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HEKTEON ENTERIC AGAR (HEA) This is both a selective and differential culture medium which is used to isolate and

to differentiate the lactose fermenting from non-lactose fermenting Gram (-) bacilli. Inhibitors bile salts and citrate pH indicator bromthymol blue Hydrogen sulfide indicator ferric ammonium citrate with !arbohydrates lactose" sucrose" salicin #lue to green colonies without blac$ center of non-lactose fermenting and nonhydrogen sulfide producing Gram(-) enteric bacilli li$e% Providencia rettgeri Morganella morganii Shigella dysenteriae #lue to green colonies with black center of non-lactose fermenting but hydrogen sulfide producing Gram(-) enteric bacilli li$e% Salmonella type Proteus vulgaris Proteus mirabilis &in$ to red colonies with black center of late lactose fermenting Salmonella li$e% Salmonella arizonae 'ellow to colorless colonies with black center of non-lactose fermenting Salmonella li$e% Salmonella typhi Salmonella typhimurium a thiosulfate

XYLOSE LYSINE DEOXYCHOLATE AGAR (XLD) This is a selective and differential medium which is used to isolate and differentiate the Salmonella and Shigella species from other coliforms. Inhibitor deo(ycholate pH indicator phenol red Hydrogen sulfide indicator ferric ammonium citrate with sodium thiosulfate !arbohydrate (ylose )mino acid - lysine 'ellow colonies with black center of (ylose fermenting and hydrogen sulfide producing Gram(-) enteric bacilli li$e% Citrobacter freundii Proteus mirabilis Proteus vulgaris *'ellow colonies without blac$ center of (ylose fermenting but non-hydrogen sulfide producing Gram(-) bacilli li$e% Escherichia coli Serratia marcescens Klebsiella pneumoniae Enterobacter aerogenes Providencia rettgeri +ed colonies without blac$ center of non-(ylose fermenting Gram (-) enteric bacilli li$e% Shigella dysenteriae *+ed colonies with black center of (ylose fermenting" hydrogen sulfide producing but lysine decarbo(ylase positive Gram(-) enteric bacilli li$e% Salmonella typhi

SAL ONELLA SHIGELLA AGAR (SSA) This is a highly selective and differential medium which is used to isolate and differentiate the species of Salmonella and Shigella. Inhibitors brilliant green" bile salts" citrate pH Indicator neutral red Hydrogen sulfide indicator ferric ammonium citrate with sodium thiosulfate !arbohydrate lactose 'ellow to orange colonies with black center of lactose fermenting and hydrogen sulfide producing Gram(-) enteric bacilli li$e% Citrobacter freundii Salmonella arizonae *'ellow to orange colonies without blac$ center of lactose fermenting but nonhydrogen sulfide producing Gram(-) enteric bacilli li$e% Escherichia coli Klebsiella pneumoniae Enterobacter aerogenes Shigelle sonnei Serratia marcescens &in$ to red colonies without blac$ center of late lactose fermenting Shigella li$e% Shigella sonnei 'ellow to colorless colonies without blac$ center of non lactose fermenting Shigella li$e% Shigelle dysenteriae Shigelle flexneri Shigelle boydii

!IS "TH S"L#ITE AGAR (!SA) This is a highly selective medium for the isolation of Salmonella typhi. Inhibitors bismuth sulfite and brilliant green !arbohydrate glucose !lack with metallic sheen colonies of Salmonella typhi ,ar$ green colonies of uninhibited Gram(-) enteric bacilli

THIOS"L#ATE CITRATE !ILE SALTS S"CROSE AGAR (TC!S) This is a highly selective medium which is used to isolate and to differentiate the sucrose fermenting from non-sucrose fermenting species of Vibrio. Inhibitors thiosulfate" citrate" bile salts" and high pH pH indicator bromthymol blue !arbohydrate - sucrose 'ellow colonies of sucrose fermenting Vibrio li$e Vibrio cholerae and Vibrio alginolyticus #lue green colonies of non-sucrose fermenting Vibrio li$e Vibrio parahaemolyticus

ANNITOL SALT AGAR ( SA) This is both a selective and differential culture medium which is used to isolate and to differentiate the mannitol fermenting from non-mannitol fermenting species of Staphylococcus Inhibitor -../ a!l pH indicator phenol red !arbohydrate mannitol 'ellow colonies of mannitol fermenting Staphylococcus li$e% Staphyloccocus aureus &in$ colonies of non-mannitol fermenting Staphylococcus li$e% Staphylococcus epidermidis Staphylococcus lugdunensis Staphylococcus saprophyticus

+ed pigmented colonies of Serratia marcescens on 0ac!on$ey agar

***1warming colonies of Proteus vulgaris*** ***#lue-green pigmented colonies of Pseudomonas aeruginosa on nutrient agar plate (,iffusable pigments blue pyocyanin2 green pyoverdin)***

&in$ to purple with greenish metallic sheen colonies of lactose fermenting Escherichia coli

3arge pin$ mucoid colonies of lactose fermenting Klebsiella penumoniae

EOSIN

ETHYLENE !L"E AGAR (E !)

This is both a selective and differential culture medium which is used to isolate and differentiate the lactose fermenting from non-lactose fermenting Gram(-) bacilli. Inhibitors eosin and methylene blue pH indicators eosin and methylene blue !arbohydrate - lactose

&in$ to purple colonies with $ark center (fish eye colonies) of lactose fermenting Enterobacter aerogenes

!olorless non-lactose fermenting colonies on 40# plate li$e those of Salmonella typhi Shigella dysenteriae Providencia species" Morganella species" etc.

ACCONKEY AGAR This is both a selective and differential culture medium which is used to isolate and to differentiate the lactose fermenting from non-lactose fermenting Gram (-) bacilli. Inhibitors crystal violet and bile salts pH indicator neutral red !arbohydrate - lactose &in$ to red colonies of lactose fermenting Gram (-) enteric bacilli 'ellow to colorless colonies of non-lactose fermenting Gram (-) enteric bacilli

"REA !ROTH This li5uid medium is used to identify Gram (-) enteric bacilli based on their ability to produce the en6yme urease withing 7 to 8 hours (rapid) or within 78 hours (late) of incubation. pH indicator phenol red 1ubstrate urea If the organism produces urease" this splits urea into ammonia" carbon dio(ide" and water. )mmonia is then converted into an al$aline compound ammonium carbonate which changes the color of the medium to pin$-red. +apid urease producers (positive within 7 to 8 hours of incubation)% Proteus vulgaris Proteus mirabilis Citrobacter freundii Klebsiella pneumonia Morganella morganii Providencia rettgeri !ersinia enterocolitica Serratia marcescens.

3ate urease producers (positive within 78 hours of incubation)%

ETHYL RED% &OG"ES 'ROSKA"ER ( R%&') !ROTH This li5uid medium is also $nown as peptone glucose broth and is used to identify the Gram (-) enteric bacilli based on the ability of the organisms to ferment glucose to pyruvic acid by one of two pathways" mi(ed acid fermentation pathway which is tested by the 04TH'3 +4, T41T and the #utylene Glycol &atch which is tested by the 9:G;41 &+:1<);4+ T41T. ETHYL RED TEST SET%"' 04TH'3 +4, T41T if the organism would ferment glucose via the mi(ed acid fermentation pathway" more acids are produced li$e lactic" acetice" formic" succinic acids" which decrease the pH of the medium to 8.8 or lower hence" upon the addition of the indicator" methyl red" the broth becomes red in color. If the organism would not use this pathway" the pH of the medium increases to ... or high hence" upon the addition of the indicator" methyl red" the broth becomes yellow in color. &OG"ES 'ROSKA"ER TEST SET%"' 9:G;41 &+:<);4+ T41T If the organism would ferment glucose via the butylene glycol pathway" an intermediate product" acetyl methyl carbinol or acetoin which is neutral" is converted to diacetyl upon the addition of the 9&-+4)G4 T-# (8=/ <:H with =.>/ creatine) in the presence of 9&-+4)G4 T-) (./ alpha-naphthol in absolute methyl alcohol). ,iacetyl is red in color. egative (-) is yellow color.

SI

ONS CITRATE AGAR (SCA)

This tubed medium is used to identify the Gram (-) enteric bacilli base on the ability of the organism to utili6e citrate as the sole source of carbon (!itrate utili6ation test). The organism which utili6es citrate as its source of carbon degrades it to ammonia and then subse5uently converts it to ammonium hydro(ide. The pH of the medium is then increased and this is indicated by a change in color from green to blue.

pH indicator bromthymol blue 1ource of carbon sodium citrate o carbohydrate or protein component

LYSINE IRON AGAR (LIA) This tubed medium is used to identify the Gram (-) enteric bacilli based on the following biochemical characteristics% 3ysine deamination indicated by red slant 3ysine decarbo(ylation and glucose fermentation indicated by purple butt Glucose fermentation only indicated by yellow butt Hydrogen sulfide production indicated by blackenin( of the medium pH indicator bromcresol purple Hydrogen sulfide indicator ferric ammonium citrate with sodium thiosulfate !arbohydrate glucose )mino acid lysine &eptone

TRI'LE S"GAR IRON AGAR (TSI) This tubed medium is used to identify the Gram (-) enteric bacilli based on the following biochemical characteristics% Glucose fermentation indicated by yellow butt 3actose fermentation indicated by yellow slant Hydrogen sulfide production indicated by blackenin( of the medium Gas production indicated by presence of a crac$" bubble or gas space pH indicator phenol red Hydrogen sulfide indicator ferric ammonium citrate with sodium thiosulfate !arbohydrate ?/ glucose" ?=/ lactose" ?=/ sucrose &eptone

!ACTERIOLOGIC ANALYSIS O# )ATER 0ost &robable umber 0ethod (0& ) or 0ultiple Tube Techni5ue &resumptive Test ) set of three tubes of #rilliant Green #ile 3actose #roth (#G#3#) and water sample Tube no. ? with ?= ml (double strength) #G#3# and ?=m3 of water sample Tube no. 7 with ?= ml (single strength) #G#3# and ?.=m3 of water sample Tube no. > with ?= m3 (single strength) #G#3# and =.?m3 of water sample (small tube is called ,urham@s fermentation tube)

TY'ICAL REACTIONS ON S"L#IDE INDOLE ?. Hydrogen sulfide positive Indole (A) 0otile &ossible organism% Proteus vulgaris 7. Hydrogen sulfide (-) Indole positive 0otile &ossible organisms% Citrobacter "oseri Escherichia coli Providencia rettgeri

OTILITY

EDI"

(SI )

Morganella morganii >. Hydrogen sulfide (-) Indole (-) on motile &ossible organisms% Klebsiella pneumonia Shigella sonnei S"L#IDE INDOLE OTILITY EDI" (SI )

This semisolid medium is used to identify Gram (-) enteric bacilli based on the following characteristics% Hydrogen sulfide production indicated by blackenin( of the medium Indole production indicated by red color of the medium 0otility indicated by diffusion of growth away from the point of inoculation

COAG"LASE TEST SET%"' !oagulase test differentiates the pathogenic Staphylococcus aureus from the nonpathogenic Staphylococcus epidermidis and Staphylococcus saprophyticus 1lide test plasma plus specimen &ositive (A) visible clumping egative (-) no visible clumping Tube test plasma plus specimen &ositive (A) visible clot or coagulum egative (-) no clot or coagulum &athogenic Staphylococcus aureus &ositive (A) on-pathogenic Staphyloccocus saprophyticus egative (-) epidermidis and Staphylococcus

CHOCOLATE AGAR )n enriched medium that can be used to isolate organisms re5uiring comple( nutrients such as #aemophilus

4nriching substance is heated blood.

!LOOD AGAR 'LATE (!A') This is an enriched medium that can be used to isolate both Gram positive and Gram (-) cocci and bacilli. The enriching substance is preferable unheated ./ defibrinated sheep blood. This is also classified as differential culture medium because it differentiates organisms based on their hemolytic patterns e(hibited on this medium. )lpha hemolytic if colony is surrounded by an incomplete 6one of hemolysis which is translucent and green in color #eta hemolytic if colony is surrounded by a complete 6one of hemolysis which is clear and transparent Gamma hemolytic if colony is not surrounded by any 6one of hemolysis2 there is no change in the surrounding medium.

CATALASE TEST SET%"' !atalase test uses >/ hydrogen pero(ide. The positive reaction is indicated by bubbling or effervescence. &ositive (A) reaction is given by% )ll Staphylococcus species )ll Gram (-) enteric bacilli e(cept Shigella dysenteriae

egative (-) reaction is given by% )ll Streptococcus species