scholarly journals Prevalence and molecular detection of fluoroquinolone-resistant genes (qnrA and qnrS) in Escherichia coli isolated from healthy broiler chickens

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.


2006 ◽  
Vol 55 (9) ◽  
pp. 1271-1275 ◽  
Author(s):  
Andrej Trampuz ◽  
Kerryl E. Piper ◽  
James M. Steckelberg ◽  
Robin Patel

Gamma irradiation is widely used for sterilization; however, its effect on elimination of amplifiable DNA, an issue of relevance to molecular diagnostic approaches, has not been well studied. The effect of gamma irradiation on the viability of Staphylococcus epidermidis and Escherichia coli (using quantitative cultures) and on their DNA (using quantitative 16S rRNA gene PCR) was evaluated. Viability was abrogated at 2.8 and 3.6 kGy for S. epidermidis and E. coli, respectively. The radiation dose required to reduce viable bacteria by one log10 (D 10 value) was 0.31 and 0.35 kGy for S. epidermidis and E. coli, respectively. D 10 values for amplifiable DNA extracted from bacteria were 2.58 and 3.09 kGy for S. epidermidis and E. coli, respectively, whereas D 10 values for amplifiable DNA were significantly higher for DNA extracted from irradiated viable bacterial cells (22.9 and 52.6 kGy for S. epidermidis and E. coli, respectively; P<0.001). This study showed that gamma irradiation of DNA in viable bacterial cells has little effect on amplifiable DNA, was not able to eliminate amplifiable 16S rRNA genes at a dose of up to 12 kGy and cannot therefore be used for elimination of DNA contamination of PCR reaction components or laboratory equipment when this DNA is present in microbial cells. This finding has practical implications for those using molecular diagnostic techniques in microbiology.


2017 ◽  
Vol 14 (2) ◽  
pp. 289-295
Author(s):  
M. M. Islam ◽  
S. Ahamed ◽  
M. Y. Arafat ◽  
I. Hasan ◽  
M. Rahman ◽  
...  

This study was designed to determine the shiga toxin producing genes and to investigate antibiotic sensitivity or resistant patterns of the Escherichia coli isolated from diarrheic children at Mymensingh Medical College Hospital, Bangladesh. A total of 83 stool samples were collected and screened for the detection of E. coli on the basis of cultural, staining and biochemical properties followed by molecular detection by Polymerase Chain Reaction (PCR) using genus specific 16SrRNA primers. Antimicrobial susceptibility pattern of E. coli was determined by disc diffusion method against 9 antimicrobial agents. In this study, 27 (32.53%) out of 83 samples, were confirmed as E. coli. Overall prevalence of shiga toxin producing E. coli (STEC) among the examined children was 1.20% (n=1/83).  Further, 27 E. coli isolates were analyzed for the presence of Stx-1 and Stx-2 genes by duplex-PCR.  The STEC isolate was confirmed to be positive for the presence of the Stx-2 gene only. Highest susceptibility of the E. coli isolates was found against Gentamicin (92.59%), followed by Ciprofloxacin (48.14%) and Moxifloxacin (33.33%). More than 77.78% of the isolates were resistant to more than three antibiotics thus defined as multi-drug resistant (MDR). In conclusion, Gentamicin and Ciprofloxacin can be recommended as the effective drugs successful treatment of STEC infections in children.


2020 ◽  
Vol 6 (1) ◽  
pp. 13
Author(s):  
Nelson Phiri ◽  
Geoffrey Mainda ◽  
Mercy Mukuma ◽  
Ntazana N. Sinyangwe ◽  
Luke J. Banda ◽  
...  

Objective: Salmonella species and Escherichia coli are major bacterial enteropathogens of worldwide public health importance that cause devastating foodborne diseases, thereby contributing to increased human morbidity and mortality. Both pathogens have also been found to contribute towards the spread of antimicrobial resistance through the food chain, especially in poultry. This study aimed to determine the occurrence of antibiotic-resistant Salmonella spp. and E. coli in broiler chickens at farm level, abattoirs, and open markets in selected districts of Zambia.Methods: A cross-sectional study was undertaken in seven districts of Zambia to determine the resistance profiles of Salmonella spp. and E. coli obtained from broiler chickens at farms, abattoirs, and open markets. A total of 470 samples were collected which include; litter, cloacal swabs, and carcass swabs. Samples were inoculated into buffered peptone water and incubated for 24 hours then sub-cultured onto MacConkey and Xylose Lysine Deoxycholate agar plates. Identification of Salmonella spp. and E. coli was done using the API-20E kit and confirmation by 16S rDNA sequencing. Confirmed isolates were tested against a panel of 09 antibiotics using the Kirby-Bauer disc diffusion method and interpreted according to the Clinical Laboratory Standards Institute guidelines. Data analysis of the antibiotic sensitivity test results was done using WHONET 2018 software.Results: Overall, 4 Salmonella spp. and 280 E. coli were isolated. One of the Salmonella spp. was resistant to ampicillin (25%), amoxicillin/clavulanic acid (25%), and cefotaxime (25%). E. coli antibiotic resistance was highest to tetracycline (81.4%) and 100% susceptibility to imipenem. The antibiotic susceptibility profile revealed 75.7% (237/280) multidrug-resistant (MDR). The highest MDR profile was observed in 8.2% (23/280) isolates in which 6 out of the 9 classes of antibiotics tested were resistant. Out of the 280 isolates, 11.4% (32/280) exhibited Extensive Drug resistance (XDR).Conclusion: The study found antimicrobial resistance to E. coli and Salmonella spp. in market-ready broiler chickens which were resistant to important antibiotics and is of public health concern.


2000 ◽  
Vol 66 (2) ◽  
pp. 844-849 ◽  
Author(s):  
G. Sabat ◽  
P. Rose ◽  
W. J. Hickey ◽  
J. M. Harkin

ABSTRACT A set of PCR primers targeting 16S rRNA gene sequences was designed, and PCR parameters were optimized to develop a robust and reliable protocol for selective amplification of Escherichia coli 16S rRNA genes. The method was capable of discriminatingE. coli from other enteric bacteria, including its closest relative, Shigella. Selective amplification of E. coli occurred only when the annealing temperature in the PCR was elevated to 72°C, which is 10°C higher than the optimum for the primers. Sensitivity was retained by modifying the length of steps in the PCR, by increasing the number of cycles, and most importantly by optimizing the MgCl2 concentration. The PCR protocol developed can be completed in less then 2 h and, by using Southern hybridization, has a detection limit of ca. 10 genomic equivalents per reaction. The method was demonstrated to be effective for detectingE. coli DNA in heterogeneous DNA samples, such as those extracted from soil.


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


2010 ◽  
Vol 76 (24) ◽  
pp. 8192-8200 ◽  
Author(s):  
D. O. Krause ◽  
S. K. Bhandari ◽  
J. D. House ◽  
C. M. Nyachoti

ABSTRACT Postweaning diarrhea in pigs is frequently caused by enterotoxigenic Escherichia coli K88 (ETEC). The aim of this study was to test the efficacy of E. coli probiotics (PRO) in young pigs challenged with E. coli K88. We also tested the synbiotic interaction with raw potato starch (RPS), which can be used as a prebiotic. Forty 17-day-old weaned piglets were randomly assigned to four treatments: treatment 1, positive-control diet (C), no probiotics or RPS but containing in-feed antibiotics; treatment 2, probiotic (PRO), no feed antibiotics plus a 50:50 mixture of probiotic E. coli strains UM-2 and UM-7; treatment 3, 14% RPS, no antibiotics (RPS); treatment 4, 14% RPS plus a 50:50 mixture of probiotic E. coli strains UM-2 and UM-7, no antibiotics (PRO-RPS). The pigs were challenged with pathogenic E. coli K88 strains on day 7 of the experiment (24-day-old pigs) and euthanized on day 10 of the experiment (35-day-old pigs). Probiotic and pathogenic E. coli strains were enumerated by selective enrichment on antibiotics, and microbial community analysis was conducted using terminal restriction length polymorphism analysis (T-RFLP) of 16S rRNA genes. The combination of raw potato starch and the probiotic had a beneficial effect on piglet growth performance and resulted in a reduction of diarrhea and increased microbial diversity in the gut. We conclude that the use of E. coli probiotic strains against E. coli K88 in the presence of raw potato starch is effective in reducing the negative effects of ETEC in a piglet challenge model.


2017 ◽  
Vol 14 (2) ◽  
pp. 203-208 ◽  
Author(s):  
F. Begum ◽  
M. M. Islam ◽  
M. Sohidullah ◽  
S. M. L. Kabir ◽  
M. Islam ◽  
...  

The present study was designed for the cultural, biochemical characterization and molecular detection of E. coli from apparently healthy and diarrheic goats in and around BAU campus including their antibiogram study. A total of 50 fecal samples were collected among which 13 originated from diarrheic goat and 37 from apparently healthy goats. Out of 50 samples, 35 were found positive for E. coli i.e., overall 70% occurrence. Occurrences of E. coli from diarrheic and apparently healthy goats were 92% and 62% respectively. Occurrences were 60%, 80% and 70% in case of BAU Goat Farm, Veterinary Teaching Hospital and Boyra respectively. On age basis 93%, 54%, 66% and 54% samples originated from 6 months, 7-12 months, 13-18 months and 19 months aged goats were found positive respectively. Occurrences of E. coli on the basis of sex were 78% for male and 62% for female. In case of breed, the occurrences were 69% in Black Bengal and 100% in for Jamunapari. Molecular detection was done by PCR and 13 out of 20 isolates tested gave the bands at the 585 bp specific for E. coli 16S rRNA gene. All the isolates (100%) were found sensitive to ciprofloxacin and norfloxacin; 100% and 35% were intermediately resistant to tetracycline and gentamicin respectively and 25% isolates were resistant to streptomycin. Ciprofloxacin and norfloxacin were found to be the best choice of antibiotics for the treatment of colibacillosis in goats in the study area.


2020 ◽  
Vol 13 (11) ◽  
pp. 2395-2400
Author(s):  
Avijit Das ◽  
Pangkaj Kumar Dhar ◽  
Avijit Dutta ◽  
Mohammad Shah Jalal ◽  
Priya Ghosh ◽  
...  

Background and Aim: The emergence of antimicrobial resistance (AMR) in commensal organism, such as Escherichia coli of food animals, is an alarming issue for global health. It increases the possibility of transmitting AMR determinant(s) to human bacterial pathogens by transferable genetic materials, particularly by plasmids. Hence, it is important to know which resistant genes are being carried by commensal organisms in food chain in a country and their level of temporal loads. As a result, pre-emptive measures can be advocated with an aim to reduce their risks in their primary source of circulation which consequently would benefit the public health. Materials and Methods: Commensal E. coli strains from broiler chickens on randomly selected 30 farms and the farm environments were examined for the frequencies of isolation of resistant strains to oxytetracycline and ciprofloxacin. Five birds were randomly selected from each farm to collect cloacal swab samples (total of 150 samples). Furthermore, a total of 150 environmental samples comprising one each from feed, water, soil, litter, and litter damping site of each farm were screened for the isolation of commensal E. coli strains. Strains thus obtained were initially tested for their resistance to oxytetracycline and ciprofloxacin by Kirby–Bauer disk diffusion method. Oxytetracycline-resistant strains were further screened for the presence of resistance determining genes, namely, tetA, tetB, and tetC by uniplex polymerase chain reactions. Risks associated with the isolation frequency of oxytetracycline- and ciprofloxacin-resistant E. coli were also assessed by univariable logistic regression analysis. Results: The results revealed that all E. coli isolates, regardless of the source of origin, were resistant to oxytetracycline, while 78.4% (95% confidence interval [CI] 69.1-85.5%) showed resistance to ciprofloxacin. All the randomly selected (20) oxytetracycline-resistant strains harbored the tetA gene, whereas tetB and tetC were reported in three and two isolates, respectively. After univariable analysis, only one variable, that is, strain 1 of broiler chickens compared to two other strains was found to be positively associated with the isolation of ciprofloxacin-resistant E. coli (odds ratio 12.75 [95% CI 1.0- 157.1], p=0.047). Conclusion: Resistance emerged against oxytetracycline and ciprofloxacin in commensal E. coli strains circulating in live poultry and farm environments in Bangladesh seems to be very high. Thus, human infection with drug-resistant E. coli strains through food chain will critically compromise the therapeutic measures currently available.


2020 ◽  
Author(s):  
Gaëlle Gruel ◽  
Arantxa Sellin ◽  
Hélène Riveiro ◽  
Matthieu Pot ◽  
Sébastien Breurec ◽  
...  

Abstract Background Selection pressure exerted by overuse of antibiotics in both human and veterinary medicine is responsible for increasing resistance to antibiotics. The objectives of this study were (i) to better understand antimicrobial use in pigs, beef cattle, and poultry on farms of Guadeloupe, French West Indies, and (ii) to acquire data on antimicrobial resistance in Escherichia coli in these food-producing animals. A cross-sectional survey was conducted in 45 farms on Guadeloupe. Practical use of antimicrobials was documented in declarative interviews between March and July 2018. Fecal samples were collected from 216 pigs, beef cattle, and broiler chickens between January 2018 and May 2019. The samples were cultured for bacterial isolation and identification, antimicrobial testing, and screening for blaCTX−M, blaTEM, and tetA resistance genes by PCR on extracted genomic DNA. Results The study shows rational use of antimicrobials consisting of occasional use for curative treatment by veterinary prescription. Tetracycline was the most commonly used antimicrobial, but this was not correlated to E. coli resistance. Extended spectrum β-lactamase (ESBL) E. coli isolates were detected in 7.3% of pigs, 14.7% of beef cattle and 35.3% of broilers. blaCTX−M−1 was the predominant gene found in ESBL E. coli isolates (68.8%), followed by blaCTX−M−15 (31.3%). Conclusion Despite rational use of antimicrobials, the rate of ESBL E. coli in food-producing animals in Guadeloupe, although moderate, is a concern. Further studies are in progress to better define the genetic background of the ESBL E. coli isolates.


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