scholarly journals Resistansi Antibiotik Secara Fenotip Dan Deteksi Gen TetA pada Sampel Hati Ayam di Pasar Dukuh Kupang Surabaya

2022 ◽  
Vol 11 (3) ◽  
pp. 284
Author(s):  
Reina Puspita Rahmaniar ◽  
Dyah Widhowati ◽  
Nurul Hidayah
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Antibiotic Sensitivity ◽  
Chicken Liver ◽  
Methyl Red ◽  
Chain Reaction ◽  
Biochemical Tests ◽  
Tetracycline Antibiotics ◽  
Polymerase Chain

Abstract The purpose of this study was to determine the resistance of several antibiotics phenotypically and genotypically to detect the tetA gene from broiler chicken liver samples at Dukuh Kupang market, Surabaya. A total of 30 samples were taken and then prepared aseptically and sterile. Isolation on Eosin Methylene Blue Agar (EMBA) media, then microscopic examination using gram staining and biochemical tests of Triple Sugar Iron Agar (TSIA), Sulfide Indole Motility (SIM), Methyl Red (MR), Voges Prouskauers (VP) and Simons Citrate Agar (SCA). The identified Escherichia coli colonies were tested for antibiotic sensitivity using the Kirby Bauer method, then isolates that were proven to be resistant to tetracycline antibiotics were followed by genetic testing using the Polymerase Chain Reaction (PCR) method. The results showed that 20 of the 30 samples were positive for Escherichia coli. Escherichia coli isolates from chicken liver samples showed resistance to 30 µg tetracycline antibiotics by 85% (17 of 20 samples) Researchers also compared with other antibiotics, the highest resistance to ampicillin 10 µg was 90% (18 out of 20 samples), gentamicin resistance was 10 µg by 50% (10 of 20 samples) and 30 µg chloramphenicol antibiotic resistance by 30% (6 of 20 samples). The isolates that were resistant to tetracycline were confirmed by Polymerase Chain Reaction to detect the tetA gene with the final product in the form of a band with a length of 210 bp. Bacterial isolates resistant to Tetracycline antibiotics did not always show TetA gene expression in the PCR test. Keywords: Antibiotic Resistance; Escherichia coli; Market; TetA gene   Abstrak Tujuan dari penelitian ini yaitu untuk mengetahui resistansi beberapa antibiotik secara fenotip dan secara genotip mendeteksi gen tetA dari sampel hati ayam broiler di pasar Dukuh Kupang Surabaya. Sebanyak 30 sampel diambil kemudian dipreparasi secara aseptis dan steril. Isolasi pada media Eosin Methilen Blue Agar (EMBA), selanjutnya dilakukan pemeriksaan mikroskopis menggunakan pewarnaan gram dan uji biokimiawi Triple Sugar Iron Agar (TSIA), Sulfide Indol Motility (SIM), Methyl Red (MR), Voges Prouskauers (VP), dan Simons Citrat Agar (SCA). Koloni Escherichia coli yang teridentifikasi dilakukan uji sensitifitas antibiotik dengan metode Kirby bauer, selanjutnya isolat yang terbukti resistan terhadap antibiotik tetrasiklin dilanjutkan pemeriksaan genetik dengan metode Polymerase Chain Reaction (PCR).  Hasil penelitian menunjukkan bahwa 20 dari 30 sampel positif Escherichia coli. Isolat Escherichia coli asal sampel hati ayam menunjukkan resistansi terhadap antibiotik Tetrasiklin 30 µg sebesar 85 % (17 dari 20 sampel) Peneliti juga melakukan perbandingan dengan antibiotik lainnya, resistensi tertinggi pada antibiotik ampisilin 10 µg sebesar 90 % (18 dari 20 sampel), resistensi gentamisin 10 µg sebesar 50 % (10 dari 20 sampel) dan resistensi antibiotik kloramfenikol 30 µg sebesar 30 % (6 dari 20 sampel). Isolat yang resisten terhadap tetrasiklin dikonfirmasi dengan Polymerase Chain Reaction untuk mendeteksi gen tetA dengan produk akhir berupa band dengan panjang 210 bp. Isolat bakteri yang resistan terhadap antibiotik Tetrasiklin tidak selalu menunjukkan ekspresi gen tetA pada uji PCR. Kata kunci: Escherichia coli; Gen TetA; Pasar; Resistansi Antibiotik.

scholarly journals Detection and characterization of verotoxin-producing strains among sorbitol non-fermenting Escherichia coli

2001 ◽  
Vol 7 (4-5) ◽  
pp. 756-762
Author(s):  
A. Jafari ◽  
M. Katouli ◽  
F. Shokouhi ◽  
S. Bouzari
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Multidrug Resistance ◽  
High Rate ◽  
Cell Adherence ◽  
Chain Reaction ◽  
Resistance Transfer ◽  
Polymerase Chain

The presence of genes for verotoxin 1 and 2 [VT1 and 2] among sorbitol non-fermenting Escherichia coli isolates from diarrhoeal cases was assessed using polymerase chain reaction assay. Of 60 [88%] positive isolates, 37 [62%] harboured VT1 and 23 [38%] both VT1 and VT2. In HeLa cell adherence assay, 48 [71%] isolates exhibited mannose-resistant adherence to HeLa cells. Multidrug resistance was observed in 56 [82%] isolates, with ampicillin, chloramphenicol, streptomycin, sulfamethoxazole-trimethoprim and tetracycline pattern being the most common. There were 13 common and 22 single biochemical phenotypes identified. Isolates belonging to common biochemical phenotypes normally had a similar pattern of adherence and VT production, but differed greatly in their pattern of antibiotic resistance, pointing to a high rate of antibiotic-resistance transfer among these isolates.

scholarly journals Diverse Escherichia coli pathovars of phylogroups B2 and D isolated from animals in Tunisia

2017 ◽  
Vol 11 (07) ◽  
pp. 549-556 ◽  
Author(s):  
Hajer Kilani ◽  
Mohamed Salah Abbassi ◽  
Sana Ferjani ◽  
Rakia Ben Salem ◽  
Riadh Mansouri ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Virulence Genes ◽  
Chain Reaction ◽  
E Coli ◽  
Class 1 ◽  
Genes Encoding ◽  
Polymerase Chain ◽  
Relationship Of

Introduction: The virulent Escherichia coli strains responsible for extraintestinal infections were mainly belonged to B2 and D phylogroups. However, no past studies have determinate via the presence of virulence genes the frequency of E. coli pathovars recovered from animals housed in farms in Tunisia. The aims of this study were to investigate 26 E. coli isolated from healthy and diarrheic animals and to determinate via the presence of virulence genes the frequency of pathovars. Methodology: Twenty-six E. coli isolates of phylogroups B2 (n = 14), B22 (n = 9), B23 (n = 5), and D2 (n = 12) were characterized. Genes encoding virulence factors (fimH,eaeA,aggC,papC, papG allele III, hlyA, east1, cnf1, exhA,stx1, stx2, iutA, fyuA, ibeA,and ipaH), and antibiotic resistance as well as class 1 and 2 integrons were searched by polymerase chain reaction (PCR). The genetic relationship of isolates was done by PFGE. Results: According to the occurrence of specific genes the 26 isolates were classified as:9 EAEC, 2 EHEC, 4 UPEC, 3 EPEC/EHEC and 1 NTEC. Therefore, 2 Ex-PEC and 5 APEC were presented amongst our strains. Some isolates (12) were clonal and the remaining was unrelated. Conclusions: Higher diversity of pathovars which carried diverse combinations of virulence genes in healthy isolates. In addition, it seems that the infections were caused by different mechanisms.

scholarly journals Antibiotic Resistance ofEscherichia coliSerotypes from Cochin Estuary

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Divya P. Sukumaran ◽  
Srinivasan Durairaj ◽  
Mohamed Hatha Abdulla
Keyword(s):  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Antimicrobial Agents ◽  
Antibiotic Sensitivity ◽  
Cochin Estuary ◽  
Chain Reaction ◽  
Monthly Basis ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Polymerase Chain

This study aimed at detecting the prevalence of antibiotic-resistant serotypes ofEscherichia coliin Cochin estuary, India.E. colistrains were isolated during the period January 2010–December 2011 from five different stations set at Cochin estuary. Water samples from five different stations in Cochin estuary were collected on a monthly basis for a period of two years. Isolates were serotyped, antibiogram-phenotyped for twelve antimicrobial agents, and genotyped by polymerase chain reaction foruidgene that codes forβ-D-glucuronidase. TheseE. colistrains from Cochin estuary were tested against twelve antibiotics to determine the prevalence of multiple antibiotic resistance among them. The results revealed that more than 53.33% of the isolates were multiple antibiotic resistant. Thirteen isolates showed resistance to sulphonamides and two of them contained thesul1 gene. Class 1 integrons were detected in twoE. colistrains which were resistant to more than seven antibiotics. In the present study, O serotyping, antibiotic sensitivity, and polymerase chain reaction were employed with the purpose of establishing the present distribution of multiple antibiotic-resistant serotypes, associated withE. coliisolated from different parts of Cochin estuary.

scholarly journals Identification, characterization and molecular epidemiology of Escherichia coli isolated from lamb and goat kids with diarrhoea

2013 ◽  
Vol 82 (4) ◽  
pp. 357-362 ◽  
Author(s):  
Süheyla Türkyılmaz ◽  
Seza Eskiizmirliler ◽  
Serra Tunaligil ◽  
Bulent Bozdogan
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Small Ruminants ◽  
Multi Drug Resistance ◽  
Enterobacterial Repetitive Intergenic Consensus ◽  
Chain Reaction ◽  
E Coli ◽  
Neonatal Diarrhoea ◽  
Polymerase Chain

Neonatal diarrhoea is a serious health problem on commercial farms. EnterovirulentEscherichia coliis a significant aetiological agent of neonatal diarrhoea. In this work, identification and classification ofE. coliisolates obtained from lambs and goat kids with diarrhoea were studied along with antibiotic resistance and clonal relationships of enterovirulent strains. A total of 107E. colistrains isolated from animals on 43 farms were investigated. Specific virulence genes were determined by multiplex and uniplex polymerase chain reaction. Testing of antibiotic susceptibility was carried out by the Vitek II compact system. The relationship ofE. coliisolates was determined by enterobacterial repetitive intergenic consensus polymerase chain reaction. A total of 39 (36.4%) enterovirulentE. colistrains were identified and of this 19 (48.7%) were shiga toxigenic, 12 (30.8%) enterotoxigenic and 8 (20.5%) enteropathogenic. Three isolates (7.7%) were found to be positive for extended spectrum beta lactamase; 10 (25.6%) isolates showed multi-drug resistance to antimicrobials. A total of 28 types were detected by enterobacterial repetitive intergenic consensus polymerase chain reaction. Twenty strains had distinct types while 5 types were common for 2 strains and 3 types were common for 3 strains. This is the first current determination of types, clonality and antibiotic resistance of enterovirulentE. coliisolated from small ruminants with diarrhoea. The results of this study showed that the rates of shiga toxigenic, enterotoxigenic and enteropathogenic isolates ofE. coliare high in the western part of Turkey. Although these isolates were not clonal, presence of multidrug resistant isolates may cause public health problems.

scholarly journals Bacteriological and molecular study of Pseudomonas aeruginosa strains isolated from different clinical cases in Erbil and Kurkuk

2018 ◽  
Vol 41 (2) ◽  
pp. 124-130
Author(s):  
Asif Hasan Abdul Razzaq
Keyword(s):  
Molecular Weight ◽  
Polymerase Chain Reaction ◽  
Virulence Genes ◽  
Antibiotic Sensitivity ◽  
Molecular Study ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Biochemical Tests ◽  
Antibiotic Sensitivity Test ◽  
Polymerase Chain

     This study included isolates of bacteria from 125 clinical samples in Erbil and Kirkuk Hospital including (burns, wounds, urine and sputum); 38 isolates were identified as P. aeruginosa after conducting microscopic and biochemical tests. The results of antibiotic sensitivity test showed that all isolates of P. aeruginosa  were different in resistance to Pipracillin, Erythromycin with rate of (100%) and to the Nalidixic acid (94.73%) while the lowest resistant antibiotics were to Co-trimoxazole, Ceftazidime and Ciprofloxacin, which amounted to (26.31%, 23.68 and 21.05%) respectively. For molecular diagnosis of P. aeruginosa some virulence genes the alg D and exo A were amplified through Polymerase Chain Reaction technique. The results showed that in 38 isolates cases only 22 (57.9%) were positive for algD gene by amplification of 520 bp band. While in urinary tract infection; 6 samples (60%) had alg D gene, and 8 (57.14%) isolates had alg D gene in wounds samples; also 7(70%) isolates from burns had that gene, while the sputum samples showed only one with alg D gene which was the lowest ratio; but in amplification of exo A, the results showed the presence of only one isolate from burns with molecular weight 396 bp with no appearance in others. 

scholarly journals Polymerase Chain Reaction: An Innovative Tool in Periodontal Diagnosis

2013 ◽  
Vol 4 (1) ◽  
pp. 60-66
Author(s):  
Suvarna H Patil ◽  
Kishore G Bhat ◽  
Paresh S Lotlekar ◽  
Laxmi V Hombal
Keyword(s):  
Polymerase Chain Reaction ◽  
Bacterial Species ◽  
Antibiotic Sensitivity ◽  
Periodontal Pathogen ◽  
Current Status ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Biochemical Tests ◽  
Innovative Tool ◽  
Polymerase Chain

ABSTRACT Bacterial species colonizing the surfaces of the human oral cavity play an important role in oral health and disease and thus an accurate means of identification is crucial. Traditionally, identification has been based on microscopy, biochemical tests, immunofluorescence staining and antibiotic sensitivity. However, these tests are labor-intensive and costly, providing sometimes inconsistent results that make identification rather tentative. Recently, molecular DNA-based techniques have been used to identify bacteria directly from clinical samples. Development of a microbiological diagnostic kit using this technology therefore requires the ability to extract the bacterial DNA from the plaque sample and amplify the specific DNA sequence of the target periodontal pathogen. Polymerase chain reaction has emerged as the most powerful tool for the amplification of the genes and their RNA transcripts. The focus of this review is to describe the current status of the DNA-based method PCR which has become a standard diagnostic and research tool in dentistry. How to cite this article Patil SH, Bhat KG, Lotlekar PS, Hombal LV. Polymerase Chain Reaction: An Innovative Tool in Periodontal Diagnosis. World J Dent 2013;4(1):60-66.

Molecular Analysis of Uropathogenic E.coli Isolates from Urinary Tract Infections

2019 ◽  
Vol 19 (3) ◽  
pp. 322-326 ◽  
Author(s):  
Hassan Valadbeigi ◽  
Elham Esmaeeli ◽  
Sobhan Ghafourian ◽  
Abbas Maleki ◽  
Nourkhoda Sadeghifard
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Urinary Tract ◽  
Urinary Tract Infections ◽  
Multiplex Polymerase Chain Reaction ◽  
Virulence Genes ◽  
Uropathogenic Escherichia Coli ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Tract Infections

Introduction: The aim of the current study was to investigate the prevalence of virulence genes in uropathogenic Escherichia coli (UPEC) isolates in Ilam. Materials and Methods: For this purpose, a total of 80 UPEC isolates were collected for patients with UTIs during a 6 months period. The multiplex polymerase chain reaction (multiplex PCR) was used to detect the papEF, fimH, iucD, hlyA, fyuA, and ompT genes. Results: The prevalence of fimH, papEF, iucD, fyuA, hlyA, hlyA, and ompT genes were 87.5%, 47.5%, 60%, 67.5%, 27.5%, 47.5% and 71.2%, respectively. Among all of the isolates, 27 profiles were obtained. Conclusion: Our findings demonstrated that the most prevalence was found for fimH, and different distribution of virulence genes suggested different ability of pathogenicity.

scholarly journals Occurrence of toxigenic Escherichia coli in raw milk cheese in Brazil

2007 ◽  
Vol 59 (2) ◽  
pp. 508-512 ◽  
Author(s):  
B.R. Paneto ◽  
R.P. Schocken-Iturrino ◽  
C. Macedo ◽  
E. Santo ◽  
J.M. Marin
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Antimicrobial Agents ◽  
Raw Milk ◽  
Diffusion Method ◽  
Chain Reaction ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Polymerase Chain ◽  
Raw Milk Cheese

The occurrence of toxigenic Escherichia coli in raw milk cheese was surveyed in Middle Western Brazil. Fifty samples of cheese from different supermarkets were analyzed for E.coli. The isolates were serotyped and screened for the presence of verotoxigenic E. coli (VTEC) and enterotoxigenic E. coli (ETEC) by Polymerase Chain Reaction (PCR). The susceptibility to thirteen antimicrobial agents was evaluated by the disk diffusion method. E.coli were recovered from 48 (96.0%) of the samples. The serogroups identified were O125 (6.0%), O111 (4.0%), O55 (2.0%) and O119 (2.0%). Three (6.0%) and 1(2.0%) of the E.coli isolates were VTEC and ETEC, respectively. Most frequent resistance was observed to the following antimicrobials: cephalothin (60.0%), nalidixic acid (40.0%), doxycyclin (33.0%), tetracycline (31.0%) and ampicillin (29.0%).

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