Background and Aim: Pathogenic Escherichia coli contamination along the broiler meat supply chain is a serious public health concern. This bacterial infection with multidrug-resistant can lead to treatment failure. Several studies have revealed that avian pathogenic E. coli (APEC) and human extraintestinal pathogenic E. coli (ExPEC) showed a close genetic relationship and may share virulence genes. This study aimed to determine the phylogenetic group and virulence gene profiles in colistin-resistant E. coli obtained from the broiler meat supply chain in Bogor, West Java, Indonesia. Materials and Methods: Fifty-eight archive isolates originated from the cloacal swab, litter, drinking water, inside plucker swab, fresh meat at small scale poultry slaughterhouses, and traditional markets were used in this study. All the isolates were characterized by a polymerase chain reaction to determine the phylogenetic group (A, B1, B2, or D) and virulence gene profiles with APEC marker genes (iutA, hlyF, iss, iroN, and ompT). Results: Phylogenetic grouping revealed that the isolates belong to A group (34.48%), D group (34.48%), B1 group (17.24%), and B2 group (13.79%). The virulence gene prevalence was as follows: iutA (36%), hlyF (21%), ompT (21%), iroN (10%), and iss (9%). The B2 group presented with more virulence genes combinations. iroN, hlyF, and ompT genes were positively associated with the B2 group (p≤0.05). Conclusion: Our results highlight the role of colistin-resistant E. coli originated from the broiler meat supply chain as a potential reservoir for human ExPEC virulence genes.
Colistin is the last drug choice for dealing with the carbapenem-resistant Enterobacteriaceae bacteria; hence, this drug is very crucial to human health. The discovery of a plasmid-mediated colistin-resistant gene, the mobilized colistin resistance-1 (mcr-1), signals a significant global health threat. Colistin sulfate is an antimicrobial agent which has been approved for use in broilers in Indonesia. Thus, this study aimed to measure the prevalence of colistin-resistant E. coli and to detect the mcr-1 colistin-resistant gene in E. coli, and E. coli O157:H7 in the entire supply chain of broilers in Bogor Regency, West Java Province, Indonesia. Samples were taken from 47 flocks that used colistin sulfate (47 samples of cloacal swabs, 47 samples of drinking water, and 47 samples of litters), seventy fresh meat samples and seven samples plucker swabs from seven small-scale poultry slaughterhouses, seventy fresh meat samples from seven traditional markets, and seventy cooked meat samples from seven small restaurants. The isolation of E. coli was done on each of the 358 samples, and 493 isolates were obtained. All the E. coli isolates were then tested for their susceptibility to colistin sulfate by using the agar dilution method. The detection of the mcr-1 gene from the colistin-resistant isolates (minimum inhibitory concentration > 2 µg/mL) was conducted using the polymerase chain reaction (PCR). The prevalence value of colistin-resistant E. coli in all the isolates was at 11.76% (CI 95%; CL 9.21–14.91%), and the prevalence of mcr-1 gene was at 10.55% CI 95%; CL 8.13–13.57%).  A very good agreement correlation existed between the colistin-resistant phenotype and the mcr-1 gene (ĸ = 0.939). The mcr-1 gene was found in 89.66% colistin-resistant E. coli isolates. The two colistin-resistant and mcr-1 carrying gene isolates were identified as E. coli O157:H7 serotype. This research was the first study attempt on the mcr-1 gene in Indonesia, covering the entire supply chain of broiler meat from farms to consumers. The results indicated the necessity to reduce the use of colistin sulfate in broiler management and to improve biosecurity measures, not only in farms but also in the entire supply chain of broiler meat production.
Colistin sulphate is the ultimate antimicrobial choice for the treatment of multidrug resistance gram negative bacteria infections with in human. The purposes of this study were to detect the presence of colistin resistant E. coli and mcr-1 gene in broiler and to transfer the mcr-1 gene to Salmonella enteritidis ATCC 13076. A total of 54 one day old broilers were divided into three groups that consists of 18 chicks broiler per group and raised up to 40 days old. The first group was used as control. The first treatment group was given colistin sulphate 5 ìg/g feed for 40 days and broilers in second treatment group was given 80.000 IU/kg body weight for first three days. Swab cloaca samples were taken every 10 days from each broiler. At age 40 days all chickens were slaughtered and meat samples were collected. Samples of cloacal swabs, fresh and cooked meat were examined for the presence of colistin resistant E. coli and mcr-1 gene. Susceptibility to colistin sulfate was conducted by agar dilution method, and detection of mcr-1 gene was conducted using polymerase chain reaction. The results showed that no colistin resistant E. coli was detected in the control group. Colistin resistant E. coli (27.78%) and mcr-1 gene (20.00%) were detected in animals in the first treatment group, respectively. Whilst 11.11% colistin resistant E. coli and 5.56% were carriying mcr-1 gene in the second treatment group. Colistin resistant E. coli were found 5.56% from raw meat samples and 3.70% had mcr-1 gene. Transfer of mcr-1 gene from colistin resistant E. coli to Salmonella enteritidis ATCC 13076 was success. These results showed the necessity of limitation usage of colistin sulphate in food animal.
Mutant prevention concentration (MPC) is an in vitro test used to determine the lowest drug concentration needed to inhibit the growth of a single-step-mutant bacterial subpopulation. The purpose of this study was to determine the MPC value of ciprofloxacin against pathogenic Escherichia coli to obtained the range of mutant selection windows (MSW) of ciprofloxacin. Ciprofloxacin is a quinolone group that is included in the Highest Priority Critically Important Antimicrobials for Human Medicine but is also used for the treatment of bacterial infections in production animals. Twenty-four of pathogenic E. coli isolates sensitive to ciprofloxacin were tested to obtain MPC values and minimum inhibitory concentration (MIC) values. Test the MPC and MIC values to get the MSW range is done by the method of agar dilution. Mueller-Hinton agar containing standard ciprofloxacin was inoculated with 1010 cfu E. coli for the MPC test and 104 for the MIC test. Based on the MPC test results, the MPC value of ciprofloxacin was 4-64 μg / mL (22.96 ± 19.07 μg / mL) and there was one isolate which had an MPC> 256 μg / mL. These results give a wide range of MSW with a lower limit of the MIC value of 0.25 - 2 µg / mL (0.55 ± 0.37 µg / mL) to the upper limit of the MPC value of 4-64 µg / mL (22.96 ± 19.07 μg / mL). Based on the results of this MPC assessment it can be concluded that the dose of ciprofloxacin in production animals has a wide range of MSW that is allow for single-step mutants.
The risk assessment of antimicrobial resistance is very important to determine the risk of decreasing antimicrobial efficacy can be used as a basis for policymakers in allowing or prohibiting the use of an antimicrobial. This study aims to assess the risk of using colistin against E. coli resistance in the broiler flock. Risk assessment is carried out qualitatively using primary data, interviews, and secondary data. To obtain primary data various studies were carried out including monitoring the prevalence of colistin-resistant E. coli and mcr-1 also mcr-2 genes in broiler flocks, mcr-1 gene transfer from E. coli to Salmonella Enteritidis, mcr-1 gene sequencing, mutant selection windows of colistin against E. coli, and also multiresistant of E. coli colistin-resistant. Assessment of the risk of E. coli colistin-resistant in the broiler flocks through direct contact with live broiler flock environment with the resulting assessment is a medium risk with low uncertainty. Since colistin sulfate is very critically important for humans, the reduced use of colistin sulfate in animal production is necessary to reduce the risk of resistance. Reducing the use of colistin sulfate requires the collaboration of various sectors such as the government, veterinary drugs industries, farmers, and consumers.
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