This document summarizes a study that investigated the occurrence of extended-spectrum beta-lactamases (ESBLs) in clinical isolates of Acinetobacter baumannii in Najaf, Iraq. 12 A. baumannii isolates were recovered from 770 clinical samples. All isolates were resistant to ampicillin and amoxicillin. Various tests found that 8-10 isolates were ESBL producers. PCR testing identified genes for SHV (in 3 isolates) and TEM (in 1 isolate) beta-lactamase types, but not VEB type. Isolates containing SHV and TEM genes also showed multidrug resistance. The study concludes there is dissemination of ESBL-producing
Emergence of ESBL worldwide has become a threat to successful treatment of noocomial infections. This deals with detection and treatment of ESBL infetions.
Isolation of Mutant
Made by :Shveta Arya
B.Pharm
Positive Selection
Positive selection entails growing the culture on a medium that will allow for the growth of only the mutant colonies.
If, for example, we want to find mutants that resistant to penicillin, we grow the culture on a medium that contains penicillin. Only those colonies that are resistant to penicillin will grow and we can identify them directly.
Negative Selection:
Negative selection is used to identify mutants that have lost the ability to perform a certain function that their parents had.
Auxotrophic mutants, for example, are bacteria that have lost the ability to synthesize an essential nutrient.
The replica-plating technique is used to identify mutants by negative selection.
the replica-plating technique can be used, for example, to identify mutants that have lost the ability to synthesize the amino acid histidine. Therefore, mutants are His- and require histidine in order to survive.
Inoculate a histidine enriched medium with bacteria. Incubate so that cells can form colonies. This is the master plate.
Press a sterile velvet surface into the colonies of the master plate. Some cells from each of the colonies adhere to the velvet.
Prepare two mediums, one with histidine, the other without histidine.
Transfer cells from the velvet to each plate.
Compare growth on the two plates after incubation. Colonies that grow on the histidine enriched medium but not on the medium lacking histidine are His- mutants
Ames test
The Ames Test utilizes a histidine auxotroph of Salmonella to determine if a chemical agent is a mutagen. Though some spontaneous back mutations (a reversion back to the strain of Salmonella that can synthesize histadine) can occur, if many colonies of the microbe appear on the minimal plate after the addition of the test chemical, this is an indication the the chemical is a mutagen.
THANK YOU
International Journal of Pharmaceutical Science Invention (IJPSI) inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
The occurrence of AmpC β-lactamase and ESBL producing Gram-negative bacteria ...iosrjce
IOSR Journal of Dental and Medical Sciences is one of the speciality Journal in Dental Science and Medical Science published by International Organization of Scientific Research (IOSR). The Journal publishes papers of the highest scientific merit and widest possible scope work in all areas related to medical and dental science. The Journal welcome review articles, leading medical and clinical research articles, technical notes, case reports and others.
Emergence of ESBL worldwide has become a threat to successful treatment of noocomial infections. This deals with detection and treatment of ESBL infetions.
Isolation of Mutant
Made by :Shveta Arya
B.Pharm
Positive Selection
Positive selection entails growing the culture on a medium that will allow for the growth of only the mutant colonies.
If, for example, we want to find mutants that resistant to penicillin, we grow the culture on a medium that contains penicillin. Only those colonies that are resistant to penicillin will grow and we can identify them directly.
Negative Selection:
Negative selection is used to identify mutants that have lost the ability to perform a certain function that their parents had.
Auxotrophic mutants, for example, are bacteria that have lost the ability to synthesize an essential nutrient.
The replica-plating technique is used to identify mutants by negative selection.
the replica-plating technique can be used, for example, to identify mutants that have lost the ability to synthesize the amino acid histidine. Therefore, mutants are His- and require histidine in order to survive.
Inoculate a histidine enriched medium with bacteria. Incubate so that cells can form colonies. This is the master plate.
Press a sterile velvet surface into the colonies of the master plate. Some cells from each of the colonies adhere to the velvet.
Prepare two mediums, one with histidine, the other without histidine.
Transfer cells from the velvet to each plate.
Compare growth on the two plates after incubation. Colonies that grow on the histidine enriched medium but not on the medium lacking histidine are His- mutants
Ames test
The Ames Test utilizes a histidine auxotroph of Salmonella to determine if a chemical agent is a mutagen. Though some spontaneous back mutations (a reversion back to the strain of Salmonella that can synthesize histadine) can occur, if many colonies of the microbe appear on the minimal plate after the addition of the test chemical, this is an indication the the chemical is a mutagen.
THANK YOU
International Journal of Pharmaceutical Science Invention (IJPSI) inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
The occurrence of AmpC β-lactamase and ESBL producing Gram-negative bacteria ...iosrjce
IOSR Journal of Dental and Medical Sciences is one of the speciality Journal in Dental Science and Medical Science published by International Organization of Scientific Research (IOSR). The Journal publishes papers of the highest scientific merit and widest possible scope work in all areas related to medical and dental science. The Journal welcome review articles, leading medical and clinical research articles, technical notes, case reports and others.
The prevalence of extended spectrum beta-lactamases (ESBLs) among Escherichia...Open Access Research Paper
The prevalence of extended spectrum β-lactamases among 246 clinical isolates from Abia State University teaching Hospital patients was investigated. The isolates were made up of 134 Escherichia coli and 112 Klebsiella species. Antimicrobial susceptibility of the isolates was determined by the disc diffusion method. ESBL phenotypes were determined by the double disc synergy method using ceftazidime, cefotaxime, ceftriaxone and co-amoxiclav. Out of the 246 isolates, 125 (50.8%) were ESBL producers, made up of 62(50.8%) E. coli and 63 (50.4%) Klebsiella isolates. Seventeen (54.8%) of the ESBL producing E. coli isolates were from in-patients while 45 (47.9%) were from out-patients. For the ESBL positive Klebsiella spp., 14 (45.2%) and 49 (52.1%) were from in-patients and out-patients respectively. ESBL producing isolates were also found to be more prevalent among the female patients (72.8%) than among the male patients (27.2%). The isolates also expressed high rates of resistance to other classes of antibiotics tested. However, Amikacin was found to have excellent performance against the urinary isolates tested and therefore is recommended for the treatment of infections caused by Escherichia coli and Klebsiella species. This study shows high prevalence of ESBL producing E. coli and Klebsiella isolates clinical samples of patients attending the Abia State University Teaching Hospital Aba, Abia State Nigeria.
A study of antibiotic resistance of Extended-Spectrum Beta-Lactamases produci...Premier Publishers
Background: Extended-Spectrum Beta-Lactamases - producing Enterobacteriaceae are common in hospitals. This study aims to describe the antibiotic resistance of these bacteria and their associated demographic and clinical factors. Methods: It was a prospective study of 73 isolates of Extended-Spectrum Beta-Lactamases - producing Enterobacteriaceae for a period of six months from July to December 2019 in the laboratory of Befelatanana. Results: This study showed 73 (6.3%) isolates of Extended-Spectrum Beta-Lactamases- producing Enterobacteriaceae, represented by 25 (34.2%) isolates of Klebsiella spp, 24 (32.9%) isolates of Escherichia coli, 22 (30.1%) isolates of Enterobacter spp and 2 (2.7%) isolates of Proteus spp. The antibiotic resistance of these bacteria varied from 0% to 100% for all of the antibiotics tested. Resistance to aminoglycosides ranged from 0% (amikacin) to 69.9% (gentamycin). Resistance to quinolones ranged from 43.8% (levofloxacin) to 76.7% (nalidixic acid). Similarly, 60 (82.2%) isolates were resistant to cotrimoxazole and 25 (34.2%) isolates to chloramphenicol. Patients under 20 years (57.1%) (p=0.03), men (52.2%)(p=0.11; NS), patients with respiratory samples (83.3%)(p=0.004), with pus (61.9%)(p=0.02) and hospitalized in surgery and intensive care units (68.4%)(p=0.0009) were the most affected by these enterobacteria. Conclusion: Extended-Spectrum Beta-Lactamases - producing Enterobacteriaceae are responsible for severe infections and the majorities are multi-resistant bacteria.
Keywords: Beta-lactamase, Enterobacteriaceae, antibiotic resistance, amikacin, imipenem.
Antibiotic Susceptibility Pattern of Pyogenic Bacterial Isolates in Sputum.IOSR Journals
Drugs Have Been Used For The Treatment Of Infectious Diseases Since 17th Century , However
Chemotherapy As A Science Has Began With Paul Ehrlich In The First Decade Of 20th Century . Paul Ehrlich
(1854-1915) Was One Of The Earliest Pioneers In The Field Of Antimicrobial Chemotherapy .1Ehrlich
Formulated The Principles Of “Selective Toxicity” ,I.E; Selective Inhibition Of The Growth Of Microorganisms
Without Damage To The Host.2 Resistance Has Been Documented Not Only Against Antibiotics Of
Natural And Semi- Synthetic Origin , But Also Against Purely Synthetic Compounds (Flouroquinolone) Or
Those Which Do Not Even Enter The Cells (Vancomycin) .3 However , The Euphoria Over The Potential
Conquest Of Infectious Diseases Was Short-Lived .Almost As Soon As Antibacterial Drugs Were Deployed ,
Bacteria Responded By Manifesting Various Forms Of Resistance.4 Considered As “Wonder Drugs”
Antibiotics Are Often Prescribed Inappropriately And Inadequately And Have Thus Became One Of The
Highly Abused Agents.5
Background: The widespread use of antibiotics has resulted in emergence of community-acquired antibiotic resistance among uropathogens in outpatient’s population. This constitutes an impediment in the management of urinary tract infection (UTI) in both community and hospital settings. Objective: The aim of this study was to determine the current antibiotic resistance trends, extended spectrum beta-lactamase (ESBL) production and plasmid profile of uropathogens from outpatients. Methods: A total of 370 mid-stream urine samples were collected and cultured by standard methods. Isolated uropathogens were identified using appropriate biochemical methods. The modified Kirby Bauer disk method was used for antibiotic susceptibility test. The ESBL-producing uropathogens were identified and their plasmid DNA extraction and curing were carried out by standard methods. Results: About 35.7% and 32.7% of uropathogens were multi-drug resistant and ESBL-producing respectively. There was higher prevalence of ESBL-production among isolates from female patients (62.5%) when compared to that from male patients (37.5%). The isolated uropathogens were most resistant to Cefotaxime, and most sensitive to Imipenem. Resistance to antibiotics by ESBL-producing uropathogens was found to be plasmid-mediated. Conclusion: Community acquired Uropathogens from outpatients were multidrug resistant due to ESBL production localized on plasmids, a probable cause of treatment failures experienced in Uyo.
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Title: Sense of Taste
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the structure and function of taste buds.
Describe the relationship between the taste threshold and taste index of common substances.
Explain the chemical basis and signal transduction of taste perception for each type of primary taste sensation.
Recognize different abnormalities of taste perception and their causes.
Key Topics:
Significance of Taste Sensation:
Differentiation between pleasant and harmful food
Influence on behavior
Selection of food based on metabolic needs
Receptors of Taste:
Taste buds on the tongue
Influence of sense of smell, texture of food, and pain stimulation (e.g., by pepper)
Primary and Secondary Taste Sensations:
Primary taste sensations: Sweet, Sour, Salty, Bitter, Umami
Chemical basis and signal transduction mechanisms for each taste
Taste Threshold and Index:
Taste threshold values for Sweet (sucrose), Salty (NaCl), Sour (HCl), and Bitter (Quinine)
Taste index relationship: Inversely proportional to taste threshold
Taste Blindness:
Inability to taste certain substances, particularly thiourea compounds
Example: Phenylthiocarbamide
Structure and Function of Taste Buds:
Composition: Epithelial cells, Sustentacular/Supporting cells, Taste cells, Basal cells
Features: Taste pores, Taste hairs/microvilli, and Taste nerve fibers
Location of Taste Buds:
Found in papillae of the tongue (Fungiform, Circumvallate, Foliate)
Also present on the palate, tonsillar pillars, epiglottis, and proximal esophagus
Mechanism of Taste Stimulation:
Interaction of taste substances with receptors on microvilli
Signal transduction pathways for Umami, Sweet, Bitter, Sour, and Salty tastes
Taste Sensitivity and Adaptation:
Decrease in sensitivity with age
Rapid adaptation of taste sensation
Role of Saliva in Taste:
Dissolution of tastants to reach receptors
Washing away the stimulus
Taste Preferences and Aversions:
Mechanisms behind taste preference and aversion
Influence of receptors and neural pathways
Impact of Sensory Nerve Damage:
Degeneration of taste buds if the sensory nerve fiber is cut
Abnormalities of Taste Detection:
Conditions: Ageusia, Hypogeusia, Dysgeusia (parageusia)
Causes: Nerve damage, neurological disorders, infections, poor oral hygiene, adverse drug effects, deficiencies, aging, tobacco use, altered neurotransmitter levels
Neurotransmitters and Taste Threshold:
Effects of serotonin (5-HT) and norepinephrine (NE) on taste sensitivity
Supertasters:
25% of the population with heightened sensitivity to taste, especially bitterness
Increased number of fungiform papillae
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TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Verified Chapters 1 - 19, Complete Newest Version.pdf
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Artificial intelligence (AI) refers to the simulation of human intelligence processes by machines, especially computer systems. It encompasses tasks such as learning, reasoning, problem-solving, perception, and language understanding. AI technologies are revolutionizing various fields, from healthcare to finance, by enabling machines to perform tasks that typically require human intelligence.
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New Directions in Targeted Therapeutic Approaches for Older Adults With Mantl...i3 Health
i3 Health is pleased to make the speaker slides from this activity available for use as a non-accredited self-study or teaching resource.
This slide deck presented by Dr. Kami Maddocks, Professor-Clinical in the Division of Hematology and
Associate Division Director for Ambulatory Operations
The Ohio State University Comprehensive Cancer Center, will provide insight into new directions in targeted therapeutic approaches for older adults with mantle cell lymphoma.
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Mantle cell lymphoma (MCL) is a rare, aggressive B-cell non-Hodgkin lymphoma (NHL) accounting for 5% to 7% of all lymphomas. Its prognosis ranges from indolent disease that does not require treatment for years to very aggressive disease, which is associated with poor survival (Silkenstedt et al, 2021). Typically, MCL is diagnosed at advanced stage and in older patients who cannot tolerate intensive therapy (NCCN, 2022). Although recent advances have slightly increased remission rates, recurrence and relapse remain very common, leading to a median overall survival between 3 and 6 years (LLS, 2021). Though there are several effective options, progress is still needed towards establishing an accepted frontline approach for MCL (Castellino et al, 2022). Treatment selection and management of MCL are complicated by the heterogeneity of prognosis, advanced age and comorbidities of patients, and lack of an established standard approach for treatment, making it vital that clinicians be familiar with the latest research and advances in this area. In this activity chaired by Michael Wang, MD, Professor in the Department of Lymphoma & Myeloma at MD Anderson Cancer Center, expert faculty will discuss prognostic factors informing treatment, the promising results of recent trials in new therapeutic approaches, and the implications of treatment resistance in therapeutic selection for MCL.
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Learning Objectives
1.) Identify clinical and biological prognostic factors that can guide treatment decision making for older adults with MCL
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Anti ulcer drugs and their Advance pharmacology ||
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These lecture slides, by Dr Sidra Arshad, offer a quick overview of physiological basis of a normal electrocardiogram.
Learning objectives:
1. Define an electrocardiogram (ECG) and electrocardiography
2. Describe how dipoles generated by the heart produce the waveforms of the ECG
3. Describe the components of a normal electrocardiogram of a typical bipolar leads (limb II)
4. Differentiate between intervals and segments
5. Enlist some common indications for obtaining an ECG
Study Resources:
1. Chapter 11, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
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The prostate is an exocrine gland of the male mammalian reproductive system
It is a walnut-sized gland that forms part of the male reproductive system and is located in front of the rectum and just below the urinary bladder
Function is to store and secrete a clear, slightly alkaline fluid that constitutes 10-30% of the volume of the seminal fluid that along with the spermatozoa, constitutes semen
A healthy human prostate measures (4cm-vertical, by 3cm-horizontal, 2cm ant-post ).
It surrounds the urethra just below the urinary bladder. It has anterior, median, posterior and two lateral lobes
It’s work is regulated by androgens which are responsible for male sex characteristics
Generalised disease of the prostate due to hormonal derangement which leads to non malignant enlargement of the gland (increase in the number of epithelial cells and stromal tissue)to cause compression of the urethra leading to symptoms (LUTS
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Molecular detection of extended spectrum beta- lactamases in clinical isolates of acinetobacter baumannii
1. Journal of Biology, Agriculture and Healthcare www.iiste.org
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online)
Vol.3, No.7, 2013
32
Molecular Detection of Extended-Spectrum Beta- Lactamases in
Clinical Isolates of Acinetobacter baumannii
Azhar A.L. AL-Thahab
Biology Department, Collage of Science, Babylon University,Iraq
* E-mail of the corresponding author: azharammran@yahoo.com
Abstract:
Resistance to third-generation oxyimino cephalosporins is emerging, and it is considered a problem in medical
field. Extended spectrum β-lactamase (ESBL) producing Acinetobacter baumannii have been noticed to be
important cause of hospital infections. This study aimed to undertaken and determine the occurrence of ESBLs
especially SHV, TEM and VEB β-lactamase types. A total of 770 clinical samples were collected from February
to June, 2011. The A. baumannii isolates were identified according to API 20NE system. Phenotypic detection of
ESBL was performed by using the combination disk, disk approximation methods then confirmed by culturing
on ESBL CHROM agar medium. The isolates were subjected to polymerase chain reaction (PCR) assays with
specific primers for blaSHV, blaTEM and blaVEB. Twelve (1.5%) A. baumannii isolates were recovered from
clinical infections. All of them were β-lactam resistant (resistant to both ampicillin and amoxicillin). Of the β-
lactam resistant isolates, 8/12 (66.7%) were found to be disk combination test positive, and 10 (83.3%) isolates
were confirmed as ESBL producers and gave heavy growth on ESBL CHROM agar. In PCR experiments using
specific primers for blaSHV, blaTEM and blaVEB genes, out of 12 A. baumannii isolates, three (25%) were harbored
blaSHV gene and only one (8.3%) isolate gave positive PCR results for blaTEM gene. This study demonstrate all
isolates were blaVEB negative. The present study concluded that the emerging of dissemination of ESBL
producing A. baumannii in Najaf hospitals.
Keywords: key words, Acinetobacter baumannii , Extended-Spectrum Beta- Lactamases, Molecular Detection
1 . Introduction
Acinetobacter baumannii has emerged over the last decade as a significant opportunistic pathogen. Although it
is generally associated with benign colonization of hospitalized patients, it is responsible for about 10% of
nosocomial infection in intensive care unit (ICU) patients, causing a wide range of infections (Levin et al, 2003;
Poirel et al, 1999). They are usually considered to be opportunistic pathogens, and of recent have been reported
to cause a number of outbreaks of nosocomial infections in hospitalized patients like septicaemia, pneumonia,
wound sepsis, endocarditis, meningitis and urinary tract infection (UTI) (Towner 1997). In Acinetobacter-
associated nosocomial infection, the major problem encountered by ICU clinicians relates to the readily
transferable antimicrobial resistance expressed by this organism (Bergogne- Berezin 2001). In addition to
intrinsic resistance, A. baumannii has the ability to acquire resistance to many major classes of antibiotics
including newer _- lactams (Perilli et al, 1996). The presence of resistance plasmids (R-plasmids) is a significant
feature of this organism, and plasmid profiling has been proposed as a method of epidemiological typing for
Acinetobacter (Joshi 1998). Although A. baumannii colonizes hospitalized patients, approximately 30% of
isolates are associated with frank infection in ICU patients and, in this setting, tend to demonstrate variable
susceptibility profiles (Dy et al, 1999).
Over the last 20 years many new β-lactam antibiotics, specifically designed to resist known β-lactamases, have
been developed . However, almost invariably new β- lactamases have emerged to combat each new class of β-
lactams. Plasmid-mediate ESBLs emerged in Gram-negative bacilli in Europe in the 1980s (Zeba 2005 ). ESBL-
producing bacteria are typically resistant to penicillins, first-and second-generation cephaloporins as well as the
third-generation oxyimino cephalosporins (Jacoby &Medeiros 1991).
Typically, ESBLs are plasmid encoded but also present on chromosomes, often in association with integrons.
These enzymes are derivatives, predominantly, of class A and D β-lactamases. Classical ESBLs evolved from
class A, TEM (from TEM-1 or TEM-2) and SHV (from SHV-1) enzymes, and these remain the most prevalent
types of ESBLs, though class D ESBLs (OXA family) have also been known for some time (Bradford 2001).
Hence the fundamental aim of this study is to identify the occurrence of ESBL in A. baumannii isolates
recovered from Hospital settings in Najaf.
2. Materials and Methods
2.1 Isolation and Identification
A total of 770 clinical samples included (sputum (n= 450), , urine (n= 210), and burn wounds (n= 110)) were
collected from patients in three separate hospitals (Al-Sader Medical City, Al-Hakeem General Hospital, Al-
Furat Teaching Hospital) in Najaf over five months period starting from February to June, 2011. Isolates were
2. Journal of Biology, Agriculture and Healthcare www.iiste.org
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online)
Vol.3, No.7, 2013
33
recovered from clinical samples after culturing on MacConkey's agar (Himedia, India) and incubated for
overnight at 37o
C, non lactose fermenting bacteria (colorless or slightly beige) were subcultured and incubated
for additional overnights. Suspected bacterial isolates which their cells are Gram negative coccobacillary or
diplobacillus and negative to oxidase which further identified by the traditional biochemical test according to
Holt et al. (1994) and MacFaddin (2000). Isolates were confirmed by API20 NE multi-test systems
(BioMerieux ,France).
2.2 Screening Test for β-lactam and Antibiotics Resistance
Ampicillin and amoxicillin were added separately, from stock solution, to the cooled Muller-Hinton agar at final
concentration of 100 and 50 µg/ml, respectively. The medium poured into sterilized Petri dishes, then stored at
4˚C. Isolates were cultured on Mueller–Hinton agar and their susceptibilities to different antibiotics were tested
by disk diffusion method (NCCLs, 2003 ). Results were compared with E.coli ATCC 25922 as negative control.
2.3 Phenotypic Detection of ESBL
2.3.1 Disk Combination Test (Recommended by CLSI 2010)
The phenotypic confirmation of potential ESBL-producing isolates was performed by using disk diffusion
method. Cefotaxime alone and in combination with clavulanic acid were tested. Inhibition zone of ≥ 5 mm
increase in diameter for antibiotic tested in combination with clavulanic acid versus its zone when tested alone
confirms an ESBL producing isolate (Cantarelli et al, 2007).
2.3.2 Disk Approximation Test
All β-lactamase producing isolates tested according to Batchoun et al. (2009). Antibiotic disks of cefotaxime
(30µg), ceftazidime (30µg), ceftriaxione (30µg), and azetreonam (30µg) were placed 15 mm (edge to edge)
around a central disk of amoxi-clav (20µg amoxicillin plus 10 µg clavulanate) on Muller-Hinton agar plates
seeded with organism being tested for ESBL production. Plates were incubated aerobically at 37◦
C for 24 hr .
Any augmentation (increase in diameter of inhibition zone) between the central amoxi-clav disk and any of the
β-lactam antibiotic disks showing resistance or intermediate susceptibility was recorded, and the organism was
thus considered as an ESBL producer.
2.3.3 CHROM agar Technique phenotype detection of ESBLs producing isolates
Extended spectrum β-lactamase CHROM agar plates were streaked in the same day of preparation by overnight
growth of A. baumannii. The plates were incubated at 37o
C for 24 hr according to manufacturer procedure.
Growth of blue colonies indicated to ESBL producer. The reference strain of E. coli ATCC 25922 was inhibited
and used as negative control.
2.4 Detection of ESBL bla Genes by Polymerase Chain Reaction
2.4.1 DNA extraction
Extraction of DNA from bacterial cells was performed by salting out method (Pospiech & Neumann, 1995) with
some modification to prepare templet DNA.
2.4.2 Polymerase Chain Reaction Protocols
2.4.2.1 PCR Mixture and thermo cycling conditions
The DNA template extracted from A. baumannii isolates were subjected to bla genes by PCR, the protocol was
used depending on manufacturer's instruction and the right PCR cycling program parameters conditions were
installed as in Table (1).
2.4.2.2 Agarose Gel Electrophoresis
All requirements, technical and preparations of agarose gel electrophoresis for DNA detection and analysis were
performed by Bartlett & Stirling (1998). Finally, the gel was photographed using Biometra gel documentation
system.
3. Results
Among the 770 clinical samples were collected during study period (Table 2), only 12 (1.5%) isolates had been
identified as A. baumannii, all isolates (100%) were resistant to both ampicillin and amoxycillin. Fifty percent of
A. baumannii isolates were recoverd from urine followed by 4(0.88%) from sputum and 2(1.8%) from burn
wound.Production of ESBL was confirmed by three different methods, disk combination , disk approximation
tests and ESBL CHROM agar (Table 3). Only 8 (66.7%) isolates demonstrated enhancement of inhibition zone,
suggesting production of ESBL by disk combination test, while no remarkable distinct change was noticed when
using disk approximation test. In same time most isolate 10 (83.3%) were identified as ESBL producer by
CHROM agar.In PCR experiments using specific primers for blaSHV and blaTEM and blaVEB , the results of table
(4) show three (25%) isolates were harbored blaSHV gene and only one (8.3%) isolate gave positive PCR results
for blaTEM gene (Figure 1 and 2). This study demonstrates all isolates were blaVEB negative.Consequently, table
(5) show all isolates that harbored SHV and TEM types gene in their genotype appeared phenotypicaly as
multidrug resistant isolates (resistant for more than three antibiotic classes).
3. Journal of Biology, Agriculture and Healthcare www.iiste.org
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4. Discussion
A. baumannii is an important causes of nosocomial infections and has been associated with a wide variety of
illness in hospitalized patients, especially patients in the intensive care units (Sinha et al, 2007). During a few
decades A. baumannii tend to be multidrug resistance (MDR) due to their ability to develop antibiotic resistance
(Kusradoze et al, 2011). Also, the extensive use of antimicrobial chemotherapy within hospital has contributed
to the emergence and procreation of A. baumannii strains which are resistance to a wide range of antibiotic
including broad spectrum β- lactams, aminoglycosides and flouroquinolones(Parisa et al, 2012).
The main mechanisms of resistance to β- lactams in A. baumannii is enzymatic degradation by β- lactamase
including the extended spectrum β- lactamase (blaTEM, blaSHV, blaVEB and blaPER) and metalo-beta-lactamase
(blaOXA51,23,24,and 58) (Shahcheraghi et al, 2011). As shown in our results, 83% of isolates showed positive results
for CHROM agar media compared with disc disc combination test (66.7%). This may due to the sensitivity of
CHROM agar for the detection of enzyme compared with other methods. Also, our results showed no correlation
between the existence of bla genes and phenotypic resistance against β- lactam antibiotics in A. baumannii. This
results was in an agreement with other studies that confirmed a specific correlation between genotypic and
phenotypic properties of β- lactam resistance among A. baumannii (Soroush et al, 2010; Srinivasan et al, 2009;
Yan et al.,2009). Several studies reported that resistance to β- lactam antibiotic was largely due to existence of
carbapenemase, ESBLs and metalo-beta-lactamase (Taherikalani et al, 2009; Lin et al.,2010; Srinivasan et
al.,2009). The vast majority of ESBLs are acquired enzyme encoded by plasmids, this confirmed our results
which showed no correlation between genotype and phenotype properties(Parisa et al ,2012; Taherikalani et
al ,2008 ; Papa et al ,2009).
The acquired ESBLs are expressed at various levels and differ significantly in biochemical characteristics such
as activity against specific β- lactams ( cefotaxim, ceftazidime and aztereonam) (Canton et al , 2012 : Kusradoz
et al , 2010 ).
On the other hand, a high distribution of multiple antibiotic resistance was found in β- lactamase resistance A.
baumannii (Table 5), this may due to the co-presence of other resistance mechanisms ( other β- lactamase,
effluex, altered permeability)( Parisa et al ,2012; Magiorakos et al , 2012; Uma et al ,2009)
References:
Bartlett, J.M.S. & Stirling, D. (1998). “PCR Protocols: Methods in Molecular Biology”. 2nd
. Humana Press Inc.
Totowa. NJ.
Batchoun, R.G., Swedan, S.F. & Shurman A.M. (2009). “ Extended spectrum β-lactamases among Gram-
negative bacterial isolates from clinical specimens in three major hospitals in Northern Jordan”. Int.J. of
Microbiol. Res. Article . ID 513874.
Bergogne-Berezin, E. (2001). The increasing role of Acinetobacter species as nosocomial pathogens. Curr Infect
Dis Rep 3, 440–444.
Bradford, P.A. (2001). “Extended-spectrum β-lactamases in t21st
century: characterization, epidemiology, and
detection of this important resistance threat”. Clin .Microbiol. Rev 14, 933-51.
Cantarelli, V.V., Teresa, E.I., Brodt, C.Z., Secchi, C., Cavalcante, B.C. & Pereira, F.S. (2007). “Utility of the
ceftazidime-imipenem antagonism test (CIAT) to detect and confirm the presence of inducible AmpC β-
lactamases among Enterobacteriaceae”.The Brazilian J. of Infect. Dis 11(2),237-239.
Cantón R, Akóva M, Carmeli Y, Giske CG, Glupczynski Y, et al.(2012). “Rapid evolution and spread of
carbapenemases among Enterobacteriaceae in Europe ”. Clin Microbiol Infect 18(5),413-31
Dy, M. E., Nord, J. A., LaBombardi, V. J. & Kislak, J. W. (1999). “The emergence of resistant strains of
Acinetobacter baumannii: clinical and infection control implications”. Infect Control Hosp Epidemiol 20, 565–
567.
Holt, J.G., Krieg, N.R., Sneath, H. A., Stanley, J. T. and Williams, S.T. (1994). “ Bergeys manual of
determinative bacteriology”. 9th ed., Baltimore; Wiliams and Wilkins, USA .
Jacoby, G.A. and Medeiros, A.A. (1991). “More extended spectrum β-lactamases”. J. Antimicrob. Agents and
Chemother 35, 1697–1704.
Joshi, S. G. (1998). “ Assignment of antibiotic resistance to naturally occurring plasmids from clinical isolates of
Acinetobacter species”. PhD Thesis, University of Pune, Pune, India.
Kusradoze I, Diene SM, Goderdzishvili M, Rolai JM . (2011). “Molecular detection of OXA carbapenemase
genes in multidrug-resistant Acinetobacter baumannii isolates from Iraq and Georgia”. Int. J. Antimicrob. Agent
38, 164-168.
Levin, A. S., Levy, C. E., Manrique, A. E. I., Medeiros, E. A. S. & Costa, S. F. (2003). “Severe nosocomial
infections with imipenem resistant Acinetobacter baumannii treated with ampicillin/sulbactam”. Int J Antimicrob
Agents 21, 58–62.
MacFaddin, J.F. (2000). “Biochemical tests for identification of medical bacteria”. 3rd
ed. Lippincott Williams
4. Journal of Biology, Agriculture and Healthcare www.iiste.org
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online)
Vol.3, No.7, 2013
35
and Wilkins, USA.
Magiorakos AP, Srinivasan A, Carey RB, Carmeli Y, Falagas ME, Giske CG, Harbarth S, Hindler JF, Kahlmeter
G, Olsson-Liljequist B, Paterson DL, Rice LB,Stelling J, Struelens MJ, Vatopoulos A, Weber JT, Monnet
DL .(2012). “Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international
expert proposal for interim standard definitions for acquired resistance”. Clin. Microbiol. Infect. 18, 268-281.
National Committee for Clinical Laboratory Standards (NCCLS). (2003). “Performance standards for
antimicrobial disc susceptibility testing”. Disc diffusion. 8th
ed. Informational supplement. M100-S13. NCCLS,
Wayne, Pa.
Papa A, Koulourida V, Souliou E. Molecular . (2009) .“Epidemiology of carbapenem-resistant Acinetobacter
baumannii in a newly established Greek hospital”. Microb Drug Resist. 15,257–60.
Parisa A. , Mahdi A. , Setareh S. , Morovat T., Khairollah A. , Kourosh S. , Abbas M. , Mohammad H., Parviz
K. , Mohammad E.(2012). “Antimicrobial resistance patterns and their encoding genes among Acinetobacter
baumannii strains isolated from burned patients.Burns.pathogens”.Curr. Infect Dis. Rep. 3,440-444.
Perilli, M., Felici, A., Oratore, A., Cornaglia, G.,Bonfiglio, G., Rossolini, G. M. & Amicosante,G. (1996).
“ Characterization of thechromosomal cephalosporinases produced byAcinetobacter lwoffii and Acinetobacter
baumannii clinical isolates”. Antimicrob Agents Chemother .40, 715–719.
Poirel, L., Karim, A., Mercat, A., Le Thomas, I., Vahaboglu, H., Richard, C. & Nordmann, P. (1999).
“Extended-spectrum lactamaseproducing strain of Acinetobacter baumannii isolated from a patient in France”. J .
Antimicrob Chemother 43, 157–158.
Pospiech, T. and Neumann, J. (1995). “In genomic DNA isolation. Kieser eds. John Innes Center”. Norwich
NR4 7UH. U.K.
Shahcheraghi F, Abbasalipour M, Feizabadi MM, Ebrahimipour GH, Akbari N. (2011). “Isolation and genetic
characterization of metallo-β-lactamase and carbapenamase producing strains of Acinetobacter baumannii from
patients at Tehran hospitals”. Iranian J. Microbiol 3(2), 68-74.
Sinha M, Srinivasa H, Macaden R. (20007). “Antibiotic resistance 1. profile and extended spectrum beta-
lactamase (ESBL) production in Acinetobacter species”. Ind J Med Res 126, 63-67.
Soroush S, Haghi-Ashtiani MT, Taheri-Kalani M, Emaneini M, Aligholi M, Sadeghifard N, et al. (2010) .
“Antimicrobial resistance of nosocomial strain of Acinetobacter baumannii in Children’s Medical Center of
Tehran: a 6-year prospective study”. Acta Med Iran 48, 178–84.
Srinivasan VB, Rajamohan G, Pancholi P, Stevenson K, Tadesse D, Patchanee P, et al.(2009). “Genetic
relatedness and molecular characterization of multidrug resistant Acinetobacter baumannii isolated in central
Ohio, USA”. Ann Clin Microbiol Antimicrob 8, 21.
Taherikalani M, Etemadi G, Geliani KN, Fatollahzadeh B, Soroush S, Feizabadi MM. (2008). “Emergence of
multi and pan-drug resistance Acinetobacter baumannii carrying blaOXA-type- carbapenemase genes among
burn patients in Tehran”. Iran Saudi Med J 29,623–4.
Towner KJ. (1997) .Clinical importance and antibiotic resistance of Acinetobacter spp. J . Med Microbio 46,721-
46.
Uma K., Srinivasa R. , Sahoo S, Shashikala P, Kanungo R, Jayachandran S. (2009). “Phenotypic and genotypic
assays for detecting the prevalence of metallo-beta-lactamases in clinical isolates of Acinetobacter baumannii
from a South Indian tertiary care hospital”. J .Med. Microbiol 58,430–5.
Yan ZQ, Shen DX, Cao JR, Chen R, Wei X, Liu LP, et al.(2009). “ Susceptibility patterns and molecular
epidemiology of multidrug-resistant Acinetobacter baumannii strains from three military hospitals in China”.
Int .J .Antimicrob Agents 35, 269–73.
Zeba, B. (2005). “Overview of β-lactamase incidence on bacterial drug resistance”. African J. Bacteriol 4,
1559-1562.
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Figure 1. Ethidium bromide-stained agarose gel of PCR amplified products from extracted DNA of A. baumannii
isolates and amplified with blaSHV gene primers. The electrophoresis was performed at 70 volt for 1.5 hr. Lane
(L), DNA molecular size marker (l0000-bp ladder), Lane (A7, A10, A12) of A. baumannii isolates show
positive results with (753bp), Lanes (A1-A6 and 8, 9 ) show negative results.
Figure 2 . Ethidium bromide-stained agarose gel of PCR amplified products from extracted DNA of A.
baumannii isolates and amplified with blaTEM gene primers. The electrophoresis was performed at 70 volt for 1.5
hr. Lane (L), DNA molecular size marker (l0000-bp ladder), Lane (A3) of A. baumannii isolate show positive
result with (822bp).
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Table 1. Programs of PCR thermocycling conditions
gene
Temperature(o
C ) / Time
Cycle
numberInitial
denaturation
Cycling condition
Final
extension
denaturation annealing extension
blaSHV 94/30 sec 94/30 sec 60/1 min 72/1 min 72/10 min 35
blaTEM 94/30 sec 94/30 sec 45/1 min 72/1 min 72/10 min 35
blaVEB 93/3 min 93/1 min 55/1 min 72/1 min 72/7 40
Table 2 . Number and percentage of β-lactam resistant A. baumannii isolates collected from clinical samples
Table 3 . Phenotypic detection of ESBL production in A. baumannii isolates
Type of
sample
No. of
β-lactam resistant
A. baumannii
isolate
No. (%) of phenotypic confirmed ESBL producer isolates
Disk Combination
Test
Disk
Approximation
Test
ESBL* CHROM
agar
technique
Sputum 4 2 (50%) 0 (0%) 4 (100%)
Urine 6 4 (66.7%) 0 (0%) 4(66.7%)
Burn wound 2 2 (100%) 0 (0%) 2 (100%)
Total 12 8 (66.7%) 0(0%) 10(83.3%)
L.S.D. (0.05) Samples = 9.143, Methods = 8.94, *ESBL: Extended spectrum β-lactamase
Clinical sample No.
No. (%) of β-lactam resistant
A. baumannii isolate
Sputum 450 4 (0.88%)
Urine 210 6 (2.8%)
Burn swab 110 2 (1.8%)
Total 770 12 (1.5%)
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