scholarly journals Molecular detection of beta-lactamase blaCTX-M group 1 in Escherichia coli isolated from drinking water in Khartoum State

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.

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.


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.


2015 ◽  
Vol 13 (1) ◽  
pp. 45-51 ◽  
Author(s):  
AKMA Islam ◽  
M Rahman ◽  
A Nahar ◽  
A Khair ◽  
MM Alam

Molecular technique was used to investigate the prevalence of virulent diarrheic genes in pathogenic Escherichia coli and their antibiotic sensitivity patterns. A hundred samples from 100 different diarrheic calves from mid-north-western part of Bangladesh were screened for the presence of virulence factors associated with diarrhea. Following isolation and identification on the basis of cultural, morphological and biochemical properties, the presence of the virulence genes such as eaeA, bfpA, elt, est, stx1 and stx2 were examined using PCR. Antimicrobial susceptibility of 57 E. coli was determined by agar disk diffusion method for 8 antimicrobial agents. Out of 100 samples 57 (57%) were found to be positive for E. coli and their distribution rates according to their age, breed and sex were  66.7% ( 6 days old ), 85.7% (Sahiwal breed) and in  64.2 % (female calves) respectively. Among 57 E. coli isolates, only 16 isolates were analyzed for the detection of the said genes. Among them, only eaeA gene was detected in 2 E. coli isolates (12.5 %). Antibiotic resistance patterns revealed that Oxacillin, Rifampicin and Penicillin were  100% resistant followed by Erythromycin which was more than 80% resistant. In case of Amoxicillin and Tetracycline, about 59.65% and 61.40% were found to be resistant respectively whereas all 57 E. coli isolates showed moderately susceptible (30%) to Cefuroxime, a second generation Cephalosporin. Therefore, none of the eight antimicrobials studied can not be recommended as single best therapeutic agent for the treatment of neonatal calf diarrhea. In addition, this study indicated that diarrhea in calves in these locations can be ascribed to mainly Enteropathogenic E. coli (EPEC) which was atypical (only contained the eaeA genes but not bfpA). However, further studies are necessary to characterize the isolated eaeA gene positive E. coli by serotyping, tissue culture assay and other molecular techniques to find out the potentiality of those virulent genes contributing pathogenicity of E. coli causing diarrhea in calves.DOI: http://dx.doi.org/10.3329/bjvm.v13i1.23716Bangl. J. Vet. Med. (2015). 13 (1): 45-51


2006 ◽  
Vol 4 (3) ◽  
pp. 289-296 ◽  
Author(s):  
Maggy N. B. Momba ◽  
Veronica K. Malakate ◽  
Jacques Theron

In order to study the prevalence of enteric pathogens capable of causing infection and disease in the rural communities of Nkonkobe, bacterial isolates were collected from several surface water and groundwater sources used by the community for their daily water needs. By making use of selective culture media and the 20E API kit, presumptive Escherichia coli, Salmonella spp. and Vibrio cholerae isolates were obtained and then analysed by polymerase chain reaction assays (PCR). The PCR successfully amplified from water samples a fragment of E. coli uidA gene that codes for β-D-glucuronidase which is a highly specific characteristic of enteropathogenic E. coli, enterotoxigenic E. coli and entero-invasive E. coli. The PCR also amplified the epsM gene from water samples containing toxigenic V. cholerae. Although E. coli was mostly detected in groundwater sources, toxigenic V. cholerae was detected in both surface and groundwater sources. There was a possibility of Salmonella typhimurium in Ngqele and Dyamala borehole water samples. The presence of these pathogenic bacteria in the above drinking water sources may pose a serious health risk to consumers.


2019 ◽  
Author(s):  
omid zarei ◽  
Leili Shokoohizadeh ◽  
Hadi Hossainpour ◽  
Mohammad Yousef Alikhani

Abstract Objective: Shiga toxin producing Escherichia coli (STEC) has known as a crucial zoonotic food borne pathogen. A total of 257 row chicken meat samples were collected from different markets in Hamadan city from January 2016 to May 2017. Samples were cultured on selective and differential culture media, and the virulence genes of E. coli isolates were analyzed by PCR assay. The antibiotic resistance patterns of E. coli isolates were determined by disk diffusion method. The genetic relatedness of STEC isolates were analyzed by ERIC-PCR. Results: Totally, 93(36%) of isolates were identified as E. coli in this current study. According serological and microbiological tests, 5(5.3%), 31(33.3%) and 7(7.5%) of E. coli isolates, characterized as Enterohemorrhagic E. coli (EHEC), STEC and attaching and effacing E. coli (AEEC) strains, respectively. High level resistance to tetracycline (89.8), ampicillin (82.8%) and sulfametoxazole-trimotoprime (71%) were detected among E. coli isolates. Analysis of ERIC-PCR results showed five different ERIC types among EHEC isolates. Based on our findings, chicken meat identified as a sources of STEC strains, therefore, the controlling and checkup the chicken meats for the resistance and virulent strains of E. coli should be consider as a crucial issues in public health.


Author(s):  
T. S. P. J. Jayaweera ◽  
J. L. P. C. Randika ◽  
H. G. C. L. Gamage ◽  
N. N. Udawatta ◽  
W. U. N. T. S. Ellepola ◽  
...  

Aims: Mastitis is one of the very important and most common diseases among dairy cattle globally which leads to severe economical losses in the dairy industry. For the sustainability of the dairy sector it is critical that efficient, economically feasible treatment regime is available for clinical cases of mastitis as a part of the control program with minimum risk for residues in milk. Antimicrobials are the most common drugs of choice for controlling and preventing this devastating condition. But the frequent use of antibiotics leads to the development of resistant bacteria which could have an adverse effect on human health as well. To mitigate this destructive constraint in the industry, identifying the etiology and their susceptibilities to remedial measures are of paramount importance. Hence this study was aimed at isolating and identifying the common bacterial etiology Escherichia coli, Klebsiella spp. and Staphylococcus spp. of mastitis and evaluating the antimicrobial susceptibility of the isolates in order to develop mastitis control strategies in the area. Study Design: Milk samples were collected from mastitic cows in different stages including subclinical and clinical cases based on the results of California Mastitis Test Place and Duration of Study: Samples were collected from dairy farms in Nuwera Eliya District, Sri Lanka and Laboratory investigations were carried out in the Laboratory of Livestock Production, Faculty of Agricultural Sciences, Sabaragamuwa University of Sri Lanka, Between Aug. 2017 and Nov. 2017. Methodology: E.coli, Klebsiella spp. and Staphylococcus spp. were isolated from 31 milk samples and susceptibility to commonly used antibiotics (Trimethoprim, Oxytetracycline, Chloramphenicol, Cephalexin, Enrofloxacin and Ciprofloxacin) was determined by Kirby Bauer disk diffusion method. Results: The study revealed that the most common isolate was the Klebsiella spp. and it is 54.8% and other two organisms Staphylococcus spp. had 51.6% and Escherichia coli 41.9%. Of all isolated pathogen, 97.1% exhibited resistant to Cephalexin and it was the highest while lowest resistance was to Chloramphenicol (31.4%). Among the other antibiotics, 54.3% of total isolates showed resistance to Trimethoprim followed by 42.9% to Oxytetracycline and Enrofloxacin, 34.3% was resistant to Ciprofloxacin. Resistance to at least one antibiotic was observed for the isolated microorganisms. All the three isolated pathogens are more resistant to Cephalexin. Both E. coli and Klebsiella spp. show 100% resistance to Cephalexin while Staphylococci had 92.9% resistance. This further revealed that E. coli (10%) and Klebsiella spp. (27.3%) are showing the least resistance to Chloramphenicol, None of Staphylococcus spp. (0%) isolated show resistance to Enrofloxacin. Conclusion: Most common organisms isolated were Klebsiella spp. followed by Staphylococcus spp., E. coli and there is a resistance of isolated organisms to some commonly used antibiotics.


2020 ◽  
Vol 8 (1) ◽  
pp. 52-60 ◽  
Author(s):  
D.B. Thapa ◽  
A. Chapagain

A cross-sectional study was conducted at National Avian Disease Investigation Laboratory, Chitwan to determine antibiogram of Escherichia coli isolated from avian colibacillosis cases of broilers and layers in Chitwan. One hundred and sixty (95 from broilers and 65 from layers) liver samples were collected aseptically during postmortem. Samples were taken purposively from dead birds showing lesions perihepatitis, pericarditis, air-saculitis, omphalitis and egg peritonitis. Isolation and identification were made by examination of cultural characteristics of E. coli in MacConkey’s agar, Eosin methylene blue (EMB) agar, Gram’s staining and biochemical tests. Antibiogram of identified E. coli isolate was evaluated against six antibiotics of six different groups by disk diffusion method following CLSI guidelines. One hundred and three E. coliisolates (73 from broilers and 30 from layers) were isolated from one hundred and sixty samples. Highest resistance was observed against Ampicillin (100%) followed by Co-trimoxazole (86.40%), Doxycycline (46.60%), Levofloxacin (45.63%), Nitrofurantoin (26.21%) and Amikacin (10.68%). Nearly about all (96.12%) isolates from 103 isolated E. coli isolates showed multidrugs resistance to two or more than two antimicrobials. All multidrug resistance isolates showed 16 different patterns with each isolate being resistance to at least two drugs. The multiple antibiotic resistance indexing ranged from 0.2 to 0.8 and proportion of isolates with MAR index greater than 0.2 was 96.12%. Int. J. Appl. Sci. Biotechnol. Vol 8(1): 52-60    


Pathogens ◽  
2019 ◽  
Vol 8 (2) ◽  
pp. 86 ◽  
Author(s):  
Samer Swedan ◽  
Heba Abu Alrub

The study investigated the prevalence of potentially pathogenic and drug resistant Escherichia coli among drinking water sources in Jordan. A total of 109 confirmed E. coli isolates were analyzed. Antimicrobial susceptibility testing was done using the Kirby Bauer disk diffusion method. Phenotypic identification of extended spectrum beta-lactamase (ESBL) and carbapenemase production was done using the double disk synergy test and the modified Hodge test, respectively. Isolates’ plasmid profiles were determined by gel electrophoresis. PCR was used for detection of virulence and resistance genes. Overall, 22.0% of the isolates were potentially intestinal pathogenic E. coli (IPEC); namely enteroaggregative E. coli (16.5%), enteropathogenic E. coli (2.8%), enteroinvasive E. coli (1.8%), and enterohemorrhagic E. coli (0.9%). A third of the isolates were multi-drug resistant. The highest rates of antimicrobials resistance were observed against ampicillin (93.6%) and sulfamethoxazole/trimethoprim (41.3%). All isolates were susceptible to imipenem, meropenem, doripenem and tigecycline. The prevalence of ESBL and carbapenemase producers was 54.1% and 2.8%, respectively. BlaVIM was the most prevalent resistance gene (68.8%), followed by blaCTX (50.5%), blaTEM (45.9%), blaNDM (11%), blaKPC (4.6%), and blaSHV (0.9%). Fifty-eight (53.2%) isolates contained one or more plasmid ranging from 1.0 to 8.0 kbp. Overall, high prevalence of potentially pathogenic and resistant isolates was observed.


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.


2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Laura Marchetti ◽  
Daniel Buldain ◽  
Lihuel Gortari Castillo ◽  
Andrea Buchamer ◽  
Manuel Chirino‐Trejo ◽  
...  

The close contact between dogs and humans creates the best bridge for interspecies transmission of antimicrobial-resistant bacteria. The surveillance of its resistance including the detection of extended-spectrum beta-lactamases (ESBLs) in Escherichia coli as indicator bacteria is an important tool to control the use of antimicrobials. The aim of this research was to evaluate the E. coli resistance in strains by phenotypic methods, isolated from pet and stray dogs of La Plata city, Argentina. Faecal samples were collected using rectal swabs from 50 dogs with owners (home dogs = HD) and 50 homeless dogs (stray dogs = SD). They were cultured in 3 MacConkey agar plates, with and without antibiotics (ciprofloxacin and cefotaxime). 197 strains were isolated, of which only 95 strains were biochemically identified as E. coli, 46 strains were from HD, and 49 were from SD. Antimicrobial susceptibility was evaluated by the Kirby–Bauer disk diffusion method. The most prevalent resistance was for tetracycline, streptomycin, and ampicillin. In both groups, the level of resistance to 3rd generation cephalosporins was high, and there were multiresistant strains. There was a higher level of antimicrobial resistance in strains from SD compared to HD. There were 8% of strains suspected of being ESBLs among samples of HD and 36% of SD. One (2%) of the strains isolated from HD and 11 (22%) from SD were phenotypically confirmed as ESBL. Pets and stray dogs are a potential source of E. coli antibiotic resistance in Argentina; therefore, its surveillance must be guaranteed.


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