Gram negative rods. Oxidase negative. Catalase positive. Facultative anaerobes. Ferment glucose. Usually cause GI diseases. Main difference is the ability to ferment Lacotose or Non lactose fermenting. Lac positive are usually normal flora and cause opportunistic infections nosocomial. Where as lac negative are normally outright pathogens.
Esherichia coli O157:H7 fermentation
Cannot ferment sorbital. Main difference between normal Escherichia coli. this can be distinguished by a SMAC plate. If the E.coli does not ferment the sorbital in the plate it is O156H7
Brilliant green agar (BGA)
Isolation of most Salmonella species. Selective agents. Brilliant green inhibits growth of Gram positives and most Gram negatives highly selective for Salmonella. DIfferential agents Lactose and sucrose bacteria that ferment these sugars produce yellowish colonies with yellow halo. Salmonella spp. do not ferment lac or suc colonies appear white with red halo
Eosin-methylene blue (EMB) agar
"Primary plating medium for Gram negative enterics. Selective agents Eosin & methylene blue dyes: select for growth of Gram negatives
Hektoen-enteric (HE) agar
"Primary plating medium for Gram negative enterics. Selective agents Bile salts inhibit growth of many Gram positives and non-enteric Gram negatives. Selective agents Sugars (lactose sucrose salicin): bacteria that ferment 1 or more of these sugars produce yellow/orange colonies
H2S production: bacteria that produce H2S form colonies with black centers. Often used for isolation of Salmonella and Shigella species H2S production is the differentiating factor
MacConkey agar (MAC)
Primary plating medium for Gram negative enterics. Selective agents Bile salts and crystal violet inhibit growth of most Gram positives. Differential agents. Lactose: bacteria that ferment lactose produce bright pink colonies. E. coli (and some other enterics) precipitate bile salts and produce cloudy zones around the colonies
SMAC (MAC + sorbitol) agar
Isolation and identification of E. coli O157H7. Exactly same as MAC plate but has Sorbitol bacteria that ferment sorbitol produce bright pink colonies. Since O157H7 can not ferment this it will have buff colored colonies.
Salmonella-Shigella (SS) agar
"Isolation and identification of Salmonella and Shigella species. Selective agents Biles salts and various dyes inhibit growth of most Gram positives.Differential agents Lactose bacteria that ferment lactose produce reddish colonies. H2S production bacteria that produce H2S form colonies with black centers. Salmonella and Shigella will form colorless colonies Salmonella is positive for H2S
Triple sugar iron (TSI) agar
"Glucose lactose and sucrose fermentation pattern Ferrous iron forms black precipitate if H2S is produced. Yellow butt red slant only glucose fermented.Yellow butt yellow slant glucose and lactose OR sucrose fermented
Xylose-lysine-deoxycholate (XLD) agar
"Primary plating medium for Gram negative enterics especially useful for Shigella isolation/identification. Selective agent Deoxycholate (bile) inhibits most Gram positives. Selective agents Xylose bacteria that ferment xylose produce yellow coloniesH2S production bacteria that produce H2S form colonies with black centers. Shigella does not ferment xylose and appears red Salmonella ferments xylose and appears yellow
After prolonged incubation Salmonella will decarboxylate lysine and turn red can then be distinguished from Shigella by black center
Used primarily for Lac negative Enterobacteriacae. Are the sugars Lysine. Orithine. Arnthenine. and a Control. Must use mineral oil on top to create anaerobic environment for fermentation. Works by removing carboxylic acids and can use them for an energy source making the solution basic. The tubes contain glucose plus one of the amino acids. If the bacteria can use the AA the solution will turn basic by breaking down the AA to an amine. The solution will turn back to its original blue color. yellow is a negative test.
Is an enzyme that removes an amine from phenylalanine. Uses teh enzyme deaminase to create phenyl pyruvic acid and an amine. To test this add ferric choloride to the slant and look for a BRIGHT GREEN COLOR.
TSI A/AG-. h2s negative. IDOLE POSITIVE. MOTILITY POSITIVE. MR POSITIVE. vp negative. citrate negative. urease positive. maltose negative. Main test SMAC PLATE WITH DULL COLORED COLINES. Not orange like normal e. coli.
A/AG-. h2s negative. Indole negative. motility negative. mr negative. VP POSITIVE. UREASE POSITVE. CITRATE POSITVE. MALONATE POSITVE. to differentiate between citrobacter and klebisella use molilty and that citrobacter is MR positve. Remeber k. pneumonia is different from k. oxytoca by indole
A/AG-. h2s negative. ((INDOLE POSITIVE)). motility negative. mr negative. VP POSITIVE. UREASE POSITVE. CITRATE POSITVE. MALONATE POSITVE. to differentiate between citrobacter and klebisella use molilty and that citrobacter is MR positve. Remeber k. pneumonia is different from k. oxytoca by indole
PROTEUS IS A SWARMER AND HIGHLY MOTILE. ALL PHEN. DEAMINASE POSITIVE. MOTILITY POSITIVE. Proteus stuartii is the only H2S NEGATIVE. Proteus mirabolis is K/AG+ and indole and sucrose negative but citrate postive ornithine Positve. Proteus vulgaris is A/Ag+ and ferments sucrose. Is indole negative and cirtrate negative. ornithine negative.
A/A- INDOLE POSITIVE. Sulfide negative. MOTILITY POSITIVE. MR POSITIVE. VP negative. Citrate negative. UREASE POSITIVE. Phen. deaminase negative. Lysine decarb negative. Arginine decarb negative. ORNITHINE POSITIVE. SUCROSE POSITIVE. IS NON MOTILE AT 35 DEGREE BUT MOTILE AT ROOM TEMP.
Split up lac negative enterbactericea
If it is PHEN. DEAMINASE POSITVE AND MOTILE (SWARMING) PROTEUS.. Negative everything except MR or One decarb AA is SHIGELLA. ALL POSITVE DECARBS IS SALMONELLA TYPHIMURIUM. If it is H2S negative and Indole positve and phen. negative but sucrose postive is Yersina enterocolitica.
Differentiate between Protues
Look for H2S production. Non H2S is Y. stuartii. Only H2S positive with A/AG+ is P. vulgaris and this is because of SUCROSE. P. mirabilis is positive for citrate and negative indole and positive ornithine.
Campylobacter general information
Gram negative s shaped. can be helical. OXIDASE POSITIVE. Microaerohilic. When staining it is hard to do secondary stain and must use carbofusion instead of safranin.
Isolation and growth of Campylobacter especially C. jejuni. Selective agents Antibiotics (cefoperazone vancomycin amphotericin B) inhibit growth of other bacteria. Contains sheep blood
Charcoal-based selective media (CSM)
Isolation and growth of Campylocbacter species. Antibiotics (cefoperazone vancomycin cyclohexamide) inhibit growth of other bacteria. Blood-free medium charcoal acts as a quencher of reactive O2 species
Marine agar (MAR)
Isolation and enumeration of marine bacteria. selective High salt concentration: selects for halophilic bacteria. No differential agent Vibrio cholerae is not halophilic and will not grow! All other Vibrio species should grow
Rimler-Shotts (RS) media
"Isolation and identification of Aeromonas species. Maltose colonies that ferment maltose appear yellow. Thiosulfate detection of H2S production. Contains novobiocin. Lysine and ornithine bacteria that decarboxylate lysine or ornithine turn media yellow.
Thiosulfate-citrate-bile salts-sucrose (TCBS) agar
"Primary isolation of Vibrio species particularly V. cholerae. Bile salts inhibit Gram positive organisms
Alkaline pH enhances growth of Vibrio and discourages growth of other enterics. Thiosulfate and ferric ammonium citrate detection of H2S production Sucrose colonies that ferment sucrose appear yellow. Good for differentiating Vibrio species based on sucrose fermentation
NALIDIXIC ACID SENSITIVE. (disk placed on campy CVA agar) cephalothin resistant (disk on campy CVA). GROWTH AT 42 POSITIVE. HIPPURATE POSITIVE.
Chemical test that use the indicator dye phenyl red to indicate if fermentation of these sugars can be conducted. Test used for vibrio species
Yellow colonies on TCBS agar (sucrose ferm). BETA hemolysis. SENSITIVE TO O129 disk. GROWTH AT 0 NACL. GROWTH AT 1 NACL. GROWTH AT 3 PERCENT NACL. No growth from 6-10 NaCL. Lysine positivite. ORINTHINE POSITIVE. pr arabinose negative. PR GALACTOSE POSITIVE. Urease negative.
Light blue or green colonies on TCBS agar (no sucrose ferm). Alpha hemolysis. NO GROWTH AT 0 NACL. GROWTH FROM 1 TO 6 PERCENT NACL. No growth at 10 percent bile. ORNITHINE POSITIVE. PR ARABINOSE PR GALACTOSE. ((((UREASE POSITIVE)))
Yellow colonies (sucorse ferm on TCBS). Alpha beta hemolysis. O129 SENSITIVE. No growth at 0 NaCl. GROWTH FROM 1 TO 6 PERCENT NACL. No growth at 10 percent. ORINTHINE NEGATIVE. Arginine negative. Urease negative. Similar to V. cholera but can not grow without NaCl.
Yellow colonies on TCBS. O129S. No growth at 0 NaCl. GROWTH FROM 1 TO 10 PERCENT NACL. HALOPHILE. NEGATIVE ORNITHINE. NEGATIVE ARGININE. NEGATIVE PR ARABINOSE AND PR GALACTOSE. Negative urease.
If growth on TCBS will be blue colonies. YELLOW COLONIES WITH BLACK CENTERS (H2S) ON RM (rimler shots media maltose fermentation. thiosulfate). (((O129 RESISTANT))). GROWTH FROM 0 TO 3 PERCENT NACL. No growth from 6 to 10 percent. LYSINE POSITIVE. Ornithine negative. ARGININE POSITVE. PR ARABINOSE AND PR GALACTOSE POSTIVE. Urease negative
Differentiate between Virbrio species.
For Vibrio cholera use the growth at 0 percent NaCl. With the beta hemolysis and growth on TCBS media. For Vibrio parahemolyticus you can use the blue colonies on TCBS media and with its ability to be urease positve and additionally it is both PR arabinose and galactose positive. For Vibrio vulnificus you can look at the no growth at 0 percent NaCl along with growth up to 6 percent NaCl. For Vibrio alginolyticus you can use the fact it is a halophile and will grow up to 10 percent NaCl also it does not use both arabinose and galactose.
Differentiate between vibrio and aeromonas hydrophila
Use the fact that Vibrio is sensitive to O129 disks where Aermonas is not. Aeromonas will grow on RS media which will show yellow colonies with H2S production along with maltose fermentation. Aeromonas is additonally arabinose and galactose positive.
General info about vibrio
Mainly intestional pathogens. A comma shaped bacteria that is gram negative and OXIDASE POSITIVE. Most need NaCl to grow except for cholera.
General info on Gram negative non-fermenters
HAVE OXIDATIVE METABOLISM. Strict aerobes unless you give a different final acceptor like Nitrate. TSI/ IS K/N-. Found in normal flora. solil. water. associated with plants. can cause wound infections. Bacteria are pseudomonas aeruginosa. pseudomonas fluorescens. Acinetobacter baumanii. Alcaligenese faecalis. Stenotrophomonas maltophilia. Shewanella putrifaceins.
Cetrimide (CET) agar
Selective isolation of Pseudomonas aeruginosa. Selective agents Cetrimide a cationic surfactant that inhibits growth of normal flora. Differential agents Cetrimide enhances pigment production by P. aeruginosa. Pigments produced by P. aeruginosa include pyocyanin (blue-green) and fluorescein (green-yellow)
Stimulates production of fluorescein pigment and inhibits production of pyocyanin pigment. Differential agent Magnesium sulfate: stimulates fluorescein production. Pseudomonas aeruginosa appears yellow-green due to fluorescein production
Stimulates production of pyocyanin pigment and inhibits production of fluorescein pigment. Magnesium chloride and potassium sulfate stimulate pyocyanin production. Pseudomonas species that produce pyocyanin will form blue pigment
Oxidase positive. Beta hemolysis. TSI K/N-. P AGAR POSITIVE. F AGAR POSITIVE. NITRATE REDUCTION (N2). UREASE POSITIVE. GLUCONATE POSITIVE. GELATIN HYDROLYSIS. O/F GLUCOSE. O/F MALTOSE. Growth at 5 celciuss. GROWTH AT 42 POSITIVE. Lac negative on MAC. GROWTH ON CET GREEN.
Oxidase positive. Weak beta hemolysis. K/N- P agar negative. F AGAR POSITIVE. Nitrate reduction negative. UREASE POSITVE. GLUCONATE POSITVE. MOTILITY POSITIVE. GELATIN HYDROLYSIS. O/F GLUCOSE POSITIVE. O/F maltose negative. (((GROWTH AT 5 DEGREES))). No growth 42 degree. Lac - MAC. Growth on CET white colonies..
OXIDASE NEGATIVE. Weakly beta hemolytic. K/N-. P agar negative. F agar negative. Nitrate negative. urease negative. gluconate negative. motility negative. gelatin hydrolysis negative. o/f glucose negative. o/f maltose negative. growth at 5 degree negative. GROWTH AT 42 degree. GROWTH ON MAC PLATE WITH SLIGHT PINK COLONIES Growth on CET white colonies.
Oxidase positive. K/N- . P agar negative. F agar negative. Nitrate reduction negative. urease negative. glucontate negative. ((MOTILITY POSITIVE)). gelatin hydrolysis negative. o/f glucose negative. o/f maltose negative. growth at 5 degree negative. growth at 42 growth positive. apperance on MAC lac negative. Growth on CET white colonies.
OXIDASE NEGATIVE. K/N-. P agar negative. F agar negative. (((NITRATE REDUCTION POSITIVE))). UREASE POSITIVE. gluconate negative. MOTILITY POSITVE. GELATIN HYDROLYSIS POSITIVE. O/F GLUCOSE POSTIVE. (((O/F MALTOSE))). no growth at 5 or 42 degree. Apperance on MAC lac -. Growth on CET white colonies.
Oxidase positive. gamma hemolysis. (((K/N+))). SULFIDE POSITVE. p agar negative. f agar negative. NITRATE REDUCTION POSITIVE (N2). UREASE POSITVE. Gluconate negative. MOTILITY POSITVE. GELATIN HYDROLYSIS POSITIVE. O/F glucose negative. O/Fmaltose negative. No growth at 5 or 42 degree. Mac plate is G-. ((NO GROWTH ON CET PLATE)).
General information on Neisseria
Gram negative cocci in pairs. Diplococci. Can be kidney shaped. N. mennigitis and N. gonorrhea are fastidious and only survive in humans and 48 hours in lab media. Grow in increased CO2. OXIDASE POSITIVE. all aerobic. Specific ones N. gonorrheae. N. meningitidis. N. sicca. N. muscosa. and Moraxella catarrhalis which is a coccobacillus that has similarities to neisseria. OXIDATIVE ATTACK OF SUGARS!! USE CTA SUGARS TO CHECK FOR OXIDATION
Modified Thayer martin mTm
Chocolate agar base but has antibiotics (vancomyocin. cholistin. and nistatin in to inhibit other bacterial growth).
Blood loving bacterai. Gram negative coccobacillus. Grow with increased CO2. Fastidious. REQUIRMENT ARE CO FACTORS.
X factor requried. Do not require heme will be positive. No substrate. ALA substrate porphobilinogen precursor of protoporphyrin which will turn to heme. Detects porphyrin UV light.
Porphyrin synthesis negative. CAS POSITIVE. X negative. V negative. CHOCOLATE POSTIVE. TSA Plate no growth X only or Y only (X+Y GROWTH)
PRORPHYIN SYNTHESIS POSITIVE. CAS +. x negative. V POSITIVE. CHOCOLATE POSITVE. TSA plate x negative (V AND X+V POSITIVE.) NEEDS V ONLY!!!!!
Has 4 quadrants. I has BA which is x/y used for hemolysis. II is x only. III is V only. IV is chocolate agar used as a control.
BA with satellite colonies
Will streak a BA plate with S. aureus to cause hemolyosis and release X cofactor. Resease of heme.
pre reduced anaerobically sterilized media. Prepared and stored under oxygen free conditions.
Bacteroides bile esculin
Isolation and differntiatino of anaerobes particular Bacteroides. Has Gentamiicin inhibits facultative anaerobes. Bile selects for resistant anaerobes such as Bacteroides. DIFFERENTIAL AGENTS. Esculin hydrolysis of esculin produces blackening of medium around colonies.
Brucella BA. Used for isolation and cultivation of obilgate anaerobes from clincial specimens. Sheep blood for hemolysis reaciton. Non selective media. general purpose.
Brucella leaked vancomycin and kanamycin. Used for identification of obiligately anaerobic Gram negative bacilli. Selective agents are vancomycin and kanamycin. DIFFERENTIAL AGENT vitiamin K1 which enhances pigment production by Prevotella. Blood is laked or lysed to release heme.
CCFA cycloserine cefoxitin fructose agar
Used for the isolation and identification of Clostridium difficile. Selective agents are cefoxitin and cycloserine inhibt most fecal flora. DIFFERENTIAL. fructose adn ph indictator. If fructose fermenetatin occurs media will turn yellow that fluoresce golden under UV light.
Egg yolk agar EYA
Cultivation and isolation of anaerobes. NO selective agents. DIFFERENTIAL AGENTS. egg yolk. lecithinase will appear as opaque precipitate around coloniesl Lipase iridescent sheen on colonies. Proteolysis clearing of medium around colonies.
PEA BA penylethyl alcohol blood agar
cultivation and isolation of anaerobes. Phenylethyl alcohol inhibits gorwth of gram negative facultatively anaerobic bacteria. prevents swarming of proteus. NO differential agents. Useful for isolation of anaerboes from mixed culturs taht might contain Proteus.
Growth of anaerobes no selective agents. DIFFerntial agent is sodium thioglycollate creates oxygen gradient from top to bottom of tube. Obligate anaerobes will grow at the bottom where there is no oxygen.
Gamma hemolytic. Lecithiniase negative. LIPASE POSITIVE. Disk sensitivity Colistin resistant Kanamyicin Sensitive Vancomycin Sensitive. (RSS) NO LITMUS MILK REACTION
Negative lecithinase. Negative lipase. POSITIVE PROTEASE. Weak beta. LITMUS MILK CURD. Semisolid from curd production from coagulation of caesin. Disks Colistin Resistant. kanamycin Sensitive. Vancomycin Sensitive. (RSS)