Hektoen_Enteric_Agar_TD

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Manufacturer Address: A- 902A RIICO Industrial Area Phase III Bhiwadi-301019. Page 1 www.tmmedia.in PRODUCT DATA SHEET INTENDED USE Hektoen Enteric Agar For differential selective isolation of Salmonella and Shigella from enteric pathological specimens. COMPOSITION Ingredients Gm\Ltr. Agar 15.000 Proteose peptone 12.000 Lactose 12.000 Sucrose 12.000 Bile salts mixture 9.000 Sodium chloride 5.000 Sodium thiosulphate 5.000 Yeast extract 3.000 Salicin 2.000 Ferric ammonium citrate 1.500 Acid fuchsin 0.100 Bromothymol blue 0.065 PRODUCT SUMMARY AND EXPLANATION Hektoen Enteric Agar was developed in 1967 by King and Metzger of the Hektoen Institute in order to increase the frequencies of isolation of Shigella and Salmonella organisms when compared with their recovery on other media frequently utilized in clinical laboratories at that time. Sodium deoxycholate has been replaced by bile salts in reduced concentration. This allows growth of Shigella as well as the Salmonellae. The peptone concentrations have been increased in order to offset the inhibitory effects of the bile salts. Hektoen Enteric Agar is currently recommended as one of several plating media for the culture of Enterobacteriaceae from stool specimens. Foods containing poultry eggs or dairy products are the most frequent vehicles for foodborne Salmonellosis and a variety of procedures have been developed using Hektoen Enteric Agar as part of the multi-step procedure to isolate Salmonella. This medium is recommended by United States Pharmacopoeia 2009 for testing the presence of Salmonella in dietary supplements. This medium is recommended in testing of Salmonella in food sample by various standards. HEKTOEN ENTERIC AGAR ISO 21567 TM 121

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Manufacturer Address: A- 902A RIICO Industrial Area Phase III Bhiwadi-301019. Page 2

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Manufacturer Address: A- 902A RIICO Industrial Area Phase III Bhiwadi-301019. Page 3 www.tmmedia.in PRODUCT DATA SHEET PRINCIPLE The increased concentration of carbohydrate and proteose peptone helps to reduce the inhibitory effect of bile salts and indicators and allows good growth of Salmonella and Shigella species while inhibiting the normal intestinal flora. The medium contains three carbohydrates i. e. lactose sucrose and salicin for differentiation of enteric pathogens. The higher lactose concentration aids in the visualization of enteric pathogens and minimizes the problem of delayed lactose fermentation. Salicin is fermented by many coliforms including those that do not ferment lactose and sucrose. Combination of ferric ammonium citrate and sodium thiosulphate in the medium enables the detection of hydrogen sulfide production thereby aiding in the differentiation process due to the formation of black centered colonies. The indicator system consisting of acid fuchsin and bromothymol blue has lower toxicity as compared to other enteric media resulting in improved recovery of enteric pathogens. Hoben et al further enhanced the selectivity of the medium by addition of novobiocin at a concentration of 15 mg/litre which inhibits Citrobacter and Proteus species. INSTRUCTION FOR USE 1. Dissolve 76.67 grams in 1000 ml distilled water. 2. Heat to boiling to dissolve the medium completely. 3. Do not autoclave. 4. Cool to 45-50°C. 5. Mix well and pour into sterile Petri plates QUALITY CONTROL SPECIFICATIONS Appearance of dehydrated Powder: Cream to yellow colour with tancast homogeneous free flowing powder Appearance of prepared medium: Green colour clear to slightly opalescent gel pH : 7.5± 0.2 INTERPRETATION: Cultural characteristics observed after incubation at 35-37°C for 18-24 hours Organism ATCC Inoculum CFU Growth Recovery Colour of colony Salmonella enteritidis 13076 50-100 Luxuriant ≥ 50 Greenish blue may have black centres H 2S production Salmonella typhi 6539 50-100 Luxuriant ≥ 50 Greenish blue may have black centres H 2S production Salmonella typhimurium 14028 50-100 Luxuriant ≥ 50 Greenish blue may have black centres H 2S

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Manufacturer Address: A- 902A RIICO Industrial Area Phase III Bhiwadi-301019. Page 4 www.tmmedia.in PRODUCT DATA SHEET production Shigella flexneri 12022 50-100 Luxuriant 50 Greenish blue Escherichia coli 25922 50-100 Fair 20-30 Orange may have bile precipitate Escherichia coli 8739 50-100 Fair 20-30 Orange may have bile precipitate STORAGE STABILITY Dehydrated powder hygroscopic in nature store in a dry place in tightly-sealed containers below 25°C and protect from direct Sunlight. Under optimal conditions the medium has a shelf life of 4 years. When the container is opened for the first time note the time and date on the label space provided on the container. After the desired amount of medium has been taken out replace the cap tightly to protect from hydration. REFERENCES 1. King S. and W.I. Metzger. 1968. A new plating medium for the isolation of enteric pathogens. I. Hektoen enteric agar. Appl. Microbiol. 16:577-578. 2. King S. and W.I. Metzger. 1968. A new plating medium for the isolation of enteric pathogens. II. Comparison of Hektoen enteric agar with SS and EMB agar. Appl. Microbiol. 16:579-581. 3. MacFaddin J. F. 1985 Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria Vol. I. Williams Wilkins Baltimore Md. 4. Bopp C.A. Brenner F.W. Fields P.I. Wells J.G. and N.A. Strockbine. 2003. Escherichia Shigella and Salmonella. In: Murray P. R. E. J. Baron J.H. Jorgensen M. A. Pfaller and R. H. Yolken ed.. Manual of clinical microbiology 8th ed. American Society for Microbiology Washington D.C. 5. Thomson R.B. and J.M. Miller. 2003. Specimen collection transport and processing: bacteriology. In: Murray P. R. E. J. Baron J.H. Jorgensen M. A. Pfaller and R. H. Yolken ed.. Manual of clinical microbiology 8th ed. American Society for Microbiology Washington D.C. 6. Kist M. et al. 2000. Infektionen des Darmes. In: MIQ – Qualitätsstandards in der mikrobiologisch-infektiologischen Diagnostik. Vol 9. Urban und Fischer München Jena. 7. Giannella R. A. 1996 Salmonella. In: Barons Medical Microbiology Baron S et al eds. 4th Ed. Univ. of Texas Medical Branch Hoben D.A. Ashton D.H.A. and Peterson A.C. 1973 Appl. Microbiol. 21:126. 8. Hoben D. A. Ashton D. H. A. and Peterson A. C. 1973 Appl. Microbiol. 21. 126-129.

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Manufacturer Address: A- 902A RIICO Industrial Area Phase III Bhiwadi-301019. Page 5 www.tmmedia.in PRODUCT DATA SHEET NOTE: Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices. https://www.tmmedia.in/content/hektoen-enteric-agar-iso-21567

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Manufacturer Address: A- 902A RIICO Industrial Area Phase III Bhiwadi-301019. Page 6

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