A Review of Slaughter Practices and Their Effectiveness to Control Microbial - esp. Salmonella spp. - Contamination of Pig Carcasses.

The BIOPIGEE project (part of the One Health European Joint Programme under Horizon 2020) aimed to identify relevant measures to effectively control Salmonella, and another zoonotic pathogen, hepatitis E virus (HEV) within the pig meat food chain. The aim of this study was to identify biosecurity measures or management practices that are relevant for limiting Salmonella and/or HEV occurrence and spread within pig slaughterhouses. This was with the final goal of compiling a list of biosecurity measures for different processes and operations along the slaughter line with evidence of their effectiveness. To achieve this, a literature review was conducted on studies estimating the effectiveness of measures applied in slaughterhouses to reduce the microbial contamination of pig carcasses. Results of this literature search are discussed and presented in summary tables that could be used as a source of information for the pig slaughter industry to further develop their guidelines on hygienic slaughter.

Assessing pig farm biosecurity measures for the control of Salmonella on European farms.

Salmonella spp. is a common zoonotic pathogen, causing gastrointestinal infections in people. Pigs and pig meat are a major source of infection. Although farm biosecurity is believed to be important for controlling Salmonella transmission, robust evidence is lacking on which measures are most effective. This study enrolled 250 pig farms across nine European countries. From each farm, 20 pooled faecal samples (or similar information) were collected and analysed for Salmonella presence. Based on the proportion of positive results, farms were categorised as at higher or lower Salmonella risk, and associations with variables from a comprehensive questionnaire investigated. Multivariable analysis indicated that farms were less likely to be in the higher-risk category if they had '<400 sows'; used rodent baits close to pig enclosures; isolated stay-behind (sick) pigs; did not answer that the hygiene lock/ anteroom was easy to clean; did not have a full perimeter fence; did apply downtime of at least 3 days between farrowing batches; and had fully slatted flooring in all fattener buildings. A principal components analysis assessed the sources of variation between farms, and correlation between variables. The study results suggest simple control measures that could be prioritised on European pig farms to control Salmonella.

Detection of HEV RNA Using One-Step Real-Time RT-PCR in Farrow-to-Finish Pig Farms in Bulgaria.

(1) Background: HEV is a zoonotic, foodborne pathogen. It is spread worldwide and represents a public health risk. The aim of this study was to evaluate the presence of HEV RNA in farrow-to-finish pig farms in different regions of Bulgaria; (2) Methods: Isolation of HEV RNA from pooled samples of feces was performed using a QIAamp® Viral RNA Mini Kit followed by HEV RNA detection using a single-step real-time RT-PCR with primers and probes targeting the ORF 3 HEV genome; (3) Results: HEV RNA was detected in 12 out of 32 tested farms in Bulgaria (37.5%). The overall percentage of HEV-positive pooled fecal samples was 10.8% (68 of 630 samples). HEV was detected mostly in pooled fecal samples from finisher pigs (66/320, 20.6%) and sporadically from dry sows (1/62, 1.6%) and gilts (1/248, 0.4%); (4) Conclusions: Our results confirm that HEV circulates in farrow-to-finish pig farms in Bulgaria. In our study, we found HEV RNA in pooled fecal samples from fattening pigs (4-6-months age), shortly before their transport to the slaughterhouse indicating a potential risk to public health. The possible circulation of HEV throughout pork production requires monitoring and containment measures.

Cross-sectional study of hepatitis E virus (HEV) circulation in Italian pig farms.

Foodborne transmission is considered the main way of spreading zoonotic hepatitis E virus (HEV) infection in Europe. In recent years, the human cases of hepatitis E in subjects without history of travel in endemic areas have raised, suggesting that domestic HEV transmission is increasing. Pork products with or without liver, are often indicated as the source of many human foodborne HEV cases as well as small outbreaks. Pigs are recognized as the main reservoir of the zoonotic HEV-3 genotype, the most frequently detected in human cases in the EU. In the absence of a harmonized surveillance of HEV circulation, data on prevalence are heterogeneous but confirm a widespread circulation of HEV-3 in pig herds across EU. HEV-3 can pass through the food chain from farm to fork when infected animals are slaughtered. In Italy, several studies reported the circulation of HEV-3 in pig farms, but results are heterogeneous due to different methodologies applied. In the present study, we performed a survey over 51 pig herds belonging to three main types of farms: breeding, fattening and farrow-to-finish. HEV-RNA was analyzed by broad range Real-time RT-PCR on 20 samples for each farm, obtained by pooling together feces from 10 individuals. Overall, HEV RNA was confirmed on 150 fecal pooled samples out of 1,032 (14.5%). At least one positive pooled sample was detected from 18 farms out of 51 tested (35.3%). By lowering the number of infected pigs at primary production, the risk of HEV-3 entering into the food chain can be reduced. Hence, information on HEV circulation in herds is highly relevant for choosing preventive measures and deserves development of a monitoring program and further investigations.

Prioritization of pig farm biosecurity for control of Salmonella and hepatitis E virus infections: results of a European expert opinion elicitation.

BACKGROUND: In the literature, there is absent or weak evidence on the effectiveness of biosecurity measures to the control of Salmonella spp. and hepatitis E virus (HEV) on pig farms. Therefore, the present study aimed to collect, weigh, and compare opinions from experts on the relevance of several biosecurity measures. An online questionnaire was submitted to selected experts, from multiple European countries, knowledgeable on either HEV or Salmonella spp., in either indoor or outdoor pig farming systems (settings). The experts ranked the relevance of eight biosecurity categories with regards to effectiveness in reducing the two pathogens separately, by assigning a score from a total of 80, and within each biosecurity category they scored the relevance of specific biosecurity measures (scale 1-5). Agreement among experts was analysed across pathogens and across settings. RESULTS: After filtering for completeness and expertise, 46 responses were analysed, with 52% of the experts identified as researchers/scientists, whereas the remaining 48% consisted of non-researchers, veterinary practitioners and advisors, governmental staff, and consultant/industrial experts. The experts self-declared their level of knowledge but neither Multidimensional Scaling nor k-means cluster analyses produced evidence of an association between expertise and the biosecurity answers, and so all experts' responses were analysed together without weighting or adaptation. Overall, the top-ranked biosecurity categories were pig mixing; cleaning and disinfection; feed, water and bedding; and purchase of pigs or semen, while the lowest ranked categories were transport, equipment, animals (other than pigs and including wildlife) and humans. Cleaning and disinfection was ranked highest for both pathogens in the indoor setting, whereas pig mixing was highest for outdoor settings. Several (94/222, 42.3%) measures across all four settings were considered highly relevant. Measures with high disagreement between the respondents were uncommon (21/222, 9.6%), but more frequent for HEV compared to Salmonella spp. CONCLUSIONS: The implementation of measures from multiple biosecurity categories was considered important to control Salmonella spp. and HEV on farms, and pig mixing activities, as well as cleaning and disinfection practices, were perceived as consistently more important than others. Similarities and differences in the prioritised biosecurity measures were identified between indoor and outdoor systems and pathogens. The study identified the need for further research especially for control of HEV and for biosecurity in outdoor farming.

What is a biosecurity measure? A definition proposal for animal production and linked processing operations.

While biosecurity, a central component of the One Health concept, is clearly defined, a harmonized definition of the term ´biosecurity measure´ (BSM) is missing. In turn, particularly at the farm and policy level, this leads to misunderstandings, low acceptance, poor implementation, and thus suboptimal biosecurity along the food animal production chain. Moreover, different views on BSMs affects making comparisons both at the policy level as well as in the scientific community. Therefore, as part of the One Health EJP BIOPIGEE project, a work group i) collected and discussed relevant inclusion and exclusion criteria for measures to be considered in the context of biosecurity and ii) conducted a systematic literature review for potentially existing definitions for the term BSM. This exercise confirmed the lack of a definition of BSM, underlining the importance of the topic. In the pool of articles considered relevant to defining the term BSM, specific research themes were identified. Based on these outcomes, we propose a definition of the term BSM: "A biosecurity measure (BSM) - is the implementation of a segregation, hygiene, or management procedure (excluding medically effective feed additives and preventive/curative treatment of animals) that specifically aims at reducing the probability of the introduction, establishment, survival, or spread of any potential pathogen to, within, or from a farm, operation or geographical area." The definition provides a basis for policymakers to identify factual BSMs, highlights the point of implementation and supports to achieve the necessary quality standards of biosecurity in food animal production. It also enables clear, harmonized, cross-sectoral communication of best biosecurity practices to and from relevant stakeholders and thus contribute to improving biosecurity and thereby strengthen the One Health approach.

Are There Effective Intervention Measures in Broiler Production against the ESBL/AmpC Producer Escherichia coli?

Extended-spectrum beta-lactamase (ESBL) and AmpC beta-lactamase (AmpC) producing Enterobacteriaceae occur frequently in livestock animals and the subsequent stages of the meat production chain and are therefore considered a risk for human health. Strict biosecurity measures and optimal farm management should reduce or even prevent poultry flock colonization at farm level. This review summarizes and evaluates published information on the effectiveness of specific intervention measures and farm management factors aiming to reduce the occurrence and spread of ESBL/AmpC producing or commensal or pathogenic E. coli in broiler chicken farms. In this systematic literature review, a total of 643 publications were analyzed, and 14 studies with significant outcome about the effectiveness of specific measures against E. coli and ESBL/AmpC producing E. coli in broiler chicken farms were found. Different feed additives seem to have an impact on the occurrence of those microorganisms. The measures 'cleaning and disinfection' and 'competitive exclusion' showed strong effects in prevention in some studies. In summary, some intervention measures showed potential to protect against or eliminate ESBL/AmpC-producing, commensal or pathogenic E. coli at farm level. Due to the high variability in the outcome of the studies, more specific, detailed investigations are needed to assess the potential of the individual intervention measures.

Antibiotic Resistance in Escherichia coli from Broiler Chickens After Amoxicillin Treatment in an Experimental Environment.

Groupwise antibiotic treatments are common in broiler chicken production. They induce selection for antibiotic resistance in commensal Escherichia coli. This study aimed to investigate antibiotic resistance after individual (I, drenching) or groupwise treatment (G, by water) with amoxicillin, and after contact with I or G (KI or KG), compared with untreated broilers without contact with treated broilers (C), and pretreatment values. Finally, we compared antibiotic resistance from broilers (G) after a second treatment, with a treatment in the contact animals (KG), and a first treatment in the control animals (C). Resistance to ampicillin and other antibiotics was significantly increased in groups G and I within 2 days, suggesting (co-)selection of resistance. The increase was lower in groups KI, KG, and C during the first treatment (days 1-5). The increased resistance in group C was interpreted as a change in the microbiota after initial moving and first feeding. After treatment, resistance rates decreased to initial or lower values in all groups. During the second treatment period (days 34-38), all three groups' (G, KG, and C) resistance levels increased to equally high levels. Cephalosporin resistance was low, and did not change over the experimental period. On days 3 and 38, resistance rates of E. coli from duodenum, jejunum, and cecum did not differ between segments and treatment routes. Overall, the baseline levels of antibiotic resistance in E. coli were high. Amoxicillin triggered an increase in resistance levels, irrespective of the mode of treatment. Substantial resistance dynamics in untreated controls warrant further investigation.

Antibiotic resistance in Escherichia coli from pigs from birth to slaughter and its association with antibiotic treatment.

The purpose of this longitudinal study was to describe the occurrence of antibiotic resistance in faecal Escherichia coli isolated from pigs between birth and slaughter and its association with antibiotic treatment. Four objectives were addressed: comparison of antibiotic resistance in isolates from a) treated vs. non-treated pigs, b) follow-up vs. initial samples of treated and non-treated pigs, c) pigs receiving treatments via different administration routes and d) sows and their piglets. Each comparison addressed the following antibiotic groups used for treatment: beta-lactams, tetracyclines, polymyxins and macrolides, and the susceptibility of E. coli isolates to the respective agents: ampicillin, tetracycline, colistin and azithromycin. Between 2014 and 2016, 406 focal animals from 29 commercial breeding herds were followed from birth to the end of the relevant fattening periods. All antibiotic treatments in these pigs were documented. Faecal samples were collected from the focal pigs once while suckling, once after weaning and three times during fattening, and from their dams once around farrowing. Escherichia coli isolated from these samples was tested for antibiotic susceptibility. In total, 264 animals from 19 breeding herds were treated with an antibiotic at least once during their lifetime. Beta-lactams, tetracyclines and colistin were used most frequently. Piglets were treated individually by injection (n = 108 treatments) or via drench (9); weaners via feed (192) or water (56) and fatteners via feed (30) or injection (15). Resistance to ampicillin and tetracycline in E. coli was already common prior to antibiotic treatment. Resistance proportions were higher for beta-lactam-, tetracycline-, colistin- and macrolide-treated pigs compared to untreated pigs at different sampling periods (p < 0.05; Fisher's exact test). In the logistic analysis, the difference was confirmed for beta-lactam-treated vs. untreated pigs. In E. coli from macrolide-untreated pigs, resistance to azithromycin was more frequent compared to pre-treatment values. Route of application did not affect rates of antibiotic resistance in the logistic analysis even though Fisher's exact test indicated associations for beta-lactams (feed/water vs. injection), tetracyclines (feed/water vs. non-treatment) and macrolides (tulathromycin-injection vs. tylosin in feed). Piglets were more likely to carry an E. coli resistant to ampicillin or azithromycin if their dams did so as well. Our results suggest further research on resistance effects by administration routes is required. Reducing antibiotic resistance in sows might lead to a lower level of beta-lactam or macrolide-resistant E. coli among their progeny. To preserve treatment options for bacterial infections, antibiotic use should be restricted to necessary cases.

Risk Factors for Antimicrobial Resistance in Escherichia coli in Pigs Receiving Oral Antimicrobial Treatment: A Systematic Review.

The aim of this literature review was to identify risk factors in addition to antimicrobial treatment for antimicrobial resistance (AMR) occurrence in commensal Escherichia coli in pigs. A variety of studies were searched in 2014 and 2015. Studies identified as potentially relevant were assessed against eligibility criteria such as observation or experiment (no review), presentation of risk factors in addition to (single dosage) antimicrobial use, risk factors for but not resulting from AMR, and the same antimicrobial used and tested. Thirteen articles (nine on observational, four on experimental studies) were finally selected as relevant. It was reported that space allowance, production size/stage, cleanliness, entry of animals and humans into herds, dosage/frequency/route of administration, time span between treatment and sampling date, herd size, distance to another farm, coldness, and season had an impact on AMR occurrence. Associations were shown by one to four studies per factor and differed in magnitude, direction, and level of significance. The risk of bias was unclear in nearly half of the information of observational studies and in most of the information from experimental studies. Further research on the effects of specific management practices is needed to develop well-founded management advice.