scholarly journals Antibiotic Resistance and Plasmid Profiling of Escherichia coli Isolated from Human Sewage Samples

2021 ◽  
Vol 14 ◽  
pp. 117863612110168
Author(s):  
Sanjida Akter ◽  
A. M. Masudul Azad Chowdhury ◽  
Sohana Akter Mina

In developing countries, the occurrence of antibiotic resistance is increasing day by day and antibiotic resistant microorganisms are being found in almost every environmental setting. Plasmids are considered as the main vector in the procurement and propagation of antibiotic resistance in many microorganisms such as Escherichia coli ( E. coli). The goal of this study was to examine the antibiotic resistance and screening of plasmid in E. coli strains which were previously identified from human sewage samples. During this study antibiotic susceptibility of E. coli isolates were determined by Kirby-Bauer disk diffusion method against 5 antibiotics (ampicilin, ceftriaxone, amoxicillin, ciprofloxacin, azithromycin). Furthermore, plasmid extraction of each isolate was done according to the protocol of FavorPrepTMPlasmid Mini Kit and plasmid profiling was done by agarose gel electrophoresis. In antibiotic sensitivity test, all E. coli strains showed resistance to ampicilin, amoxicillin, and ceftriaxone. In the plasmid profiling, it was revealed that all the isolates of E. coli harbored plasmids. The plasmid sizes ranged from approximately 1.5 to 15 kb. The findings of this study prove the consequences of antibiotic resistance as well as relationship of plasmid with antibiotic resistance which necessitates proper surveillance on antibiotic usage in the developing countries.


2021 ◽  
Vol 8 (1) ◽  
pp. 24-27
Author(s):  
Wanikda Eh Chuan ◽  
Akrimah Yusof ◽  
Aweng Eh Rak ◽  
Shareef Mohd Hafiz Mani ◽  
Seong Wei Lee

In the present study, antibiogram of Escherichia coli isolated from farmed Asian clam, Corbiculafluminea was characterised. Asian clam or locally known as ‘etak’ is processed to became smokedclam and consumed as snack by Kelantanese. However, there are many diarrhoea cases afterconsuming smoked clam. Furthermore, there are also insufficient information about the E.coli levelin Asian clam farm and effective antibiotic in controlling the bacteria in the litereature. Hence, thisstudy was carried out to provide information of antibiogram of E.coli to be reference in the future.Eosin Methylene Blue agar (EMB) was used to isolate E. coli. A total of 100 isolated bacteria weresubjected to antibiotic sensitivity test using disk diffusion method. A total of 18 types of antibioticsnamely novobiocin (30µg/disk), fosfomycin (50 µg/disk), tetracycline (30µg/disk), lincomycin(15µg/disk), flumequine (30µg/disk), sulphamethoxazole (25 µg/disk), amoxycillin (25 µg/disk),chloramphenicol (30 µg/disk), oleandomycin (15 µg/disk), spiramycin (100 µg/disk), ampicillin(10 µg/disk), oxytetracycline (30 µg/disk), doxycycline (30 µg/disk), nalidixic acid (30 µg/disk),florfenicol (30 µg/disk), erythromycin (15 µg/disk), kanamycin (30 µg/disk) and oxolinic acid (2µg/disk). The findings of the present study showed total plate count of E. coli was 6.45 x 103 colonyforming unit (CFU/100g) of sampled Asian clam. Hence, the clam is needed to be under cleansingtreatment before can consider safe for human consumption. Antibiotic results showed 51 % wasrecorded as antibiotic resistance case, 44 % antibiotic sensitive case and 5 % as antibioticintermediary sensitive case. None of the tested antibiotics was successfully inhibited the growth ofthe present bacterial isolates indicating more antibiotics are needed to be screen in the future studyto find out the most effective antibiotic in controlling isolated E. coli.



2018 ◽  
Vol 4 (1) ◽  
pp. 36-43 ◽  
Author(s):  
Md Rashed Kamal ◽  
Md Fakhruzzaman ◽  
Mir Rowshan Akter ◽  
Md Atiqul Haque

Enterovirulent Escherichia coli remain as an important etiological agent of goat diarrhoea in Bangladesh. The present study was designed with a view to isolate and identifies E. coli from field cases. For this purpose, a total of 135 faecal samples (85 from diarrhoeic and 50 from apparently healthy goat) were collected during the period from January 2012 to July 2012 from different areas in Rangpur District. It was found that the prevalence of E. coli was higher (18.82 %) in diarrhoeic goats while it was lower (14.00 %) in non diarrhoeic goats. Age wise distribution of E. coli isolates were 26.42% in day old to 1 year, 10.53% in 1-2 years and 11.36% in above 2 years age of goat respectively. All the isolates of E. coli revealed greenish black colony with metallic sheen in Eosine methylene blue agar, bright pink color smooth transparent colony in MacConkey agar and slight pinkish smooth colony in Salmonella-Shigella agar. Gram stain and hanging drop techniques were performed with the cultured bacteria. Biochemical properties of the isolates were studied, and antibiotic sensitivity test was done by agar disk diffusion method. In Gram stain, the organisms revealed Gram negative, small rod shaped, occurs singly or paired. Biochemically, all of the isolates showed fermentation of dextrose, sucrose and maltose with the production of acid and gas, negative result to Voges-Proskauer test, positive result to Methylred test and differential result to Indol test. All the isolates of E. coli were highly sensitive to ciprofloxacin and gentamicin while moderately sensitive to colistin, livofloxacin and azithromycin and less sensitive to ceftraexon and tetracyclin and resistant to amoxycillin, ampicillin, erythromycin, and neomycin. Therefore, ciprofloxacin and gentamicin may be the antibiotics of first choice, and colistin, livofloxacin and azithromycin may be the second choice among the test antibiotics for the treatment of illness caused by these bacteria.Asian J. Med. Biol. Res. March 2018, 4(1): 36-43



Animals ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 396 ◽  
Author(s):  
Michaela Sannettha van den Honert ◽  
Pieter Andries Gouws ◽  
Louwrens Christiaan Hoffman

Studies have shown that antibiotic resistance among wild animals is becoming a public health concern, owing to increased contact and co-habitation with domestic animals that, in turn, results in increased human contact, indirectly and directly. This type of farming practice intensifies the likelihood of antibiotic resistant traits in microorganisms transferring between ecosystems which are linked via various transfer vectors, such as rivers and birds. This study aimed to determine whether the practice of wildlife supplementary feeding could have an influence on the antibiotic resistance of the bacteria harboured by the supplementary fed wildlife, and thus play a potential role in the dissemination of antibiotic resistance throughout nature. Escherichia coli and Enterococcus were isolated from the faeces of various wildlife species from seven different farms across South Africa. The Kirby-Bauer disk diffusion method was used according to the Clinical and Laboratory Standards Institute 2018 guidelines. The E. coli (F: 57%; N = 75% susceptible) and Enterococcus (F: 67%; N = 78% susceptible) isolates from the supplementary fed (F) wildlife were in general, found to be more frequently resistant to the selection of antibiotics than from those which were not supplementary fed (N), particularly towards tetracycline (E. coli F: 56%; N: 71%/Enterococcus F: 53%; N: 89% susceptible), ampicillin (F: 82%; N = 95% susceptible) and sulphafurazole (F: 68%; N = 98% susceptible). Interestingly, high resistance towards streptomycin was observed in the bacteria from both the supplementary fed (7% susceptible) and non-supplementary fed (6% susceptible) wildlife isolates. No resistance was found towards chloramphenicol and ceftazidime.



2015 ◽  
Vol 9 (05) ◽  
pp. 496-504 ◽  
Author(s):  
Divya Sukumaran ◽  
Abdulla A Mohamed Hatha

Introduction: Escherichia coli strains can cause a variety of intestinal and extraintestinal diseases. Extraintestinal pathogenic E. coli (ExPEC) strains have the ability to cause severe extraintestinal infections. Multidrug resistance among ExPEC could complicate human infections. Methodology: Escherichia coli strains were isolated during the period of January 2010 to December 2012 from five different stations set at Cochin estuary. Susceptibility testing was determined by the disk-diffusion method using nine different antimicrobial agents. A total of 155 strains of Escherichia coli were screened for the presence of virulence factor genes including papAH, papC, sfa/focDE, iutA,and kpsMT II associated with ExPEC. Results: Among the 155 E. coli isolates, 26 (16.77%), carried two or more virulence genes typical of ExPEC. Furthermore, 19.23% of the ExPEC isolates with multidrug resistance were identified to belong to phylogenetic groups B2 and D. Statistically significant association of iutA gene in ExPEC was found with papC (p < 0.001) and kpsMT II (p < 0.001) genes. ExPEC isolates were mainly resistant to ampicillin (23.07%), tetracycline (19.23%), co-trimoxazole (15.38%), and cefotaxime (15.38%). The adhesion genes papAH and sfa/focDE were positively associated with resistance to gentamicin, chloramphenicol, and cefotaxime (p < 0.05). Conclusions: Co-occurrence of virulence factor genes with antibiotic resistance among ExPEC poses considerable threat to those who use this aquatic system for a living and for recreation.



Author(s):  
Nasrin Bahmani ◽  
Noshin Abdolmaleki ◽  
Afshin Bahmani

Background and Objectives: Urinary tract infection (UTI) is one of the most frequent infectious diseases which is caused by Gram-negative bacteria especially Escherichia coli. Multiple resistance to antimicrobial agents are increasing quickly in E. coli isolates and may complicate therapeutic strategies for UTI. The propose of this study was to determine the antibiotic resistance patterns and the multidrug-resistance (MDR) phenotypes in uropathogenic E. coli (UPEC). Materials and Methods: A total of 153 UPEC isolates were collected from both hospitalized patients (95 isolates) and outpatients (58 isolates) from March to October 2018. In order to determine the MDR among UPEC isolates, we have tested 15 antimicrobial agents on Muller Hinton agar by the disk diffusion method. Results: The percentage of MDR isolates (resistant to at least three drug classes such as fluoroquinolones, penicillins and cephalosporins) was 55.5% in the hospitalized patients and the outpatients. Antibiotic resistance to ampicillin, ceftazidime, nalidixic acid and trimethoprim/ sulfamethoxazole was higher than 60%. Meropenem, Imipenem and norfloxacin indicated markedly greater activity (93.3%, 80% and 85.6%, respectively) than other antimicrobial agents. Conclusions: Urinary tract infection due to MDR E. coli may be difficult to treat empirically due to high resistance to commonly used antibiotics, so, empirical antibiotic treatment should be reviewed periodically at local studies.



Author(s):  
Farzad Esavand Heydari ◽  
Mojtaba Bonyadian ◽  
Hamdallah Moshtaghi ◽  
Masoud Sami

Background and Objectives: Enterohemorrhagic Escherichia coli (EHEC) causes bloody and non-bloody diarrhea, intestinal infection and extraintestinal complications in humans. This study aimed to isolate and evaluate the prevalence of E. coli O157: H7 and other Shiga toxin-producing E. coli (STEC) and identify the virulence genes (stx1, stx2, hly and eaeA) from patients with diarrhea. Also, the antibiotic resistance profile of the isolated strains was evaluated. Materials and Methods: A total of 100 stool samples were collected from patients with acute diarrhea referring to the hospital and clinics in Isfahan County, Iran. Phenotypic tests and PCR assay were used for detection of E. coli O157: H7 and other Shiga toxin-producing E. coli. The presence of virulence genes (stx1, stx2, hly and eaeA) were identified by PCR. The antibiotic resistance profile of the isolates was determined using the agar disk diffusion method. The results were analyzed descriptively by Sigma stat version 4 software. Results: Seventy - eight out of 100 samples (78%) were contaminated with E. coli. E. coli O157 was isolated from five samples (6.4%), of which only two strains (2.56%) were identified as E. coli O157: H7. According to the results, out of two E. coli O157: H7 isolates, one (50%) isolate contained eaeA and two isolates (100%) contained Stx1, Stx2, hlyA genes. Out of three (3.84%) E. coli O157: HN, one of the isolate (33.3%) contained stx1 and, two isolates (66.7%) were positive for hlyA genes. Also, the results revealed that six strains (7.69%) were non-O157: H7 STEC, of which two isolates (33.3%) contained stx1 and four isolates (66.7%) were positive for stx2 and hlyA genes. The results of antibiogram tests revealed that all of the STEC isolates (100%) were sensitive to imipenem followed by kanamycin, gentamicin and nitrofurantoin (91%). High resistance (54.5%) to ampicillin and ciprofloxacin was observed among the STEC isolates. Conclusion: The results of the current study showed that although the prevalence of E. coli O157: H7 was low among patients with diarrhea, the other STEC strains with relative resistance to antibiotics are more prevalent.



2020 ◽  
Vol 13 (6) ◽  
pp. 1037-1044
Author(s):  
Mona A. A. AbdelRahman ◽  
Heba Roshdy ◽  
Abdelhafez H. Samir ◽  
Engy A. Hamed

Aim: Antimicrobial resistance is a global health threat. This study investigated the prevalence of Escherichia coli in imported 1-day-old chicks, ducklings, and turkey poults. Materials and Methods: The liver, heart, lungs, and yolk sacs of 148 imported batches of 1-day-old flocks (chicks, 45; ducklings, 63; and turkey poults, 40) were bacteriologically examined for the presence of E. coli. Results: We isolated 38 E. coli strains from 13.5%, 6.7%, and 5.4% of imported batches of 1-day-old chicks, ducklings, and turkey poults, respectively. They were serotyped as O91, O125, O145, O78, O44, O36, O169, O124, O15, O26, and untyped in the imported chicks; O91, O119, O145, O15, O169, and untyped in the imported ducklings; and O78, O28, O29, O168, O125, O158, and O115 in the imported turkey poults. The E. coli isolates were investigated for antibiotic resistance against 16 antibiotics using the disk diffusion method and were found resistant to cefotaxime (60.5%), nalidixic acid (44.7%), tetracycline (44.7%), and trimethoprim-sulfamethoxazole (42.1%). The distribution of extended-spectrum β-lactamase (ESBL) and ampC β-lactamase genes was blaTEM (52.6%), blaSHV (28.9%), blaCTX-M (39.5%), blaOXA-1 (13.1%), and ampC (28.9%). Conclusion: Imported 1-day-old poultry flocks may be a potential source for the dissemination of antibiotic-resistant E. coli and the ESBL genes in poultry production.



2020 ◽  
Vol 23 (6) ◽  
pp. 517-524
Author(s):  
Khayam ul Haq ◽  
Shazia Noreen ◽  
Sheikh A. Sehgal ◽  
Rana A. Tahir ◽  
Amjad Essa ◽  
...  

Background: Escherichia coli various strains can cause alarmingly serious infections. Countries like Pakistan harbour the class of bacteria with one of the highest rates of resistance, but very little has been done to explore their genetic pool. Objectives: This study was designed to find out the frequency of virulence genes of Uropathogenic E. coli and their association with antibiotic resistance along with the evolutionary adaptation of the selected gene through the phylogenetic tree. Methods: Isolates from 120 urinary tract infected patients were collected. Antibiotic sensitivity was detected by the disk diffusion method and DNA extraction was done by the boiling lysis method followed by PCR-based detection of virulence genes. The final results were analysed using the chi-square test. Results: The isolates were found to be least susceptible to nalidixic acid, followed by ampicillin, cotrimoxazole, cefotaxime, ciprofloxacin, aztreonam, amoxicillin, gentamycin, nitrofurantoin and imipenem. The iucC was the most common virulence gene among the resistant isolates. About 86% of the collected samples were found to be multi-drug resistant. Statistical analysis revealed a significant association between the iucC gene and resistance to ampicillin (P=0.03) and amoxicillin (P=0.04), and also between fimH and resistance to aztreonam (P=0.03). Conclusion: This study unravels the uncharted virulence genes of UPEC in our community for the very first time. We report a high frequency of the iucC and fimH virulence genes. This, along with their positive association with resistance to beta-lactam antibiotics in the studied community, indicates their important role in the development of complicated UTIs.



2019 ◽  
Vol 63 (3) ◽  
pp. 1-8
Author(s):  
M. Sciberras ◽  
M. Pipová ◽  
I. Regecová ◽  
P. Jevinová ◽  
S. Demjanová

Abstract The purpose of this study was to detect the antibiotic resistance of forty-one Escherichia coli isolates from the intestinal contents of slaughtered broiler chickens using the disk diffusion method according to Kirby-Bauer. Mueller-Hinton agar plates were inoculated with 0.1 ml overnight broth cultures of individual E. coli isolates and the disks with the following concentrations of antibiotics were applied onto them: ampicillin (10 μg), cefotaxime (30 μg), gentamicin (10 μg), streptomycin (10 μg), azithromycin (15 μg), tetracycline (30 μg), ciprofloxacin (30 μg) and levofloxacin (3 μg). After the incubation at 37 °C for 16—18 hours, the inhibition zones were measured and interpreted in accordance with the Clinical and Laboratory Standard Institute (CLSI) zone diameter breakpoints. Almost all E. coli isolates showed resistance to tetracycline (92.68 %), most of them were resistant to gentamicin (75.61 %) and levofloxacine (70.73 %). Phenotypic resistance to tetracycline was further confirmed with the help of the Polymerase Chain Reaction (PCR) procedure focused on the presence of specific tet(A) and tet(B) genes. These genes were detected in all 41 E. coli isolates. On the contrary, E. coli isolates were highly susceptible to both azithromycin and streptomycin. In conclusion, the study highlighted the role of commensal E. coli bacteria isolated from the intestines of broiler chickens as an important reservoir of tetracycline resistance genes.



Author(s):  
D.W. Lukman ◽  
D.Y. Sari ◽  
H. Pisestyani

Background: Kebab is one of the popular ready-to-eat foods in Indonesia and sold as a street food. The purpose of this study was to determine occurrence and antibiotic resistance of Escherichia coli in street-vended kebab in Dramaga, Bogor, Indonesia. Methods: Totally, 43 samples of kebab meat were collected from street food kebab vendors. Examinations on Total Plate Count (TPC), and total E. coli using Most Probable Number (MPN) method were referred to the Indonesia National Standard. The antibiotic resistance test was carried out using the Kirby-Bauer disk diffusion method. Results: The mean of TPC in kebab samples was 5.3 log10 Colony Forming Unit (CFU)/g. Based on TPC, 13 out of 43 (30.2%) of kebab samples did not comply with the Indonesia National Standard with maximum acceptable level of 5 log10 CFU/g. E. coli was identified in 5 out of 43 samples (11.6%) with mean of 39.2 MPN/g ranged from 7.4 to 150 MPN/g which were higher than standard level (0.5 log10 CFU/g). Four out of 5 E. coli isolated from kebab samples were resistant to gentamycin. All E. coli isolates were susceptible to amoxicillin/clavulanate and chloramphenicol. Conclusion: The occurrence of antibiotic resistant E. coli in ready-to-eat kebab in this area of Indonesia could cause the health problem in consumers. The local government should conduct the monitoring and surveillance on the occurrence of pathogens and the antibiotic resistance in food chain. 



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