ANTIMICROBIAL RESISTANCE PROFILING OF SELECTED E. coli ISOLATES AND DETECTION OF ESBL/PAMPC-ENCODING GENES IN BROILER FLOCKS IN BOSNIA AND HERZEGOVINA USING REAL-TIME PCR

Irrational use of antimicrobials in humans, livestock, and poultry is often cited as the key driver of the accelerated emergence of antimicrobial resistance (AMR) in humans and animals. In Pakistan, the use of antimicrobials in livestock and poultry is not well-regulated, owing to limited and/or ineffective implementation of existing legislations and lack of coordination amongst the key stakeholders. To gather data and information pertinent to the supply chain mapping and the usage of antibiotics in the livestock and poultry sectors of Pakistan, a national workshop for selected influential stakeholders was held in Islamabad, Pakistan during March 2020 to map the supply chain and the usage of antibiotics in the country. Participants from all relevant organizations identified the supply chain and discussed the status, challenges, and the way forward to enhance data collection and monitoring of antimicrobial usage (AMU) in livestock and poultry sectors to contain the emergence of AMR. The pre-workshop questionnaire was designed to gather relevant information on AMU and its distribution among diverse markets and users from the workshop participants, utilizing open-ended questions. A chart depicting the relative magnitude of an antimicrobial use (AMU) supply chain was constructed to illustrate the flow of antimicrobials from import and production to end-use at the farm level. This chart was shared with participants to gather their professional opinions and potential corrections. It also presented a list of agencies and their roles in regulating AMU, along with the types of AMU data available at each level. Specific recommendations were made at the end of the workshop to review and update legislation to cover the entire AMU supply chain, enhance regulations to restrict the use of antimicrobial growth promoters, build an integrated national AMU database system, and raise awareness about the responsible use of antimicrobials in the livestock and poultry sectors. It was concluded that the AMU supply chain in the veterinary sector of Pakistan is fragmented and is co-regulated by various federal and provincial stakeholders. There are some drugs, such as antibiotic growth promoters, which are not regulated at all. The approach and findings from this study can serve as a model to validate the use and management of antibiotics in other, similar countries currently grappling with serious antimicrobial resistance (AMR) crises.

Antimicrobials are widely used in agriculture and livestock, nevertheless their large-scale and indiscriminate use contributes considerably to the development of microbial resistance. The intense commercialization of agricultural products accelerates the dissemination of pathogens that present resistance profiles, which characterize them as extremely harmful to plants and animals. Thus, there is an urgent need to control the use of antimicrobials in animal and vegetable food products in order to find new agents that are less able to develop resistance. It is essential the guidance of the professionals and the population about the rational use of antimicrobials in agriculture and livestock for the maintenance and protection of the human healthy and environment.

Data on the use of antimicrobials in humans and livestock may provide evidence to guide policy changes to mitigate the risk of antimicrobial resistance (AMR). However, there is limited information available about antimicrobial use in livestock in low- and middle-income countries, even though these nations are most vulnerable to the impact of AMR. This study aimed to assess the consumption of veterinary antimicrobials in Bhutan and identify areas for improvement to reduce the use of antimicrobials in livestock. National data on livestock numbers and annual procurement of veterinary antimicrobials over five years (2017-2021) were used to calculate rates of antimicrobial consumption and annual national expenditure on veterinary antimicrobials in Bhutan. The rate of antimicrobial consumption in Bhutan was 3.83 mg per population correction unit, which is lower than most countries in Europe, comparable with the rates of consumption in Iceland and Norway, and approximately 120-fold lower than published rates of antimicrobial consumption in South Asian countries, including Nepal and Pakistan. The low rates of antimicrobial consumption by the animal health sector in Bhutan could be attributable to stronger governance of antimicrobial use in Bhutan, higher levels of compliance with regulation, and better adherence to standard guidelines for antimicrobial treatment of livestock.

Veterinary RecordVolume 169, Issue 19 p. 479-480 News … Report Call to phase out prophylactic use of antimicrobials in livestock First published: 05 November 2011 https://doi.org/10.1136/vr.d7090Read the full textAboutPDF ToolsRequest...

SummaryThe urgency to reduce the use of antimicrobials in the livestock sector has become part of the European Farm‐to‐Fork agenda. To develop innovations and pave the way for prudent antimicrobial use (AMU), the H2020 four‐year‐long ROADMAP project implemented the approach of Living Labs (LL). It stands for a process that involves a heterogeneous group of stakeholders in co‐creation, testing and evaluating innovations in real‐world contexts. The use of LLs has gained momentum to accelerate transformation of European agriculture by developing innovations through end‐user (farmer) involvement. However, the LL‐approach is new in the livestock sector. During the project, 11 LLs were established in seven European countries, each with its own settings but linked to the project's overarching goals. Some LLs developed a common understanding of AMU within the sector or between farmers and veterinarians, while others focussed on policy development or better management practices. During the ROADMAP project, we learned about the potential and limitations of the LL approach in the livestock sector. Using the approach of social innovations, we developed recommendations on how to realise the potential of LLs in a research context to meet the goals of the EU Farm‐to‐Fork strategy of prudent AMU.

Manure compost has been thought of as a potential important route of transmission of antimicrobial-resistant bacteria (ARB) and antimicrobial resistance genes (ARGs) from livestock to humans. To clarify the abundance of ARB and ARGs, ARB and ARGs were quantitatively determined in tetracycline-resistant Escherichia coli (harboring the tetA gene)-spiked feces in simulated composts. In the simulated composts, the concentration of spiked E. coli decreased below the detection limit at day 7. The tetA gene remained in manure compost for 20 days, although the levels of the gene decreased. Next, to clarify the field conditions of manure compost in Japan, the quantities of tetracycline-resistant bacteria, tetracycline resistance genes, and residual tetracyclines were determined using field-manure-matured composts in livestock farms. Tetracycline-resistant bacteria were detected in 54.5% of tested matured compost (6/11 farms). The copy number of the tetA gene and the concentrations of residual tetracyclines in field manure compost were significantly correlated. These results suggest that the use of antimicrobials in livestock constitutes a selective pressure, not only in livestock feces but also in manure compost. The appropriate use of antimicrobials in livestock and treatment of manure compost are important for avoiding the spread of ARB and ARGs.

Data on antimicrobial use in livestock: Lessons from Uganda

Interventions to change antimicrobial use in livestock: A scoping review and an impact pathway analysis of what works, how, for whom and why.

The development of antimicrobial resistance (AMR) due to over or misuse of antibiotics/antimicrobials is a globally increasing public health concern. This study was conducted to assess the current knowledge, attitudes, and practices (KAP) of swine and poultry producers on antimicrobial use (AMU) and antimicrobial resistance (AMR). A total of 254 swine and poultry producers of 5 provinces in Cambodia were purposively surveyed using validated KAP questionnaires, a useful tool for promoting the rational use of antimicrobials in livestock. Collectively, this study found out that livestock producers in Cambodia have a low level of knowledge, neutral rather than positive attitudes, and employed poor practices on the AMU and AMR. Thus, there is a need to promote among producers the rational and responsible use of antimicrobials. Furthermore, public awareness of the AMR implications on public health can contribute to the prevention or reduction of AMU and AMR in Cambodia.

Reducing antimicrobial use (AMU) in livestock may be one of the keys to limit the emergence of antimicrobial resistance (AMR) in bacterial populations, including zoonotic pathogens. This study assessed the temporal association between AMU in livestock and AMR among Campylobacter isolates from human infections in the Netherlands between 2004 - 2020. Moreover, the associations between AMU and AMR in livestock and between AMR in livestock and AMR in human isolates were assessed. AMU and AMR data per antimicrobial class (tetracyclines, macrolides and fluoroquinolones) for Campylobacter jejuni and Campylobacter coli from poultry, cattle, and human patients were retrieved from national surveillance programs. Associations were assessed using logistic regression and the Spearman correlation test. Overall, there was an increasing trend in AMR among human C. jejuni/coli isolates during the study period, which contrasted with a decreasing trend in livestock AMU. In addition, stable trends in AMR in broilers were observed. No significant associations were observed between AMU and AMR in domestically produced broilers. Moderate to strong positive correlations were found between the yearly prevalence of AMR in broiler and human isolates. Reducing AMU in Dutch livestock alone may therefore not be sufficient to tackle the growing problem of AMR in Campylobacter among human cases in the Netherlands. More insight is needed regarding the population genetics and the evolutionary processes involved in resistance and fitness among Campylobacter.

Herd typologies based on multivariate analysis of biosecurity, productivity, antimicrobial and vaccine use data from Danish sow herds

Background: Campylobacteriosis is the leading zoonotic disease in developed countries with C. jejuni and C. coli being the two predominant causative pathogens. It has been shown that quinolone consumption in livestock is associated with increased quinolone resistance of Campylobacter isolates from food producing animals and infected patients. However, susceptibility testing of clinical isolates is not commonly performed and, consequently, resistance rates of human C. jejuni and C. coli isolates in areas of high consumption of antimicrobials in livestock may be undervalued. A strong association between C. jejuni infections and patients` age and gender has been highlighted by several authors. However, there is still little information on the demographic pattern in C. coli infections.Methods: 1135 C. jejuni and 156 C. coli human isolates were obtained within a rural region of Germany. The study area was characterised by intensive swine and poultry farming involving high consumption of clinically important antimicrobials. Isolates were analysed for susceptibility to amoxicillin, ciprofloxacin, tetracycline and erythromycin using the EUCAST disc diffusion method. Furthermore, data were stratified with respect to patients age and gender.Results: Contrary to male-biased distribution in C. jejuni isolates, C. coli was the predominant species in female patients with a maximum female surplus in young children and middle-aged adults. Resistance rates of C. coli vs. C. jejuni to amoxicillin, ciprofloxacin, tetracycline and erythromycin were 46.2% vs. 48.3%, 62.8% vs. 64.5%, 68.6% vs. 35.2% and 14.7 vs. 0.6%, respectively. Resistance rates were found to correlate with usage of these antimicrobials in livestock.Conclusion: The high prevalence of C. coli in female patients may point to sex-specific behavioural or physiological aspects. The observed high to moderate resistance rates of Campylobacter isolates warrant prudent use of antimicrobials in livestock as well as routine susceptibility testing of human isolates to ensure efficacy of antimicrobial therapy.

A total of 3246 swine slaughter and 3147 swine diagnostic Salmonella isolates from the National Antimicrobial Resistance Monitoring System (NARMS) for the years 1997-2003 were tested for antimicrobial resistance. No resistance was detected in any Salmonella isolates for ciprofloxacin or amikacin. For all other antimicrobics tested, more resistance was observed in diagnostic isolates compared to slaughter isolates. In 2003, more than 50% of diagnostic isolates were resistant to 4 antibiotics: tetracycline (77.4%), streptomycin (75.5%), sulfamethoxazole (65.5%), and ampicillin (57.1%). Tetracycline (30.8%) was the antimicrobial with the most resistance in slaughter isolates. Continued monitoring of both slaughter and diagnostic isolates from swine are needed to demonstrate if changes industry use patterns of antimicrobials lead to changes in resistance patterns of Salmonella isolates from swine. Introduction Public health concerns associated with the use of antimicrobials in livestock led to the development of the National Antimicrobial Resistance Monitoring System (NARMS) in 1996. The program was developed in part as a mechanism to monitor the effect of the use of veterinary antimicrobials on changes in antimicrobial resistance patterns in bacteria associated with farm animals. Non-typhoid Salmonella were selected as the sentinel organism and have been continuously tested for since 1997. This paper focuses on the monitoring of resistance changes in Salmonella isolated from diagnostic and slaughter samples from swine from 1997–2003. Materials and Methods Slaughter samples were collected and cultured by the USDA, FSIS, and diagnostic samples were collected and cultured by state veterinary laboratories around the U.S or the USDA, APHIS, National Veterinary Services Laboratory (NVSL). Slaughter and diagnostic isolates were shipped to USDA, ARS in Athens, GA. All Salmonella isolates were serotyped at the NVSL in Ames, IA. After serotyping, 3246 slaughter and 3147 diagnostic isolates were tested for antimicrobial resistance using custom made plates containing up to 16 antimicrobics in a semiautomated minimal inhibitory concentration format system (SensititerTM, Trek Diagnostic) at the USDA, ARS in Athens, GA. Isolates were classified as susceptible, intermediate, or resistant based on Clinical Laboratory Standards Institute (CLSI) (formerly National Committee for Clinical Laboratory Standards) established breakpoints, where available. Results No resistance was detected in any Salmonella isolates for ciprofloxacin or amikacin. For all other antimicrobics tested, more resistance was observed in diagnostic isolates than in slaughter isolates. In 2003, more than 50% of diagnostic isolates were resistant to tetracycline (77.4%), streptomycin (75.5%), sulfamethoxazole (65.5%), and ampicillin (57.1%). For slaughter isolates, tetracycline (30.8%) was the antimicrobial with the most resistance. Also, in 2003, amoxicillin, cefoxitin, ceftiofur, cephalothin, and chloramphenicol resistance in diagnostic isolates was 10–20% greater than in any previous year. An analysis of antimicrobial resistance profiles of the eight most frequently identified serotypes of Salmonella suggests that S. Derby, Typhimurium var. Copenhagen, Typhimurium, Heidelberg, and Agona were more resistant to tetracycline, sulfamethoxazole, and streptomycin, with little difference observed between slaughter and diagnostic isolates. More ampicillin resistance was observed with slaughter and diagnostic isolates of S. Typhimurium var Copenhagen, Typhimurium, and diagnostic isolates of Agona. S. Heidelberg appears to be developing more resistance to ampicillin and ceftiofur in diagnostic isolates and to streptomycin and tetracycline in slaughter isolates. Multiple drug resistance will be reported in a separate paper. O R A L P R E S E N T A T IO N S 203 SafePork 2005 Discussion A limited number of studies have closely examined the development of antibiotic resistance in Salmonella from swine samples. Cray et al. (1999) reported on the swine NARMS samples collected in 1998 and as in this larger study reported that more resistance was seen in diagnostic samples than in slaughter samples and that most resistance was to tetracycline, streptomycin and sulfamethoxazole. Farrington and co-workers (2001) determined antibiotic resistance patterns of 365 Salmonella isolates from lymph nodes and cecal content of market-age swine at slaughter, and reported that although no subtherapeutic antibiotics were used on the farm in the past five years that resistance to chlortetracycline, penicillin G, streptomycin, and sulfisoxazole was common. hese authors suggest that a correlation exists between the somatic serogroup or serovar of a Salmonella isolate and its antimicrobial resistance status. Although specific serotypes of Salmonella may possess different susceptibilities to antibiotic resistance, it is also possible that the source, diagnostic versus slaughter, may contribute more to the development of resistance than does the specific serotype. Conclusion Continued monitoring of both slaughter and diagnostic isolates from swine are needed to demonstrate if changes in industry use patterns of antimicrobials lead to changes in resistance patterns of Salmonella isolates from swine. More directed studies are needed to determine if development in resistance characteristics if individual Salmonella isolates is influence more by serotype or more by sample source.

There are growing concerns over the threat to human health from the unregulated use of antimicrobials in livestock. Broiler production is of great economic and social importance in Indonesia. This study used a structured questionnaire approach to explore the human behaviours and economic drivers associated with antimicrobial use in small commercial broiler systems in Indonesia (n = 509). The study showed that antimicrobial use was high with farmers easily able to access antimicrobials through local animal medicine, however, it was difficult for farmers to access veterinary advice on responsible antimicrobial use. The most significant finding was that the relative cost of antimicrobials was low, and farmers observed improvements in productivity rates from routine antimicrobial administration. However, farmers seldom kept detailed records on farm productivity or economic costs; this is a hurdle to undertaking a more detailed economic analysis of antimicrobial use. There is a need for further research on the cost-effectiveness of alternative methods of preventing disease and ensuring that feasible alternatives are easily available. Farm-level economics and securing the food supply chain need to be central to any future policy interventions to reduce antimicrobial use in broiler systems in Indonesia and this observation is relevant at a regional and global level.

In dairy intensification, mastitis infections become prevalent and induce frequent Antimicrobial Use (AMU), sometimes inappropriately. This poses public health risks because of growing Antimicrobial Resistance (AMR), which calls for stewardship programs informed by Knowledge, Attitude and Practices (KAPs) about AMU and AMR to halt or reverse the worrying trend. Data was obtained in cross sectional survey conducted in four peri-urban wards around Nakuru city in Kenya. Randomly selected sample farmers (n=124) with free-grazing, semi-zero-grazing or zero-grazing dairy management, representing increasing dairy intensification levels provided data on the KAPs. Chi-square test statistics was fitted to establish associations between KAPs and dairy intensification levels. Among the sample farmers, six in ten (58.8 percent) had intensified dairy production, at least six in ten were marketing milk through informal outlets and were using antimicrobial drugs. Compliance with the withdrawal period was high and increased (p<0.05) with increasing intensification from free-grazing to zero-grazing. Within antibiotic withdrawal period, at least seven in ten farmers did not sell milk, fewer than four in ten consumed their milk at home and fewer than three in ten fed the milk to calves. Though independent of dairy intensification level (p>0.05), using antimicrobials for mastitis treatment increased while sourcing information on antimicrobial use from extension and veterinary officers decreased, with increasing intensification level. Farmers with some training on prudent antimicrobial use and with positive attitudes that milk from antimicrobial treated cows is unsafe, antimicrobial resistant pathogens and residues can be passed from milk to humans, mastitis can be treated without antimicrobial drugs, and antimicrobial residues can end up accumulating in the soils increased (p>0.05) with increasing dairy intensification levels. These results show that regarding AMU and AMR, farmers become more knowledgeable, with positive attitudes and good practices as they intensify their dairy management. The implication is that intensification of dairy management motivates farmers to gain more knowledge, acquire positive attitudes and apply good practices towards responsible prudent use of antimicrobials in livestock. Therefore, strengthening stewardship with targeted training and sensitization can foster prudent and responsible antimicrobial use.