SELECTIVE and DIFFERENTIAL MEDIA Principle and Purpose Microorganisms Are Generally Studied After Being Grown on Or in a Microbiological Medium
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Enterobacteriaceae Family (CRE), Is of Utmost Importance for the Enterobacteriaceae Management of Infected Or Colonized Patients
2013 iMedPub Journals THE INTERNATIONAL ARABIC JOURNAL Vol. 3 No. 3:5 Our Site: http://www.imedpub.com/ OF ANTIMICROBIAL AGENTS doi: 10.3823/737 Evaluation of Rula Al-Dawodi1,3, Rawan Liddawi1,3, Raed Ghneim1,3, Randa Kattan1,3, Issa Siryani1,3, Afaf Abu-Diab1, Riyad Meropenem, Ghneim1, Madeleine Zoughbi1,3, Abed-El-Razeq Issa1,3, Randa Al Qass1,3, Sultan Turkuman1, Hiyam Marzouqa1, and Imipenem and Musa Hindiyeh1,2,3* Ertapenem 1 Caritas Baby Hospital, 3 Palestinian Forum for Medical * Correspondence: Bethlehem, Palestine; Research (PFMR), Ramallah, Impregnated 2 Bethlehem University, Palestine Bethlehem, Palestine; [email protected] MacConkey * Musa Y Hindiyeh, Caritas Baby Hospital Bethlehem Agar Plates for Palestine. the Detection of Carbapenem Abstract Resistant Background: Rapid detection of carbapenem resistant bacteria, in particular, members of the Enterobacteriaceae family (CRE), is of utmost importance for the Enterobacteriaceae management of infected or colonized patients. Methods: Three carbapenems; meropenem, imipenem and ertapenem, with two different concentrations (0.5 mg/ml and 1.0 mg/ml), were impregnated in Mac- Conkey agar. The carbapenem impregnated MacConkey agar plates; ([Mac-Mem], [Mac-Imp] and [Mac-Ert]), were then evaluated for the detection of carbapenem resistant Gram-negative bacteria in particular the blaKPC producing Enterobacteria- ceae. The Limit of Detection (LOD) of the plates was determined in triplicate after serial logarithmic dilution of the bacterial strains in saline. This was followed by inoculating the plates and counting the colonies that grew after 24 hours of incu- bation. The specificity and the shelf-life of the plates were determined by testing the plates with six Extended Spectrum β-lactamases (ESBL) producing members of the Enterobacteriaceae family and one genus with the blaAmpC phenotype. -
Macconkey Agar, CS, Product Information
MacCONKEY AGAR, CS (7391) Intended Use MacConkey Agar, CS is used for the isolation and differentiation of Gram-negative enteric bacilli from specimens containing swarming strains of Proteus spp. in a laboratory setting. MacConkey Agar, CS is not intended for use in the diagnosis of disease or other conditions in humans Product Summary and Explanation MacConkey Agar is based on the bile salt-neutral red-lactose agar of MacConkey.1 The original MacConkey medium was used to differentiate strains of Salmonella typhosa from members of the coliform group. Formula modifications improved growth of Shigella and Salmonella strains. These modifications include the addition of 0.5% sodium chloride, decreased agar content, altered bile salts, and neutral red concentrations. The formula modifications improved differential reactions between enteric pathogens and coliforms. MacConkey Agar, CS (“Controlled Swarming”) contains carefully selected raw materials to reduce swarming of Proteus spp., which could cause difficulty in isolating and enumerating other Gram-negative bacilli. Principles of the Procedure Enzymatic Digest of Gelatin, Enzymatic Digest of Casein, and Enzymatic Digest of Animal Tissue are the nitrogen and vitamin sources in MacConkey Agar, CS. Lactose is the fermentable carbohydrate. During Lactose fermentation a local pH drop around the colony causes a color change in the pH indicator, Neutral Red, and bile precipitation. Bile Salts and Crystal Violet are the selective agents, inhibiting Gram-positive cocci and allowing Gram-negative organisms to grow. Sodium Chloride maintains the osmotic environment. Agar is the solidifying agent. Formula / Liter Enzymatic Digest of Gelatin .................................................... 17 g Enzymatic Digest of Casein ................................................... 1.5 g Enzymatic Digest of Animal Tissue....................................... -
Eosin Methylene Blue Agar for the Isolation, Cultivation and Differentiation of Gram Negative Enteric Bacilli from Clinical and Other Specimens Product Description
FT-A2WQP0 Eosin Methylene Blue Agar For the isolation, cultivation and differentiation of gram negative enteric bacilli from clinical and other specimens Product Description Name : Eosin Methylene Blue Agar Catalog Number : A2WQP0, 500 g Storage: 2-25°C - Once opened keep powdered medium closed to avoid hydration. Directions for use Formula • Bacteriological Peptone 10 • Eosin Y 0.4 • Lactose 5 • Methylene Blue 0.065 • Sucrose 5 • Bacteriological Agar 13.5 • Dipotassium Phosphate 2 Final pH 7.2 ± 0.2 at 25ºC Preparation Suspend 36 grams of medium in one liter of distilled water. Mix well and dissolve by heating with frequent agitation. Boil for one minute until complete dissolution. Sterilize in autoclave at 121ºC for 15 minutes. Cool to 45-50ºC, mix well, avoiding the formation of bubbles and dispense carefully into Petri Dishes. DO NOT OVEARHEAT. The prepared medium should be stored at 8-15°C. The color is tournasol blue. Sterilization reduces the methylene blue, leaving the medium orange in color. The normal purple may be restored by gently mixing. The reduced medium should be shaken to oxidize the methylene blue; otherwise a dark zone from the top extending downwards will gradually appear. The dehydrated medium should be homogeneous, free flowing and purple-rose flocculent precipitate in color. If there are any physical changes, discard the medium. Uses EOSIN METHYLENE BLUE AGAR is a differential medium similar to Levine EMB Agar (Cat. 1050) and is used for the isolation of Enterobacteria. The use of Eosin Y and Methylene Blue enable differentiation between lactose- fermenting and non-fermenting organisms. -
XL Agar Base • XLD Agar
XL Agar Base • XLD Agar clinical evaluations have supported the claim for the relatively Intended Use high efficiency of XLD Agar in the primary isolation ofShigella XL (Xylose Lysine) Agar Base is used for the isolation and and Salmonella.5-9 differentiation of enteric pathogens and, when supplemented with appropriate additives, as a base for selective enteric media. XLD Agar is a selective and differential medium used for the isolation and differentiation of enteric pathogens from clinical XLD Agar is the complete Xylose Lysine Desoxycholate Agar, specimens.10-12 The value of XLD Agar in the clinical laboratory a moderately selective medium recommended for isolation and is that the medium is more supportive of fastidious enteric organ- differentiation of enteric pathogens, especially Shigella species. isms such as Shigella.12 XLD Agar is also recommended for the XLD Agar meets United States Pharmacopeia (USP), European testing of food, dairy products and water in various industrial Pharmacopoeia (EP) and Japanese Pharmacopoeia (JP)1-3 standard test methods.13-17 General Chapter <62> of the USP performance specifications, where applicable. describes the test method for the isolation of Salmonella from nonsterile pharmaceutical products using XLD Agar as the solid Summary and Explanation culture medium.1 A wide variety of media have been developed to aid in the selective isolation and differentiation of enteric pathogens. Due Principles of the Procedure to the large numbers of different microbial species and strains Xylose is incorporated into the medium because it is fermented with varying nutritional requirements and chemical resistance by practically all enterics except for the shigellae. -
EOSIN METHYLENE BLUE AGAR (LEVINE) - for in Vitro Use Only - Catalogue No
EOSIN METHYLENE BLUE AGAR (LEVINE) - For in vitro use only - Catalogue No. PE60 Our Eosin Methylene Blue Agar (Levine) is a Recommended Procedure selective and differential medium used in the isolation of gram-negative enteric organisms from 1. Allow medium to reach room temperature. a variety of samples. 2. Using an inoculum from the specimen, streak The Levine formulation of EMB Agar is a the plate as to obtain isolated colonies. slight modification of Holt-Harris and Teague’s 3. Incubate aerobically at 35°C. original recipe from 1916. Unlike the Holt-Harris 4. Examine after 24 hours. and Teague formulation, which contains two 5. Incubate an additional 24 hours if no growth carbohydrate sources, the Levine formulation is observed. contains only one, lactose. This is beneficial since lactose fermenters can be differentiated from non-lactose fermenters. Pancreatic digest of Interpretation of Results gelatin provides a source of carbon, nitrogen, and other essential growth factors. The dyes, eosin Y On EMB Agar (Levine), colony and methylene blue, act both as differential differentiation is due to the uptake of dyes by indictors and inhibitors in the medium; the uptake lactose fermenting organisms. The dyes, eosin of dyes during the growth cycle by some bacteria and methylene blue, react to form a dark allows for differentiation between lactose precipitate in an acid environment, therefore fermenters and non-fermenters. Eosin Y is lactose fermenters take up the dyes giving inhibitory to most gram-positive organisms colonies their typically blue-black coloration. although only to a limited degree; therefore some Additionally, some rapid lactose fermenters, such streptococci, staphylococci and yeasts may grow as E. -
Detection of Staphylococcus Aureus and Other Coagulase Positive Staphylococci in Bovine Raw Milk in Khartoum State by Ikhtyar Ah
View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by KhartoumSpace Detection of Staphylococcus aureus and other Coagulase Positive Staphylococci in Bovine Raw Milk in Khartoum State By Ikhtyar Ahmed Hassan Ali B.C.Sc (2003) Supervisor Prof. Mohammed Taha Shigiddi Department of Microbiology Faculty of Veterinary medicine A dissertation submitted to University of Khartoum in partial fulfillment for the requirement of the Degree of Master of Science in Microbiology Department of Microbiology Faculty of veterinary Medicine University of Khartoum 2010 Dedication to my father, mother, brothers and sisters with love I Table of Contents Subject Page Dedication………………………………………………………. I Table of Contents………………………………………………. II List of Figures…………………………………………………… VII List of Table…………………………………………………….. VIII Acknowledgments………………………………………………. IX Abstract…………………………………………………………. X Abstract (Arabic)……………………………………………… XI Introduction…………………………………………………… 1 Chapter One: Literature Review…………………………….. 3 1.1. Health Hazards of Raw Milk…………………………………… 4 1.2. Pathogenic bacteria in milk........................................................ 5 1.3. Microbial quality of raw milk.................................................... 6 1.4. Staphylococci........................................................................... 7 1.4.1. Coagulase positive staphylococci (CPS)……………………… 8 1.4.2. Coagulase negative staphylococci (CNS)……………………… 10 1.5. Staphylococcus aureus………………………………………… 10 1.5.1. Virulence characteristics of S. -
OXOID MANUAL PRELIMS 16/6/06 12:18 Pm Page 1
OXOID MANUAL PRELIMS 16/6/06 12:18 pm Page 1 The OXOID MANUAL 9th Edition 2006 Compiled by E. Y. Bridson (substantially revised) (former Technical Director of Oxoid) Price: £50 OXOID MANUAL PRELIMS 16/6/06 12:18 pm Page 2 The OXOID MANUAL 9th Edition 2006 Compiled by E. Y. Bridson (substantially revised) (former Technical Director of Oxoid) 9th Edition 2006 Published by OXOID Limited, Wade Road, Basingstoke, Hampshire RG24 8PW, England Telephone National: 01256 841144 International: +44 1256 841144 Email: [email protected] Facsimile National: 01256 463388 International: +44 1256 463388 Website http://www.oxoid.com OXOID SUBSIDIARIES AROUND THE WORLD AUSTRALIA DENMARK NEW ZEALAND Oxoid Australia Pty Ltd Oxoid A/S Oxoid NZ Ltd 20 Dalgleish Street Lunikvej 28 3 Atlas Place Thebarton, Adelaide DK-2670 Greve, Denmark Mairangi Bay South Australia 5031, Australia Tel: 45 44 97 97 35 Auckland 1333, New Zealand Tel: 618 8238 9000 or Fax: 45 44 97 97 45 Tel: 00 64 9 478 0522 Tel: 1 800 331163 Toll Free Email: [email protected] NORWAY Fax: 618 8238 9060 or FRANCE Oxoid AS Fax: 1 800 007054 Toll Free Oxoid s.a. Nils Hansen vei 2, 3 etg Email: [email protected] 6 Route de Paisy BP13 0667 Oslo BELGIUM 69571 Dardilly Cedex, France PB 6490 Etterstad, 0606 Oxoid N.V./S.A. Tel: 33 4 72 52 33 70 Oslo, Norway Industriepark, 4E Fax: 33 4 78 66 03 76 Tel: 47 23 03 9690 B-9031 Drongen, Belgium Email: [email protected] Fax: 47 23 09 96 99 Tel: 32 9 2811220 Email: [email protected] GERMANY Fax: 32 9 2811223 Oxoid GmbH SPAIN Email: [email protected] Postfach 10 07 53 Oxoid S.A. -
Macconkey Agar Base
MACCONKEY AGAR BASE INTENDED USE Remel MacConkey Agar Base is a solid medium recommended for use in qualitative procedures for ithe cultivation of gram-negative bacilli. SUMMARY AND EXPLANATION In 1900, MacConkey first described a neutral red bile salt medium for cultivation and identification of enteric organisms.1 A detailed description of the selective and differential properties of the medium was published in 1905.2 Over the years, MacConkey’s original formula has been modified; the agar content has been reduced, the concentration of bile salts and neutral red has been adjusted, and sodium chloride has been added.3 MacConkey Agar Base is used with added carbohydrate to differentiate enteric gram-negative bacilli based on fermentation reactions. PRINCIPLE Peptones provide nitrogenous nutrients and amino acids necessary for bacterial growth. Sodium chloride supplies essential electrolytes and maintains osmotic equilibrium. Crystal violet and bile salts are selective agents which inhibit most gram-positive organisms. MacConkey Agar Base is used with added carbohydrate to differentiate enteric gram-negative bacilli based on fermentation reactions. When the carbohydrate is fermented, a local pH drop around the colony causes bile preciptitation in the agar around the colony. Neutral red is an indicator which turns colonies pink when the carbohydrate is fermented. Agar is a solidifying agent. REAGENTS (CLASSICAL FORMULA)* Gelatin Peptone .............................................................. 17.0 g Meat Peptone ..................................................................1.5 -
Salmonella Enterica and the Specific Interaction with Lactuca Sativa
Quantitative detection of Salmonella enterica and the specific interaction with Lactuca sativa Michel M. Klerks 2007 Promotor: Prof. Dr. Ir. A. H. C. van Bruggen Hoogleraar in de Biologische Landbouwsystemen Wageningen Universiteit Co-promotor: Dr. Ir. C. Zijlstra Clusterleider Moleculaire Phytopathologie Wageningen UR, Plant Research International BV Promotie commissie: Prof. Dr. Ir. P. J. G. M. de Wit, Wageningen Universiteit Prof. Dr. Ir. M. H. Zwietering, Wageningen Universiteit Prof. Dr. Ir. J. D. van Elsas, Rijksuniversiteit Groningen Dr. Ir. H.J. M. Aarts, RIKILT, Instituut voor voedselveiligheid Dit onderzoek is uitgevoerd binnen de C.T. de Wit Onderzoekschool ‘Productie Ecologie en Beheer van Natuurlijke Hulpbronnen’ Quantitative detection of Salmonella enterica and the specific interaction with Lactuca sativa Michel M. Klerks Proefschrift ter verkrijging van de graad van doctor op gezag van de rector magnificus van Wageningen Universiteit Prof. Dr. M. J. Kropff, in het openbaar te verdedigen op woensdag 20 juni 2007 des namiddags te 13.30 uur in de Aula Michel M. Klerks (2007) Quantitative detection of Salmonella enterica and the specific interaction with Lactuca sativa. Doctoral thesis - Biological Farming Systems Group - Wageningen University - The Netherlands. Subject headings: Salmonella enterica, Escherichia coli, detection, lettuce, plant response, pathogenicity, route of infection ISBN: 978-90-8504-674-5 Contents Abstract Chapter 1 General introduction 9 Chapter 2 Comparison of real-time PCR methods for detection of 39 -
Mannitol Salt Agar, Product Information
MANNITOL SALT AGAR (7143) Intended Use Mannitol Salt Agar is used for the isolation of staphylococci in a laboratory setting. Mannitol Salt Agar is not intended for use in the diagnosis of disease or other conditions in humans. Conforms to Harmonized USP/EP/JP Requirements.1,2,3 Product Summary and Explanation Chapman formulated Mannitol Salt Agar to isolate staphylococci by inhibiting growth of most other bacteria with a high salt concentration.4 Chapman added 7.5% Sodium Chloride to Phenol Red Mannitol Agar, and noted pathogenic strains of staphylococci (coagulase-positive staphylococci) grew luxuriantly and produced yellow colonies with yellow zones. Nonpathogenic staphylococci produced small red colonies with no color change to the surrounding medium. Mannitol Salt Agar is highly selective, and specimens from heavily contaminated sources may be streaked onto this medium without danger of overgrowth.5 Mannitol Salt Agar is recommended for isolating pathogenic staphylococci from specimens, cosmetics, and microbial limit tests.1,2,3,5,6 Principles of the Procedure Enzymatic Digest of Casein, Enzymatic Digest of Animal Tissue, and Beef Extract provide the nitrogen, vitamins, and carbon in Mannitol Salt Agar. D-Mannitol is the carbohydrate source. In high concentrations, Sodium Chloride inhibits most bacteria other than staphylococci. Phenol Red is the pH indicator. Agar is the solidifying agent. Bacteria that grow in the presence of a high salt concentration and ferment mannitol produce acid products, turning the Phenol Red pH indicator from red to yellow. Typical pathogenic staphylococci ferment mannitol and form yellow colonies with yellow zones. Typical non-pathogenic staphylococci do not ferment mannitol and form red colonies. -
Macconkey Agar Plate
MacConkey Agar Plate MPH081 Intended Use Recommended for selective isolation and differentiation of E.coli and other enteric bacteria from pharmaceutical products in accordance with the microbial limit testing by harmonized methodology of USP/EP/BP/JP. Composition** Ingredients Gms / Litre Gelatin peptone # 17.000 HMC peptone ## 3.000 Lactose monohydrate 10.000 Sodium chloride 5.000 Bile salts 1.500 Neutral red 0.030 Crystal violet 0.001 Agar 13.500 pH after sterilization ( at 25°C) 7.1±0.2 **Formula adjusted, standardized to suit performance parameters # Pancreatic digest of gelatin ## Peptones (meat and casein) Directions Either streak, inoculate or surface spread the test inoculum (50-100 CFU) aseptically on the plate. Principle And Interpretation MacConkey Agar is the earliest selective and differential medium for cultivation of coliform organisms (8,9). Subsequently MacConkey Agar and Broth have been recommended for use in microbiological examination of foodstuffs (10) and for direct plating / inoculation of water samples for coliform counts (1). This medium is also accepted by the Standard Methods for the Examination of Milk and Dairy Products (12). It is recommended in pharmaceutical preparations and is in accordance with the harmonized method of USP/EP/BP/JP (11,2,3,6). Gelatin peptone and HMC peptone provide the essential nutrients, vitamins and nitrogenous factors required for growth of microorganisms. Lactose monohydrate is the fermentable source of carbohydrate. The selective action of this medium is attributed to crystal violet and bile salts, which are inhibitory to most species of gram-positive bacteria. Sodium chloride maintains the osmotic balance in the medium. -
TS-4015414.Pdf
Biolife f4d8-8c9d-e216-7f3b 2020/12 page 1 / 3 INSTRUCTIONS FOR USE . HEKTOEN ENTERIC AGAR Dehydrated culture medium 1-INTENDED USE In vitro diagnostic. Selective and differential medium for the isolation of Gram- negative enteric pathogens, especially Salmonella and Shigella, from clinical and non clinical specimens. 2- COMPOSITION - TYPICAL FORMULA * (AFTER RECONSTITUTION WITH 1 L OF WATER ) Tryptose 12.000 g Yeast Extract 3.000 g Bile salts n° 3 9.000 g Lactose 12.000 g Sucrose 12.000 g Salicin 2.000 g Sodium chloride 5.000 g Sodium thiosulphate 5.000 g Fe-ammonium citrate 1.500 g Bromothymol blue 0.065 g Acid fuchsin 0.100 g HEA: Salmonella colonies with large black centre and Agar 15.000 g yellow-orange K.pneumoniae colonies. *The formula may be adjusted and/or supplemented to meet the required performances criteria. 3 - PRINCIPLE OF THE METHOD AND EXPLANATION OF THE PROCEDURE In the first half of the twentieth century, several culture media were developed and proposed for the isolation of enteric pathogens from faeces and other materials. Some of them were moderately selective and allowed the growth of faecal contaminants, others showed excessive toxicity for the growth of pathogens, especially of Shigella .1 Sylvia King and William I. Metzger, working at the Hektoen Institute in Chicago, formulated HE agar in 1968 2 with the goal to increase the recovery of Shigella species from mixed cultures. They enriched SS Agar formulation, evaluated in 1941 by Catherine Mayfield and Maud Gober 3, with extra amounts of carbohydrates and peptones to offset the inhibitory effects of the bile salts.