SlideShare a Scribd company logo
1 of 75
Biochemical Tests for
Identification of Bacteria
Dr. Suprakash Das
Assist. Prof.
WorkflowforBacteriology
Laboratory
1. Patient (D1)
2. Sample
Collection
& Transport
(D1)
3. Direct
Smear from
Samples and
Staining-
Gram
/ Z-N/ Albert
(D1)
4. Selection of Appropriate
Culture Media (D1)
5. ID- Colony Characteristics (D2)
6. Gram Stain from Colony
in culture plate (D2)
6. Selection of Appropriate
BIOCHEMICAL TESTS (D2)
7. AST (D2/D3)
Tests Used most commonly used for
Identification of Gram Positive Cocci
1. Catalase
2. Coagulase- Slide & Tube
3. Hemolysis
4. Sensitivity to Bacitracin/ Furazolidone/ Polymyxin B/Novobiocin
5. PYR Test
6. MR
7. VP
8. Bile Esculin Hydrolysis
9. Bile Solubility
10. DNAse
11. Hippurate/ Ornithin Hydrolysis
12. CAMP test
13. Growth on selective media (e.g. Mannitol salt agar)
Catalase Negative
Organisms
Streptococcus spp.
Enterococcus spp.
Coagulase Positive Organism other than
S. aureus S. hycus/ S. intermedius
Algorithm for Detection of
Common Gram Positive
Pathogens
Tests most commonly used for Identification of
Gram Negative Organisms
1. Indole production
2. Catalase
3. Nitrate reduction
4. MR/VP
5. Citrate utilization
6. Urea Hydrolysis
7. Motility
8. TSI reactions
9. O/F Tests
10. Hydrolysis of- Lysin/ Ornithine/ Arginine
11. Sensitivity to Ampicillin/ O-129/ etc.
12. Growth on MAC
13. Oxidase test
14. Pigment Production.
Indole Positive
Organisms
 Aeromonas
hydrophila,
 Aeromonas punctata,
 Bacillus alvei,
 Edwardsiella sp.,
 Escherichia coli,
 Flavobacterium sp.,
 Klebsiella oxytoca,
 Proteus vulgaris
With the exception of a few
species, Salmonella, Edwardsiella,
Citrobacter, Klebsiella, Enterobacte
r, Serratia, and Providencia usually
give a positive reaction, and
Escherichia, Shigella, Morganella,
and Yersinia give a negative
reaction. Proteus is a citrate
variable.
Oxidation and Fermentation of Medium
(CDC Method)
Purpose :
 This test is used to differentiate microorganisms based on the ability to oxidize or ferment
specific carbohydrates.
Principle:
 This test is used to determine whether an organism uses carbohydrate substrates to
produce acid by products. Non-fermentative bacteria are routinely tested for their ability
to produce acid from six carbohydrates (glucose, xylose, mannitol, lactose, sucrose, and
maltose).
 In addition to the six tubes containing carbohydrates, a control tube containing the OF
base without carbohydrate is also inoculated.
 Triple sugar iron (TSI) agar is also used to determine whether an organism can ferment
glucose.
 OF glucose is used to determine whether an organism ferments or oxidizes glucose.
Oxidation and Fermentation of Medium
(CDC Method)
 If no reaction occurs in either the TSI or OF glucose, the organism is considered a
non–glucose utilizer.
 Hugh and Leifson’s formula uses a low peptone-to-carbohydrate ratio and a
limiting amount of carbohydrate.
 The reduced peptone limits the formation of alkaline amines that may mask acid
production resulting from oxidative metabolism.
 Two tubes are required for interpretation of the OF test. Both are inoculated, and
one tube is overlaid with mineral oil, producing an anaerobic environment.
 Production of acid in the overlaid tube results in a color change and is an
indication of fermentation.
 Acid production in the open tube and color change is the result of oxidation.
 Media: Pancreatic digest of casein (2 g), glycerol (10.0 mL), phenol red (King
method) (0.03 g), agar (3 g) per 1000 mL, pH 7.3.
Oxidation and Fermentation of Medium
(CDC Method)
Method
1. To determine whether acid is produced from carbohydrates, inoculate agar
deeps, each containing a single carbohydrate, with bacterial growth from
an 18- to 24-hour culture by stabbing a needle four to five times into the
medium to a depth of 1 cm.
Note: Two tubes of OF dextrose are usually inoculated; one is overlaid with
either sterile melted petrolatum or sterile paraffin oil to detect fermentation.
2. Incubate the tubes at 35°C to 37°C in ambient air for up to 7 days.
Note: If screwcap tubes are used, loosen the caps during incubation to allow
for air exchange. Otherwise, the control tube and tubes containing
carbohydrates that are not oxidized might not become alkaline.
Oxidation and Fermentation of Medium
(CDC Method)
• Expected Results
 Positive: Acid production (A) is indicated by the color indicator changing to
yellow in the carbohydrate-containing deep.
 Weak-positive (Aw): Weak acid formation can be detected by comparing the
tube containing the medium with carbohydrate with the inoculated tube
containing medium with no carbohydrate.
 Most bacteria that can grow in the OF base produce an alkaline reaction in
the control tube.
 If the color of the medium in a tube containing carbohydrate remains about
the same as it was before the medium was inoculated and if the inoculated
medium in the control tube becomes a deeper red (i.e., becomes alkaline),
the culture being tested is considered weakly positive, assuming the amount
of growth is about the same in both tubes.
Oxidation and Fermentation of Medium
(CDC Method)
 Negative: Red or alkaline (K) color in the deep with carbohydrate equal to the color of the
inoculated control tube.
 No change (NC) or neutral (N): There is growth in the media, but neither the
carbohydrate-containing medium nor the control base turns alkaline (red).
 Note: If the organism does not grow at all in the OF medium, mark the reaction as no
growth (NG).
Limitations
 Slow-growing organisms may not produce results for several days.
Quality Control
 Note: Appropriate organisms depend on which carbohydrate has been added to the basal
medium. Glucose is used as an example.
 Fermenter: Escherichia coli (ATCC25922)
 Oxidizer: Pseudomonas aeruginosa (ATCC27853)
VP Positive bacteria
 Viridans group streptococci
(except Streptococcus
vestibularis),
 Listeria,
 Enterobacter,
 Klebsiella,
 Serratia marcescens,
 Hafnia alvei,
 Vibrio eltor,
 Vibrio alginolyticus
Algorithm for Detection of
Common Gram Negative
Pathogens
Identification of Common Pathogens by
Colony Characteristics and
Biochemical Tests
Staphylococcus aureus
E. coli
I= Indole, M= Methyl Red, Vi= VP Test, C= Citrare
(IMViC Tests)
TSI agar= A/A with
Gas
Urea
Hydrolysis
I= Indole, M= MR Test, Vi= VP Test, C= Citrate Test
(IMViC Tests)
Urea
Hydr
olysis
TSI Agar= A/A with
Gas
 Salmonella sp. after 24 hours growth on XLD agar.
 Xylose Lysine (XL) agar is used when trying to culture
and isolate Gram-negative enteric bacilli.
 When XL agar is supplemented with sodium thiosulfate,
ferric ammonium citrate, and sodium deoxycholate, it is
then termed XLD agar, and is then an even more selective
medium than XL alone.
 The presence of any black colored area indicates the
deposition of hydrogen sulfide, (H2S) under alkaline
conditions.
Citrobacter freundii
I= Indole, M= MR Test, Vi= VP Test, C= Citrate Test
(IMViC Tests)
Urea
Hydr
olysis
TSI Agar= K/A
without gas
Motility
Semisolid
Agar
Proteus mirabilis
On
MacConkey Agar
-NLF colonies.
Proteus mirabilis
On
Blood Agar showing
SWARMING growth.
I= Indole, M= MR Test, Vi= VP Test, C= Citrate Test
(IMViC Tests)
Urea
Hydr
olysis
TSI Agar= K/A with
H2S gas production.
Motility
Semisolid
Agar
P. aeruginosa showing
Production of soluble
Pyocyanin pigment on
NA
2-3 mm, flat, smooth,
non-lactose fermenting
colonies with regular
margin and Alligator
skin like from top view
I= Indole, M= MR Test, Vi= VP Test, C= Citrate Test
(IMViC Tests)
Urea
Hydr
olysis
TSI Agar= K/K
without gas.
Motility
Semisolid
Agar
NOTE- Urea Hydrolysis
can be variable
Oxidase Test
Positive
Acinetobacter baumannii
I= Indole, M= MR Test, Vi= VP Test, C= Citrate Test
(IMViC Tests)
Urea
Hydr
olysis
TSI Agar= K/K
without gas.
Motility
Semisolid
Agar
NOTE- Urea Hydrolysis
can be variable
Oxidase Test Negative
Automated ID & AST
Thank You

More Related Content

What's hot

Mac conkey agar presentation
Mac conkey agar presentationMac conkey agar presentation
Mac conkey agar presentationAsmita Dhakal
 
How to read a culture plate
How to read a culture plateHow to read a culture plate
How to read a culture plateImtiyazz Wanii
 
Diagnostic methods of Parasites
Diagnostic methods of ParasitesDiagnostic methods of Parasites
Diagnostic methods of ParasitesSHRUTHI VASAN
 
Identification of Yeasts & Candida species
Identification of Yeasts & Candida speciesIdentification of Yeasts & Candida species
Identification of Yeasts & Candida speciesDr Shams Afridi
 
Automated microbial identification system
Automated microbial identification systemAutomated microbial identification system
Automated microbial identification systemHanu Pratap
 
Clinical Microbiology in Laboratory
Clinical Microbiology in LaboratoryClinical Microbiology in Laboratory
Clinical Microbiology in LaboratoryBhoj Raj Singh
 
Automated system for bacterial identification
Automated system for bacterial identificationAutomated system for bacterial identification
Automated system for bacterial identificationDEEKSHANT KUMAR
 
Biochemical tests for identification of bacteria
Biochemical tests for identification of bacteriaBiochemical tests for identification of bacteria
Biochemical tests for identification of bacteriaRavi Kant Agrawal
 
Medical Microbiology Laboratory (biochemical tests - iii)
Medical Microbiology Laboratory (biochemical tests - iii)Medical Microbiology Laboratory (biochemical tests - iii)
Medical Microbiology Laboratory (biochemical tests - iii)Hussein Al-tameemi
 
Clinical Microbiology Practical - 1
Clinical Microbiology Practical - 1Clinical Microbiology Practical - 1
Clinical Microbiology Practical - 1Sijo A
 
Conventional methods for identification and characterization of pathogenic ba...
Conventional methods for identification and characterization of pathogenic ba...Conventional methods for identification and characterization of pathogenic ba...
Conventional methods for identification and characterization of pathogenic ba...Bhoj Raj Singh
 

What's hot (20)

Mac conkey agar presentation
Mac conkey agar presentationMac conkey agar presentation
Mac conkey agar presentation
 
How to read a culture plate
How to read a culture plateHow to read a culture plate
How to read a culture plate
 
Diagnostic methods of Parasites
Diagnostic methods of ParasitesDiagnostic methods of Parasites
Diagnostic methods of Parasites
 
Oxidase Test Microbiology
Oxidase Test MicrobiologyOxidase Test Microbiology
Oxidase Test Microbiology
 
Identification of Yeasts & Candida species
Identification of Yeasts & Candida speciesIdentification of Yeasts & Candida species
Identification of Yeasts & Candida species
 
Indole test
Indole testIndole test
Indole test
 
Automated microbial identification system
Automated microbial identification systemAutomated microbial identification system
Automated microbial identification system
 
Clinical Microbiology in Laboratory
Clinical Microbiology in LaboratoryClinical Microbiology in Laboratory
Clinical Microbiology in Laboratory
 
Automated system for bacterial identification
Automated system for bacterial identificationAutomated system for bacterial identification
Automated system for bacterial identification
 
Biochemical tests for identification of bacteria
Biochemical tests for identification of bacteriaBiochemical tests for identification of bacteria
Biochemical tests for identification of bacteria
 
Medical Microbiology Laboratory (biochemical tests - iii)
Medical Microbiology Laboratory (biochemical tests - iii)Medical Microbiology Laboratory (biochemical tests - iii)
Medical Microbiology Laboratory (biochemical tests - iii)
 
2 specimen collection
2 specimen collection2 specimen collection
2 specimen collection
 
Klebsiella spp
Klebsiella sppKlebsiella spp
Klebsiella spp
 
Lecture 5
Lecture 5Lecture 5
Lecture 5
 
identification of bacteria
identification of bacteriaidentification of bacteria
identification of bacteria
 
Clinical Microbiology Practical - 1
Clinical Microbiology Practical - 1Clinical Microbiology Practical - 1
Clinical Microbiology Practical - 1
 
Salmonella
SalmonellaSalmonella
Salmonella
 
Conventional methods for identification and characterization of pathogenic ba...
Conventional methods for identification and characterization of pathogenic ba...Conventional methods for identification and characterization of pathogenic ba...
Conventional methods for identification and characterization of pathogenic ba...
 
Oxidase test
Oxidase testOxidase test
Oxidase test
 
Spore staining
Spore stainingSpore staining
Spore staining
 

Similar to Biochemical tests for bacterial identification

Medical Microbiology Laboratory (biochemical tests - ii)
Medical Microbiology Laboratory (biochemical tests - ii)Medical Microbiology Laboratory (biochemical tests - ii)
Medical Microbiology Laboratory (biochemical tests - ii)Hussein Al-tameemi
 
ISOLATION AND IDENTIFICATION OF NLF BACTERIA IN VARIOUS SAMPLES.
ISOLATION AND IDENTIFICATION OF NLF BACTERIA IN VARIOUS SAMPLES.ISOLATION AND IDENTIFICATION OF NLF BACTERIA IN VARIOUS SAMPLES.
ISOLATION AND IDENTIFICATION OF NLF BACTERIA IN VARIOUS SAMPLES.Daisy Saini
 
Bacterial identification
Bacterial identificationBacterial identification
Bacterial identificationChota Alexander
 
Practical 40 biochemical test microbiology.pdf
Practical 40 biochemical test microbiology.pdfPractical 40 biochemical test microbiology.pdf
Practical 40 biochemical test microbiology.pdfssuser668f10
 
Biochemical tests and physiological tests for various groups of Bacteria
Biochemical tests and physiological tests for various groups of BacteriaBiochemical tests and physiological tests for various groups of Bacteria
Biochemical tests and physiological tests for various groups of BacteriaHARISH J
 
⁨فحوصات كيميائيه_بكتيريه_240920220229⁩
 ⁨فحوصات كيميائيه_بكتيريه_240920220229⁩ ⁨فحوصات كيميائيه_بكتيريه_240920220229⁩
⁨فحوصات كيميائيه_بكتيريه_240920220229⁩Osman Guure
 
Biochemical identification of bacteria
Biochemical identification of bacteriaBiochemical identification of bacteria
Biochemical identification of bacteriaBianca Isaguirre
 
Biochemical tests (1st part)
Biochemical tests (1st part)Biochemical tests (1st part)
Biochemical tests (1st part)Aman Ullah
 
PPT_B.Voc. MLT_Sem 3_MLT 7_Ch 16_v 1.3_Shweta.pptx
PPT_B.Voc. MLT_Sem 3_MLT 7_Ch 16_v 1.3_Shweta.pptxPPT_B.Voc. MLT_Sem 3_MLT 7_Ch 16_v 1.3_Shweta.pptx
PPT_B.Voc. MLT_Sem 3_MLT 7_Ch 16_v 1.3_Shweta.pptxAdityasonawane32
 
Medical Microbiology Laboratory (biochemical tests - i)
Medical Microbiology Laboratory (biochemical tests - i)Medical Microbiology Laboratory (biochemical tests - i)
Medical Microbiology Laboratory (biochemical tests - i)Hussein Al-tameemi
 
New Microsoft PowerPoint Presentation.pptx
New Microsoft PowerPoint Presentation.pptxNew Microsoft PowerPoint Presentation.pptx
New Microsoft PowerPoint Presentation.pptxPratikShinde189184
 
Enterobacteriaceae.ppt.jjkn.kjkhjjbjbhbhbh
Enterobacteriaceae.ppt.jjkn.kjkhjjbjbhbhbhEnterobacteriaceae.ppt.jjkn.kjkhjjbjbhbhbh
Enterobacteriaceae.ppt.jjkn.kjkhjjbjbhbhbhssuser06f49d
 
biochemical reactions(gnb)-4.pptx
biochemical reactions(gnb)-4.pptxbiochemical reactions(gnb)-4.pptx
biochemical reactions(gnb)-4.pptxVaisHali822687
 
biochemicaltestsforidentificationofbacteria-191123064938 (2).pdf
biochemicaltestsforidentificationofbacteria-191123064938 (2).pdfbiochemicaltestsforidentificationofbacteria-191123064938 (2).pdf
biochemicaltestsforidentificationofbacteria-191123064938 (2).pdfNimraBarkaat
 
enumeration and detection of food born diseases and detection of salmon ella ...
enumeration and detection of food born diseases and detection of salmon ella ...enumeration and detection of food born diseases and detection of salmon ella ...
enumeration and detection of food born diseases and detection of salmon ella ...Praveen kumar verma
 
Biochemical reactions PPT.pptx
Biochemical reactions PPT.pptxBiochemical reactions PPT.pptx
Biochemical reactions PPT.pptxMonishabasavaraj
 
Biochemical Identification of bacteria 2023 sk.ppt
Biochemical Identification of bacteria 2023 sk.pptBiochemical Identification of bacteria 2023 sk.ppt
Biochemical Identification of bacteria 2023 sk.pptSandhya Kulkarni
 

Similar to Biochemical tests for bacterial identification (20)

Medical Microbiology Laboratory (biochemical tests - ii)
Medical Microbiology Laboratory (biochemical tests - ii)Medical Microbiology Laboratory (biochemical tests - ii)
Medical Microbiology Laboratory (biochemical tests - ii)
 
ISOLATION AND IDENTIFICATION OF NLF BACTERIA IN VARIOUS SAMPLES.
ISOLATION AND IDENTIFICATION OF NLF BACTERIA IN VARIOUS SAMPLES.ISOLATION AND IDENTIFICATION OF NLF BACTERIA IN VARIOUS SAMPLES.
ISOLATION AND IDENTIFICATION OF NLF BACTERIA IN VARIOUS SAMPLES.
 
Bacterial identification
Bacterial identificationBacterial identification
Bacterial identification
 
Practical microbiology dr.nada khazal k. hendi
Practical microbiology       dr.nada khazal k. hendiPractical microbiology       dr.nada khazal k. hendi
Practical microbiology dr.nada khazal k. hendi
 
Practical 40 biochemical test microbiology.pdf
Practical 40 biochemical test microbiology.pdfPractical 40 biochemical test microbiology.pdf
Practical 40 biochemical test microbiology.pdf
 
Biochemical tests and physiological tests for various groups of Bacteria
Biochemical tests and physiological tests for various groups of BacteriaBiochemical tests and physiological tests for various groups of Bacteria
Biochemical tests and physiological tests for various groups of Bacteria
 
⁨فحوصات كيميائيه_بكتيريه_240920220229⁩
 ⁨فحوصات كيميائيه_بكتيريه_240920220229⁩ ⁨فحوصات كيميائيه_بكتيريه_240920220229⁩
⁨فحوصات كيميائيه_بكتيريه_240920220229⁩
 
Biochemical identification of bacteria
Biochemical identification of bacteriaBiochemical identification of bacteria
Biochemical identification of bacteria
 
Biochemical tests (1st part)
Biochemical tests (1st part)Biochemical tests (1st part)
Biochemical tests (1st part)
 
PPT_B.Voc. MLT_Sem 3_MLT 7_Ch 16_v 1.3_Shweta.pptx
PPT_B.Voc. MLT_Sem 3_MLT 7_Ch 16_v 1.3_Shweta.pptxPPT_B.Voc. MLT_Sem 3_MLT 7_Ch 16_v 1.3_Shweta.pptx
PPT_B.Voc. MLT_Sem 3_MLT 7_Ch 16_v 1.3_Shweta.pptx
 
Medical Microbiology Laboratory (biochemical tests - i)
Medical Microbiology Laboratory (biochemical tests - i)Medical Microbiology Laboratory (biochemical tests - i)
Medical Microbiology Laboratory (biochemical tests - i)
 
Biochemical reaction
Biochemical reactionBiochemical reaction
Biochemical reaction
 
Unknown organism
Unknown organismUnknown organism
Unknown organism
 
New Microsoft PowerPoint Presentation.pptx
New Microsoft PowerPoint Presentation.pptxNew Microsoft PowerPoint Presentation.pptx
New Microsoft PowerPoint Presentation.pptx
 
Enterobacteriaceae.ppt.jjkn.kjkhjjbjbhbhbh
Enterobacteriaceae.ppt.jjkn.kjkhjjbjbhbhbhEnterobacteriaceae.ppt.jjkn.kjkhjjbjbhbhbh
Enterobacteriaceae.ppt.jjkn.kjkhjjbjbhbhbh
 
biochemical reactions(gnb)-4.pptx
biochemical reactions(gnb)-4.pptxbiochemical reactions(gnb)-4.pptx
biochemical reactions(gnb)-4.pptx
 
biochemicaltestsforidentificationofbacteria-191123064938 (2).pdf
biochemicaltestsforidentificationofbacteria-191123064938 (2).pdfbiochemicaltestsforidentificationofbacteria-191123064938 (2).pdf
biochemicaltestsforidentificationofbacteria-191123064938 (2).pdf
 
enumeration and detection of food born diseases and detection of salmon ella ...
enumeration and detection of food born diseases and detection of salmon ella ...enumeration and detection of food born diseases and detection of salmon ella ...
enumeration and detection of food born diseases and detection of salmon ella ...
 
Biochemical reactions PPT.pptx
Biochemical reactions PPT.pptxBiochemical reactions PPT.pptx
Biochemical reactions PPT.pptx
 
Biochemical Identification of bacteria 2023 sk.ppt
Biochemical Identification of bacteria 2023 sk.pptBiochemical Identification of bacteria 2023 sk.ppt
Biochemical Identification of bacteria 2023 sk.ppt
 

More from Suprakash Das

Mycobacterium tuberculosis
Mycobacterium tuberculosisMycobacterium tuberculosis
Mycobacterium tuberculosisSuprakash Das
 
Hepatitis viruses a & e
Hepatitis viruses a & eHepatitis viruses a & e
Hepatitis viruses a & eSuprakash Das
 
Coccidian parasites -Cyclospora cayetanensis
Coccidian parasites -Cyclospora cayetanensisCoccidian parasites -Cyclospora cayetanensis
Coccidian parasites -Cyclospora cayetanensisSuprakash Das
 
Coccidian parasites- Cryptosporidiosis
Coccidian parasites- CryptosporidiosisCoccidian parasites- Cryptosporidiosis
Coccidian parasites- CryptosporidiosisSuprakash Das
 
Vibrio practical notes
Vibrio practical notesVibrio practical notes
Vibrio practical notesSuprakash Das
 
Leptospira weil's disease
Leptospira weil's diseaseLeptospira weil's disease
Leptospira weil's diseaseSuprakash Das
 
Infective endocarditis
Infective endocarditisInfective endocarditis
Infective endocarditisSuprakash Das
 
Borrelia lyme disease
Borrelia lyme diseaseBorrelia lyme disease
Borrelia lyme diseaseSuprakash Das
 
Laboratory diagnosis of Blood Stream Infections (BSIs)
Laboratory diagnosis of Blood Stream Infections (BSIs)Laboratory diagnosis of Blood Stream Infections (BSIs)
Laboratory diagnosis of Blood Stream Infections (BSIs)Suprakash Das
 
Transplant and Cancer Immunology
Transplant and Cancer ImmunologyTransplant and Cancer Immunology
Transplant and Cancer ImmunologySuprakash Das
 

More from Suprakash Das (20)

Mycobacterium tuberculosis
Mycobacterium tuberculosisMycobacterium tuberculosis
Mycobacterium tuberculosis
 
HIV/AIDS
HIV/AIDSHIV/AIDS
HIV/AIDS
 
Cestodes
CestodesCestodes
Cestodes
 
Hepatitis viruses a & e
Hepatitis viruses a & eHepatitis viruses a & e
Hepatitis viruses a & e
 
Coccidian parasites -Cyclospora cayetanensis
Coccidian parasites -Cyclospora cayetanensisCoccidian parasites -Cyclospora cayetanensis
Coccidian parasites -Cyclospora cayetanensis
 
Coccidian parasites- Cryptosporidiosis
Coccidian parasites- CryptosporidiosisCoccidian parasites- Cryptosporidiosis
Coccidian parasites- Cryptosporidiosis
 
E. histolytica
E. histolyticaE. histolytica
E. histolytica
 
Vibrio practical notes
Vibrio practical notesVibrio practical notes
Vibrio practical notes
 
Food poisoning
Food poisoningFood poisoning
Food poisoning
 
Leptospira weil's disease
Leptospira weil's diseaseLeptospira weil's disease
Leptospira weil's disease
 
Infective endocarditis
Infective endocarditisInfective endocarditis
Infective endocarditis
 
Borrelia lyme disease
Borrelia lyme diseaseBorrelia lyme disease
Borrelia lyme disease
 
Leishmaniasis
LeishmaniasisLeishmaniasis
Leishmaniasis
 
Trypanosoma
TrypanosomaTrypanosoma
Trypanosoma
 
Malaria
MalariaMalaria
Malaria
 
Filarisis
FilarisisFilarisis
Filarisis
 
Laboratory diagnosis of Blood Stream Infections (BSIs)
Laboratory diagnosis of Blood Stream Infections (BSIs)Laboratory diagnosis of Blood Stream Infections (BSIs)
Laboratory diagnosis of Blood Stream Infections (BSIs)
 
Transplant and Cancer Immunology
Transplant and Cancer ImmunologyTransplant and Cancer Immunology
Transplant and Cancer Immunology
 
Gram staining
Gram stainingGram staining
Gram staining
 
Gram stain viva 1
Gram stain viva 1Gram stain viva 1
Gram stain viva 1
 

Recently uploaded

Failure to thrive in neonates and infants + pediatric case.pptx
Failure to thrive in neonates and infants  + pediatric case.pptxFailure to thrive in neonates and infants  + pediatric case.pptx
Failure to thrive in neonates and infants + pediatric case.pptxclaviclebrown44
 
SEMESTER-V CHILD HEALTH NURSING-UNIT-1-INTRODUCTION.pdf
SEMESTER-V CHILD HEALTH NURSING-UNIT-1-INTRODUCTION.pdfSEMESTER-V CHILD HEALTH NURSING-UNIT-1-INTRODUCTION.pdf
SEMESTER-V CHILD HEALTH NURSING-UNIT-1-INTRODUCTION.pdfSachin Sharma
 
Emerging Markets Documentation for Drug Regulatory Affairs
Emerging Markets Documentation for Drug Regulatory AffairsEmerging Markets Documentation for Drug Regulatory Affairs
Emerging Markets Documentation for Drug Regulatory AffairsDimple Marathe
 
Part I - Anticipatory Grief: Experiencing grief before the loss has happened
Part I - Anticipatory Grief: Experiencing grief before the loss has happenedPart I - Anticipatory Grief: Experiencing grief before the loss has happened
Part I - Anticipatory Grief: Experiencing grief before the loss has happenedbkling
 
High Purity 99% PMK Ethyl Glycidate Powder CAS 28578-16-7
High Purity 99% PMK Ethyl Glycidate Powder CAS 28578-16-7High Purity 99% PMK Ethyl Glycidate Powder CAS 28578-16-7
High Purity 99% PMK Ethyl Glycidate Powder CAS 28578-16-7grandmotherprocess99
 
Gross Anatomy and Histology of Tongue by Dr. Rabia Inam Gandapore.pptx
Gross Anatomy and Histology of Tongue by Dr. Rabia Inam Gandapore.pptxGross Anatomy and Histology of Tongue by Dr. Rabia Inam Gandapore.pptx
Gross Anatomy and Histology of Tongue by Dr. Rabia Inam Gandapore.pptxDr. Rabia Inam Gandapore
 
VIP ℂall Girls Electronic City Bangalore 9079923931 WhatsApp: Me All Time Ser...
VIP ℂall Girls Electronic City Bangalore 9079923931 WhatsApp: Me All Time Ser...VIP ℂall Girls Electronic City Bangalore 9079923931 WhatsApp: Me All Time Ser...
VIP ℂall Girls Electronic City Bangalore 9079923931 WhatsApp: Me All Time Ser...shriyaguptaescortp#11
 
VIP ℂall Girls Arekere Bangalore 6378878445 WhatsApp: Me All Time Serviℂe Ava...
VIP ℂall Girls Arekere Bangalore 6378878445 WhatsApp: Me All Time Serviℂe Ava...VIP ℂall Girls Arekere Bangalore 6378878445 WhatsApp: Me All Time Serviℂe Ava...
VIP ℂall Girls Arekere Bangalore 6378878445 WhatsApp: Me All Time Serviℂe Ava...deepakkumar115120
 
💚Mature Women / Aunty Call Girl Chennai Escorts Service 💯Call Us 🔝 6367492432...
💚Mature Women / Aunty Call Girl Chennai Escorts Service 💯Call Us 🔝 6367492432...💚Mature Women / Aunty Call Girl Chennai Escorts Service 💯Call Us 🔝 6367492432...
💚Mature Women / Aunty Call Girl Chennai Escorts Service 💯Call Us 🔝 6367492432...chaddanishu186
 
ℂall Girls Chandigarh 💚8868886958 💚ℂall Girls Serviℂe Available Day and Night
ℂall Girls Chandigarh  💚8868886958  💚ℂall Girls Serviℂe Available Day and Nightℂall Girls Chandigarh  💚8868886958  💚ℂall Girls Serviℂe Available Day and Night
ℂall Girls Chandigarh 💚8868886958 💚ℂall Girls Serviℂe Available Day and Nightminkseocompany
 
Difference Between Skeletal Smooth and Cardiac Muscles
Difference Between Skeletal Smooth and Cardiac MusclesDifference Between Skeletal Smooth and Cardiac Muscles
Difference Between Skeletal Smooth and Cardiac MusclesMedicoseAcademics
 
Vesu * HiFi ℂall Girls Surat Phone No 8849756361 Elite ℂall Serviℂe Available...
Vesu * HiFi ℂall Girls Surat Phone No 8849756361 Elite ℂall Serviℂe Available...Vesu * HiFi ℂall Girls Surat Phone No 8849756361 Elite ℂall Serviℂe Available...
Vesu * HiFi ℂall Girls Surat Phone No 8849756361 Elite ℂall Serviℂe Available...Dharvi Mehra$I11
 
Unsatisfied Bhabhi ℂall Girls Surat Book Sneha 8849756361 Top Class ℂall Girl...
Unsatisfied Bhabhi ℂall Girls Surat Book Sneha 8849756361 Top Class ℂall Girl...Unsatisfied Bhabhi ℂall Girls Surat Book Sneha 8849756361 Top Class ℂall Girl...
Unsatisfied Bhabhi ℂall Girls Surat Book Sneha 8849756361 Top Class ℂall Girl...Dharvi Mehra$I11
 
CONGENITAL HYPERTROPHIC PYLORIC STENOSIS by Dr M.KARTHIK EMMANUEL
CONGENITAL HYPERTROPHIC PYLORIC STENOSIS  by Dr M.KARTHIK EMMANUELCONGENITAL HYPERTROPHIC PYLORIC STENOSIS  by Dr M.KARTHIK EMMANUEL
CONGENITAL HYPERTROPHIC PYLORIC STENOSIS by Dr M.KARTHIK EMMANUELMKARTHIKEMMANUEL
 
Top 10 Most Beautiful Russian Pornstars List 2024
Top 10 Most Beautiful Russian Pornstars List 2024Top 10 Most Beautiful Russian Pornstars List 2024
Top 10 Most Beautiful Russian Pornstars List 2024locantocallgirl01
 
Report Back from SGO: What’s the Latest in Ovarian Cancer?
Report Back from SGO: What’s the Latest in Ovarian Cancer?Report Back from SGO: What’s the Latest in Ovarian Cancer?
Report Back from SGO: What’s the Latest in Ovarian Cancer?bkling
 
TEST BANK For Nursing Leadership, Management, and Professional Practice for t...
TEST BANK For Nursing Leadership, Management, and Professional Practice for t...TEST BANK For Nursing Leadership, Management, and Professional Practice for t...
TEST BANK For Nursing Leadership, Management, and Professional Practice for t...rightmanforbloodline
 
Borivali East * HiFi ℂall Girls Mumbai Phone No 9004268417 Elite ℂall Serviℂe...
Borivali East * HiFi ℂall Girls Mumbai Phone No 9004268417 Elite ℂall Serviℂe...Borivali East * HiFi ℂall Girls Mumbai Phone No 9004268417 Elite ℂall Serviℂe...
Borivali East * HiFi ℂall Girls Mumbai Phone No 9004268417 Elite ℂall Serviℂe...Dharvi Mehra$I11
 
VIP ℂall Girls Miyapur Hyderabad 7014168258 WhatsApp: Me All Time Serviℂe Ava...
VIP ℂall Girls Miyapur Hyderabad 7014168258 WhatsApp: Me All Time Serviℂe Ava...VIP ℂall Girls Miyapur Hyderabad 7014168258 WhatsApp: Me All Time Serviℂe Ava...
VIP ℂall Girls Miyapur Hyderabad 7014168258 WhatsApp: Me All Time Serviℂe Ava...anshu khanak#N06
 
Creeping Stroke - Venous thrombosis presenting with pc-stroke.pptx
Creeping Stroke - Venous thrombosis presenting with pc-stroke.pptxCreeping Stroke - Venous thrombosis presenting with pc-stroke.pptx
Creeping Stroke - Venous thrombosis presenting with pc-stroke.pptxYasser Alzainy
 

Recently uploaded (20)

Failure to thrive in neonates and infants + pediatric case.pptx
Failure to thrive in neonates and infants  + pediatric case.pptxFailure to thrive in neonates and infants  + pediatric case.pptx
Failure to thrive in neonates and infants + pediatric case.pptx
 
SEMESTER-V CHILD HEALTH NURSING-UNIT-1-INTRODUCTION.pdf
SEMESTER-V CHILD HEALTH NURSING-UNIT-1-INTRODUCTION.pdfSEMESTER-V CHILD HEALTH NURSING-UNIT-1-INTRODUCTION.pdf
SEMESTER-V CHILD HEALTH NURSING-UNIT-1-INTRODUCTION.pdf
 
Emerging Markets Documentation for Drug Regulatory Affairs
Emerging Markets Documentation for Drug Regulatory AffairsEmerging Markets Documentation for Drug Regulatory Affairs
Emerging Markets Documentation for Drug Regulatory Affairs
 
Part I - Anticipatory Grief: Experiencing grief before the loss has happened
Part I - Anticipatory Grief: Experiencing grief before the loss has happenedPart I - Anticipatory Grief: Experiencing grief before the loss has happened
Part I - Anticipatory Grief: Experiencing grief before the loss has happened
 
High Purity 99% PMK Ethyl Glycidate Powder CAS 28578-16-7
High Purity 99% PMK Ethyl Glycidate Powder CAS 28578-16-7High Purity 99% PMK Ethyl Glycidate Powder CAS 28578-16-7
High Purity 99% PMK Ethyl Glycidate Powder CAS 28578-16-7
 
Gross Anatomy and Histology of Tongue by Dr. Rabia Inam Gandapore.pptx
Gross Anatomy and Histology of Tongue by Dr. Rabia Inam Gandapore.pptxGross Anatomy and Histology of Tongue by Dr. Rabia Inam Gandapore.pptx
Gross Anatomy and Histology of Tongue by Dr. Rabia Inam Gandapore.pptx
 
VIP ℂall Girls Electronic City Bangalore 9079923931 WhatsApp: Me All Time Ser...
VIP ℂall Girls Electronic City Bangalore 9079923931 WhatsApp: Me All Time Ser...VIP ℂall Girls Electronic City Bangalore 9079923931 WhatsApp: Me All Time Ser...
VIP ℂall Girls Electronic City Bangalore 9079923931 WhatsApp: Me All Time Ser...
 
VIP ℂall Girls Arekere Bangalore 6378878445 WhatsApp: Me All Time Serviℂe Ava...
VIP ℂall Girls Arekere Bangalore 6378878445 WhatsApp: Me All Time Serviℂe Ava...VIP ℂall Girls Arekere Bangalore 6378878445 WhatsApp: Me All Time Serviℂe Ava...
VIP ℂall Girls Arekere Bangalore 6378878445 WhatsApp: Me All Time Serviℂe Ava...
 
💚Mature Women / Aunty Call Girl Chennai Escorts Service 💯Call Us 🔝 6367492432...
💚Mature Women / Aunty Call Girl Chennai Escorts Service 💯Call Us 🔝 6367492432...💚Mature Women / Aunty Call Girl Chennai Escorts Service 💯Call Us 🔝 6367492432...
💚Mature Women / Aunty Call Girl Chennai Escorts Service 💯Call Us 🔝 6367492432...
 
ℂall Girls Chandigarh 💚8868886958 💚ℂall Girls Serviℂe Available Day and Night
ℂall Girls Chandigarh  💚8868886958  💚ℂall Girls Serviℂe Available Day and Nightℂall Girls Chandigarh  💚8868886958  💚ℂall Girls Serviℂe Available Day and Night
ℂall Girls Chandigarh 💚8868886958 💚ℂall Girls Serviℂe Available Day and Night
 
Difference Between Skeletal Smooth and Cardiac Muscles
Difference Between Skeletal Smooth and Cardiac MusclesDifference Between Skeletal Smooth and Cardiac Muscles
Difference Between Skeletal Smooth and Cardiac Muscles
 
Vesu * HiFi ℂall Girls Surat Phone No 8849756361 Elite ℂall Serviℂe Available...
Vesu * HiFi ℂall Girls Surat Phone No 8849756361 Elite ℂall Serviℂe Available...Vesu * HiFi ℂall Girls Surat Phone No 8849756361 Elite ℂall Serviℂe Available...
Vesu * HiFi ℂall Girls Surat Phone No 8849756361 Elite ℂall Serviℂe Available...
 
Unsatisfied Bhabhi ℂall Girls Surat Book Sneha 8849756361 Top Class ℂall Girl...
Unsatisfied Bhabhi ℂall Girls Surat Book Sneha 8849756361 Top Class ℂall Girl...Unsatisfied Bhabhi ℂall Girls Surat Book Sneha 8849756361 Top Class ℂall Girl...
Unsatisfied Bhabhi ℂall Girls Surat Book Sneha 8849756361 Top Class ℂall Girl...
 
CONGENITAL HYPERTROPHIC PYLORIC STENOSIS by Dr M.KARTHIK EMMANUEL
CONGENITAL HYPERTROPHIC PYLORIC STENOSIS  by Dr M.KARTHIK EMMANUELCONGENITAL HYPERTROPHIC PYLORIC STENOSIS  by Dr M.KARTHIK EMMANUEL
CONGENITAL HYPERTROPHIC PYLORIC STENOSIS by Dr M.KARTHIK EMMANUEL
 
Top 10 Most Beautiful Russian Pornstars List 2024
Top 10 Most Beautiful Russian Pornstars List 2024Top 10 Most Beautiful Russian Pornstars List 2024
Top 10 Most Beautiful Russian Pornstars List 2024
 
Report Back from SGO: What’s the Latest in Ovarian Cancer?
Report Back from SGO: What’s the Latest in Ovarian Cancer?Report Back from SGO: What’s the Latest in Ovarian Cancer?
Report Back from SGO: What’s the Latest in Ovarian Cancer?
 
TEST BANK For Nursing Leadership, Management, and Professional Practice for t...
TEST BANK For Nursing Leadership, Management, and Professional Practice for t...TEST BANK For Nursing Leadership, Management, and Professional Practice for t...
TEST BANK For Nursing Leadership, Management, and Professional Practice for t...
 
Borivali East * HiFi ℂall Girls Mumbai Phone No 9004268417 Elite ℂall Serviℂe...
Borivali East * HiFi ℂall Girls Mumbai Phone No 9004268417 Elite ℂall Serviℂe...Borivali East * HiFi ℂall Girls Mumbai Phone No 9004268417 Elite ℂall Serviℂe...
Borivali East * HiFi ℂall Girls Mumbai Phone No 9004268417 Elite ℂall Serviℂe...
 
VIP ℂall Girls Miyapur Hyderabad 7014168258 WhatsApp: Me All Time Serviℂe Ava...
VIP ℂall Girls Miyapur Hyderabad 7014168258 WhatsApp: Me All Time Serviℂe Ava...VIP ℂall Girls Miyapur Hyderabad 7014168258 WhatsApp: Me All Time Serviℂe Ava...
VIP ℂall Girls Miyapur Hyderabad 7014168258 WhatsApp: Me All Time Serviℂe Ava...
 
Creeping Stroke - Venous thrombosis presenting with pc-stroke.pptx
Creeping Stroke - Venous thrombosis presenting with pc-stroke.pptxCreeping Stroke - Venous thrombosis presenting with pc-stroke.pptx
Creeping Stroke - Venous thrombosis presenting with pc-stroke.pptx
 

Biochemical tests for bacterial identification

  • 1. Biochemical Tests for Identification of Bacteria Dr. Suprakash Das Assist. Prof.
  • 3. 1. Patient (D1) 2. Sample Collection & Transport (D1) 3. Direct Smear from Samples and Staining- Gram / Z-N/ Albert (D1)
  • 4. 4. Selection of Appropriate Culture Media (D1) 5. ID- Colony Characteristics (D2) 6. Gram Stain from Colony in culture plate (D2) 6. Selection of Appropriate BIOCHEMICAL TESTS (D2) 7. AST (D2/D3)
  • 5. Tests Used most commonly used for Identification of Gram Positive Cocci 1. Catalase 2. Coagulase- Slide & Tube 3. Hemolysis 4. Sensitivity to Bacitracin/ Furazolidone/ Polymyxin B/Novobiocin 5. PYR Test 6. MR 7. VP 8. Bile Esculin Hydrolysis 9. Bile Solubility 10. DNAse 11. Hippurate/ Ornithin Hydrolysis 12. CAMP test 13. Growth on selective media (e.g. Mannitol salt agar)
  • 7.
  • 8. Coagulase Positive Organism other than S. aureus S. hycus/ S. intermedius
  • 9.
  • 10.
  • 11.
  • 12.
  • 13. Algorithm for Detection of Common Gram Positive Pathogens
  • 14.
  • 15.
  • 16.
  • 17.
  • 18. Tests most commonly used for Identification of Gram Negative Organisms 1. Indole production 2. Catalase 3. Nitrate reduction 4. MR/VP 5. Citrate utilization 6. Urea Hydrolysis 7. Motility 8. TSI reactions 9. O/F Tests 10. Hydrolysis of- Lysin/ Ornithine/ Arginine 11. Sensitivity to Ampicillin/ O-129/ etc. 12. Growth on MAC 13. Oxidase test 14. Pigment Production.
  • 19. Indole Positive Organisms  Aeromonas hydrophila,  Aeromonas punctata,  Bacillus alvei,  Edwardsiella sp.,  Escherichia coli,  Flavobacterium sp.,  Klebsiella oxytoca,  Proteus vulgaris
  • 20.
  • 21.
  • 22.
  • 23. With the exception of a few species, Salmonella, Edwardsiella, Citrobacter, Klebsiella, Enterobacte r, Serratia, and Providencia usually give a positive reaction, and Escherichia, Shigella, Morganella, and Yersinia give a negative reaction. Proteus is a citrate variable.
  • 24.
  • 25.
  • 26.
  • 27.
  • 28.
  • 29.
  • 30.
  • 31.
  • 32. Oxidation and Fermentation of Medium (CDC Method) Purpose :  This test is used to differentiate microorganisms based on the ability to oxidize or ferment specific carbohydrates. Principle:  This test is used to determine whether an organism uses carbohydrate substrates to produce acid by products. Non-fermentative bacteria are routinely tested for their ability to produce acid from six carbohydrates (glucose, xylose, mannitol, lactose, sucrose, and maltose).  In addition to the six tubes containing carbohydrates, a control tube containing the OF base without carbohydrate is also inoculated.  Triple sugar iron (TSI) agar is also used to determine whether an organism can ferment glucose.  OF glucose is used to determine whether an organism ferments or oxidizes glucose.
  • 33. Oxidation and Fermentation of Medium (CDC Method)  If no reaction occurs in either the TSI or OF glucose, the organism is considered a non–glucose utilizer.  Hugh and Leifson’s formula uses a low peptone-to-carbohydrate ratio and a limiting amount of carbohydrate.  The reduced peptone limits the formation of alkaline amines that may mask acid production resulting from oxidative metabolism.  Two tubes are required for interpretation of the OF test. Both are inoculated, and one tube is overlaid with mineral oil, producing an anaerobic environment.  Production of acid in the overlaid tube results in a color change and is an indication of fermentation.  Acid production in the open tube and color change is the result of oxidation.  Media: Pancreatic digest of casein (2 g), glycerol (10.0 mL), phenol red (King method) (0.03 g), agar (3 g) per 1000 mL, pH 7.3.
  • 34. Oxidation and Fermentation of Medium (CDC Method) Method 1. To determine whether acid is produced from carbohydrates, inoculate agar deeps, each containing a single carbohydrate, with bacterial growth from an 18- to 24-hour culture by stabbing a needle four to five times into the medium to a depth of 1 cm. Note: Two tubes of OF dextrose are usually inoculated; one is overlaid with either sterile melted petrolatum or sterile paraffin oil to detect fermentation. 2. Incubate the tubes at 35°C to 37°C in ambient air for up to 7 days. Note: If screwcap tubes are used, loosen the caps during incubation to allow for air exchange. Otherwise, the control tube and tubes containing carbohydrates that are not oxidized might not become alkaline.
  • 35. Oxidation and Fermentation of Medium (CDC Method) • Expected Results  Positive: Acid production (A) is indicated by the color indicator changing to yellow in the carbohydrate-containing deep.  Weak-positive (Aw): Weak acid formation can be detected by comparing the tube containing the medium with carbohydrate with the inoculated tube containing medium with no carbohydrate.  Most bacteria that can grow in the OF base produce an alkaline reaction in the control tube.  If the color of the medium in a tube containing carbohydrate remains about the same as it was before the medium was inoculated and if the inoculated medium in the control tube becomes a deeper red (i.e., becomes alkaline), the culture being tested is considered weakly positive, assuming the amount of growth is about the same in both tubes.
  • 36. Oxidation and Fermentation of Medium (CDC Method)  Negative: Red or alkaline (K) color in the deep with carbohydrate equal to the color of the inoculated control tube.  No change (NC) or neutral (N): There is growth in the media, but neither the carbohydrate-containing medium nor the control base turns alkaline (red).  Note: If the organism does not grow at all in the OF medium, mark the reaction as no growth (NG). Limitations  Slow-growing organisms may not produce results for several days. Quality Control  Note: Appropriate organisms depend on which carbohydrate has been added to the basal medium. Glucose is used as an example.  Fermenter: Escherichia coli (ATCC25922)  Oxidizer: Pseudomonas aeruginosa (ATCC27853)
  • 37.
  • 38.
  • 39.
  • 40.
  • 41.
  • 42.
  • 43.
  • 44.
  • 45. VP Positive bacteria  Viridans group streptococci (except Streptococcus vestibularis),  Listeria,  Enterobacter,  Klebsiella,  Serratia marcescens,  Hafnia alvei,  Vibrio eltor,  Vibrio alginolyticus
  • 46.
  • 47.
  • 48. Algorithm for Detection of Common Gram Negative Pathogens
  • 49.
  • 50.
  • 51.
  • 52. Identification of Common Pathogens by Colony Characteristics and Biochemical Tests
  • 55. I= Indole, M= Methyl Red, Vi= VP Test, C= Citrare (IMViC Tests) TSI agar= A/A with Gas Urea Hydrolysis
  • 56.
  • 57. I= Indole, M= MR Test, Vi= VP Test, C= Citrate Test (IMViC Tests) Urea Hydr olysis TSI Agar= A/A with Gas
  • 58.  Salmonella sp. after 24 hours growth on XLD agar.  Xylose Lysine (XL) agar is used when trying to culture and isolate Gram-negative enteric bacilli.  When XL agar is supplemented with sodium thiosulfate, ferric ammonium citrate, and sodium deoxycholate, it is then termed XLD agar, and is then an even more selective medium than XL alone.  The presence of any black colored area indicates the deposition of hydrogen sulfide, (H2S) under alkaline conditions.
  • 59.
  • 60.
  • 61.
  • 62.
  • 63.
  • 65. I= Indole, M= MR Test, Vi= VP Test, C= Citrate Test (IMViC Tests) Urea Hydr olysis TSI Agar= K/A without gas Motility Semisolid Agar
  • 67. Proteus mirabilis On Blood Agar showing SWARMING growth.
  • 68. I= Indole, M= MR Test, Vi= VP Test, C= Citrate Test (IMViC Tests) Urea Hydr olysis TSI Agar= K/A with H2S gas production. Motility Semisolid Agar
  • 69. P. aeruginosa showing Production of soluble Pyocyanin pigment on NA
  • 70. 2-3 mm, flat, smooth, non-lactose fermenting colonies with regular margin and Alligator skin like from top view
  • 71. I= Indole, M= MR Test, Vi= VP Test, C= Citrate Test (IMViC Tests) Urea Hydr olysis TSI Agar= K/K without gas. Motility Semisolid Agar NOTE- Urea Hydrolysis can be variable Oxidase Test Positive
  • 73. I= Indole, M= MR Test, Vi= VP Test, C= Citrate Test (IMViC Tests) Urea Hydr olysis TSI Agar= K/K without gas. Motility Semisolid Agar NOTE- Urea Hydrolysis can be variable Oxidase Test Negative