P1 Bacterial Cultivation (LECTURE) Flashcards
prelims
Refers to the increase in the number of bacteria rather than in size
- bacteria grow in number and do not grow in size
- This growth is affected by various factors such as optimum growth requirements, dynamics of growth, including the use of a medium that can be artificially prepared in the laboratory
Bacterial Growth
Bacterial Growth Requirements
- Nutrition
- Environmental
NUTRITIONAL REQUIREMENTS:
(C.ES.ES.N.W.ME.S.A/SGR.GR.TR.PHR)
- Carbon
- Energy Source
- Electron Source
- Nitrogen
- Water/Moisture/Humidity
- Mineral Elements
- Salt
- Additional/ Special Growth Requirements
- Gaseous Requirement
- Temperature Requirement
- pH Requirement
Needed for the synthesis of cellular components
Carbon
Carbon is at least ____ of weight
50%
Carbon Dioxide from the air
Autotroph
Organic compounds in the culture media (glucose)
Heterotroph
Energy source is ______ of weight?
4%
Light?
Phototroph
Chemical Energy?
Chemotroph
Electron Source
(I.O.N.F) “INOFF”
- Inorganic molecule (Fe2+)
- Organic molecule
- NADH reduced from NAD
- FADH2 reduced from FAD
- Inorganic molecule - litotroph
- Organic molecule - organotroph
Needed for the synthesis of proteins
Nitrogen
- Free nitrogen from the air
- Nitrogenous compounds in the culture media (e.g., peptone, yeast, beef extract)
Nitrogen is (___) of weight?
14%
Water/ Moisture/ Humidity is (_________) of bacteria
70%
Needed as co-factors in various metabolic process of the bacteria
Mineral Elements
Bacteria can tolerate salt concentration below (____)?
6%
certain bacteria that can survive high salt concentration environment
Halophilic bacteria or Halophiles (salt loving)
Bacterias that are Halophilic or Halophiles (salt loving):
(S.E.V.B)
- Staphylococcus spp.
- Enterococcus spp.
- Vibrio spp. (except: Vibrio cholerae & Vibrio mimicus)
- Bacillus spp.
Bacteria that are very difficult to grow
Fastidious
- requires special or additional requirements to grow in culture media
Example of blood loving organism?
Haemophilus spp.
Requires both X & V factor
Haemophilus spp.
Additional/ Special Growth Requirements
Culture media:
* intact RBC
* x factor only
Blood Agar Plate
Culture media: Blood Agar Plate
1st priority:
5% defribinated sheep’s blood
Culture media: Blood Agar Plate
2nd priority:
horse blood
Culture media: Blood Agar Plate
3rd priority:
Rabbit’s blood
Culture media: Blood Agar Plate
Least priority:
Human blood
(must be type O – no presence of antigens)
Since blood agar plate lacks V factor, use (____)?
Chocolate Agar Plate
contains NAD – v factor
* lysed RBC (heat)
* has v and x factor
Chocolate Agar Plate
hydrolyzes the NAD
NADase
hydrolyzed NAD means?
NAD = Nicotinamide Adenine Dinucleotide
No V factor
Degradation product of Hgb (hemoglobin)
X Factor (Hemin/ Hematin)
Gaseous Requirement
Bacteria that grow, live, and survive in the presence of oxygen
Aerobe
Gaseous Requirement
Absolutely requires oxygen to grown, live, and survive
Strict/Obligate Aerobe
Gaseous Requirement
Example of Strict/Obligate Aerobe bacteria?
(M.M.P.N.B.F.B.L) - spp.
- Micrococcus spp.
- Mycobacterium spp.
- Pseudomonas spp.
- Neisseria spp.
- Brucella spp.
- Francisella spp.
- Bordetella spp.
- Leptospira spp.
Gaseous Requirement
Bacteria that have the ability to grow, line and survive in small concentration of oxygen environment
Facultative Anaerobe
Gaseous Requirement
Example of Facultative Anaerobe bacteria?
(S.S.FE)
- Staphylococcus spp.
- Streptococcus spp.
- Family Enterobacteriaceae
Gaseous Requirement
Bacteria that prefers small concentration of oxygen environment
* approximately (_____)?
Microaerophilic
- 2%-10%
Gaseous Requirement
Example of Microaerophilic bacteria?
(C.H.A.S) - spp.
- Campylobacter spp.
- Helicobacter spp.
- Arcobacter spp.
- and some Streptococcus spp.
Gaseous Requirement
Bacteria that grow, live and survive in the absence of oxygen
Anaerobe
Gaseous Requirement
Absolutely do not requires oxygen to grown, live, and survive
Strict/ Obligate Anaerobe
Gaseous Requirement
Example of Strict/Obligate Anaerobe bacteria?
(C/CN.B.FB.PS.PR) - spp.
- many Clostridium spp. (Clostridium noyvi)
- Most Bacteroides spp.
- Fusobacterium spp.
- Peptostreptococcus spp.
- Porphyromonas spp.
Gaseous Requirement
Bacteria that do not require oxygen but may tolerate or withstand limited exposure to oxygen
Aerotolerant Anaerobe
Gaseous Requirement
Example of Aerotolerant Anaerobe bacteria?
(C [CP,BF])
(PB & LB)
- Some Clostridium spp. (Clostridium perfringens, Bacteroides fragilis)
- Most strains of Proprionibacterium & Lactobacillus
Gaseous Requirement
Bacteria that requires 5%-10% CO2 to grow
Capnophilic
Gaseous Requirement: Capnophilic
N.H.A.CB.E.K.SP GROUP
- Neisseria spp.
- Haemophilus spp.
- Aggregatibacter spp.
- Cardiobacterium spp.
- Eikenella spp.
- Kingella spp.
- Streptococcuse pneumoniae
Most pathogenic bacteria would grow at temperature between 35-37C
- hence incubator in the laboratory is usually set and maintained within this temperature range for routine isolation of pathogens
Temperature Requirement
Temperature Requirement
- Mesophilic
- Psychrophilic/Cryophilic
- Thermophilic
- Hyperthermophilic/ Extremely Thermophilic:
- Eurithermophilic:
- Sternothermophilic:
- 20-40C
- 0-20C
- 50-60C
- 80-110C
- wide range
- narrow range
most pathogenic bacteria can grow in a neutral or slightly alkaline environment
hence most culture media used in routine isolation of pathogens is adjusted to this (____)
pH Requirement
(pH range)
pH requirement (___)
pH 7.0 – 7.5
pH Requirement
Acid loving bacteria
Acidophilic
pH Requirement
Example of acid loving bacteria?
(LB.A)
Lactobacillus acidophilus
normal flora of vagina
pH Requirement
Alkali loving bacteria
Alkaliphilic
pH Requirement
Example of alkali loving bacteria?
(GV)
Gardnerella vaginalis
pH Requirement
Osmophilic bacteria
High Osmotic Pressure
pH Requirement
Example of osmophilic bacteria
(AB spp.)
Archaebacteria spp.
Bacterial Growth Phase
- Generation time
- Fast Growing Bacteria
- Slow Growing Bacteria
- Time to replicate & Refers to the stages of bacterial growth
- 20 minutes
- 24 hours
4 Bacterial Growth Phase:
- Lap Phase
- Log/Logarithmic/Exponential Phase
- Maximum stationary/Plateau phase
- Decline/Death phase
Bacterial Growth phase:
Bacteria are still adjusting to the new environment hence there is no cell division that occurs in this phase
they are activity synthesizing DNA & proteins that are necessary for cell division
Lag Phase
Bacterial Growth phase
The number of generation per hour
growth rate constant
Bacterial Growth Phase
Phase where there is a sudden increase of bacteria because of rapid generation or doubling time.
- also the phase where bacteria are most metabolically active hence most susceptible to the action of antimicrobial agents
Log/Logarithmic/Exponential phase
Bacterial Growth phase
After essential nutrients are depleted, toxic products accumulate or oxygen becomes limiting, the rate of cell division equates the rate of cell death
Maximum stationary/Plateau phase
Bacterial Growth Phase
Due to unfavorable environment for growth, bacterial cell division decreases while cell death becomes more accelerated
Decline/Death phase
Bacterial Growth phase: answer whether it has C.D and Increase in Number
Lag phase
C.D = Cell Division
(YES OR NO)
- Cell Division: NO
- Increase in Number: NO
Bacterial Growth phase: answer whether it has C.D and Increase in Number
Log/Logarithmic/Exponential phase
C.D = Cell Division
(YES OR NO)
- Cell Division: YES
- Increase in Number: YES
Bacterial Growth phase: answer whether it has C.D and Increase in Number
Maximum stationary/Plateau phase
C.D = Cell Division
(YES OR NO)
- Cell Division: YES
- Increase in Number: NO
Bacterial Growth phase: answer whether it has C.D and Increase in Number
Decline/Death phase
C.D = Cell Division
(YES OR NO)
- Cell Division: YES
- Increase in Number: NO
artificial preparation in the laboratory which contains basic foundation of nutrients and a solidifying agent (if needed) to support the growth of microorganisms
- Additional substances may be added to enrich the media for growth of microorganisms that are very difficult to grow (FASTIDIOUS)
CULTURE MEDIA
TERMINOLOGIES: Culture Media
CULTURE
* Noun:
* Verb:
- growth of microorganisms
- to growth/ to cultivate microorganism
TERMINOLOGIES: Culture Media
Introducing the microorganism to the culture media
Inoculate/Plant/Cultivate
TERMINOLOGIES: Culture Media
Transfer of microorganisms from one culture media to another
Transplant/ Subculture
Classification of Culture Media
Composed of known and exact amounts of pure chemical substances
* according to Composition
(S/CD)
Synthetic/Chemically Defined
Classification of Culture Media
Composed of complex materials that are rich in vitamins and nutrients that are not usually represented by a chemical formula such as peptones, beef or yeast extract, plant extracts etc.
* according to Composition
(N-S, NCD, C)
Non-Synthetic/Non-Chemically Defined/Complex
Classification of Culture Media
Live cells harvested from organs of humans and animals that supports the growth of obligate intracellular organisms that cannot grow in artificially prepared culture media
* according to Composition
(TCM)
Tissue Culture Media
Tissue Culture Media used in Microbiology Laboratory:
(VCL, MCCCL, CE, AC, HCL, H2CL)
- Vero Cell Line
- Mc Coy Cell Line
- Chicken Embryo
- A549 Cells
- HELA Cell Line
- Hep-2 Cell Line
Tissue Culture Media used in Microbiology Laboratory
Kidney cells of an African Green Monkey
source
Vero Cell Line
Tissue Culture Media used in Microbiology Laboratory
Mouse cell line
source
Mc Coy Cell Line
Tissue Culture Media used in Microbiology Laboratory
Fertilized chicken egg
source
Chicken Embryo
Tissue Culture Media used in Microbiology Laboratory
Human lung carcinoma
source
A549 Cells
Tissue Culture Media used in Microbiology Laboratory
Human cervical carcinoma
source
HELA Cell Line
Tissue Culture Media used in Microbiology Laboratory
Human epithelial cells of larynx carcinoma
source
Hep-2 Cell Line
Classification of Culture Media according to Physical State/Consistency
a culture medium that doesn’t contain a solidifying agent
Liquid
Classification of Culture Media according to Physical State/Consistency
A culture medium that contains 0.5% - 1% agar
Semi-solid
Classification of Culture Media according to Physical State/Consistency
Used for observation of hydrogen sulfide gas production, indole production and motility
under semi-solid
(SIMM)
Sulfide Indole Motility (SIM) Medium
Classification of Culture Media according to Physical State/Consistency
A culture medium which contains 1.5% - 3%T (2%-3%) agar
Solid
Classification of Culture Media according to Manner of:
Usually contained in a container that can be made of glass (pyrex) or disposable plastic (Petridish)
Dispensing/Formation
Plated
Classification of Culture Media according to Manner of:
Usually, container in glass tubes such as Wassermann tubes with different volume capacity (3mL, 5mL, 10mL) or in a tube with a flat bottom and a screw cap
Dispensing/Formation
Tubed
Classification of Culture Media according to Manner of:
Culture media contained in a glass bottle that is usually used for blood culture
Dispensing/Formation
Bottled
Classification of Culture Media according to Function/ Use
Contains basic nutritional requirements to support the growth of non-fastidious microorganisms
This is also used a base medium in the preparation of other culture media
General Purpose/Primary/Basic/Basal/ Supportive/General Isolation Culture Media
Classification of Culture Media according to Function/ Use
Contains the basic nutritional requirement to support the growth of non-fastidious microorganisms with additives, enriching substances, or supplements to support the growth of fastidious microorganisms
(e.g. Blood Agar & Chocolate Agar Plate)
Enriched Culture Media
Classification of Culture Media according to Function/ Use
A primary media used to support or favor the selective growth of pathogens in a specimen, such as stool or sputum, where the number of normal flora outnumber the pathogens
* Use commonly stool specimen
(EB)
Enrichment Broth
Examples of Enrichment Broth:
(APW, SFB, THIOB, GNB,T-HB)
- Alkaline Peptone Water (APW)
- Selenite F Broth
- Thioglycolate Broth (THIO)
- GN Broth (Gram Negative Broth)
- Todd-Hewitt Broth
Used to selectively favor the growth of Vibrio while inhibiting all other normal intestinal flora due its high pH
Alkaline Peptone Water (APW)
Used to selectively favor the growth of Salmonella while inhibiting all other normal intestinal flora
Selenite F Broth
An enrichment broth for anaerobic bacteria but should be used solely in the isolation of anaerobic bacteria since it can also grow aerobes and facultative anaerobes
(THIOB)
Thioglycolate Broth (THIO)
Used to selectively favor the growth of Salmonella and Shigella while inhibiting all other normal intestinal flora
GN Broth (Gram Negative Broth)
A liquid enrichment recommended for the production of Streptococcal haemolysin and the cultivation of Streptococci prior to serological grouping
Todd-Hewitt Broth
A primary isolation culture media which maintains the viability of bacteria allowing rapid multiplication if there is an anticipated delay in bringing the specimen collected bedside or remotely to the laboratory
(TCM)
Transport Culture Media (TCM)
Favors the growth of the organism of interest using inhibitors added in the culture media
(SCM)
Selective Culture Media
Contains indicators which changes in color as a result of a product produced be a chemical reaction in the components of the media such as glucose
(DCM)
Differential Culture Media
NOTE:
it is important to remember and understand that not all selective culture media are differential but all differential culture media are selective
INHIBITORS IN CULTURE MEDIA
- Dyes:
- Chemicals:
- Antibiotics:
Inhibitors for Gram Positive Bacteria
(CV,E,MB,BG,ETC)
(BMS,BS[SD],TS,C,ETC.)
(V)
- Crystal violet, eosin, methylene blue, brilliant green, etc.
- Bismuth sulfite, bile salts (sodium desoxycholate), thiosulfate, citrate, etc.
- Vancomycin
INHIBITORS IN CULTURE MEDIA
- Dyes:
- Chemicals:
- Antibiotics:
Inhibitors for Gram Negative Bacteria
(BF & T [BA])
- Basic fuchsin and
- thionine for Brucella abortus
INHIBITORS IN CULTURE MEDIA
- Antibiotics:
Inhibitors for Fungi
(N.A.AB)
- Nystatin
- Anisomycin
- Ampothericin B.
INHIBITORS IN CULTURE MEDIA
- Indicators in Culture Media:
- Dyes and chemical substances such as pH indicators
Eosin Methylene Blue (EMB) Agar
- Original Color:
- Selective for:
- Inhibitors:
- Differential Indicators:
(DV)
(GNEB)
(E&MB)
(E&MB)
MOST COMMONLY AND ROUTINELY USED SELECTIVE & DIFFERENTIAL CULTURE MEDIA IN THE LABORATORY
- Dark violet
- Gram negative enteric bacilli
- Eosin and Methylene Blue
- Eosin and Methylene Blue
RLF: Rapid Lactose Fermenter
Number of hrs of incubation
able to ferment lactose for 18-24 hours incubation
LLF: Late Lactose Fermenter
number of hrs of incubation
able to ferment lactose for 36-72 hours incubation
RLF
medium sized dark violet colonies with greenish metallic sheen
(EC)
Differentiation of Family Enterobacteriaceae based on lactose fermentation
Escherichia
(E. coli)
RLF
Appears medium sized dark violet colonies with/without dark center
(EB)
Differentiation of Family Enterobacteriaceae based on lactose fermentation
Enterobacter
RLF
Large sized mucoid dark violet colonies with or without dark center
(KS)
Differentiation of Family Enterobacteriaceae based on lactose fermentation
Klebsiella
LLF bacteria:
(HSCB,SA,SS,YEC)
- Hafnia, Serratia, Citrobacter
- Salmonella arizonae
- Shigella sonnei
- Yersinia enterocolitica
NLF (appear colorless colonies) bacteria:
(PPME)
- All Salmonella except S. arizonae
- All Shigella except S. sonnei
- All Yersinia except enterocolitica
- Proteus, Providencia, Morganella, Edwardsiella
Mc Conkey (MAC) Agar
- Original Color:
- Selective for:
- Inhibitors:
- Differential Indicators:
(LP)
(GNEB)
(CV, BS, C)
(NR)
- Light pink
- Gram negative enteric bacilli
- Crystal violet, bile salts, Citrate
- Neutral Red
Salmonella-Shigella (SSA) Agar
- Orginal color:
-
Selective
For:
Inhibitors: -
Differential:
Indicators: pH indicators:
H2S indicator:
Sulfur Source:
(LO)
(S & SG spp.)
(BG, BS, C)
(NR)
(FAC)
(ST)
- Orginal color: Light orange
- For: Salmonella and Shigella spp.
- Inhibitors: Brilliant Green, Bile Salts, Citrate
- Indicators: pH indicator: Neutral red
- H2S Indicator: Ferric ammonium citrate
- Sulfur Source: Sodium thiosulfate
Hektoen Enteric (HEA) Agar
- Orginal color:
-
Selective
For:
Inhibitors: -
Differential:
Indicators: pH indicators:
H2S indicator:
Sulfur Source:
(DG)
(GNEB)
(BS, C)
(BTB)
(FAC)
(ST)
- Orginal color: Dark Green
- For: Gram negative Enteric bacilli
- Inhibitors: Bile Salts, Citrate
- Indicators: pH indicator: Bromthymol Blue (BTB)
- H2S Indicator: Ferric ammonium citrate
- Sulfur Source: Sodium thiosulfate
Bismuth Sulfite (BSA) Agar
-
Selective
For:
Inhibitors:
CHO Incorporated:
(S spp)
(BMS)
(GL)
- For: Salmonella spp. (Salm. Typhi has distinct appearance)
- Inhibitors: Bismuth sulfite
- CHO Incorporated: Glucose
Salmonella typhi colonies appear as black colonies with silver metallic sheen
Brilliant Green (BGA) Agar
-
Selective
For:
Inhibitors:
CHO Incorporated:
(S spp/S t)
(BG)
(LC)
- For: Salmonella spp. except for Salmonella typhi
- Inhibitors: Brilliant green
- CHO Incorporated: Lactose
Salmonella spp., colonies appear as white colonies resembling a snowflake surrounded by brilliant red medium
Thiosulfate Citrate Bile Salts Sucrose (TCBS) Agar
- Orginal color:
-
Selective
For:
Inhibitors: -
Differential:
Indicators:
(LG/OG)
(VB spp.)
(TS, C, BS)
(BTB)
- Original Color: Light Green/ Olive Green
- For: Vibrio spp.
- Inhibitors: Thiosulfate, Citrate, Bile Salts
- Indicators: Bromthymol Blue (BTB)
Mannitol Salt (MSA) Agar
- Orginal color:
-
Selective
For:
Inhibitors: -
Differential:
Indicators:
(L/SP)
(SP spp.)
(^ s 7.5%)
(PR)
- Original Color: Light/ Salmon Pink
- For: Staphylococcus spp.
- Inhibitors: High concentration of salts (7.5%)
- Indicators: Phenol Red
Lowenstein Jensen (LJ) Medium
- Orginal color:
-
Selective
For:
Inhibitors:
(LG)
(MB spp.)
(MG)
- Original Color: Light Green
- For: Mycobacterium spp.
- Inhibitors: Malachite Green
Lowenstein Jensen (LJ) Medium
Sputum Sample needs to be:
(DC/DG)
(DCT)
- Decongested/ Digested
- Decontaminated
Lowenstein Jensen (LJ) Medium
To dissolve the thick mucus/mucin that might be trapping the bacteria in the sample.
N-acetyl-L-cysteine (NALC) is usually used
(DC/DG)
Decongested/Digested
Lowenstein Jensen (LJ) Medium
To eliminate normal flora that contaminates the sample.
NaOH is usually used
Decontaminated
Selective Medium for Neisseria spp.
(N. spp - GNC)
(CHA-AB)
- Neisseria spp., Gram negative cocci
- Usually composed of chocolate agar base with antibiotics
Selective Medium for Neisseria spp.
- Gram + Inhibitor:
- Gram - Inhibitor:
- Fungal Inhibitor:
- Proteus spp. Inhibitor
Culture media: Thayer Martin Agar (TMA)
(V,C,N, -)
- Vancomycin
- Colistin
- Nystatin
- no proteus spp. inhibitor
Selective Medium for Neisseria spp.
- Gram + Inhibitor:
- Gram - Inhibitor:
- Fungal Inhibitor:
- Proteus spp. Inhibitor
Culture media: Modified Thayer Martin Agar (MTMA)
- Vancomycin
- Colistin
- Nystatin
- Trimethoprim
Selective Medium for Neisseria spp.
- Gram + Inhibitor:
- Gram - Inhibitor:
- Fungal Inhibitor:
- Proteus spp. Inhibitor
Culture media: Martin-Lewis Agar (MLA)
- Vancomycin
- Colistin
- Anisomycin
- Trimethoprim
Selective Medium for Neisseria spp.
- Gram + Inhibitor:
- Gram - Inhibitor:
- Fungal Inhibitor:
- Proteus spp. Inhibitor
Culture media: New York City Agar (NYCA)
- Vancomycin
- Colistin
- Amphotericin B
- Trimethoprim
Selective Medium for Neisseria spp.
- Gram + Inhibitor:
- Gram - Inhibitor:
- Fungal Inhibitor:
- Proteus spp. Inhibitor
Culture media: GC-LECT
- Decreased concentration of Vancomycin; added w/ Lincomycin
- Colistin
- Amphotericin B
- Trimethoprim
CULTURE MEDIA FOR ANTIBIOTIC SUSCEPTIBILITY/SENSITIVITY TESTING (AST)
(MB, HP spp., MB spp.)
- Most bacteria
- Haemophilus spp.
- Mycobacterium spp.
CULTURE MEDIA FOR ANTIBIOTIC SUSCEPTIBILITY/SENSITIVITY TESTING (AST)
Mueller Hinton Agar (MHA) and Mueller Hinton Broth (MHB)
Most Bacteria
CULTURE MEDIA FOR ANTIBIOTIC SUSCEPTIBILITY/SENSITIVITY TESTING (AST)
Mueller Hinton with Chocolate Agar Base or Haemophilus Test Medium (HTM) Agar
(fastidious organism – difficult to grow)
Haemophilus spp.
CULTURE MEDIA FOR ANTIBIOTIC SUSCEPTIBILITY/SENSITIVITY TESTING (AST)
Middlebrook 7H10 or 7H11 Medium
Mycobacterium spp.
Characteristic/Biochemical Culture Media
(SIMM,MR,VP,SCA,TSIA,LIA,MB,SUB/CUA)
- Sulfide Indole Motility Medium (SIM)
- Methyl Red (MR)
- Voges-Proskauer (VP)
- Simmon Citrate Agar (SCA)
- Triple Sugar Ion Agar (TSI)
- Lysine Iron Agar (LIA)
- Moeller’s Broth
- Stuart’s Urea Broth or Christensen Urea Agar
Characteristic/ Biochemical Culture Media
Used for observation of hydrogen sulfide gas production
Sulfide Indole Motility Medium (SIM)
Characteristic/ Biochemical Culture Media
there will be presence of black precipitate
Sulfide Indole Motility Medium (SIM)
(HSGP)
hydrogen sulfide gas production
Characteristic/ Biochemical Culture Media
can be observed by the appearance of red or pink ring formation
Sulfide Indole Motility Medium (SIM)
(IP)
indole production
Characteristic/ Biochemical Culture Media
presence of turbidity or spread of colonies along the side of inoculation
Sulfide Indole Motility Medium (SIM)
(M)
motility
Characteristic/ Biochemical Culture Media
Used for the detection of bacterial pathogen that metabolize glucose using the Mixed Acid pathway
- Add (__) as reagent
Methyl Red (MR)
Characteristic/ Biochemical Culture Media
Used for the detection of bacterial pathogen that metabolizes glucose using the Butylene Glycol Pathway
- Add alpha-naptol as reagent
Voges-Proskauer (VP)
Note:
* Positive MR = Negative VP
* Negative MR = Positive VP
(to determine what pathway of glucose metabolism)
Characteristic/ Biochemical Culture Media
Used for the detection of bacterial pathogen that can utilize citrate as a sole source of carbon
- Green slanted media (original color)
- Positive result (blue)
Simmon Citrate Agar (SCA)
Characteristic/ Biochemical Culture Media
Used for the determination of bacterial pathogen’s ability to ferment glucose, sucrose or lactose.
- It can also detect sulfide
production (blackening of the agar) - gas production (+ result = bubbles, cracks or spaces on the culture media)
Triple Sugar Ion Agar (TSIA)
Characteristic/ Biochemical Culture Media
- Red/red:
- Control
- Red/yellow:
- Yellow/yellow:
- Red/yellow with H2S:
Triple Sugar Ion Agar (TSI)
- no sugar fermentation
- control
- Glucose fermented but lactose and sucrose not fermented
- Glucose fermented. Lactose and/or sucrose fermented
- Red/yellow with H2S
Triple Sugar Ion Agar (TSI): percentage/parts
- dextrose:
- sucrose:
- lactose:
the “parts” are SIMPLIFIED VERSION
- 0.1% (1 part)
- 1.0% (10 parts)
- 1.0% (10 parts)
Characteristic/ Biochemical Culture Media
Used for the determination of bacterial pathogen’s ability to decarboxylate or deaminate lysine.
- It can also detect glucose fermentation, sulfide production and gas production
Lysine Iron Agar (LIA)
Characteristic/ Biochemical Culture Media
Used to detect lysine decarboxylation, ornithine decarboxylation, and arginine dihydrolysis
Moeller’s Broth
Characteristic/ Biochemical Culture Media
Used to detect bacterial pathogen that hydrolyze urea substrate
Stuart’s Urea Broth or Christensen Urea Agar