DOCSLIB.ORG

Enterobacteriaceae Family (CRE), Is of Utmost Importance for the Enterobacteriaceae Management of Infected Or Colonized Patients

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
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. Finally, the 0.5 mg/ml Mac-Mem plates were further challenged by incorporating them in the routine active surveillance program for the detection of carbapenem resistant bacteria. Results: Of the three carbapenems impregnated plates, Mac-Ert plates gave the lowest LOD; however, its specificity was poor as it failed to inhibit the growth of some of the ESBL producing strains. The Mac-Imp plate LOD and specificity were both unsatisfactory for several strains of the bacterial isolates tested. On the other hand the 0.5 mg/ml Mac-Mem plates showed low LOD for the blaKPC producing bacteria and had excellent specificity as it prevented the growth of all the ESBL and blaAmpC positive isolates tested. Not only that, but the shelf-life of the 0.5mg/ ml Mac-Mem plate was long, up to one month at 4oC. Conclusions: The high specificity and the good selectivity, in addition to the long shelf-life allowed the 0.5µg/ml Mac-Mem agar to be used as a cost effective selec- tive medium for the isolation of carbapenem resistant Gram-negative bacteria, in particular the blaKPC producing members of the Enterobacteriaceae family. This article is available from: Key words: blaKPC, blaOXA-48, Meropenem Agar (Mac-Mem), Carbapenemase, www.iajaa.org Palestine © Under License of Creative Commons Attribution 3.0 License 1 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 Introduction and utilized for the rapid detection of blaKPC producing bac- teria. Chromogenic agar, CHROMagarTM KPC (CHROMagar, Rapid detection of carbapenem resistant bacteria, in par- France), has been evaluated for the detection of blaKPC posi- ticular, members of the Enterobacteriaceae family (CRE) is of tive bacteria. However, this agar base medium still missed utmost importance for the management of infected or colo- certain strains of blaKPC positive Enterobacteriaceae (1, 13, TM nized patients (1). Carbapenems are widely used to treat se- 14). Similar performance between the CHROMagar KPC vere bacterial infections that are often hard to manage. Sever- and MacConkey Agar impregnated with 2 µg/ml imipenem al carbapenem resistance mechanisms have been implicated was noted by Adler et al. (15). More recently, the Drigalski in how bacterial strains become resistant to carbapenems. agar-based culture medium was reported to have excellent Based on the molecular properties of the β–lactamases, four sensitivity and specificity for the detection of bacteria carry- classes of carbapenemases have been identified (2). These ing the blaKPC resistance mechanism (14). include the molecular classes (A, C, and D) β-lactamases with the amino acid serine at their active site, and the molecular In this study, we report the evaluation of cost effective MacCo- class B where the β-lactamases are all metalloenzymes with nkey Agar plates impregnated with one of the carbapenems an active-site zinc (2). Class A enzymes, an Ambler molecu- (meropenem, imipenem and ertapenem) for the detection of CRE’s. In addition, the specificity, analytical sensitivity and lar class enzymes (ex: blaKPC), have been the most common detected carbapenemases with worldwide distribution (1). the plates’ shelf-life were also evaluated. The concentrations of the antibiotics evaluated were 1.0 µg/mL and 0.5 µg/mL. For any health care facility, it is important to understand the These concentrations were chosen after careful evaluation level of CRE spreading in its different medical departments. It of the carbapenems lowest minimal inhibitory concentration has been recommended by the American Centers of Disease (MIC) interpretation guidelines published by the Clinical and Control and Prevention (CDC), that in areas which are not Laboratory Standards Institute (CLSI) (17). endemic for CRE, the acute care facilities should review all the microbiology records for the preceding 6-12 months to determine if CRE’s have been isolated at the institution (3). Materials and Methods Upon the identification of CRE cases, surveillance cultures from high risk areas should be performed to identify any Reagents unrecognized cases. Meropenem trihydrate (Catalogue # M2574) and imipenem CRE’s active surveillance cultures (perianal/rectal swabs), tak- monohydrate (Catalogue # I0160) were supplied by Sig- en from patients upon admission are of great importance for ma-Aldrich, USA. Ertapenem sodium, Invanz (Catalogue # the identification of unrecognized CRE-colonized patients, in 7290004997960) was supplied by Merck Sharp and Dohme order to take immediate and appropriate isolation precau- Corp, USA. All reagents were stored and diluted according tions (4, 5). Studies have shown that in areas where CRE’s to the manufacturer’s guidelines. are endemic, up to 5.4% of hospitalized patients can be car- riers of CRE’s, of which only one-third had a positive clinical Bacterial Strains culture (4). Moreover, surveillance swabs have been reported to considerably lower the cost of patient hospitalization (6). Carbapenem resistant Enterobacteriaceae members (N=9), Active surveillance cultures taken from patients coming from Extended Spectrum β–Lactamases (ESBL) producing Entero- highly CRE endemic medical settings upon admission have bacteriaceae strains (N= 6), blaAmpC β-lactamase (N=1) and been used as part of a comprehensive strategy to combat the multi-drug resistant Acinetobacter baumannii (N=1), all iso- spread of CRE’s. Studies have shown a 4.7-fold reduction in lated from patients’ samples at Caritas Baby Hospital (CBH) the incidence of CRE infection following the activation of the laboratory, were used to evaluate the performance of the active CRE surveillance program (7, 8). carbapenems impregnated MacConkey Agar. The carbap- enem resistant isolates included: 3 Klebsiella pneumoniae Several highly sensitive Polymerase Chain Reaction (PCR) as- blaKPC positive; 2 Enterobacter species blaKPC positive; 2 E. says and Real-Time PCR assays have been developed and coli blaKPC positive; 1 Citrobacter species blaKPC positive and 1 Klebsiella pneumoniae bla positive. The ESBL positive validated for the rapid detection and identification of blaKPC OXA-48 positive bacteria (9-12). However, one of the disadvantages of isolates were (K. pneumoniae, Salmonella species, Shigella these assays was the need for specialized equipment and well sonnei, E. coli , Proteus species and Citrobacter species); while trained personnel. Consequently, screening agar plates, that the blaAmpC positive bacteria was an Enterobacter species. have incorporated one of the commonly used carbapenems Moreover, 14 Klebsiella pneumoniae blaOXA-48 positive and (ertapenem, meropenem or imipenem), have been developed 2 E. coli blaOXA-48 positive were used to evaluate the analyti- 2 © Under License of Creative Commons Attribution 3.0 License 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 cal sensitivity of the 0.5 µg/ml Mac-Mem agar plates for the Determination of the specificity and Shelf-Life of detection of low level carbapenemase producing blaOXA-48 the different Carbapenems Impregnated Plates Enterobacteriaceae. MacConkey agar plates (Mac-Mem, Mac-Imp, and Mac-Ert) All bacterial strains were identified according to the Clinical impregnated with the two different concentrations (0.5 µg/ Microbiology Procedures Handbook, and their antimicrobial ml and 1.0 µg/ml) of the three carbapenems, were prepared susceptibility testing was performed in duplicate according and stored at 4-8oC. The plates were inoculated with colonies to the Clinical and Laboratory Standard Institute
Load more

Recommended publications
  • 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.......................................
    [Show full text]
  • 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
    [Show full text]
  • 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.
    [Show full text]
  • Macconkey Agar SM081
    MacConkey Agar SM081 Recommended for selective isolation and differentiation of coliform organisms and other enteric pathogens. Composition** Ingredients Gms / Litre Peptic digest of animal tissue 1.500 Casein enzymic hydrolysate 1.500 Pancreatic digest of gelatin 17.000 Lactose 10.000 Bile salts 1.500 Sodium chloride 5.000 Crystal violet 0.001 Neutral red 0.030 Agar 15.000 **Formula adjusted, standardized to suit performance parameters Directions MacConkey Agar is a ready to use solid media in glass bottle. The medium is pre-sterilized; hence it does not need sterilization. Medium in the bottle can be melted either by using a pre-heated water bath or any other method. Slightly loosen the cap before melting. When complete melting of medium is observed dispense the medium as desired and allowed to solidify. Principle And Interpretation MacConkey agars are slightly selective and differential plating media mainly used for the detection and isolation of gram- negative organisms from clinical (1), dairy (2), food (3,4), water (5) pharmaceutical (6, 14) and industrial sources (7). It is also recommended for the selection and recovery of the Enterobacteriaceae and related enteric gram-negative bacilli. USP recommends this medium for use in the performance of Microbial Limit Tests (6). These agar media are selective since the concentration of bile salts, which inhibit gram-positive microorganisms, is low in comparison with other enteric plating media. The medium M081, which corresponds with, that recommended by APHA can be used for the direct plating of water samples for coliform bacilli, for the examination of food samples for food poisoning organisms (3) and for the isolation of Salmonella and Shigella species in cheese (2).
    [Show full text]
  • Prepared Culture Media
    PREPARED CULTURE MEDIA 121517SS PREPARED CULTURE MEDIA Made in the USA AnaeroGRO™ DuoPak A 02 Bovine Blood Agar, 5%, with Esculin 13 AnaeroGRO™ DuoPak B 02 Bovine Blood Agar, 5%, with Esculin/ AnaeroGRO™ BBE Agar 03 MacConkey Biplate 13 AnaeroGRO™ BBE/PEA 03 Bovine Selective Strep Agar 13 AnaeroGRO™ Brucella Agar 03 Brucella Agar with 5% Sheep Blood, Hemin, AnaeroGRO™ Campylobacter and Vitamin K 13 Selective Agar 03 Brucella Broth with 15% Glycerol 13 AnaeroGRO™ CCFA 03 Brucella with H and K/LKV Biplate 14 AnaeroGRO™ Egg Yolk Agar, Modified 03 Buffered Peptone Water 14 AnaeroGRO™ LKV Agar 03 Buffered Peptone Water with 1% AnaeroGRO™ PEA 03 Tween® 20 14 AnaeroGRO™ MultiPak A 04 Buffered NaCl Peptone EP, USP 14 AnaeroGRO™ MultiPak B 04 Butterfield’s Phosphate Buffer 14 AnaeroGRO™ Chopped Meat Broth 05 Campy Cefex Agar, Modified 14 AnaeroGRO™ Chopped Meat Campy CVA Agar 14 Carbohydrate Broth 05 Campy FDA Agar 14 AnaeroGRO™ Chopped Meat Campy, Blood Free, Karmali Agar 14 Glucose Broth 05 Cetrimide Select Agar, USP 14 AnaeroGRO™ Thioglycollate with Hemin and CET/MAC/VJ Triplate 14 Vitamin K (H and K), without Indicator 05 CGB Agar for Cryptococcus 14 Anaerobic PEA 08 Chocolate Agar 15 Baird-Parker Agar 08 Chocolate/Martin Lewis with Barney Miller Medium 08 Lincomycin Biplate 15 BBE Agar 08 CompactDry™ SL 16 BBE Agar/PEA Agar 08 CompactDry™ LS 16 BBE/LKV Biplate 09 CompactDry™ TC 17 BCSA 09 CompactDry™ EC 17 BCYE Agar 09 CompactDry™ YMR 17 BCYE Selective Agar with CAV 09 CompactDry™ ETB 17 BCYE Selective Agar with CCVC 09 CompactDry™ YM 17 BCYE
    [Show full text]
  • Tularemia – Epidemiology
    This first edition of theWHO guidelines on tularaemia is the WHO GUIDELINES ON TULARAEMIA result of an international collaboration, initiated at a WHO meeting WHO GUIDELINES ON in Bath, UK in 2003. The target audience includes clinicians, laboratory personnel, public health workers, veterinarians, and any other person with an interest in zoonoses. Tularaemia Tularaemia is a bacterial zoonotic disease of the northern hemisphere. The bacterium (Francisella tularensis) is highly virulent for humans and a range of animals such as rodents, hares and rabbits. Humans can infect themselves by direct contact with infected animals, by arthropod bites, by ingestion of contaminated water or food, or by inhalation of infective aerosols. There is no human-to-human transmission. In addition to its natural occurrence, F. tularensis evokes great concern as a potential bioterrorism agent. F. tularensis subspecies tularensis is one of the most infectious pathogens known in human medicine. In order to avoid laboratory-associated infection, safety measures are needed and consequently, clinical laboratories do not generally accept specimens for culture. However, since clinical management of cases depends on early recognition, there is an urgent need for diagnostic services. The book provides background information on the disease, describes the current best practices for its diagnosis and treatment in humans, suggests measures to be taken in case of epidemics and provides guidance on how to handle F. tularensis in the laboratory. ISBN 978 92 4 154737 6 WHO EPIDEMIC AND PANDEMIC ALERT AND RESPONSE WHO Guidelines on Tularaemia EPIDEMIC AND PANDEMIC ALERT AND RESPONSE WHO Library Cataloguing-in-Publication Data WHO Guidelines on Tularaemia.
    [Show full text]
  • Enterobacteriaceae Biochemical Reactions
    Gram-negative rods Enterobacteriaceae Biochemical Reactions Manal AL khulaifi Bacteria Gram positive Gram negative Cocci Bacilli Cocci Rods Manal AL khulaifi Characters of Enterobacteriaceae All Enterobacteriaciae – Gram-negative rods – Reduce nitrates into nitrites – Oxidase negative Facultative anaerobic Motile except Shigella and Klebsiella Non-capsulated except Klebsiella Non-fastidious Grow on bile containing media (MacConkey agar) Manal AL khulaifi Enterobacteriaceae Some Enterobacteriaceae are true pathogens – Salmonella spp. – Shigella spp. – Yersinia spp. – Certain strains of E. coli (ETEC, EPEC, EIEC, EHEC) Most members of the Enterobacteriaceae are opportunistic or cause secondary infections of wounds, the urinary and respiratory tracts, and the circulatory system e.g. E. coli. Enterobacteriaceae divided into TWO main groups according to action on LACTOSE – Lactose Fermenters (LF) E. coli, Citrobacter, Klbesiella, Enterobacter – Lactose Non-Fermenters (LNF) Salmonella, Shigella, Proteus, Yersinia Manal AL khulaifi Identification of Enterobacteriaceae Gram stain – All Enterobacteriaceae are Gram-negative rods – Arranged in single Manal AL khulaifi Identification of Enterobacteriaceae Biochemical reactions Oxidase test – All members of Enterobacteriaceae are oxidase negative – Pseudomonas is oxidase positive O/F test – All members of Enterobacteriaceae are O+/F+ – Pseudomonas is O+/F- See & compare these tests under Pseudomonas Lab Manal AL khulaifi Oxidase Test: Principle: Tetramethyl p-phenylene diamine (oxidase reagent) colourless
    [Show full text]
  • Macconkey Agar Is Medium to Dark Pink to Dark Is Medium Agar Macconkey Prepared Grow but Remain Colorless with No Bile Precipitate
    Technical Specification Sheet MacConkey Agar (NCM0017) Intended Use MacConkey Agar is used for the isolation and differentiation of Gram-negative enteric bacilli in a laboratory setting. Conforms to Harmonized USP/EP/JP Requirements and FDA/BAM. MacConkey Agar is not intended for use in the diagnosis of disease or other conditions in humans. Description A medium recommended by the Harmonized USP/EP/JP for isolation and identification of Escherichia coli from non-sterile products. Conforms to USP/EP/JP performance specification. Gelatin serves as source of carbon and nitrogen. Lactose is a fermentable carbohydrate and sodium chloride maintains the osmotic balance. Bile salts and crystal violet act as selective agents inhibiting many Gram-positive bacteria. Escherichia coli can ferment lactose to produce acid which results in a pH drop. This is indicated by neutral red resulting in pink colonies. Enough acid production will cause the precipitation of bile salts resulting in a bile precipitate or halo around lactose fermenting bacteria. Non-lactose fermenting bacteria such as Salmonella spp. grow but remain colorless with no bile precipitate. According to the Harmonized USP/EP/JP, MacConkey Broth is used as a selective enrichment broth, with subculture performed onto MacConkey Agar. Typical Formulation Enzymatic Digest of Gelatin 17.0 g/L Enzymatic Digest of Casein 1.5 g/L Enzymatic Digest of Animal Tissue 1.5 g/L Lactose 10.0 g/L Bile Salts Mixture 1.5 g/L Sodium Chloride 5.0 g/L Neutral Red 0.03 g/L Crystal Violet 0.001 g/L Agar 13.5 g/L Final pH: 7.1 ± 0.2 at 25ºC Formula may be adjusted and/or supplemented as required to meet performance specifications.
    [Show full text]
  • Bacterial Cultures Care Guide SCIENTIFIC Introduction Use the Following Guide to Learn How to Properly Care for Bacterial Cultures
    Bacterial Cultures Care Guide SCIENTIFIC Introduction Use the following guide to learn how to properly care for bacterial cultures. BIO FAX! Safety Precautions After use, agar plates will likely contain viable microbes. Although the bacteria are not likely to be pathogenic, do not open the plates unnecessarily. Use sterile techniques at all times when handling bacterial cultures. When plates are done being used, they should be autoclaved or opened under a 10% bleach solution and soaked for at least one hour. Bleach solution is a corrosive liquid that may dis- color clothing and cause skin burns. Avoid contact of bleach with heat, acids and organic materials; chlorine gas will be generated. Wear chemical splash goggles, chemical-resistant gloves, and a chemical-resistant apron. Wash hands thoroughly with soap and water before leaving the laboratory. Please follow all laboratory safety guidelines. Culturing and Maintenance Prior to receiving bacterial cultures it is helpful to be aware of the required conditions for the particular strain of bacteria purchased. Each species has specific conditions that are necessary for optimal growth. Refer to the Bacterial Cultures Table below for incubation temperature and the appropriate general medium. Bacterial Cultures Table Description (Genus species) Incubation Temperature† Medium Bacillus cereus (20–35) 30 °C Nutrient agar Bacillus mycoides 30 °C Nutrient agar Bacillus megaterium (25–35) 30 °C Nutrient agar Bacillus subtilis (25–35) 25–30 °C Nutrient agar Enterobacter aerogenes (30–37) 30 °C Nutrient
    [Show full text]
  • Prepared Media Guide
    Prepared media guide For the isolation, identification, differentiation and susceptibility testing of microorganisms Your partner in ready prepared media. This is our culture for innovation. Contents Thermo Scientific™ prepared media Anaerobe Agars 4 Culture media by organism type 40 Aeromonas 41 Antimicrobial Susceptibility Bacillus cereus 41 Testing (AST) Agars 6 Bordetella 42 Burkholderia cepacia 42 Biplates 9 Campylobacter 43 Clostridium species 43 Coliforms/Escherichia coli 47 Blood Agars 17 Corynebacteria 48 Dermatophytes 48 Thermo Scientific™ Brilliance™ Chromogenic Escherichia coli O157 49 Media (Clinical) 20 Enterobacteriaceae 50 Enterococci 53 Gardnerella 54 Thermo Scientific™ Brilliance™ Chromogenic Haemophilus and Neisseria 54 Media (Food) 25 Helicobacter pylori 56 Lactobacilli/Bifidobacteria 56 Diluents, water and peptones 27 Legionella 57 Listeria 58 Dip-Slides 28 Mycoplasma/Ureaplasma 58 Pasteurella 59 Pseudomonas 59 General purpose media 29 Salmonella 60 Staphylococci/Streptococci 63 Pharmaceutical media 33 Staphylococcus aureus 64 Streptococcus agalactiae 65 ReadyBags 36 Trichomonas 65 Vibrio 65 Yeasts and molds 66 Water testing 38 Yersinia 68 Quality control 69 Empowering the people who dedicate their lives to microbiology. Those who diagnose against the clock, around the clock, research against the odds, and uncover the incredible. All of these people deserve uncompromised quality and impeccable service. And they need a partner with the drive to accelerate innovation, enhance productivity and help shape the future. Clinical microbiology Food microbiology Combining over 150 years of technical and scientific There’s very little room for error in food safety and expertise in serving the microbiology community, quality. That’s why all Thermo Scientific microbiology Remel™, Oxoid, VersaTREK™ and Sensititre™ products solutions are developed with a deep understanding of are part of the industry-leading Thermo Scientific the unique needs of the food testing laboratory.
    [Show full text]
  • BD Industry Catalog
    PRODUCT CATALOG INDUSTRIAL MICROBIOLOGY BD Diagnostics Diagnostic Systems Table of Contents Table of Contents 1. Dehydrated Culture Media and Ingredients 5. Stains & Reagents 1.1 Dehydrated Culture Media and Ingredients .................................................................3 5.1 Gram Stains (Kits) ......................................................................................................75 1.1.1 Dehydrated Culture Media ......................................................................................... 3 5.2 Stains and Indicators ..................................................................................................75 5 1.1.2 Additives ...................................................................................................................31 5.3. Reagents and Enzymes ..............................................................................................75 1.2 Media and Ingredients ...............................................................................................34 1 6. Identification and Quality Control Products 1.2.1 Enrichments and Enzymes .........................................................................................34 6.1 BBL™ Crystal™ Identification Systems ..........................................................................79 1.2.2 Meat Peptones and Media ........................................................................................35 6.2 BBL™ Dryslide™ ..........................................................................................................80
    [Show full text]
  • Optimizing Microbiome Sequencing for Small Intestinal Aspirates
    Leite et al. BMC Microbiology (2019) 19:239 https://doi.org/10.1186/s12866-019-1617-1 METHODOLOGY ARTICLE Open Access Optimizing microbiome sequencing for small intestinal aspirates: validation of novel techniques through the REIMAGINE study Gabriela Guimaraes Sousa Leite1, Walter Morales1, Stacy Weitsman1, Shreya Celly1, Gonzalo Parodi1, Ruchi Mathur1,2, Rashin Sedighi1, Gillian M. Barlow1, Ali Rezaie1,3 and Mark Pimentel1,3* Abstract Background: The human small intestine plays a central role in the processes of digestion and nutrient absorption. However, characterizations of the human gut microbiome have largely relied on stool samples, and the associated methodologies are ill-suited for the viscosity and low microbial biomass of small intestine samples. As part of the REIMAGINE study to examine the specific roles of the small bowel microbiome in human health and disease, this study aimed to develop and validate methodologies to optimize microbial analysis of the small intestine. Results: Subjects undergoing esophagogastroduodenoscopy without colon preparation for standard of care were prospectively recruited, and ~ 2 ml samples of luminal fluid were obtained from the duodenum using a custom sterile aspiration catheter. Samples of duodenal aspirates were either untreated (DA-U, N = 127) or pretreated with dithiothreitol (DA-DTT, N = 101), then cultured on MacConkey agar for quantitation of aerobic gram-negative bacteria, typically from the class Gammaproteobacteria, and on blood agar for quantitation of anaerobic microorganisms. DA-DTT exhibited 2.86-fold greater anaerobic bacterial counts compared to DA-U (P = 0.0101), but were not statistically different on MacConkey agar. DNA isolation from DA-U (N = 112) and DA-DTT (N = 43) samples and library preparation for 16S rRNA gene sequencing were also performed using modified protocols.
    [Show full text]