Laboratory Protocols Level 1 Training Course Agar Diffusion Using E-Test 4
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Meningitis Manual Text
Laboratory Methods for the Diagnosis of MENINGITIS Caused by Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae Centers for Disease Control and Prevention August, 1998 Laboratory Methods for the Diagnosis of Meningitis Caused by Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae Table of Contents Introduction………………………………………………………………………………… 1 Acknowledgments ……………………………………………………………………….. 2 I. Epidemiology of Meningitis Caused by Neisseria meningitidis, Haemophilus influenzae and Streptococcus pneumoniae,…………………………………………… 3 II. General Considerations ......................................................................................................... 5 A. Record Keeping ................................................................................................................... 5 III. Collection and Transport of Clinical Specimens ................................................................... 6 A. Collection of Cerebrospinal Fluid (CSF)............................................................................... 6 A1. Lumbar Puncture ................................................................................................... 6 B. Collection of Blood .............................................................................................................. 7 B1. Precautions ............................................................................................................ 7 B2. Sensitivity of Blood Cultures ................................................................................ -
Figure 1: Etest Gradient Configuration When an Etest Gradient Strip Is
Inoculum preparation Table 1. Recommended media, inoculum and To obtain reproducible MICs from a gradient based Use the inoculum guide in TABLE 1. Emulsify several incubation1). system, the stability of the gradient must be maintai- well-isolated colonies from an overnight agar plate in lêÖ~åáëã= ^Ö~ê= fåçÅìäìã fåÅìÄ~íáçå ned throughout the critical period when the position a suitable suspension medium to achieve the specifi ed Öêçìé ãÉÇá~ of the growth/inhibition edge for a particular bac- inoculum turbidity by comparing to a McFarland pìëéÉåëáçå qìêÄáÇáíó= qÉãéÉê~íìêÉ= ^íãçëéÜÉêÉ qáãÉ= Antimicrobial Susceptibility Testing terium/antibiotic combination is determined. Due turbidity standard. For fastidious organisms such as EjÅc~êä~åÇF Eœ=O=ø`F EÜçìêëF For In Vitro Diagnostic Use to the stability and precision of the Etest predefi ned pneumococci, streptococci, gonococci, anaerobes and gradient, MIC values have been shown to be repro- Haemophilus spp., use the suspension prepared in INTENDED USE ducible and equivalent to those of the CLSI reference broth within 15 minutes. ^ÉêçÄÉë jìÉääÉê= MKURB=k~`ä MKR================== PR=ø` ~ãÄáÉåí NSJOM Etest is a quantitative technique for determining dilution procedures. eáåíçå EN=áÑ=ãìÅçáÇF the antimicrobial susceptibility of Gram negative Inoculation and Gram positive aerobic bacteria such as En- REAGENTS Soak a sterile, non-toxic swab in the inoculum terobacteriaceae, Pseudomonas, Staphylococcus and Etest is supplied in a package of 100 or 30 (some suspension and remove excess fl uid by pressing it lop^L=lopb jìÉääÉê= MKURB=k~`ä MKR PR=ø` ~ãÄáÉåí OQ=lop^==== Enterococcus species and fastidious bacteria, such as reagents) test strips of one antimicrobial agent. -
Multicellular Oxidant Defense in Unicellular Organisms MUCHOU MA and JOHN W
Proc. Natl. Acad. Sci. USA Vol. 89, pp. 7924-7928, September 1992 Microbiology Multicellular oxidant defense in unicellular organisms MUCHOU MA AND JOHN W. EATON* Division of Experimental Pathology, Department of Pathology and Laboratory Medicine, Albany Medical College, A-81, 47 New Scotland Avenue, Albany, NY 12208 Communicated by David W. Talmage, May 8, 1992 ABSTRACT Although catalase is thought to be a major MATERIALS AND METHODS defense against hydrogen peroxide (H202), the catalase activity Reagents. Brain heart infusion broth, Todd-Hewitt broth, within individual Escherichia coil fails to protect against ex- Lennox L agar (LB agar), and Bactoagar were obtained from ogenous H202. Contrary to earlier reports, we find that dilute GIBCO/BRL. The bicinchoninic acid protein microassay suspensions, of wild-type and catalase-deficient E. colt are was from Pierce. All other enzymes and chemicals were identical in their sensitivity to H202, perhaps because even purchased from Sigma. wild-type, catalase-positive E. colU cannot maintain an inter- Bacterial Strains and Culture Conditions. A catalase- nal/externail concentration gradient of this highly diffusible deficient mutant strain of E. coli K-12 [UM1, hereafter, oxidant. However, concentrated suspensions or colonies of cat(-)] and its parent wild-type [CSH7, hereafter cat(+)] (17) catalase-positive E. colt do preferentially survive H202 chal- were provided by P. C. Loewen (University ofManitoba). E. lenge and can even cross-protect adjacent catalase-deficient coli were grown statically in brain heart infusion broth or M9 organisms. Furthermore, high-density catalase-positive-but minimal salts medium supplemented with 10 mM glucose not catalase-negative-E. colt can survive and multiply in the (M9/glucose) (25) at 370C in room air overnight (18-20 hr) to presence of competitive, peroxide-generating streptococci. -
Growth Characteristics of Escherichia Coli and Staphylococcus Aureus Bacteria on Alternative Medium Leaves of Lamtoro (Leucaena Leucocephala)
Journal of Xi'an University of Architecture & Technology ISSN No : 1006-7930 Growth Characteristics of Escherichia coli and Staphylococcus aureus Bacteria on Alternative Medium Leaves of Lamtoro (Leucaena leucocephala) Meidawati Suswandari*, Department of Primary School, Faculty of Teacher Training and Education, Universitas Veteran Bangun Nusantara, Sukoharjo, Indonesia Lamtoro leaf has a high protein content. The protein content is very suitable for bacterial growth. Because of the high cost of bacterial growth media for educational and research institutions, lamtoro leaves can be used as an alternative medium for bacterial growth in general. The purpose of this study was to determine the potential of lamtoro leaf as an alternative medium for bacterial growth in general. This research is descriptive. Alternative mediums of lamtoro leaf were tested for the growth of Escherichia coli and Staphylococcus aureus. Escherichia coli bacteria grow on three alternative medium plates. After final identification, there are Escherichia coli bacteria. Whereas the Staphylococcus aureus bacterium did not grow on seven plates of alternative medium despite being incubated for 48 hours. Lamtoro leaf has less potential as an alternative medium for bacterial growth in general. The lamtoro leaf medium can only be used as a growth medium for gram-negative bacteria. While the growth of gram-positive bacteria there is no growth due to the presence of active substances in lamtoro leaves. Key words: Leaves of Lamtoro, Alternative Media, Escherichia coli, Staphylococcus aureus Introduction Bacteria are single-celled creatures that are very small or microscopic. Hans Christian Gram divides bacteria based on the characteristics of cell walls through the Gram staining system, namely Gram Positive and Gram Negative bacteria (Elferia, et al, 1996; Elliot, 2013; Harvey, 2001; Clausen, Gildberg, and Raa, 1985). -
Laboratory Exercises in Microbiology: Discovering the Unseen World Through Hands-On Investigation
City University of New York (CUNY) CUNY Academic Works Open Educational Resources Queensborough Community College 2016 Laboratory Exercises in Microbiology: Discovering the Unseen World Through Hands-On Investigation Joan Petersen CUNY Queensborough Community College Susan McLaughlin CUNY Queensborough Community College How does access to this work benefit ou?y Let us know! More information about this work at: https://academicworks.cuny.edu/qb_oers/16 Discover additional works at: https://academicworks.cuny.edu This work is made publicly available by the City University of New York (CUNY). Contact: [email protected] Laboratory Exercises in Microbiology: Discovering the Unseen World through Hands-On Investigation By Dr. Susan McLaughlin & Dr. Joan Petersen Queensborough Community College Laboratory Exercises in Microbiology: Discovering the Unseen World through Hands-On Investigation Table of Contents Preface………………………………………………………………………………………i Acknowledgments…………………………………………………………………………..ii Microbiology Lab Safety Instructions…………………………………………………...... iii Lab 1. Introduction to Microscopy and Diversity of Cell Types……………………......... 1 Lab 2. Introduction to Aseptic Techniques and Growth Media………………………...... 19 Lab 3. Preparation of Bacterial Smears and Introduction to Staining…………………...... 37 Lab 4. Acid fast and Endospore Staining……………………………………………......... 49 Lab 5. Metabolic Activities of Bacteria…………………………………………….…....... 59 Lab 6. Dichotomous Keys……………………………………………………………......... 77 Lab 7. The Effect of Physical Factors on Microbial Growth……………………………... 85 Lab 8. Chemical Control of Microbial Growth—Disinfectants and Antibiotics…………. 99 Lab 9. The Microbiology of Milk and Food………………………………………………. 111 Lab 10. The Eukaryotes………………………………………………………………........ 123 Lab 11. Clinical Microbiology I; Anaerobic pathogens; Vectors of Infectious Disease….. 141 Lab 12. Clinical Microbiology II—Immunology and the Biolog System………………… 153 Lab 13. Putting it all Together: Case Studies in Microbiology…………………………… 163 Appendix I. -
Laboratory Manual for Diagnosis of Sexually Transmitted And
Department of AIDS Control LaborLaboraattororyy ManualManual fforor DiagnosisDiagnosis ofof SeSexxuallyually TTrransmitansmittteded andand RRepreproductivoductivee TTrractact InInffectionsections FOREWORD Sexually Transmitted Infections (STIs) and Reproductive Tract Infections (RTIs) are diseases of major global concern. About 6% of Indian population is reported to be having STIs. In addition to having high levels of morbidity, they also facilitate transmission of HIV infection. Thus control of STIs goes hand in hand with control of HIV/AIDS. Countrywide strengthening of laboratories by helping them to adopt uniform standardized protocols is very important not only for case detection and treatment, but also to have reliable epidemiological information which will help in evaluation and monitoring of control efforts. It is also essential to have good referral services between primary level of health facilities and higher levels. This manual aims to bring in standard testing practices among laboratories that serve health facilities involved in managing STIs and RTIs. While generic procedures such as staining, microscopy and culture have been dealt with in detail, procedures that employ specific manufacturer defined kits have been left to the laboratories to follow the respective protocols. An introduction to quality system essentials and quality control principles has also been included in the manual to sensitize the readers on the importance of quality assurance and quality management system, which is very much the need of the hour. Manual of Operating Procedures for Diagnosis of STIs/RTIs i PREFACE Sexually Transmitted Infections (STIs) are the most common infectious diseases worldwide, with over 350 million new cases occurring each year, and have far-reaching health, social, and economic consequences. -
Sexually Transmitted Diseases Treatment Guidelines, 2015
Morbidity and Mortality Weekly Report Recommendations and Reports / Vol. 64 / No. 3 June 5, 2015 Sexually Transmitted Diseases Treatment Guidelines, 2015 U.S. Department of Health and Human Services Centers for Disease Control and Prevention Recommendations and Reports CONTENTS CONTENTS (Continued) Introduction ............................................................................................................1 Gonococcal Infections ...................................................................................... 60 Methods ....................................................................................................................1 Diseases Characterized by Vaginal Discharge .......................................... 69 Clinical Prevention Guidance ............................................................................2 Bacterial Vaginosis .......................................................................................... 69 Special Populations ..............................................................................................9 Trichomoniasis ................................................................................................. 72 Emerging Issues .................................................................................................. 17 Vulvovaginal Candidiasis ............................................................................. 75 Hepatitis C ......................................................................................................... 17 Pelvic Inflammatory -
Comparison of the Etest and the Routine Multi-Disc Agar Diffusion Susceptibility of Staphylococcus Species
Yah SC Etest and routine agar testing of Staphylococcus Comparison of the Etest and the routine multi-disc agar diffusion susceptibility of Staphylococcus species *Yah SC1, Ching FP2 and Atuanya EI3 Abstract. Aims: The present study, tend to evaluate the validity and accuracy of Etest as a method for performing in-vitro antimicrobial susceptibility testing of Staphylococcus with comparison to the routine multi disc agar diffusion. This is because the Etest susceptibility method is not yet known as a rapid, simple reliable technique in developing countries as it combine the functions of both dilution and diffusion technique. Materials and methods: Ninety-seven Staphylococcus aureus and eighty- three Staphylococcus epidermidis isolates were obtained from wound samples and identified according to standard morphological and biochemical methods. The antibiotics susceptibility patterns were determined both by agar disc diffusion and Etest methods in accordance to NCCLS (1997) criteria and manufacturer (AB Biodisk Sweden) respectively. Results: On the Etest strips, Staph aureus was 83.5% sensitive to ciprofloxacin, 52.6% to gentamicin, 48.5% to ampicillin and 8.2% to chloramphenicol while on the multi-disc agar diffusion plates 80.4% of Staph aureus were sensitive to ciprofloxacin, 49.5% to gentamicin, 39.2% to ampicillin and 12.4% to chloramphenicol.. On the Etest strips, 80.7% of Staph epidermidis were sensitive to ciprofloxacin, 34.9% to gentamicin, 25.3% to ampicillin and 15.7% to chloramphenicol while on the multi- disc agar diffusion plates 89.2% of Staph epidermidis were sensitive to ciprofloxacin, 34.9% to gentamicin, 25.3% to ampicillin and 32.5% to chloramphenicol. -
Biomerieux Product List 2016
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Fosfomycin Drug Review
THE NEBRASKA MEDICAL CENTER FOSFOMYCIN: REVIEW AND USE CRITERIA BACKGROUND Fosfomycin is a phosphonic acid derivative, which inhibits peptidoglycan assembly, thereby disrupting cell wall synthesis.1 Its uptake into the bacterial cell occurs via active transport, by the L-α-glycerophosphate transport and hexose phosphate uptake systems. Once inside the bacteria, it competes with phosphoenolpyruvate to irreversibly inhibit the enzyme enolpyruvyl transferase that catalyzes the first step of peptidoglycan synthesis.1,2 By irreversibly blocking this enzyme, cell wall synthesis is interrupted. Fosfomycin also decreases bacteria adherence to uroepithelial cells. Fosfomycin has been approved by the Food and Drug Administration (FDA) for the treatment of uncomplicated urinary tract infection (UTI) in adult women that is caused by Escherichia coli and Enterococcus faecalis.2 Oral fosfomycin has also been used for non-FDA approved indication such as complicated UTI without bacteremia. Intravenous fosfomycin, which is not available in the US, has also been used for a variety of infections including meningitis, pneumonia, and pyelonephritis.2 Fosfomycin does not have an indication for the treatment of pyelonephritis or perinephric abscess.2,3 Fosfomycin has broad spectrum of activity against aerobic gram positive and gram negative pathogens. It was shown in vitro and in clinical studies, to have activity against ≥90% of strains of E. coli, Citrobacter diversus, C. freundii, Klebsiella oxytoca, Klebsiella pneumoniae, Enterobacter cloacae, Serratia marcescens, Proteus mirabilis, P. vulgaris, Providencia rettgeri, Pseudomonas aeruginosa, E. faecalis and E. faecium [including vancomycin resistant (VRE)] species, and Staphylococcus aureus [including Methicillin resistant Staphylococcus aureus (MRSA)] associated with UTI.2,3 Currently, the Clinical and Laboratory Standards Institute (CLSI) susceptibility breakpoints exist only for E. -
D-Mannose Treatment Neither Affects Uropathogenic Escherichia Coli
molecules Article d-Mannose Treatment neither Affects Uropathogenic Escherichia coli Properties nor Induces Stable FimH Modifications 1,2, 3,4, 1 5,6 Daniela Scribano y , Meysam Sarshar y , Carla Prezioso , Marco Lucarelli , 5 1 3,7, 7, , Antonio Angeloni , Carlo Zagaglia , Anna Teresa Palamara y and Cecilia Ambrosi y * 1 Department of Public Health and Infectious Diseases, Sapienza University of Rome, 00185 Rome, Italy; [email protected] (D.S.); [email protected] (C.P.); [email protected] (C.Z.) 2 Dani Di Giò Foundation-Onlus, 00193 Rome, Italy 3 Department of Public Health and Infectious Diseases, Sapienza University of Rome, Laboratory Affiliated to Institute Pasteur Italia-Cenci Bolognetti Foundation, 00185 Rome, Italy; [email protected] (M.S.); [email protected] (A.T.P.) 4 Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran 1316943551, Iran 5 Department of Experimental Medicine, Sapienza University of Rome, 00185 Rome, Italy; [email protected] (M.L.); [email protected] (A.A.) 6 Pasteur Institute Cenci Bolognetti Foundation, 00161 Rome, Italy 7 IRCCS San Raffaele Pisana, Department of Human Sciences and Promotion of the Quality of Life, San Raffaele Roma Open University, 00166 Rome, Italy * Correspondence: [email protected]; Tel.: +39-06-4991-4622 These authors contributed equally to this work. y Academic Editor: László Somsák Received: 19 December 2019; Accepted: 10 January 2020; Published: 13 January 2020 Abstract: Urinary tract infections (UTIs) are mainly caused by uropathogenic Escherichia coli (UPEC). Acute and recurrent UTIs are commonly treated with antibiotics, the efficacy of which is limited by the emergence of antibiotic resistant strains. -
Product List Clinical
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