A Case of Gluconacetobacter Liquefaciens Bacteremia Associated with Sugarcane Juice Ingestion
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Microbiology Laboratory Exercises Third Edition 2020
MICROBIOLOGY Laboratory Exercises Third Edition Keddis & Rauschenbach 2020 Photo Credits (in order of contribution): Diane Davis, Ines Rauschenbach & Ramaydalis Keddis Acknowledgements: Many thanks to those in the Department of Biochemistry and Microbiology, Rutgers University, who have through the years inspired our enthusiasm for the science and teaching of microbiology, with special thanks to Diane Davis, Douglas Eveleigh and Max Häggblom. Safety: The experiments included in this manual have been deemed safe by the authors when all necessary safety precautions are met. The authors recommend maintaining biosafety level 2 in the laboratory setting and using risk level 1 organisms for all exercises. License: This work is licensed under a Creative Commons Attribution- NonCommercial-NoDerivatives 4.0 International License Microbiology Laboratory Exercises Third Edition 2020 Ramaydalis Keddis, Ph.D. Ines Rauschenbach, Ph.D. Department of Biochemistry and Microbiology Rutgers, The State University of New Jersey CONTENTS PAGE Introduction Schedule ii Best Laboratory Practices Iii Working in a Microbiology Laboratory iv Exercises Preparation of a Culture Medium 1 Culturing and Handling Microorganisms 3 Isolation of a Pure Culture 5 Counting Bacterial Populations 8 Controlling Microorganisms 10 Disinfectants 10 Antimicrobial Agents: Susceptibility Testing 12 Hand Washing 14 The Lethal Effects of Ultraviolet Light 15 Selection of Fungi from Air 17 Microscopy 21 Morphology and Staining of Bacteria 26 Microbial Metabolism 30 Enzyme Assay 32 Metabolic -
Chocolate Agar Plate MP103 Intended Use for Isolation of Neisseria Gonorrhoeae from Chronic and Acute Gonococcal Infections
Chocolate Agar Plate MP103 Intended use For isolation of Neisseria gonorrhoeae from chronic and acute gonococcal infections. Composition** Ingredients Gms / Litre Proteose peptone 20.000 Dextrose 0.500 Sodium chloride 5.000 Disodium phosphate 5.000 Agar 15.000 After sterilization Sterile Lysed blood (at 80°C) 50.000 Vitamino Growth Supplement (FD025) 2 vials Final pH ( at 25°C) 7.3±0.2 **Formula adjusted, standardized to suit performance parameters Directions Either streak, inoculate or surface spread the test inoculum (50-100 CFU) aseptically on the plate. Principle And Interpretation Neisseria gonorrhoeae is a gram-negative bacteria and the causative agent of gonorrhea, however it is also occasionally found in the throat. The cultivation medium for gonococci should ideally be a rich nutrients base with blood, either partially lysed or completely lysed. The diagnosis and control of gonorrhea have been greatly facilitated by improved laboratory methods for detecting, isolating and studying N. gonorrhoeae. Chocolate Agar Base, with the addition of supplements, gives excellent growth of the gonococcus without overgrowth by contaminating organisms. G.C. Agar (M434) can also be used in place of Chocolate Agar Base, which gives slightly better results than Chocolate Agar (4). The diagnosis and control of gonorrhea have been greatly facilitated by improved laboratory methods for detecting, isolating and studying N. gonorrhoea. Interest in the cultural procedure for the diagnosis of gonococcal infection was stimulated by Ruys and Jens (9), Mcleod and co-workers (8), Thompson (7), Leahy and Carpenter (1), Carpenter, Leahy and Wilson (2) and Carpenter (10), who clearly demonstrated the superiority of this method over the microscopic technique. -
Protein Expression Profile of Gluconacetobacter Diazotrophicus PAL5, a Sugarcane Endophytic Plant Growth-Promoting Bacterium
Proteomics 2008, 8, 1631–1644 DOI 10.1002/pmic.200700912 1631 RESEARCH ARTICLE Protein expression profile of Gluconacetobacter diazotrophicus PAL5, a sugarcane endophytic plant growth-promoting bacterium Leticia M. S. Lery1, 2, Ana Coelho1, 3, Wanda M. A. von Kruger1, 2, Mayla S. M. Gonc¸alves1, 3, Marise F. Santos1, 4, Richard H. Valente1, 5, Eidy O. Santos1, 3, Surza L. G. Rocha1, 5, Jonas Perales1, 5, Gilberto B. Domont1, 4, Katia R. S. Teixeira1, 6 and Paulo M. Bisch1, 2 1 Rio de Janeiro Proteomics Network, Rio de Janeiro, Brazil 2 Unidade Multidisciplinar de Genômica, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil 3 Departamento de Genética, Instituto de Biologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil 4 Laboratório de Química de Proteínas, Departamento de Bioquímica, Instituto de Química, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil 5 Laboratório de Toxinologia, Departamento de Fisiologia e Farmacodinâmica- Instituto Oswaldo Cruz- Fundac¸ão Oswaldo Cruz, Rio de Janeiro, Rio de Janeiro, Brazil 6 Laboratório de Genética e Bioquímica, Embrapa Agrobiologia, Seropédica, Brazil This is the first broad proteomic description of Gluconacetobacter diazotrophicus, an endophytic Received: September 25, 2007 bacterium, responsible for the major fraction of the atmospheric nitrogen fixed in sugarcane in Revised: December 18, 2007 tropical regions. Proteomic coverage of G. diazotrophicus PAL5 was obtained by two independent Accepted: December 19, 2007 approaches: 2-DE followed by MALDI-TOF or TOF-TOF MS and 1-DE followed by chromatog- raphy in a C18 column online coupled to an ESI-Q-TOF or ESI-IT mass spectrometer. -
Francisella Tularensis 6/06 Tularemia Is a Commonly Acquired Laboratory Colony Morphology Infection; All Work on Suspect F
Francisella tularensis 6/06 Tularemia is a commonly acquired laboratory Colony Morphology infection; all work on suspect F. tularensis cultures .Aerobic, fastidious, requires cysteine for growth should be performed at minimum under BSL2 .Grows poorly on Blood Agar (BA) conditions with BSL3 practices. .Chocolate Agar (CA): tiny, grey-white, opaque A colonies, 1-2 mm ≥48hr B .Cysteine Heart Agar (CHA): greenish-blue colonies, 2-4 mm ≥48h .Colonies are butyrous and smooth Gram Stain .Tiny, 0.2–0.7 μm pleomorphic, poorly stained gram-negative coccobacilli .Mostly single cells Growth on BA (A) 48 h, (B) 72 h Biochemical/Test Reactions .Oxidase: Negative A B .Catalase: Weak positive .Urease: Negative Additional Information .Can be misidentified as: Haemophilus influenzae, Actinobacillus spp. by automated ID systems .Infective Dose: 10 colony forming units Biosafety Level 3 agent (once Francisella tularensis is . Growth on CA (A) 48 h, (B) 72 h suspected, work should only be done in a certified Class II Biosafety Cabinet) .Transmission: Inhalation, insect bite, contact with tissues or bodily fluids of infected animals .Contagious: No Acceptable Specimen Types .Tissue biopsy .Whole blood: 5-10 ml blood in EDTA, and/or Inoculated blood culture bottle Swab of lesion in transport media . Gram stain Sentinel Laboratory Rule-Out of Francisella tularensis Oxidase Little to no growth on BA >48 h Small, grey-white opaque colonies on CA after ≥48 h at 35/37ºC Positive Weak Negative Positive Catalase Tiny, pleomorphic, faintly stained, gram-negative coccobacilli (red, round, and random) Perform all additional work in a certified Class II Positive Biosafety Cabinet Weak Negative Positive *Oxidase: Negative Urease *Catalase: Weak positive *Urease: Negative *Oxidase, Catalase, and Urease: Appearances of test results are not agent-specific. -
Chocolate Agar W/Enrichments Catalog No.: P3025 Blood Agar/Chocolate Bi-Plate Catalog No.: T1250chocolate Agar Slant
Administrative Offices Phone: 207-873-7711 Fax: 207-873-7022 Customer Service P.O. Box 788 Phone: 1-800-244-8378 Fax: 207-873-7022 Waterville, Maine 04903-0788 227 China Road Winslow, Maine 04901 TECHNICAL PRODUCT INFORMATION Catalog No.: P1150Chocolate Agar w/Enrichments Catalog No.: P3025 Blood Agar/Chocolate Bi-plate Catalog No.: T1250Chocolate Agar Slant INTENDED USE: Chocolate Agar is recommended for the cultivation and isolation of Neisseria and Haemophilus species. CO 2 favors primary isolation. The medium is best for organisms which require X and V Factor. HISTORY/SUMMARY: Interest in the cultural procedure for the diagnosis of gonococcal infections was stimulated by Ruys and Hens McLeod et al 2, Leahy and Carpenter, Leahy and Wilson 5 and Carpenter 6 who clearly demonstrated the superiority of this method over the microscopic technique. Further studies in cooperation with Carpenter, and McLeod 7 and Herrold resulted in the development of Chocolate Agar prepared with Proteose No. 3 Agar and Hemoglobin, which proved to be satisfactory for isolating the organism from all types of gonococcal infections. Chapin and Doern found that in only 6 of 17 cases was Haemophilus influenzae recovered from sputum specimens cultured by using conventional techniques including enriched chocolate agar (CHOC) media, despite the fact that gram stained smears of sputum specimens often revealed a predominance of pleomorphic gram- negative bacilli. Observations such as these have lead to the development of selective media which inhibit upper respiratory tract microbial flora while permitting growth of Haemophilus influenzae . Approaches utilized most frequently incorporate bacitracin into various enriched basal media ( 11, 12, 13, 14 ). -
Rapid Identification Ofbacteroides Fragilis with Bile and Antibiotic Disks D
JOURNAL OF CuNICAL MICROBIOLOGY, Apr. 1977, p. 439-443 Vol. 5, No. 4 Copyright C 1977 American Society for Microbiology Printed in U.S.A. Rapid Identification ofBacteroides fragilis with Bile and Antibiotic Disks D. L. DRAPER1 AND A. L. BARRY* Section ofInfectious and Immunologic Diseases, School of Medicine, University of California, Davis, California 95616, and Clinical Microbiology Laboratories, Medical Center, Sacramento, California 95817* Received for publication 21 December 1976 A simple screening test is described for separating Bacteroides fragilis from other anaerobic gram-negative bacilli. The test utilizes filter paper disks im- pregnated with 25 mg of oxgall (Difco), tested in conjunction with antibiotic identification disks. The bile disks and antibiotic disks are placed on a supple- mented brucella blood agar plate which has been inoculated by swabbing with a standardized cell suspension. After 24 h at 350C in a GasPak jar, resistance to kanamycin and bile is taken as a presumptive identification of B. fragilis. Susceptibility to one or both disks indicates the need for further identification and additional biochemical tests are required. Those strains that produce insuf- ficient growth within 24 h are not likely to be B. fragilis. The reliability of the bile disk method was tested by comparing results with 100 clinical isolates versus results with bile in thioglycolate broth, peptone-yeast extract-glucose broth, and tryptic soy agar. All four bile test methods gave equilvalent results, but the broth media required much longer periods of incubation. Bacteroides fragilis is the anaerobic gram- would quickly identify most B. fragilis strains negative bacillus most frequently recovered could significantly reduce the number of iso- from human infections. -
Medical Bacteriology
LECTURE NOTES Degree and Diploma Programs For Environmental Health Students Medical Bacteriology Abilo Tadesse, Meseret Alem University of Gondar In collaboration with the Ethiopia Public Health Training Initiative, The Carter Center, the Ethiopia Ministry of Health, and the Ethiopia Ministry of Education September 2006 Funded under USAID Cooperative Agreement No. 663-A-00-00-0358-00. Produced in collaboration with the Ethiopia Public Health Training Initiative, The Carter Center, the Ethiopia Ministry of Health, and the Ethiopia Ministry of Education. Important Guidelines for Printing and Photocopying Limited permission is granted free of charge to print or photocopy all pages of this publication for educational, not-for-profit use by health care workers, students or faculty. All copies must retain all author credits and copyright notices included in the original document. Under no circumstances is it permissible to sell or distribute on a commercial basis, or to claim authorship of, copies of material reproduced from this publication. ©2006 by Abilo Tadesse, Meseret Alem All rights reserved. Except as expressly provided above, no part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, recording, or by any information storage and retrieval system, without written permission of the author or authors. This material is intended for educational use only by practicing health care workers or students and faculty in a health care field. PREFACE Text book on Medical Bacteriology for Medical Laboratory Technology students are not available as need, so this lecture note will alleviate the acute shortage of text books and reference materials on medical bacteriology. -
APPENDIX a Media and Reagents
APPENDIX A Media and Reagents Pauline K. w. Yu, M.S. The use of appropriate and dependable media is integral to the isolation and identification of microorganisms. Unfortunately, comparative data docu menting the relative efficacy or value of media designed for similar purposes are often lacking. Moreover, one cannot presume identity in composition of a given generic product which is manufactured by several companies because each may supplement the generic products with components, often of a proprietary nature and not specified in the product's labeling. Finally, the actual production of similar products may vary among manufacturers to a sufficient extent to affect their performance. For all of these reasons, therefore, product selection for the laboratory should not be strictly based on cost considerations and should certainly not be based on promotional materials. Evaluations that have been published in the scientific literature should be consulted when available. Alternatively, the prospective buyer should consult a recognized authority in the field. It is seldom necessary for the laboratory to prepare media using basic components since these are usually available combined in dehydrated form from commercial sources; however, knowledge of a medium's basic compo nents is helpful in understanding how the medium works and what might be wrong when it does not work. Hence, the components have been listed for each medium included in this chapter. All dehydrated media must be prepared exactly according to the manu facturers' directions. Any deviation from these directions may adversely affect or significantly alter a medium's performance. Containers of media should be dated on receipt and when opened, and the media should never be used beyond expiration dates specified by the manufacturers or recom mended by quality control programs. -
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. -
Obligate Anaerobe in Thioglycollate Medium
Obligate Anaerobe In Thioglycollate Medium Restiform and pernicious Monty addled her libertarians chastens rurally or alluding drawlingly, is Lorne ungainful? Ernst often laicises intrusively when haustellate Talbert underpeep parchedly and skiagraph her therapies. Adair uncanonizing shily as Mozartean Gifford tellurizing her unsociableness disentwining septically. Anaerobic bacteria can be identified by separate them in test tubes of thioglycollate broth. Aerobic and Anaerobic Growth Lab Exercise 6 Sp16 Canvas. Enriched Thioglycollate Broth is recommended for the isolation cultivation and identification of society wide following of obligate anaerobic bacteria Composition. How to the center of oxygen allowing the pus should not as they may live session is medium in thioglycollate are entire plate in air and aerobic respiration. We extend such an organism an aerotolerant anaerobe and set just off against an additional category of oxygen relationship Those left black the facultative. A movie free from thioglycollate for the growth of aerobic and anaerobic. Opened under a medium onto a medium on medium low, obligate anaerobe in thioglycollate medium, obligate and environmental parameters. Anaerobic Thioglycollate Medium Base Himedia. Find each pipette, obligate anaerobe growth. Incubation obligate anaerobes will grow abnormal in that portion of turkey broth for the. Obligate Anaerobes are actually poisoned by various forms of oxygen. Enriched Thioglycollate Broth M73 VWR. BBL Thioglycollate Medium Enriched with CiteSeerX. The model organism Escherichia coli is a facultative anaerobic bacterium ie it is solar to operate in both aerobic and anaerobic environments. Thioglycollate medium Difco containing 1 dextrose and B. Oxygen Requirements for Microbial Growth Microbiology. Fluid Thioglycollate Medium Flashcards Quizlet. The Thioglycolate medium other the word hand stands out fast its ability to. -
Glucanoacetobacter
GLUCANOACETOBACTER INTRODUCTION Glucanoacetobacter is a genuine in the phylum proto bacteria. It is like rod shape and circular ends. It can be classified as gram negative bacterium. The bacterium is known for stimulating plant growth and being tolerant to acetic acid with one to three lateral flagella and known to be found on sugar cane. Gluconacetobacter diazotrophicus was discovered in Brazil by Bladimir A Cavalcante and Johannna/Dobereiner. Domain: Bacteria Phylum: Proteobacteria Class :Alphaproteobacteria Order : Rhodospirillales Family: Acetobacteraceae Genus:Gluconacetobacter Species:G.diazotrophicus CHARACTERISTICS Originally found in Alagoas, Brazil, Gluconacetobacter diazotrophicus is a bacterium that has several interesting features and aspects which are important to note. The bacterium was first discovered by Vladimir A. Cavalcante and Johanna Dobereiner while analyzing sugarcane in Brazil. Gluconacetobacter diazotrophicus is a part of the Acetobacteraceae family and started out with the name, Saccharibacter nitrocaptans, however, the bacterium is renamed as Acetobacter diazotrophicus because the bacterium is found to belong with bacteria that are able to tolerate acetic acid. Again, the bacterium’s name was changed to Gluconacetobacter diazotrophicus when its taxonomic position was resolved using 16s ribosomal RNA analysis. In addition to being a part of the Acetobacter family, Gluconacetobacter azotrophicus belongs to the Proteobacteria phylum, the Alphaprotebacteria class, and the Gluconacetobacter genus while being a part of the Rhodosprillales order. Other nitrogen-fixing species in this same genus include Gluconacetobacter azotocaptans and Gluconacetobacter johannae. Gluconacetobacter diazotrophicus cells are shaped like rods, have ends that are circular or round, and have anywhere from one to three flagella that are lateral. Based on these descriptions of the cell, Gluconacetobacter diazotrophicus can be classified with the bacillus genus. -
Metaproteomics Characterization of the Alphaproteobacteria
Avian Pathology ISSN: 0307-9457 (Print) 1465-3338 (Online) Journal homepage: https://www.tandfonline.com/loi/cavp20 Metaproteomics characterization of the alphaproteobacteria microbiome in different developmental and feeding stages of the poultry red mite Dermanyssus gallinae (De Geer, 1778) José Francisco Lima-Barbero, Sandra Díaz-Sanchez, Olivier Sparagano, Robert D. Finn, José de la Fuente & Margarita Villar To cite this article: José Francisco Lima-Barbero, Sandra Díaz-Sanchez, Olivier Sparagano, Robert D. Finn, José de la Fuente & Margarita Villar (2019) Metaproteomics characterization of the alphaproteobacteria microbiome in different developmental and feeding stages of the poultry red mite Dermanyssusgallinae (De Geer, 1778), Avian Pathology, 48:sup1, S52-S59, DOI: 10.1080/03079457.2019.1635679 To link to this article: https://doi.org/10.1080/03079457.2019.1635679 © 2019 The Author(s). Published by Informa View supplementary material UK Limited, trading as Taylor & Francis Group Accepted author version posted online: 03 Submit your article to this journal Jul 2019. Published online: 02 Aug 2019. Article views: 694 View related articles View Crossmark data Citing articles: 3 View citing articles Full Terms & Conditions of access and use can be found at https://www.tandfonline.com/action/journalInformation?journalCode=cavp20 AVIAN PATHOLOGY 2019, VOL. 48, NO. S1, S52–S59 https://doi.org/10.1080/03079457.2019.1635679 ORIGINAL ARTICLE Metaproteomics characterization of the alphaproteobacteria microbiome in different developmental and feeding stages of the poultry red mite Dermanyssus gallinae (De Geer, 1778) José Francisco Lima-Barbero a,b, Sandra Díaz-Sanchez a, Olivier Sparagano c, Robert D. Finn d, José de la Fuente a,e and Margarita Villar a aSaBio.