (Rod Shaped).
• Size - 2-4 X 0.6 μm. (length X diameter)
• Non Spore forming bacterium
• Non Capsulated bacteria
• Motile bacteria – Have peritrichous flagella which help in movements.
6. Salmonella
Culture Characteristics:
• Aerobic and facultatively anaerobic
• Salmonellae Growth Conditions: Optimum temperature: 37°C (range: 15–41°C),
pH range: 6-8
• MacConkey’s Agar and Deoxycholate Citrate Agar (DCA): Colonies - Colorless due
to non-lactose fermentation (NLF)
• Wilson and Blair Bismuth Sulphite Medium: Colonies: Jet black with metallic
sheen (due to hydrogen sulphide production), Exception: S. paratyphi A and
other non-H2S-producing species form green colonies
• Xylose Lysine Deoxycholate (XLD) Agar: Colonies - Red with black centers (due to
H2S production) , H2S-negative Salmonella serotypes: Red colonies without black
centers.
• Enrichment Broths (Selenite F and Tetrathionate): Used for inoculating
specimens, especially feces
7. Salmonella
Laboratory Diagnosis:
Duration of illness Specimen, Test done
1st Weak Culture of Blood
Culture of Bone marrow aspirate
Culture of Duodenal aspirate
2nd & 3rd Weak Serum
For Antibody (Ab) detection by Widal.
For Antigen (Ag) detection.
Stool & Urine Culture
4th Weak Stool & Urine Culture
Carriers Stool & Urine Culture
Serum for detection of Abs to Vi Ag.
Sewage culture – Indirect way
8. Salmonella Laboratory Diagnosis: Continued ………………
• Specimen: Blood, Serum, Stool and urine, Bone marrow aspirate,
duodenal aspirate, sewage sample (Indirect Test).
• Gram Staining: Gram Negative (-ve) Bacilli, red color.
• Motility Test: By hanging drop method, Salmonella are motile with
peritrichous flagella.
• Culture Isolation
1. Blood & Bone marrow culture: (in the first week of illness):
in blood culture bottle (BHI broth, automated BACTEC)
9. Salmonella Laboratory Diagnosis: Continued ………………
2. Stool culture (in 3-4 weeks of illness):
• Enrichment broth - Selenite F broth, tetrathionate broth and gram-negative(-ve)
broth
• Low selective media - e.g. MacConkey Agar Media - Non-lactose fermenting
(NLF) do not ferment lactose, produces a colorless or pale colony, translucent
colony.
• Highly selective medium:
3. Urine culture (in 3-4 weeks of illness) - done on MacConkey agar
Agar Media Appearance
Deoxycholate Citrate Agar (DCA) NLF pale colonies with black center.
Xylose Lysine Deoxycholate (XLD agar) Red colonies with black center.
Shigella-Salmonella agar (SSA) Colorless colonies with black center.
Hektoen Enteric Agar (HEA) Blue Green colonies with black center.
Wilson Blair's bismuth sulphite medium jet black colonies
10. Salmonella Laboratory Diagnosis: Continued ………………
SS Agar - Colorless colonies with
black center
XLD Agar - Red colonies with black
center.
DCA - NLF pale colonies with black
center.
Hektoen Enteric Agar (HEA) - Blue
Green colonies with black center. Wilson Blair's bismuth sulphite
medium - jet black colonies
MacConkey Agar Media - colorless or
pale colony, translucent colony.
11. Salmonella Laboratory Diagnosis: Continued ………………
1. Catalase Test: Positive for catalase, Indicates the production of catalase enzyme.
2. Oxidase Test: Negative for oxidase., No cytochrome c oxidase production.
3. Nitrate Reduction: Nitrate reduced to nitrite, Presence of nitrate reductase enzyme.
4. Indole Test Negative for indole production, No indole from tryptophan. (ICUT)
5. Citrate Utilization Test (TSl): Variable result for citrate utilization, May or may not utilize
citrate as a carbon source.
6. Urease Test: Negative for urease production.
7. TSI (Triple Sugar Iron) Test: K/A (alkaline/acid) result, Gas production (except in
Salmonella Typhi), H2S production (positive in Salmonella Typhi, absent in Salmonella
Paratyphi A, abundant in Salmonella Paratyphi B).
Biochemical Test
12. Salmonella Laboratory Diagnosis: Continued ………………
• Serum Antibody Detection (Widal Test):
- Detects antibodies against O and H antigens.
- Specific antibodies for different infections.
• Typhi Dot (Rapid Test): Based on Immunochromatography (or ICT,
lateral flow assay), detect IgM & IgG.
• Antigen Detection: Serum and urine antigen detection by ELISA and
CIEP.
• Molecular Methods: PCR detecting specific genes.
• Nonspecific Findings: Neutropenia (reduced white blood cell count).
• Antimicrobial Susceptibility Testing: Determines antibiotic sensitivity.
16. Shigella
Introduction:
1. Infective Dose: low infective dose (10 Bacilli)
2. Distribution: Shigella is primarily found in the human intestinal tract.
It can also be present in contaminated environments and organic
matter.
3. Colony Appearance: Shigella forms small, smooth, moist, and
creamy colonies on culture media. Notably, it exhibits a characteristic
“swarming” ability.
4. Infection Site: Shigella primarily infects the gastrointestinal tract,
leading to severe inflammation and dysentery.
17. Shigella
Morphology:
• Gram Negative Bacilli (Rod Shaped).
• Size - 1-3 µm X 0.5 µm (length X diameter).
• Non-Spore forming bacterium.
• Non Capsulated bacteria.
• Non Motile bacteria – It Does not have flagella for movements
18. Shigella
Culture Characteristics:
1. Growth Conditions:
• Aerobes and facultative anaerobes
• Optimal temperature: 37°C & pH: 7.4
• Tolerate temperature range: 10°C to 40°C
2. Colony Characteristics:
• After overnight incubation: 2 mm diameter, circular, convex, smooth,
translucent.
• MacConkey’s agar and DCA: Colourless (non-lactose fermenting), except
Sh. sonnei (pink).
• XLD agar: Shigella colonies appear red without black centers, while H2S-
producing bacteria have black centers in colonies.
19. Shigella Culture Characteristics: Continued ………
3. Selective Media:
Salmonella-Shigella (SS) agar: Contains bile salts, sodium citrate, sodium
thiosulphate, ferric citrate, lactose, and neutral red.
- Shigella: Colourless (non-lactose fermenting), no blackening.
- Salmonella: Colourless with black centers.
Hektoen-enteric (HE) agar: Contains bile salts, lactose, sucrose, sodium
thiosulphate, ferric ammonium citrate, acid fuchsin, and thymol blue
- Shigella: Green (fading to periphery)
- Salmonella: Blue-green with black centers
20. Shigella
Laboratory Diagnosis:
1. Specimens:
- Fresh stool collection is ideal.
- Rectal swabs are not recommended.
- Direct swab of an ulcer under sigmoidoscopic examination is preferred.
2. Transport:
- Transport specimens immediately.
- Use Sach's buffered glycerol saline (pH 7.0-7.4) if delay is inevitable.
- Avoid alkaline transport media for vibrios (inhibitory for shigella).
3. Direct Microscopy:
- Saline and iodine preparation reveals pus cells, erythrocytes, and macrophages.
- Excludes parasitic causes of dysentery.
21. Shigella Laboratory Diagnosis: Continued ………
4. Culture:
- Inoculate specimen on selective media (e.g., MacConkey's agar, DCA, XLD
agar).
- Use Selenite F broth (0.4%) for enrichment.
- Subculture on solid media from Selenite F broth.
- Incubate at 37°C for 24 hours.
5. Colony Morphology and Staining:
- Colorless (NLF) colonies on MacConkey's agar. [Non-Lactose fermenting
(do not ferment lactose)].
6. Gram staining - Shigellae are Gram-negative
7. hanging drop preparation - Shigellae are non-motile.
22. Shigella Laboratory Diagnosis: Continued ………
8. Biochemical Reactions:
- Confirm non-motile bacillus (urease, citrate, H₂S, KCN negative) with
biochemical tests.
9. Slide Agglutination Test:
- Confirm shigella with polyvalent and monovalent antisera.
- Use type-specific antisera for agglutination (subgroups A, B, C).
10. Colicin Typing:
- Done for subgroup D (Sh. sonnei) strains.
23. Shigella Laboratory Diagnosis: Continued ………
XLD agar: Shigella
colonies appear red
without black centers,
SS Agar Shigella:
Colourless (non-
lactose fermenting),
no blackening
MacConkey’s agar:
Colourless (non-lactose
fermenting), SS
25. Proteus
Introduction:
1. Distribution and Habitat: Proteus bacteria are widely distributed in nature. They
occur as normal intestinal flora in humans. Found in decomposing animal matter,
sewage, manure soil, the mammalian intestine, and human and animal feces.
2. Culture Colony Appearance: Proteus bacteria exhibit a characteristic “swarming”
behaviour. This allows them to migrate across surfaces. They tend to have an
ammonia smell.
3. Infection Sites: Proteus bacteria are opportunistic pathogens. Commonly
responsible for urinary and septic infections, often nosocomial (hospital-acquired).
Associated with infection-induced renal stones in the urinary tract.
26. Proteus
Morphology:
• Gram Negative Bacilli (rod-shaped).
• Size - 1.0–3.0 μm in length and 0.4 to 0.8 μm in diameter.
• Non-Spore forming bacterium.
• Non Capsulated bacteria.
• Motile bacteria – Have peritrichous flagella which help in movements.
• Pleomorphism – It can change in shape, sometimes present as coccobacilli
and sometimes in long filamentous form based on environmental
conditions.
27. Proteus
Culture Characteristics:
• Aerobic & facultative anaerobe
• Optimum temperature for growth – 37° C,
• Optimal pH for growth is around 7 (neutral).
• Odour – Putrefactive odor (Fruity and seminal smell).
• On nutrient agar media; (NAM) or blood agar media (BAM) , Pr.
vulgaris and Pr. Mirabilis exhibit swarming behavior. Swarming does
not occur with Pr. Morganella and Pr. Providencia species.
• The exact cause of swarming is not fully established but appears
related to vigorous bacterial motility.
28. Proteus Culture Characteristics: Continued………..
• Inhibition Methods: Methods are used to inhibit swarming.
1. Increase agar concentration to 6% (instead of 1-2%).
2. Addition - chloral hydrate (1:500), sodium azide (1:500), or boric
acid (1:1000) in the medium.
3. On MacConkey’s agar, Proteus forms smooth, pale, or colorless
(NLF) colonies without swarming.
4. In liquid medium (peptone water), Proteus produces uniform
turbidity with a slight powdery deposit and an ammonical odor.
29. Proteus Culture Characteristics: Continued………..
Proteus Swarming Colony on Blood
Agar
Proteus Colony on MacConkey Agar
Proteus Translucent blue colonies
Colony on CLED
Proteus Swarming Colony on Blood
Agar
30. Proteus
Laboratory Diagnosis:
1. Specimens:
- Midstream urine sample in UTI.
- Pus in pyogenic lesions.
2. Collection:
- Collect specimen in a sterile container under aseptic conditions.
- Transport immediately.
3. Culture:
- Culture on MacConkey agar or blood agar with 6% agar to inhibit
swarming.
- Incubate at 37°C for 18-24 hours.
- Non-lactose fermenting (NLF) colonies seen on MacConkey agar.
- Inoculate peptone water.
31. Proteus Laboratory Diagnosis Continued………..
4. Gram's Staining: Gram-negative bacilli, non-capsulated and non-
sporing.
5. Hanging Drop Preparation:- Observe actively motile bacilli.
6. Biochemical Reactions: Key tests - PPA (positive in all Proteus
species) and urease (positive in all Proteus species except Providencia).
Other biochemical reactions can differentiate various species.
7. Agglutination Test:- Confirm strain using polyvalent group-specific
sera.
8. Antibiotic Susceptibility Test:- Important due to Proteus bacilli's
resistance to common antibiotics.