Assessing the Feasibility of a Two-Cohort Design to Assess the Potential of Homeopathic Medicinal Products to Reduce Antimicrobial Resistance in Turkeys (The HOMAMR Project)-Study Protocol.

BACKGROUND: Antimicrobial resistance (AMR) is a serious public health concern worldwide. The European Union requires a reduction in the use of antibiotics by 50% by 2030, with separate regulations on organic production that give preference to homeopathy and phytotherapy in organic farms before the use of conventional medicines (including antibiotics). We have therefore designed a two-phased project whose overarching aim is to investigate the potential role of homeopathic medicinal products (HMPs) in combating AMR in turkeys (the HOMAMR project): a two-cohort feasibility study using turkey-farm data that have been collected and analyzed retrospectively, followed by a prospective two-cohort study in turkey farms that would examine the impact of HMPs on changing antibiotic use. OBJECTIVE: The objective of this paper is to describe the protocol for the first phase of HOMAMR, which is a study under field conditions to assess the feasibility of collecting data retrospectively about turkeys that have been managed conventionally only or with HMPs. METHODS: Surveys performed on farms in Germany and Austria, including interviews with consenting farmers, will determine the feasibility of comparing retrospectively gathered data on antibiotic use and performance/production data in two cohorts of turkeys: (1) homeopathic treatment with conventional care (antibiotics) added if necessary or (2) conventional care (antibiotics) only. Co-primary outcomes to be studied are the amount of antimicrobial use and production period-related mortality. In addition, other production/performance parameters will be compared between the two treated cohorts. CONCLUSION: To our knowledge, this is the first feasibility study on the treatment of turkeys using homeopathy, and whose retrospectively obtained data will inform a prospective study that would examine the impact of HMPs on antibiotic use in commercial turkey raising, fattening and breeding production.

Animal welfare at German abattoirs: insights into the occurrence of violations against laws and regulations from official veterinarians and judicial decisions.

Introduction: This study investigated the occurrence of various animal welfare violations at German abattoirs by analyzing the results of an anonymous online survey and relevant German court decisions. Methods: The survey targeted official veterinarians (OVs) and other individuals responsible for enforcing animal welfare laws and regulations at German abattoirs. Participants were asked to report the five most common animal welfare violations in their workplaces during the past 3 years (2019-2021), and whether 22 specific given violations had occurred or not (in the same timeframe). The court decisions were examined to determine how many abattoirs were penalized for a specific animal welfare violation and the details about the number and species of affected animals. Results: Overall, the violations examined throughout this study fell into one of the following categories: (1) use of prohibited driving aids (e.g., illicit use of electric prods, or hitting/beating animals); (2) inadequate husbandry in lairage; (3) inappropriate handling of animals with special care needs; (4) improper restraint, stunning, and bleeding of animals; and (5) unqualified task execution/inadequate administrative work (e.g., documentation of a violation). The violations analyzed in the scope of this study most frequently fell into categories 1, 2, and 4. Regarding the five violations that survey participants reported to be the most common in the years 2019-2021, 42.6% of responses (N = 277 reported violations) fell into category 4, followed closely by category 1 with 37.9%. Of the 22 violations surveyed in the second part of the survey, those reported by 50.0% or more participants were concentrated in categories 1, 2, and 4. Regarding the 16 relevant court rulings spanning from 2015 to 2022, the most frequently documented instances of misconduct primarily fell into category 1. Discussion: The collected data suggest that there may be need for improvements regarding compliance with animal welfare legislation, especially concerning categories 1, 2, and 4. The authors suggest OVs to consider these findings when conducting monitoring activities at abattoirs and in the training and education of abattoir personnel.

Innovative e-Learning Training Modules to Improve Animal Welfare during Transport and Slaughter of Pigs: A Pretest-Posttest Study to Pre-Evaluate the General Didactical Concept.

In addition to the information on the possession of a certificate of competence, there are no concrete obligations for repetitive training for personnel handling live animals at transport and slaughter. Deficiencies in the animal-welfare-friendly handling of pigs are known. The developed pilot modules "Handling of pigs" and "Electrical stunning" were tested in a pretest-posttest study in German and Romanian using questions of knowledge before and after the implementation of the modules. In this study, 45 and 46 datasets of participants could be analyzed. The mean percentages of correctly answered questions in the posttest increased by 5.6% in the module "Handling of pigs" and by 10.6% in the module "Electrical stunning". A significant interaction was found for the language match and trend categories in the module "Handling of pigs". No Romanian native speaker had a positive trend in this module. For both modules separately, participant education level significantly interacted with the language match and the presence or absence of a certificate of competence. Comparing the percentages of the correct given answers, significant interactions in the subgroups were more common in the module "Electrical stunning". One question in "Electrical stunning" was correctly answered significantly more often in the posttest. Because of the positive mean trends of knowledge within this pre-evaluation, we assume the didactical concept was suitable for our target groups. Holders of a certificate of competence also gave more correct answers in the post-test. This underlines the importance of repetitive training. Differences in the trends of knowledge gain seem to be topic and experience related.

Opportunities to Improve Animal Welfare during Transport and Slaughter of Cattle and Pigs through Staff Training-Results of a Delphi Survey.

To improve animal welfare for cattle and pigs during transport and at slaughter, online training modules for all staff including employees in the lairage pen, the slaughter line as well as animal welfare officers are developed at Freie Universität Berlin, Germany. Before starting the development of these modules, an expert elicitation survey using a modified Delphi approach was performed to identify action points considered most relevant for animal welfare during transport and slaughter, and as having the potential for improvement through training. In total, 49 participating experts rated predetermined action points of each step in the transport and slaughter process in two survey rounds. The rating scale included numbers 0 (='not relevant', respectively 'no possibility of improvement') to 10 (='very relevant', respectively 'very high possibility of improvement'). None of the action points were rated with a median score of less than 5. Assessment of fitness for transport, unloading at the abattoir, handling at stunning and exsanguination were amongst the highest rated action points, and were therefore selected to develop online training modules. The Delphi approach was seen as a valuable method to include external expertise to select the most relevant action points for the development of online training modules.

Agar Contact Method as a Valuable Tool to Identify Slaughter Hygiene Deficiencies along the Slaughter Process by Longitudinally Sampling Pig Skin Surfaces.

Examinations of total viable counts (TVCs) and Salmonella spp. on the skin of individual pigs during the slaughter process are useful to identify abattoir-specific risk factors for (cross-)contamination. At seven process stages (lairage to before chilling), pigs were bacteriologically investigated by repeatedly sampling the same animals using the agar contact method. The mean TVC of all pigs increased significantly at the first three tested process stages (mean count, after delivery: 5.70 log cfu/cm2, after showering: 6.27 log cfu/cm2, after stunning: 6.48 log cfu/cm2). Significant mean TVC reductions occurred after scalding/dehairing (mean count: 3.71 log cfu/cm2), after singeing/flaming (2.70 log cfu/cm2), and after evisceration (2.44 log cfu/cm2) compared with the respective preceding process stages. At the end of the slaughter line and before chilling, the mean TVC was 2.33 log cfu/cm2, showing that the slaughter process reduced contamination significantly. The slaughter process effectively reduced even very high levels of incoming TVCs, since at the individual animal level, at the end of the slaughter process, there was no difference in the TVCs of animals with initially high and initially low TVCs. Additionally, 12 Salmonella spp. isolates were recovered from 12 different pigs, but only until the stage after scalding/dehairing. Overall, the agar contact method used is valuable for detecting hygiene deficiencies at slaughter, and is animal-equitable, practical, and suitable for use on live animals.

Detection of Extended-Spectrum ß-Lactamase (ESBL) E. coli at Different Processing Stages in Three Broiler Abattoirs.

The European Food Safety Authority (EFSA) identified extended-spectrum ß-lactamase/AmpC ß-lactamase (ESBL/AmpC)-producing E. coli as one of the main priority hazards for poultry. Different studies detected ESBL-producing E. coli at broiler fattening farms and in abattoirs, concluding that poultry meat is a potential source of human infection. Broiler breast skin samples taken in three abattoirs with different scalding techniques were examined for ESBL-producing Escherichia (E.) coli and their phylogenetic groups. A total of 307 ESBL-producing E. coli isolates were found, and the abattoir with conventional immersion scalding with thermal treatment of the water had the lowest incidence. Phylogroups D/E and B1 were mostly detected, while phylogroups C, D, and E were not detected. Phylogroup B2 was detected in low proportions. The phylogroups B2 and D are important as they have been associated with urinary tract infections in humans, but were only detected in low proportions at different processing stages in this study. Since the risk for the consumer of being infected via chicken meat with ESBL-producing E. coli and E. coli of highly pathogenic phylogroups cannot be excluded, good kitchen hygiene is of great importance.

Drinking Pipes and Nipple Drinkers in Pig Abattoir Lairage Pens-A Source of Zoonotic Pathogens as a Hazard to Meat Safety.

The water distribution system in the lairage pens of abattoirs could act as a route of contamination for produced meat. In this study, biofilm formation and the occurrence of specific pathogens in drinking equipment was investigated in different lairage pens in a German commercial pig abattoir. Samples of the water and the drinkers in different locations were microbiologically cultivated and examined. After new drinking equipment had been installed for one month, three months and five years, biofilm formation was detectable, and retrograde growth from the nipple drinkers was seen up to the connection with the main water distribution system. In particular, Enterobacteriaceae and Pseudomonas spp. were found in all samplings of the nipple drinkers. Zoonotic pathogens, Salmonella, pathogenic Yersinia enterocolitica and methicillin-resistant Staphylococcus aureus, were also isolated from the nipple drinkers, while Listeria monocytogenes was not detected via microbial cultivation methods in any of the samples. Since the pigs take the contaminated nipple drinkers into their mouths to drink, or drink contaminated water containing the pathogens, transmission and even infection of the pigs in the lairage can be assumed. This could consequently lead to contamination or cross-contamination of the meat during slaughter and processing and to a public health risk.

Verification of documentation plausibility in equine passports-drug documentation for geldings in comparison to self-reported veterinarian drug usage for equine castrations in Germany.

The 'positive list for equines' (Regulation (EC) No 1950/2006) was implemented in the European Union in 2006. The drugs listed are approved for use in slaughter equines under certain conditions, although those drugs are not approved for use in livestock and are not listed in Table 1 of the annex to Regulation (EU) No 37/2010. The usage of such drugs has to be documented in the equine passport and a withdrawal period of six months must be adhered to before the equine can be slaughtered for human consumption. Since the withdrawal period is long, correct documentation is particularly important. This study compared the results of two sub-studies. In sub-study 1, 116 veterinarians and nine equine clinics in Germany were surveyed about the methods and drugs used for castration of equine stallions. In sub-study 2, the documentational findings of 195 equine passports, belonging to 194 horses and one donkey, were analyzed. Regarding sub-study 1, the most commonly used method for castration was reported as 'laid down'. Drug combinations entailing at least one drug from the 'positive list' were used by 86.7% (91/105) of veterinarians castrating horse stallions 'laid down' and by 64.3% (36/56) of veterinarians utilizing this method on donkey stallions. Regarding sub-study 2, drug documentation was verified in the passports of 4.6% (9/195) of all equines and in just 12.0% (3/25) of those belonging to slaughter equine geldings. Anesthetics from the 'positive list' were documented in 4.0% (1/25) of equine passports belonging to slaughter geldings. Because of the high discrepancy of the drug combinations used by veterinarians and the documentation actually found in equine passports, we conclude that drug administration is very seldom documented in equine passports in Germany. This could result in drug residues in equine meat and poses a potential risk for consumers.

Measures and Penalties for Animal Welfare Violations at German Abattoirs: A Compilation of Current Recommendations and Practices.

Protecting animal welfare during preslaughter handling and slaughter is an important ethical concern with growing importance to consumers. However, animal welfare violations in abattoirs remain a serious problem, and the enforcement of relevant laws and regulations is often inadequate. This study investigated and compiled the measures and penalties which official veterinarians (OVs) consider appropriate for different animal welfare violations at German abattoirs, including ranges for fines. Additionally, information regarding which measures were taken in past cases, including past procedural outcomes (e.g., judicial decisions and regulatory animal welfare orders in Germany), were gathered and summarised. The aim is to provide insights into the status quo, so that flaws in law enforcement (e.g., imposing low penalties or not filing a criminal complaint when necessary) can be identified in a future study. To achieve this, the following five steps were utilised: acquiring relevant judicial decisions; conducting an anonymous online survey among German OVs; conducting semi-structured interviews with OVs; conducting a virtual colloquium with OVs; and consulting lawyers. Measures and penalties for violations of 40 relevant and frequent different provisions of the German Ordinance on the Protection of Animals in connection with Slaughter or Killing (TierSchlV), in conjunction with the Regulation (EC) No 1099/2009 and the German Animal Welfare Act, were gathered. The findings were compiled into a list of measures and penalties, which contains a separate table for all 40 violations, entailing an overview of the following information: citation(s) of legal/regulatory requirements to protect animals at the time of killing/slaughter; citation(s) of relevant regulatory and criminal penalties; special measures/penalties considered appropriate for the first and repeated offence by OVs; and information on penalties listed in judicial decisions of past similar cases. This initiative is a step towards achieving a reduction of animal welfare violations at German abattoirs.

Comparing Visual-Only and Visual-Palpation Post-Mortem Lung Scoring Systems in Slaughtering Pigs.

Respiratory diseases continue to pose significant challenges in pig production, and the assessment of lung lesions at the abattoir can provide valuable data for epidemiological investigations and disease surveillance. The evaluation of lung lesions at slaughter is a relatively simple, fast, and straightforward process but variations arising from different abattoirs, observers, and scoring methods can introduce uncertainty; moreover, the presence of multiple scoring systems complicates the comparisons of different studies, and currently, there are limited studies that compare these systems among each other. The objective of this study was to compare validated, simplified, and standardized schemes for assessing surface-related lung lesions in slaughtered pigs and analyze their reliability under field conditions. This study was conducted in a high-throughput abattoir in Italy, where two different scoring methods (Madec and Blaha) were benchmarked using 637 plucks. Statistical analysis revealed a good agreement between the two methods when severe or medium lesions were observed; however, their ability to accurately identify healthy lungs and minor injuries diverged significantly. These findings demonstrate that the Blaha method is more suitable for routine surveillance of swine respiratory diseases, whereas the Madec method can give more detailed and reliable results for the respiratory and welfare status of the animals at the farm level.

A comparative survey of veterinarians, equine owners, and equine keepers regarding the knowledge and implementation of legal requirements in Germany for the use and documentation of veterinary medicines in equines intended for slaughter.

In Europe, equines destined for human consumption (hereafter called slaughter equines) are subject to the same restrictions of usage of veterinary drugs as other food-producing animals, with amendments regulated in the so-called 'positive list', Regulation (EC) No. 1950/2006. Due to the complex legal requirements for drug administration in slaughter equines, it might be that specific knowledge regarding the legislation of slaughter equines may be insufficient among veterinarians, equine owners, and equine keepers. To study this assumption, three target group-specific surveys were conducted in 2021. Answers from 153 equine treating veterinarians, 170 equine owners, and 70 equine keepers were included in the analysis. In total 68.4% (91/133) of the participating veterinarians, the regulations of the 'positive list', Regulation (EC) No. 1950/2006, were 'rather complicated' to 'complicated'. Among the participating veterinarians, 38.4% (58/151) did not or could not answer correctly how to proceed if a slaughter equine is scheduled to receive phenylbutazone, usage of which is prohibited in all livestock by Regulation (EU) No. 37/2010. Simultaneously, 56.2% (86/153) of the participating veterinarians named phenylbutazone as the, or one of the, most often used non-steroidal anti-inflammatory drugs. Altogether, 41.2% (70/170) of participating equine owners and 42.9% (30/70) of equine keepers did not know under which circumstances an equine can legally be slaughtered for human consumption. In total, 34.3% (24/70) of the equine keepers classified their knowledge of national regulations for animal keepers regarding the documentation of drug usage in equines as 'poor' to 'nonexistent'. This lack of knowledge in all three surveyed groups, combined with the complex legal regulations regarding the usage and documentation of drugs in slaughter equines, could result in missing and false documentation, treatment of slaughter equines with prohibited substances and therefore pose a risk factor for drug residues in equine meat.

Impact of Procedures and Human-Animal Interactions during Transport and Slaughter on Animal Welfare of Pigs: A Systematic Literature Review.

Animal welfare is a high social and political priority and is enshrined in European and national legislation. This systematic literature review (funded by the German Ministry of Food and Agriculture; grant no. 2817806A18) was conducted to find animal and management-based indicators that could be influenced by changes in pig handling and management procedures on the day of slaughter and, therefore, will directly result in improved animal welfare. For this systematic literature review, following the PRISMA guidelines, we conducted a structured literature search using the databases PubMed®, Web of ScienceTM, and LIVIVO with set search terms and their combinations in German and English. Only peer-reviewed original articles from European countries from 2009 until 2022 that dealt with welfare during transport and/or slaughter of fattening pigs and offered potential measurable indicators on the basis of the animal or management, and either directly or indirectly recommended handling practices were included. We used the literature management system EndNoteTM, and after duplicate removal, a total of 1099 records were found for further analysis. After analysis and discussion in the review team, which consisted of three experts in total, 105 articles underwent full-text screening. Finally, 39 articles were included in the results for this systematic literature review. According to this systematic literature review, the following procedures have a positive impact on the animal welfare of pigs on the day of slaughter. Calm-rather than rough-handling of pigs was the most influential factor. This can be achieved by using animal-friendly driving aids. Loud noise should be avoided in general or reduced as far as possible. Transport time is not always influenceable, but shorter transport duration with stocking density as stated by the European regulations as a maximum will reduce stress in pigs. Our review showed that there are differing scientific results about lairage duration, although this should be set in relation to prior stress and transport time. Knowledge of good stunning parameters, such as gas concentration, exposure time to the gas, recommended amperage, and current flow duration is essential. If electrical stunning is performed, the correct attachment of electrodes is crucial. The control of stunning effectiveness is extremely important to avoid suffering and pain, with the proviso that it is assessed on the basis of multiple parameters. The corneal reflex, regular gasping, and the up-righting reflex could be assessed together. Certificates of competence and continuing educational programmes for the personnel form the basis of animal-friendly handling and a stressless slaughter routine. It can be assumed that the results of this systematic literature review reflect the state of current research of pig welfare during transport and slaughter in the European Union, even though it must be assumed that not all relevant points were covered by the review, which can be seen as a limitation at the same time because important non-European or older publications could not be included.

The microbiological and sensory status of dual-purpose chickens (Lohmann Dual), male Lohmann Brown Plus chickens, and conventional laying hens slaughtered in a laying hen abattoir compared to conventional broilers slaughtered in a broiler abattoir.

Alternatives to conventional chicken meat and egg production are increasingly under discussion, especially because of the common practice of killing male day-old chicks from laying lines which has been banned from the beginning of 2022 in Germany and is planned to be banned during 2022 in other countries. Production of dual-purpose chicken lines is one possible solution, as such lines combine moderate laying and growth performance. The microbiological status of products from such breeds must be comparable to existing products on the market for food safety purposes. Additionally, the production of such products will take longer because of the feeding regimes required, and again, comparability should be safeguarded for the best consumer protection. The dual-purpose chicken line, Lohmann Dual (males), was compared to males from the laying line Lohmann Brown Plus, conventional laying hens (all slaughtered and processed in the same conventional laying hen abattoir), and conventional broilers (slaughtered in a conventional broiler abattoir). Neck skin samples were taken before chilling at the end of each slaughter line to determine the microbial status of the carcasses. Additionally, fresh and cooked meat sensory analysis was performed on meat from broilers and male and female Lohmann Dual and Lohmann Brown Plus chickens (for three carcasses of each group) at the German Agricultural Society Test Center in Kassel. The focus was on the performance of male Lohmann Dual compared to the other lines. There was no difference in the Enterobacteriaceae count of the dual-purpose chicken line compared to conventional broilers, whereas laying hens had a significantly higher microbial load before chilling, as based on neck skin examinations (p<0.001). According to sensory test results, the meat from dual-purpose chickens was the best (as no defects were found) among the five chicken meat types examined. In conclusion, based on their microbial status and sensory analysis of fresh and cooked meat, Lohmann Dual males slaughtered in a laying hen abattoir can be considered as an alternative to conventionally kept broilers slaughtered in a broiler abattoir.

A European survey on post-mortem inspection of finishing pigs: Total condemnation criteria to declare meat unfit for human consumption.

Meat inspection (MI) is essential to verify compliance with legal requirements related to human and animal health and animal welfare protections. Judgement criteria applied during MI, resulting in condemnation data of importance, among other things, for livestock producers and for benchmarking reasons. However, although the Meat Inspection Regulation sets out judgement criteria, most are generic, favouring flexibility, but also subjectivity. To address the degree of variation on total condemnation (TC) criteria applied during post-mortem inspection (PMI) of finishing pigs, an online survey was prepared aiming to collect this information from several European countries. The focus was on TC criteria regarding the following PMI findings: abscesses, arthritis, cachexia, erysipelas, icterus, Mycobacterium-like lesions, osteomyelitis, peritonitis, pleuritis and pneumonia. From September to November 2020, a total of 44 completed questionnaires were obtained from 26 European countries. The results showed a substantial variation in the TC criteria in place in the participating countries. One of the main reasons for the variability seen in the respondents' reported answers was related to the indicators used to define a generalised condition related to the 10 PMI findings addressed, making harmonisation a challenge and avoiding to draw conclusions when comparing condemnation causes between abattoirs. This implies that it would make sense to look into how a generalised condition can be identified/described and how it should be judged. The results should be used as inspiration towards possible harmonisation, improving decision-making, and permitting comparative analysis between different reports to allow trend analyses and benchmarking.

Characterization of qnrB-carrying plasmids from ESBL- and non-ESBL-producing Escherichia coli.

BACKGROUND: Escherichia coli carrying clinically important antimicrobial resistances [i.e., against extended-spectrum-beta-lactamases (ESBL)] are of high concern for human health and are increasingly detected worldwide. Worryingly, they are often identified as multidrug-resistant (MDR) isolates, frequently including resistances against quinolones/fluoroquinolones. RESULTS: Here, the occurrence and genetic basis of the fluoroquinolone resistance enhancing determinant qnrB in ESBL-/non-ESBL-producing E. coli was investigated. Overall, 33 qnrB-carrying isolates out of the annual German antimicrobial resistance (AMR) monitoring on commensal E. coli (incl. ESBL-/AmpC-producing E. coli) recovered from food and livestock between 2013 and 2018 were analysed in detail. Whole-genome sequencing, bioinformatics analyses and transferability evaluation was conducted to characterise the prevailing qnrB-associated plasmids. Furthermore, predominant qnrB-carrying plasmid-types were subjected to in silico genome reconstruction analysis. In general, the qnrB-carrying E. coli were found to be highly heterogenic in their multilocus sequence types (STs) and their phenotypic resistance profiles. Most of them appeared to be MDR and exhibited resistances against up to ten antimicrobials of different classes. With respect to qnrB-carrying plasmids, we found qnrB19 located on small Col440I plasmids to be most widespread among ESBL-producing E. coli from German livestock and food. This Col440I plasmid-type was found to be highly conserved by exhibiting qnrB19, a pspF operon and different genes of unassigned function. Furthermore, we detected plasmids of the incompatibility groups IncN and IncH as carriers of qnrB. All qnrB-carrying plasmids also exhibited virulence factors and various insertion sequences (IS). The majority of the qnrB-carrying plasmids were determined to be self-transmissible, indicating their possible contribution to the spread of resistances against (fluoro)quinolones and other antimicrobials. CONCLUSION: In this study, a diversity of different plasmid types carrying qnrB alone or in combination with other resistance determinants (i.e., beta-lactamase genes) were found. The spread of these plasmids, especially those carrying antimicrobial resistance genes against highest priority critically important antimicrobial agents, is highly unfavourable and can pose a threat for public health. Therefore, the dissemination pathways and evolution of these plasmids need to be further monitored.

Seroprevalence and Associated Risk Factors of Trichinellosis and T. Solium Cysticercosis in Indigenous Pigs in Hoa Binh Province, Vietnam.

Trichinellosis and cysticercosis remain challenges to human health and animal productivity worldwide, especially in developing countries. While information on the occurrence of both diseases is infrequent, they are endemic in parts of Vietnam and mainly related to indigenous pigs kept by ethnic minorities. This study aimed to determine the seroprevalence and risk factors of both diseases in indigenous pigs and explore the perception and awareness of both human and pig trichinellosis and cysticercosis of pig farmers. A total of 352 pig sera samples from 131 holdings were collected and analyzed using ELISA antibody tests in six communes in the Da Bac districts of Hoa Binh province, Vietnam. A survey was conducted with representatives from these households to understand the knowledge and perspective on food-borne parasitic diseases. Overall, the seroprevalence of trichinellosis and T. solium cysticercosis was 13.6% (95% CI 10.2-17.7) and 1.7% (95% CI 0.6-3.7), respectively. The seroprevalence of trichinellosis was significantly higher in female and older pigs. Risk perception and knowledge of interviewed people on both human and pig trichinellosis and cysticercosis of pig farmers was poor. Risky practices, including free roaming of pigs and eating undercooked or fermented pork, were observed. Educational and awareness campaigns aligned with further research on feasible practice changes are critical to addressing these issues.

Distribution and Characteristics of Listeria spp. in Pigs and Pork Production Chains in Germany.

Listeria (L.) monocytogenes is a foodborne pathogen that can cause disease, mainly in elderly, pregnant or immunocompromised persons through consumption of contaminated food, including pork products. It is widespread in the environment and can also be found in asymptomatic carrier animals, for example, in different tissues of pigs. To learn more about their nature, 16 Listeria spp. isolates found in tonsils and intestinal content of pigs and 13 isolates from the slaughterhouse environment were characterized using next-generation sequencing (NGS). A wide distribution of clonal complexes was observed in pigs, as well as in the pork production chain, suggesting multiple sources of entry. Hypervirulent clones were found in pig tonsils, showing the potential risk of pigs as source of isolates causing human disease. The presence of closely related isolates along the production chain suggests a cross-contamination in the slaughterhouse or recontamination from the same source, strengthening the importance of efficient cleaning and disinfection procedures. The phenotypical antimicrobial resistance status of L. monocytogenes isolates was examined via broth microdilution and revealed a low resistance level. Nevertheless, genotypical resistance data suggested multiple resistances in some non-pathogenic L. innocua isolates from pig samples, which might pose a risk of spreading resistances to pathogenic species.

Dissection of Highly Prevalent qnrS1-Carrying IncX Plasmid Types in Commensal Escherichia coli from German Food and Livestock.

Plasmids are mobile genetic elements, contributing to the spread of resistance determinants by horizontal gene transfer. Plasmid-mediated quinolone resistances (PMQRs) are important determinants able to decrease the antimicrobial susceptibility of bacteria against fluoroquinolones and quinolones. The PMQR gene qnrS1, especially, is broadly present in the livestock and food sector. Thus, it is of interest to understand the characteristics of plasmids able to carry and disseminate this determinant and therewith contribute to the resistance development against this class of high-priority, critically important antimicrobials. Therefore, we investigated all commensal Escherichia (E.) coli isolates, with reduced susceptibility to quinolones, recovered during the annual zoonosis monitoring 2017 in the pork and beef production chain in Germany (n = 2799). Through short-read whole-genome sequencing and bioinformatics analysis, the composition of the plasmids and factors involved in their occurrence were determined. We analysed the presence and structures of predominant plasmids carrying the PMQR qnrS1. This gene was most frequently located on IncX plasmids. Although the E. coli harbouring these IncX plasmids were highly diverse in their sequence types as well as their phenotypic resistance profiles, the IncX plasmids-carrying the qnrS1 gene were rather conserved. Thus, we only detected three distinct IncX plasmids carrying qnrS1 in the investigated isolates. The IncX plasmids were assigned either to IncX1 or to IncX3. All qnrS1-carrying IncX plasmids further harboured a ß-lactamase gene (bla). In addition, all investigated IncX plasmids were transmissible. Overall, we found highly heterogenic E. coli harbouring conserved IncX plasmids as vehicles for the most prevalent qnr gene qnrS1. These IncX plasmids may play an important role in the dissemination of those two resistance determinants and their presence, transfer and co-selection properties require a deeper understanding for a thorough risk assessment.

A Systematic Review on the Effectiveness of Pre-Harvest Meat Safety Interventions in Pig Herds to Control Salmonella and Other Foodborne Pathogens.

This systematic review aimed to assess the effectiveness of pre-harvest interventions to control the main foodborne pathogens in pork in the European Union. A total of 1180 studies were retrieved from PubMed® and Web of Science for 15 pathogens identified as relevant in EFSA's scientific opinion on the public health hazards related to pork (2011). The study selection focused on controlled studies where a cause-effect could be attributed to the interventions tested, and their effectiveness could be inferred. Altogether, 52 studies published from 1983 to 2020 regarding Campylobacter spp., Clostridium perfringens, Methicillin-resistant Staphylococcus aureus, Mycobacterium avium, and Salmonella spp. were retained and analysed. Research was mostly focused on Salmonella (n = 43 studies). In-feed and/or water treatments, and vaccination were the most tested interventions and were, overall, successful. However, the previously agreed criteria for this systematic review excluded other effective interventions to control Salmonella and other pathogens, like Yersinia enterocolitica, which is one of the most relevant biological hazards in pork. Examples of such successful interventions are the Specific Pathogen Free herd principle, stamping out and repopulating with disease-free animals. Research on other pathogens (i.e., Hepatitis E, Trichinella spiralis and Toxoplasma gondii) was scarce, with publications focusing on epidemiology, risk factors and/or observational studies. Overall, high herd health coupled with good management and biosecurity were effective to control or prevent most foodborne pathogens in pork at the pre-harvest level.

Genomic Characterization of Multidrug-Resistant Salmonella Serovars Derby and Rissen From the Pig Value Chain in Vietnam.

Nontyphoidal Salmonella (NTS) is the most reported cause of bacterial foodborne zoonoses in Vietnam, and contaminated pork is one of the main sources of human infection. In recent years, the prevalence of NTS carrying multiple antimicrobial resistance genes (ARGs) have been increased. The genomic characterization along the pig value chain and the identification of ARGs and plasmids have the potential to improve food safety by understanding the dissemination of ARGs from the farm to the table. We report an analysis of 13 S. Derby and 10 S. Rissen isolates, collected in 2013 at different stages in Vietnamese slaughterhouses and markets. VITEK 2 Compact System was used to characterize the phenotypical antimicrobial resistance of the isolates. In addition, whole-genome sequencing (WGS) was used to detect ARGs and plasmids conferring multidrug resistance. Whole genome single nucleotide polymorphism typing was used to determine the genetic diversity of the strains and the spread of ARGs along the pig value chain. Altogether, 86.9% (20/23) of the samples were resistant to at least one antibiotic. Resistance to ampicillin was most frequently detected (73.9%), followed by piperacillin and moxifloxacin (both 69.6%). At least one ARG was found in all strains, and 69.6% (16/23) were multidrug-resistant (MDR). The observed phenotype and genotype of antimicrobial resistance were not always concordant. Plasmid replicons were found in almost all strains [95.6% (22/23)], and the phylogenetic analysis detected nine clusters (S. Derby, n = 5; S. Rissen, n = 4). ARGs and plasmid content were almost identical within clusters. We found six MDR IncHI1s with identical plasmid sequence type in strains of different genetic clusters at the slaughterhouse and the market. In conclusion, high rates of multidrug resistance were observed in Salmonella strains from Vietnam in 2013. Genomic analysis revealed many resistance genes and plasmids, which have the potential to spread along the pig value chain from the slaughterhouse to the market. This study pointed out that bioinformatics analyses of WGS data are essential to detect, trace back, and control the MDR strains along the pig value chain. Further studies are necessary to assess the more recent MDR Salmonella strains spreading in Vietnam.