scholarly journals Molecular detection of Beta-lactamase blaCTX-M group 1 gene in Escherichia coli Isolated from Drinking Water in Khartoum State

2020 ◽  
Author(s):  
Hisham N Altayeb ◽  
Eman Khalid ◽  
Ehssan h Moglad

Abstract BackgroundOne of the most common challenges facing the world today is the rapid spread of antibiotic-resistant bacteria. According to current evidence, this is mainly as a result of presumed use of antibiotics, the interaction between humans or from animal sources. Still, there is limited focus on the role of environmental factors, especially water. This study aimed to detect the blaCTX-M group 1 gene in Escherichia coli (E. coli) isolated from drinking water in Khartoum State. Methods Two hundred and eighty water samples were collected from different localities in Khartoum State and resources. Isolation and identification of E. coli performed by using lauryl tryptose broth, Brilliant Green Bile Broth, biochemical reaction, and culture characteristics on EMB media. Antimicrobial sensitivity tests were performed for all isolated bacteria using cefotaxime (30μg), gentamycin (10μg), tetracycline (25μg) and chloramphenicol (30μg) by agar diffusion method. DNA extracted by boiling method, bacterial genomic DNA used as a template to detect the blaCTX-M group 1 gene by PCR. Results The result displayed that eighty-six (30.7%) E. coli isolated out of two hundred and eighty water samples. Out of eighty-sixing isolated bacteria, 49% (42/86) were from Khartoum locality, 28% (24/86) were from Omdurman locality, and 23% (20/86) were from Khartoum North locality. Antimicrobial susceptibility testing revealed that the highest resistant percentage was 59% for tetracycline, followed by 35% for gentamycin. While for chloramphenicol and cefotaxime were 22% and 20%, respectively. CTX-M group 1 gene was detected in 40% of all isolates. Conclusion This study concludes that drinking water in Khartoum State might be contaminated with faces and be a possible source for the transmission of resistant bacteria to humans.

2020 ◽  
Vol 18 (6) ◽  
pp. 1091-1097
Author(s):  
Hisham N. Altayb ◽  
Eman Khalid Salih ◽  
Ehssan H. Moglad

Abstract This study aimed to detect the blaCTX-M group 1 in Escherichia coli (E. coli) isolated from drinking water in Khartoum State. Two hundred and eighty water samples were collected randomly from different areas, places, and sources from the state and examined for the presence of E. coli as a fecal contamination indicator. Isolation and identification of E. coli were performed using culture characteristics on different culture media and biochemical reactions. An antimicrobial sensitivity test was performed for all isolated E. coli using agar disk diffusion method. DNA was extracted by boiling method, and bacterial genomic DNA used as a template to detect blaCTX-M group 1 by PCR. Results showed 86 (30.7%) E. coli were isolated out of 280 water samples. Antimicrobial susceptibility testing revealed the highest resistant percentage was 59% for tetracycline, followed by 35% for gentamycin, while for chloramphenicol and cefotaxime was 22 and 20%, respectively. blaCTX-M group 1 was detected in about 40% of all isolates. This study concludes that drinking water in Khartoum State may be contaminated with feces and might be a possible source for transferring resistant bacteria. Thus, it may be one of the critical causes of increasing reports of antimicrobial resistance in Khartoum State.


2005 ◽  
Vol 71 (3) ◽  
pp. 1394-1404 ◽  
Author(s):  
Raida S. Sayah ◽  
John B. Kaneene ◽  
Yvette Johnson ◽  
RoseAnn Miller

ABSTRACT A repeated cross-sectional study was conducted to determine the patterns of antimicrobial resistance in 1,286 Escherichia coli strains isolated from human septage, wildlife, domestic animals, farm environments, and surface water in the Red Cedar watershed in Michigan. Isolation and identification of E. coli were done by using enrichment media, selective media, and biochemical tests. Antimicrobial susceptibility testing by the disk diffusion method was conducted for neomycin, gentamicin, streptomycin, chloramphenicol, ofloxacin, trimethoprim-sulfamethoxazole, tetracycline, ampicillin, nalidixic acid, nitrofurantoin, cephalothin, and sulfisoxazole. Resistance to at least one antimicrobial agent was demonstrated in isolates from livestock, companion animals, human septage, wildlife, and surface water. In general, E. coli isolates from domestic species showed resistance to the largest number of antimicrobial agents compared to isolates from human septage, wildlife, and surface water. The agents to which resistance was demonstrated most frequently were tetracycline, cephalothin, sulfisoxazole, and streptomycin. There were similarities in the patterns of resistance in fecal samples and farm environment samples by animal, and the levels of cephalothin-resistant isolates were higher in farm environment samples than in fecal samples. Multidrug resistance was seen in a variety of sources, and the highest levels of multidrug-resistant E. coli were observed for swine fecal samples. The fact that water sample isolates were resistant only to cephalothin may suggest that the resistance patterns for farm environment samples may be more representative of the risk of contamination of surface waters with antimicrobial agent-resistant bacteria.


Author(s):  
Reza Ranjbar ◽  
Maryam Zeynali ◽  
Nooshin Sohrabi ◽  
Asghar Ali Kamboh

BACKGROUND <br />Hospital wastewaters may contain antibiotic resistant bacteria such as Escherichia coli. These E. coli harbor integron genes that are responsible for antibiotic resistance. The purpose of the current study was to evaluate the frequency of class 1 and 2 integrons in environmental antibiotic resistant E. coli strains isolated from the hospital wastewaters in Tehran, Iran.<br /><br />METHODS<br />As a descriptive cross-sectional study, this research was performed from April to September 2015 on hospital wastewaters in Tehran. Bacterial isolation and identification was performed by standard biochemical and bacteriological procedures. Susceptibility testing was done by employing the disk diffusion method using different antibiotics. Total DNAs were extracted to evaluate the presence of class 1 and 2 integrons by using the polymerase chain reaction (PCR) method with specific primers. <br /><br />RESULTS<br />Fifty E. coli strains were isolated and identified from the wastewaters of 25 hospitals in Tehran. The phenotype results showed that 46 isolates (92%) were resistant to at least one antibiotic and 27 isolates (54%) were multidrug resistant. PCR showed that 35 (70%) and 20 (40%) of the isolates had class 1 and 2 integrons respectively and 14 isolates (28%) had both class 1 and class 2 integrons.<br /><br />CONCLUSION<br />This study has shown a considerable presence of class 1 and class 2 integrons in E. coli strains isolated from hospital wastewaters in Tehran. Proper antibiotics prescription and appropriate hospital wastewater treatment can prevent resistance genes in E. coli from circulating in the environment.


2021 ◽  
Vol 15 (5) ◽  
pp. 1080-1083
Author(s):  
Nazia Mir ◽  
Sidrah Saleem ◽  
Muhammad Imran ◽  
Ayesha Ghazal ◽  
Muhammad Usman

Background: The major faecal coliform is Escherichia coli which contaminates the drinking water from human and animal faecal waste. In developing regions drinking water is a vital source of microbiological pathogens. Multi-drug resistant (MDR) is defined as resistance to one or more antibiotic groups. An E. coli count greater than 4/dl in municipal drinking water is indicative of unacceptable fecal contamination. Aim: To find out the frequency of MDR E. coli in water system of Lahore, Pakistan. Methodology: Drinking water was collected from different towns of Lahore. The samples were processed using Membrane Filtration Technique. In the present study, Multidrug Resistant E.coli was isolated and antibiotic resistant pattern was seen against 16 most commonly antimicrobials, using Kirby Bauer disc diffusion method. Results: Total 100 water samples were collected, frequency of Escherichia coliisolated was 27% and frequency of MDR E.coli was 19%. The highest resistance showed by the organism towards Ampicillin (AMP) 21(81.48%), Augmentin (AMC), and Ceftazidime (CAZ) 14(51.85%) respectively. In this study also frequency of Extended Spectrum β-Lactamases were seen. Most common organisms isolated were E. coli 7% and Klebsiella 5%. Frequency of other coliforms isolated from drinking water other than E. coli was Klebsiella species 26%, Pseudomonas species 27%, Enterobacter 7%, Citrobacter species 8% and Acinetobacter species 5%. Conclusion: This study revealed that drinking water of Lahore city is heavilycontaminated with pathogenic microorganisms. A high proportion of antibiotic resistant is due to overuse of antibiotics, in patients with mild infections and secretion of these resistant bacteria from patients to environment. One of the reasons could be the mixing of sewage lines with drinking water supply. So, there is solely requirement for further studies for the identification of the sources for these contaminants. Keywords: Isolates, E. coli, Klebsiella, Multi-drug resistant (MDR)


2021 ◽  
Vol 15 (6) ◽  
pp. 1188-1191
Author(s):  
N. Mir ◽  
S. Saleem ◽  
M. Imran ◽  
A. Ghazal ◽  
M. Usman

Background: The major faecal coliform is Escherichia coli which contaminates the drinking water from human and animal faecal waste. In developing regions drinking water is a vital source of microbiological pathogens. Multi-drug resistant (MDR) is defined as resistance to one or more antibiotic groups. An E. coli count greater than 4/dl in municipal drinking water is indicative of unacceptable fecal contamination. Aim: To find out the frequency of MDR E. coli in water system of Lahore, Pakistan. Methodology: Drinking water was collected from different towns of Lahore. The samples were processed using Membrane Filtration Technique. In the present study, Multidrug Resistant E.coli was isolated and antibiotic resistant pattern was seen against 16 most commonly antimicrobials, using Kirby Bauer disc diffusion method. Results: Total 100 water samples were collected, frequency of Escherichia coliisolated was 27% and frequency of MDR E.coli was 19%. The highest resistance showed by the organism towards Ampicillin (AMP) 21(81.48%), Augmentin (AMC), and Ceftazidime (CAZ) 14(51.85%) respectively. In this study also frequency of Extended Spectrum β-Lactamases were seen. Most common organisms isolated were E. coli 7% and Klebsiella 5%. Frequency of other coliforms isolated from drinking water other than E. coli was Klebsiella species 26%, Pseudomonas species 27%, Enterobacter 7%, Citrobacter species 8% and Acinetobacter species 5%. Conclusion: This study revealed that drinking water of Lahore city is heavilycontaminated with pathogenic microorganisms. A high proportion of antibiotic resistant is due to overuse of antibiotics, in patients with mild infections and secretion of these resistant bacteria from patients to environment. One of the reasons could be the mixing of sewage lines with drinking water supply. So, there is solely requirement for further studies for the identification of the sources for these contaminants. Keywords: Isolates, E. coli, Klebsiella, Multi-drug resistant (MDR)


2021 ◽  
Vol 9 (2) ◽  
pp. 326
Author(s):  
Frederick Adzitey ◽  
Nurul Huda ◽  
Amir Husni Mohd Shariff

Meat is an important food source that can provide a significant amount of protein for human development. The occurrence of bacteria that are resistant to antimicrobials in meat poses a public health risk. This study evaluated the occurrence and antimicrobial resistance of E. coli (Escherichia coli) isolated from raw meats, ready-to-eat (RTE) meats and their related samples in Ghana. E. coli was isolated using the USA-FDA Bacteriological Analytical Manual and phenotypic antimicrobial susceptibility test was performed by the disk diffusion method. Of the 200 examined meats and their related samples, 38% were positive for E. coli. Notably, E. coli was highest in raw beef (80%) and lowest in RTE pork (0%). The 45 E. coli isolates were resistant ≥ 50% to amoxicillin, trimethoprim and tetracycline. They were susceptible to azithromycin (87.1%), chloramphenicol (81.3%), imipenem (74.8%), gentamicin (72.0%) and ciprofloxacin (69.5%). A relatively high intermediate resistance of 33.0% was observed for ceftriaxone. E. coli from raw meats, RTE meats, hands of meat sellers and working tools showed some differences and similarities in their phenotypic antimicrobial resistance patterns. Half (51.1%) of the E. coli isolates exhibited multidrug resistance. The E. coli isolates showed twenty-two different resistant patterns, with a multiple antibiotic resistance index of 0.0 to 0.7. The resistant pattern amoxicillin (A, n = 6 isolates) and amoxicillin-trimethoprim (A-TM, n = 6 isolates) were the most common. This study documents that raw meats, RTE meats and their related samples in Ghana are potential sources of antimicrobial-resistant E. coli and pose a risk for the transfer of resistant bacteria to the food chain, environment and humans.


2018 ◽  
pp. 1720-1724 ◽  
Author(s):  
Shahin Mahmud ◽  
K. H. M. Nazmul Hussain Nazir ◽  
Md. Tanvir Rahman

Aim: The present study was carried out to determine the prevalence and molecular detection of fluoroquinolone-resistant Escherichia coli carrying qnrA and qnrS genes in healthy broiler chickens in Mymensingh, Bangladesh, and also to identify the genes responsible for such resistance. Materials and Methods: A total of 65 cloacal swabs were collected from apparently healthy chickens of 0-14 days (n=23) and 15-35 days (n=42) old. The samples were cultured onto Eosin Methylene Blue Agar, and the isolation and identification of the E. coli were performed based on morphology, cultural, staining, and biochemical properties followed by polymerase chain reaction (PCR) targeting E. coli 16S rRNA genes. The isolates were subjected to antimicrobial susceptibility test against five commonly used antibiotics under fluoroquinolone (quinolone) group, namely gatifloxacin, levofloxacin, moxifloxacin, ofloxacin, and pefloxacin by disk diffusion method. Detection of qnrA and qnrS genes was performed by PCR. Results: Among the 65 cloacal samples, 54 (83.08%) were found to be positive for E. coli. Antibiotic sensitivity test revealed that, of these 54 isolates, 18 (33.33%) were found to be resistant to at least one fluoroquinolone antibiotic. The highest resistance was observed against pefloxacin (61.11%). By PCR, of 18 E. coli resistant to fluoroquinolone, 13 (72.22%) were found to be positive for the presence of qnrS. None of the isolates were found positive for qnrA. Conclusion: Fluoroquinolone-resistant E. coli harboring qnrS genes is highly prevalent in apparently healthy broiler chickens and possesses a potential threat to human health.


Pathogens ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 420 ◽  
Author(s):  
Mst. Sonia Parvin ◽  
Sudipta Talukder ◽  
Md. Yamin Ali ◽  
Emdadul Haque Chowdhury ◽  
Md. Tanvir Rahman ◽  
...  

Escherichia coli is known as one of the most important foodborne pathogens in humans, and contaminated chicken meat is an important source of foodborne infection with this bacterium. The occurrence of extended-spectrum β-lactamase (ESBL)-producing E. coli (ESBL-Ec), in particular, in chicken meat is considered a global health problem. This study aimed to determine the magnitude of E. coli, with special emphasis on ESBL-Ec, along with their phenotypic antimicrobial resistance pattern in frozen chicken meat. The study also focused on the determination of ESBL-encoding genes in E. coli. A total of 113 frozen chicken meat samples were purchased from 40 outlets of nine branded supershops in five megacities in Bangladesh. Isolation and identification of E. coli were done based on cultural and biochemical properties, as well as PCR assay. The resistance pattern was determined by the disc diffusion method. ESBL-encoding genes were determined by multiplex PCR. The results showed that 76.1% of samples were positive for E. coli, of which 86% were ESBL producers. All the isolates were multidrug-resistant (MDR). Resistance to 9–11 and 12–13 antimicrobial classes was observed in 38.4% and 17.4% isolates, respectively, while only 11.6% were resistant to 3–5 classes. Possible extensive drug resistance (pXDR) was found in 2.3% of isolates. High single resistance was observed for oxytetracycline (93%) and amoxicillin (91.9%), followed by ampicillin (89.5%), trimethoprim–sulfamethoxazole, and pefloxacin (88.4%), and tetracycline (84.9%). Most importantly, 89.6% of isolates were resistant to carbapenems. All the isolates were positive for the blaTEM gene. However, the blaSHV and blaCTX-M-2 genes were identified in two ESBL-non producer isolates. None of the isolates carried the blaCTX-M-1 gene. This study provided evidence of the existence of MDR and pXDR ESBL-Ec in frozen chicken meat in Bangladesh, which may pose a risk to human health if the meat is not properly cooked or pickled raw only. This emphasizes the importance of the implementation of good slaughtering and processing practices by the processors.


2015 ◽  
Vol 78 (1) ◽  
pp. 72-77 ◽  
Author(s):  
ALENA SKOČKOVÁ ◽  
KATEŘINA BOGDANOVIČOVÁ ◽  
IVANA KOLÁČKOVÁ ◽  
RENÁTA KARPÍŠKOVÁ

The occurrence of antimicrobial-resistant bacteria is an important public health issue. The aim of this study was the monitoring of resistant Escherichia coli in raw cow's milk with a focus on the detection of extended-spectrum β-lactamase (ESBL)–producing strains. In total, 263 samples of raw milk from 40 farms were collected and investigated in 2010 to 2013 in the Czech Republic. Detection of E. coli was performed and evaluated according to ISO 16649-2, and antibiotic resistance was screened by the disk diffusion method. The presence of E. coli was detected in 243 (92.4%) samples. In total, 270 isolates were obtained. Resistance to β-lactam (31.8%) and tetracycline (13.0%) antibiotics was detected most often and also multiresistant strains (5.5%) were observed. E. coli isolates found to be resistant to β-lactam, tetracycline, and quinolone antibiotics were assayed by PCR to detect selected genes encoding those resistance mechanisms. In isolates in which any bla genes were detected, a double-disk synergy test was performed. ESBL production was confirmed in 2 (0.7%) isolates. The genetic analysis identified the presence of the blaCTX-M gene and other resistance genes (tet(B) and qnrB). Both ESBL-positive isolates originated from the same farm and had an identical pulsed-field gel electrophoresis profile. The findings of our study indicate that milk can be a reservoir of bacteria carrying resistance genes with a potential for spreading through the food chain.


Water ◽  
2021 ◽  
Vol 13 (23) ◽  
pp. 3449
Author(s):  
Cristina-Mirabela Gaşpar ◽  
Ludovic Toma Cziszter ◽  
Cristian Florin Lăzărescu ◽  
Ioan Ţibru ◽  
Marius Pentea ◽  
...  

This study aimed to compare the antibiotic resistance levels of the indicator bacteria Escherichia coli in wastewater samples collected from two hospitals and two urban communities. Antimicrobial susceptibility testing was performed on 81 E. coli isolates (47 from hospitals and 34 from communities) using the disc diffusion method according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) methodology. Ten antibiotics from nine different classes were chosen. The strains isolated from the community wastewater, compared to those from the hospital wastewater, were not resistant to gentamicin (p = 0.03), but they showed a significantly higher susceptibility—increased exposure to ceftazidime (p = 0.001). Multidrug resistance was observed in 85.11% of the hospital wastewater isolates and 73.53% of the community isolates (p > 0.05). The frequency of the presumed carbapenemase-producing E. coli was higher among the community isolates (76.47% compared to 68.09%) (p > 0.05), whereas the frequency of the presumed extended-spectrum beta-lactamase (ESBL)-producing E. coli was higher among the hospital isolates (21.28% compared to 5.88%) (p > 0.05). The antibiotic resistance rates were high in both the hospital and community wastewaters, with very few significant differences between them, so the community outlet might be a source of resistant bacteria that is at least as important as the well-recognised hospitals.


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