A Study on Microbiological Profile of Vaginitis and Its
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Characterization and Antibiotic Sensitivity Profile of Bacteria Isolated from Patients with Respiratory Tract Infections in Bangladesh
Characterization and Antibiotic Sensitivity Profile of Bacteria Isolated from Patients with Respiratory Tract Infections in Bangladesh Shukla Promite1, Sajal K. Saha2, Sunjukta Ahsan1 and Marufa Zerin Akhter1 1Department of Microbiology, University of Dhaka, Dhaka, Bangladesh 2Department of General Practice, Monash University, Building 1, 270 Ferntree Gully Road, Notting Hill VIC 3168, Australia (Received: October 08, 2017; Accepted: December 15, 2017; Published (web): December 23, 2017) ABSTRACT: The study was aimed to characterize bacterial isolates from respiratory tract infections (RTI) and investigate their antibiotic sensitivity profile. Selective media and biochemical tests were used to characterize 40 bacterial isolates. Antibiotic sensitivity testing was conducted using Kirby-Bauer disc diffusion method. About 42.5% (17) RTI patients were infected by Klebsiella pneumoniae, 30% (12) by Escherichia coli and 27.5% (11) by Pseudomonas aeruginosa with no significant gender variation (p-value <0.578). Overall, 47% (out of 20) antibiotics were sensitive, whereas 48% were resistant. Surprisingly, 18% P. aeruginosa and 20% K. pneumoniae were carbapenem-resistant and 4 out of 7 cephalosporin antibiotics were highly resistant irrespective of pathogens. E. coli showed better sensitivity to nitrofurantoin (78%) and levofloxacin (89%), while K. pneumoniae was insensitive to cotrimoxazole (88%), gentamycin (77%) and piperacillin/tazobactam (66%). On the other hand, P. aeruginosa did not respond to P. aeruginosa to nalidixic acid (60%) and ciprofloxacin (60%). This study concludes that nitrofurantoin, levofloxacin, cotrimoxazole, gentamycin and piperacillin/tazobactam antibiotics could be better alternative in treating bacterial RTIs. Key words: Antibiotic sensitivity, bacterial pathogens, RTIs, Bangladesh. INTRODUCTION Antibiotic resistance (AR) is a global public The rise of AR in Bangladesh is probably due to 1 health concern. -
MIO Medium (Motility Indole Ornithine Medium) M378
MIO Medium (Motility Indole Ornithine Medium) M378 Motility Indole Ornithine Medium (MIO Medium) is used for the identification of Enterobacteriaceae on the basis of motility, indole production and ornithine decarboxylase activity. Composition** Ingredients Gms / Litre Casein enzymic hydrolysate 10.000 Peptic digest of animal tissue 10.000 Yeast extract 3.000 L-Ornithine hydrochloride 5.000 Dextrose 1.000 Bromocresol purple 0.020 Agar 2.000 Final pH ( at 25°C) 6.5±0.2 **Formula adjusted, standardized to suit performance parameters Directions Suspend 31.02 grams in 1000 ml distilled water. Heat to boiling to dissolve the medium completely. Dispense in test tubes in 5 ml amounts. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool the tubes in an upright position. Principle And Interpretation Motility, indole production and ornithine decarboxylation are routine biochemical tests employed during identification of Enterobacteriaceae . Motility can be demonstrated microscopically (hanging drop) or macroscopically (tube method), where motility is observed as a diffused zone of growth flaring out from the line of inoculation. Indole test is carried out to determine the ability of an organism to split indole from tryptophan by the tryptophanase enzyme. On reaction with Kovacs reagent, indole combines with the colour in the alcohol layer, which is visualized as a red ring (in the alcohol layer) (1). If the test organisms possess the specific decarboxylase enzyme, then ornithine is decarboxylated to putrescine, an amine, resulting in a subsequent rise in the pH of the medium towards alkalinity. This causes the pH indicator bromocresol purple to change from purple to yellow colour. -
05686 DMACA (Indol Detection Disks)
05686 DMACA (Indol Detection Disks) The DMACA Indole discs are used for Indole test to determine the ability of an organism to split tryptophan into indole and α-aminopropionic acid. The presence of indole can be detected by the addition of DMACA which results in a bluish-purple complex. With this method it is possible to differentiate Escherichia coli from Klebsiella. Composition: (1 package contains 50 disks) The discs contain p-Dimethylaminocinnamaldehyde (DMACA). Directions: Place the disc on suspected colony from e.g. UTI ChromoSelect Agar, modified (16636) or Christensen’s Urea Agar (27048) plate on filter paper. Observe for appearance of blue-purple color within 10-30 seconds. Quality control: Test Organisms (ATCC) DMACA Escherichia coli (25922) + Pseudomonas aeruginosa (27853) - Klebsiella pneumoniae (13883) - 1. Staphylococcus aureus 2. Escherichia coli Page 1 of 2 References: 1. R. Vracko, J.C. Sherris, Indole-spot test in bacteriology., Am. J. Clin. Pathol., 39, 429 (1963) 2. V.L. Sutter, W.T. Carter, Evaluation of media and reagents for indole-spot test in anaerobic bacteriology., Am. J. Clin. Pathol., 58, 335 (1972) 3. G.D. Fay, A.L. Barry, Methods for detecting indole production by gram-negative nonsporeforming anaerobes., Appl. Micro. 27, 562 (1974) 4. D.F. Welch, P.A. Ahlin, J.M. Matsen, Differentiation of Haemophilus spp. in respiratory isolate cultures by an indole spot test., J. Clin. Micro. 15, 216 (1982) 5. H.D. Isenberg, Ed., Clinical microbiology procedures handbook, Vol 1., Washington, DC, ASM (1992) 6. B.A. Forbes, D.F. Sahm, A.S. Weissfeld, Bailey and Scott's diagnostic microbiology., 10th ed., St Louis, Mosby (1998) 7. -
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. -
Escherichia Coli Isolates Causing Asymptomatic Bacteriuria in Catheterized and Noncatheterized Individuals Possess Similar Virulence Propertiesᰔ 1 2 1 2
JOURNAL OF CLINICAL MICROBIOLOGY, July 2010, p. 2449–2458 Vol. 48, No. 7 0095-1137/10/$12.00 doi:10.1128/JCM.01611-09 Copyright © 2010, American Society for Microbiology. All Rights Reserved. Escherichia coli Isolates Causing Asymptomatic Bacteriuria in Catheterized and Noncatheterized Individuals Possess Similar Virulence Propertiesᰔ 1 2 1 2 Rebecca E. Watts, Viktoria Hancock, Cheryl-Lynn Y. Ong, Rebecca Munk Vejborg, Downloaded from Amanda N. Mabbett,1 Makrina Totsika,1 David F. Looke,3 Graeme R. Nimmo,4 Per Klemm,2 and Mark A. Schembri1* School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, Queensland, Australia1; Microbial Adhesion Group, DTU Food, Technical University of Denmark, Lyngby, Denmark2; and Infection Management Services, Princess Alexandra Hospital,3 and Queensland Health Pathology Service, Department of Microbiology, Royal Brisbane and Women’s Hospital,4 Brisbane, Queensland, Australia Received 19 August 2009/Returned for modification 12 November 2009/Accepted 22 April 2010 http://jcm.asm.org/ Urinary tract infections (UTIs) are among the most common infectious diseases of humans, with Escherichia coli being responsible for >80% of all cases. Asymptomatic bacteriuria (ABU) occurs when bacteria colonize the urinary tract without causing clinical symptoms and can affect both catheterized patients (catheter- associated ABU [CA-ABU]) and noncatheterized patients. Here, we compared the virulence properties of a collection of ABU and CA-ABU nosocomial E. coli isolates in terms of antibiotic resistance, phylogenetic grouping, specific UTI-associated virulence genes, hemagglutination characteristics, and biofilm formation. CA-ABU isolates were similar to ABU isolates with regard to the majority of these characteristics; exceptions were that CA-ABU isolates had a higher prevalence of the polysaccharide capsule marker genes kpsMT II and on October 22, 2015 by University of Queensland Library kpsMT K1, while more ABU strains were capable of mannose-resistant hemagglutination. -
Spot Indole Reagent
PRODUCT DETERIORATION This product should not be used if (1) the color has changed, (2) the expiration date has passed, or (3) there are other signs of deterioration. SPOT INDOLE REAGENT SPECIMEN COLLECTION, STORAGE, TRANSPORT Specimens should be collected and handled INTENDED USE 3 Remel Spot Indole Reagent is recommended for use following recommended guidelines. in qualitative procedures to determine the ability of an organism to split indole from the tryptophan MATERIALS REQUIRED BUT NOT SUPPLIED molecule. (1) Loop sterilization device, (2) Inoculating loop, swab, collection containers, (3) Incubators, alternative SUMMARY AND EXPLANATION environmental systems, (4) Supplemental media, Vracko and Sherris, in 1963, utilized Spot Indole (5) Quality control organisms, (6) Whatman (No. 1) Reagent for the presumptive separation of the filter paper. Proteus species and Escherichia coli.1 In 1969, Lowrance, Reich, and Traub found ρ-Dimethylamino- PROCEDURE cinnamaldehyde to be the most sensitive indole Filter Paper Method: reagent, capable of detecting 3 mcg of indole per 1. Dispense 1 or 2 drops of reagent onto a piece of milliliter of medium.2 Whatman (No. 1) filter paper or equivalent. 2. Smear the growth from an actively growing pure PRINCIPLE culture onto the saturated filter paper. Intracellular enzymes (i.e., tryptophanases) mediate 3. Observe for the development of a blue color the production of indole by hydrolytic activity against within 1 to 3 minutes. the amino acid tryptophan. Indole combines with Swab Method: dimethylaminocinnamaldehyde to form a blue-green 1. Dispense 1 or 2 drops of reagent onto the tip of compound. The reaction occurs by a condensation a cotton swab. -
Biochemical Tests Indole Test
Biochemical Tests Indole test Objective: to detect the ability of organism to produce enzyme tryptophanase. Principle: Indole test is a biochemical test which differentiates the coliform from other members of Enterobacteriacee by detecting their ability to produce the enzyme tryptophanase. This enzyme hydrolyses the amino acid tryptophan into indole, pyruvic acid and ammonia. It is the intracellular enzyme (endoenzyme). Tryptophan + H2O————tryptophanase enzyme——> Indole + Pyruvic acid + Ammonia Pyruvic acid can then be used by the organism in the Kreb’s cycle or it can enter glycolysis and be used to synthesize other compounds necessary for the cell. The media that is used for indole test is SIM (sulphide, indole, motility) medium or nutrient peptone, both of those media provides sufficient amino acid, tryptophan which acts as substrate for the above reaction. Hence, the organism that are able to produce the pyruvic acid as main product and indole, ammonia as the byproduct. The reagent used for this test is kovac’s reagent; (HCI and dimenthyl aminobenzaldehyde dissolved in amyl alcohol) which reacts with the side product of the tryptophan catabolism reaction i.e indole to form Rosindole dye which is cherry red in colour. Hence the formation of cherry red colour of Rosindole dye indicates positive indole test, otherwise not Escherichia coli is positive to indole test while klebsiella is negative to it. Voges–Proskauer (VP) Test The Voges-Proskauer (VP) test is used to determine if an organism produces acetylmethyl carbinol from glucose fermentation. If present, acetylmethyl carbinol is converted to diacetyl in the presence of ∝- naphthol, strong alkali (40% KOH), and atmospheric oxygen. -
SPOT INDOLE REAGENT Σ (For in Vitro Diagnostic Use) 250
PRODUCT CODE PL.391-10 SPOT INDOLE REAGENT Σ (for in vitro diagnostic use) 250 INTENDED USE MATERIALS REQUIRED BUT NOT PROVIDED should be checked with known positive and negative control Pro-Lab’s Spot Indole Reagent is to be used in the qualitative method to 1. Inoculating loops organisms. determine the ability of an organism to split indole from the tryptophan 2. Filter paper (Whatman No. 1 or equivalent) 5. Only pure cultures of organisms are to be tested. Weakly false molecule. 3. Cotton-tipped swabs positive reactions may occur if the inoculum is a mixed culture of 4. Incubator indole positive and negative organisms, since adjacent colonies are SUMMARY AND EXPLANATION 5. Supplemental media likely to take up diffused indole4. Spot Indole Reagent was used by Vracko and Sherris in 1963 for the 6. Quality control organisms presumptive differentiation of Proteus species and Escherichia coli1. The REFERENCES work of Lowrence, Reich and Traub in 1969, indicated that p-diemethyl- PROCEDURE 1. Vracko, R. and J.C. Sherris. (1963). Am. J. Clin. Path. 39:429-432. aminocinnamaldeyde is the most sensitive indole reagent, capable of Allow the reagent to come to room temperature prior to use. 2. Lowrance, B.L., P. Reich and W.H. Traub. (1969). Appl. Microbiol. detecting 3 mcg of indole per millilitre of medium2. Filter Paper Method: 17:923-924. 1. Dispense 1 to 2 drops of Spot Indole Reagent onto a piece of filter 3. Balzevic, D.J. and G.M. Ederer. (1975). Principles of Biochemical Tests PRINCIPLE OF THE PROCEDURE paper (Whatman No. -
Research Journal of Pharmaceutical, Biological and Chemical Sciences
ISSN: 0975-8585 Research Journal of Pharmaceutical, Biological and Chemical Sciences Analysis of Important Dairy Products: Isolation and Characterization LK Attri* and Humeet Narang Desh Bhagat University, Mandi Gobindgarh-147 301, Punjab, India. ABSTRACT In the present study, different daily use dairy products were procured and isolation was carried out using standard morphological. Based upon the morphological and biochemical standard test, the isolates were confirmed to be E.coli, Pseudomonas, Lactobacillus and Staphylococcus. Different biochemical standard methods including Indole production test, Methyl-Red and Voges-Proskauer (MRVP) test, Catalase Test and Citrate Utilization Test etc. were performed for final conclusion. Therefore, the results showed the presence of harmful bacterias in daily use dairy product. Further, Slants of nutrient agar media were made and kept for solidification. After solidification pure colonies of isolates were streaked on these slants and incubated at 37° C for 2-3 days. After growth these slants were stored at 4° C for further use. Keywords: Antibacterial agents, bacterias, standard tests, nutrient medias *Corresponding author January - February 2014 RJPBCS 5(1) Page No. 698 ISSN: 0975-8585 INTRODUCTION Dairy products are generally defined as food produced from the milk of mammals (the Food Standards Agency of the United Kingdom defines dairy as "foodstuffs made from mammalian milk). They are usually high energy-yielding food products. A production plant for the processing of milk is called a dairy or a dairy factory. Apart from breastfed infants, the human consumption of dairy products is sourced primarily from the milk of cows, goats, sheep, yaks, camels, and other mammals are other sources of dairy products consumed by humans. -
Indole Test Reagents Kovacs, DMACA, Spot Test
10/20/2016 Indole Test Reagents Kovacs, DMACA, Spot test INDOLE TEST REAGENTS Cat. no. Z65 Indole Spot Reagent 15ml Cat. no. Z67 Indole Kovacs Reagent 15ml INTENDED USE Hardy Diagnostics Indole Spot Reagent and Indole Kovacs Reagent are recommended for use in determining the indole reaction of bacteria. SUMMARY The indole test is a qualitative procedure for determining the ability of bacteria to produce indole by deamination of tryptophan. Using Kovacs tube method, indole combines, in the presence of a tryptophan rich medium, with p Dimethylaminobenzaldehyde at an acid pH in alcohol to produce a redviolet compound. In the spot test, indole combines, in the filter paper matrix, at an acid pH with pDimethylaminocinnamaldehyde (DMACA) to produce a blue to bluegreen compound. Indole Spot Reagent (DMACA) has been reported to be useful in detecting indole production by members of the family Enterobacteriaceae and certain anaerobic species. REAGENT FORMULA Ingredients per liter:* Indole Spot Reagent: pDimethylaminocinnamaldehyde (DMACA) 10.0gm Hydrochloric Acid, 37% 100.0ml Deionized Water 900.0ml Indole Kovacs Reagent: pDimethylaminobenzaldehyde 50.0gm Hydrochloric Acid, 37% 250.0ml Amyl Alcohol 750.0ml * Adjusted and/or supplemented as required to meet performance criteria. STORAGE AND SHELF LIFE https://catalog.hardydiagnostics.com/cp_prod/Content/hugo/IndoleTestRgnts.htm 1/5 10/20/2016 Indole Test Reagents Kovacs, DMACA, Spot test Storage: Upon receipt store at 230ºC. Products should not be used if there are any signs of deterioration or if the expiration date has passed. The expiration dating on the product label applies to the product in its intact packaging when stored as directed. -
Department of Microbiology Quality Manual Policy # MI GEN Page 1 of 36 Version: 1.5 CURRENT Section: Bacteriology Procedures Su
Policy # MI_GEN Page 1 of 36 Department of Microbiology Quality Manual Version: 1.5 CURRENT Section: Bacteriology Procedures Subject Title: Genital Tract Culture Manual Prepared by QA Committee Issued by: Laboratory Manager Revision Date: 7/14/2021 Approved by Laboratory Director: Next Review Date: 7/14/2023 Microbiologist-in-Chief Uncontrolled When Printed TABLE OF CONTENTS INTRODUCTION ............................................................................................................................... 2 LOWER GENITAL TRACT ............................................................................................................ 4 CERVICAL (ENDOCERVICAL) SWAB .............................................................................. 4 GROUP B STREPTOCOCCUS SCREEN ............................................................................. 7 VAGINITIS SCREEN ............................................................................................................ 10 VAGINAL CULTURE ........................................................................................................... 14 URETHRAL SWAB .............................................................................................................. 18 PENIS SWAB ......................................................................................................................... 20 SEMINAL FLUID .................................................................................................................. 23 UPPER GENITAL TRACT CULTURE ....................................................................................... -
Laboratory Diagnosis of Sexually Transmitted Infections, Including Human Immunodeficiency Virus
Laboratory diagnosis of sexually transmitted infections, including human immunodeficiency virus human immunodeficiency including Laboratory transmitted infections, diagnosis of sexually Laboratory diagnosis of sexually transmitted infections, including human immunodeficiency virus Editor-in-Chief Magnus Unemo Editors Ronald Ballard, Catherine Ison, David Lewis, Francis Ndowa, Rosanna Peeling For more information, please contact: Department of Reproductive Health and Research World Health Organization Avenue Appia 20, CH-1211 Geneva 27, Switzerland ISBN 978 92 4 150584 0 Fax: +41 22 791 4171 E-mail: [email protected] www.who.int/reproductivehealth 7892419 505840 WHO_STI-HIV_lab_manual_cover_final_spread_revised.indd 1 02/07/2013 14:45 Laboratory diagnosis of sexually transmitted infections, including human immunodeficiency virus Editor-in-Chief Magnus Unemo Editors Ronald Ballard Catherine Ison David Lewis Francis Ndowa Rosanna Peeling WHO Library Cataloguing-in-Publication Data Laboratory diagnosis of sexually transmitted infections, including human immunodeficiency virus / edited by Magnus Unemo … [et al]. 1.Sexually transmitted diseases – diagnosis. 2.HIV infections – diagnosis. 3.Diagnostic techniques and procedures. 4.Laboratories. I.Unemo, Magnus. II.Ballard, Ronald. III.Ison, Catherine. IV.Lewis, David. V.Ndowa, Francis. VI.Peeling, Rosanna. VII.World Health Organization. ISBN 978 92 4 150584 0 (NLM classification: WC 503.1) © World Health Organization 2013 All rights reserved. Publications of the World Health Organization are available on the WHO web site (www.who.int) or can be purchased from WHO Press, World Health Organization, 20 Avenue Appia, 1211 Geneva 27, Switzerland (tel.: +41 22 791 3264; fax: +41 22 791 4857; e-mail: [email protected]). Requests for permission to reproduce or translate WHO publications – whether for sale or for non-commercial distribution – should be addressed to WHO Press through the WHO web site (www.who.int/about/licensing/copyright_form/en/index.html).