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.

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.

Use of meat juice and blood serum with a miniaturised protein microarray assay to develop a multi-parameter IgG screening test with high sample throughput potential for slaughtering pigs.

BACKGROUND: Serological screening of pig herds at the abattoir is considered a potential tool to improve meat inspection procedures and herd health management. Therefore, we previously reported the feasibility of a miniaturised protein microarray as a new serological IgG screening test for zoonotic agents and production diseases in pigs. The present study investigates whether the protein microarray-based assay is applicable for high sample throughput using either blood serum or meat juice. MATERIAL AND METHODS: Microarrays with 12 different antigens were produced by Abbott (formerly Alere Technologies GmbH) Jena, Germany in a previously offered 'ArrayTube' platform and in an 'ArrayStrip' platform for large-scale use. A test protocol for the use of meat juice on both microarray platforms was developed. Agreement between serum and meat juice was analysed with 88 paired samples from three German abattoirs. Serum was diluted 1:50 and meat juice 1:2. ELISA results for all tested antigens from a preceding study were used as reference test to perform Receiver Operating Characteristic analysis for both test specimens on both microarray platforms. RESULTS: High area under curve values (AUC > 0.7) were calculated for the analysis of T. gondii (0.87), Y. enterocolitica (0.97), Mycoplasma hyopneumoniae (0.84) and Actinobacillus pleuropneumoniae (0.71) with serum as the test specimen and for T. gondii (0.99), Y. enterocolitica (0.94), PRRSV (0.88), A. pleuropneumoniae (0.78) and Salmonella spp. (0.72) with meat juice as the test specimen on the ArrayStrip platform. Cohens kappa values of 0.92 for T. gondii and 0.82 for Y. enterocolitica were obtained for the comparison between serum and meat juice. When applying the new method in two further laboratories, kappa values between 0.63 and 0.94 were achieved between the laboratories for these two pathogens. CONCLUSION: Further development of a miniaturised pig-specific IgG protein microarray assay showed that meat juice can be used on microarray platforms. Two out of twelve tested antigens (T. gondii, Y. enterocolitica) showed high test accuracy on the ArrayTube and the ArrayStrip platform with both sample materials.

High Predictive Power of Meat Juice Serology on the Presence of Hepatitis E Virus in Slaughter Pigs.

Hepatitis E virus (HEV) as a zoonotic agent can be responsible for an acute hepatitis in humans, which is usually self-limiting. Progression toward a chronic stage is possible, especially in immunocompromised patients. In the past decade, the number of hepatitis E cases in humans in Germany has increased enormously to 3491 cases in 2018. Domestic pigs have been identified as a main animal reservoir and the consumption of raw and undercooked pork products, that is, livers or liver products, meat or meat products, is known as a potential risk of foodborne HEV infection. The aim of this study was to determine whether serological tests are appropriate to predict the occurrence of HEV in the liver and muscle of domestic pigs in Germany. In 2018, samples of meat juice, liver, and ham muscle were collected from 250 fattening pigs at an abattoir in North West Germany. Samples were analyzed for the presence of HEV antibodies using enzyme-linked immunosorbent assay respectively for the presence of HEV RNA using real-time reverse transcription-polymerase chain reaction. In total, 62% (155/250) of the meat juice samples were positive for HEV antibodies at a single animal basis. At herd level, 72% (18/25) of the herds were seropositive. The HEV prevalence in the liver was 17.2% (43/250). Each positive liver sample originated from seropositive herds respectively from HEV seropositive pigs. This study demonstrates for the first time the significant correlation between a positive HEV serology and the occurrence of HEV RNA in the liver of slaughter pigs (χ2 = 31.83; p < 0.001), highlighting the significant predictive power of positive serological results on the occurrence of HEV RNA in the liver.

Towards a Standardized Method for Broth Microdilution Susceptibility Testing of Haemophilus parasuis.

Currently, there is no agreed method available for broth microdilution susceptibility testing of Haemophilus parasuis, one of the most important bacterial pathogens in pig production. Therefore, the aim of this study was to develop a method that could be easily performed by diagnostic laboratories and that appears suitable for a harmonized susceptibility testing. Growth determinations using one type strain and three field isolates revealed no visible growth of H. parasuis in media which have proven to be suitable for susceptibility testing of fastidious organisms. Therefore, a new medium, cation-adjusted Mueller-Hinton broth (CAMHB) plus NADH and sterile filtered heat-inactivated chicken serum, was developed. The reproducibility of MICs obtained in this medium was evaluated and statistically analyzed, considering a model with two different variables (precondition of five identical MICs and MIC mode accepting a deviation of ±1 dilution step, respectively). No significant differences for both variables were seen between two time points investigated and between results obtained with the recently proposed test medium broth (TMB). Nearly all MICs of quality control strains were in the acceptable range. Subsequently, 47 H. parasuis isolates representing 13 serovars were tested with the newly developed medium and TMB. Statistical analysis of all isolates and 15 antimicrobial agents and antimicrobial combinations showed no significant difference between MICs obtained in supplemented CAMHB and TMB. Because of a simplified implementation in routine diagnostic and a lower chance of interference between medium components and antimicrobial agents, supplemented CAMHB is recommended with an incubation time of 24 h.

A simplified evaluation system of surface-related lung lesions of pigs for official meat inspection under industrial slaughter conditions in Germany.

BACKGROUND: European and national administrative legislation require objective evaluation systems for organ lesions at pig slaughter. These results can be used as basis for herd health improvement programs by farmers and their consulting veterinarians. Various studies have shown that the current evaluation and recording of lesions by authorized meat inspectors are not reliable and produce significant inter-rater disagreement especially for lung lesions in pigs. The objectives of this study were to increase the usability of official meat inspection data by a developed and validated scheme and to analyze potential improvements in the reliability of the proposed system under industrialized slaughter conditions. RESULTS: A simplified evaluation scheme for surface-related lung lesions was developed based on morphometric evaluations of unaffected lungs with quantitative relationships of each lobe to the whole lung ("Rule of Tens"). Furthermore, a theoretical as well as a hands-on training program for meat inspectors was developed and applied. Based on 5,183 lungs, the authors established a baseline of the inter-rater reliability of current routine assessments of lung lesions as documented by meat inspectors compared with the assessments of an independent veterinarian using the developed simplified evaluation scheme. Most frequent inter-rater disagreements greater than 75% were found for moderate pneumonia. Sources of the deviations most frequently included misinterpretations of technical artifacts, which were erroneously assessed by the meat inspectors as pneumonic lung lesions. Results of the post-training investigation based on 4,646 lungs showed a significantly improved reliability of lung lesion evaluation and the inter-rater agreement increased in all respects. Especially the disagreement of recording moderate cases of pneumonia decreased in total to 15% deviations from reference. CONCLUSIONS: The presented simplified lung evaluation scheme showed its capability to standardize the evaluation of lung lesions according to administrative legislation under industrialial slaughter conditions. The reliability of official meat inspections can be significantly increased with the help of the presented scheme to assess surface-related lung lesions of slaughter pigs. Continuous standardization and optimization can be achieved by personalized training programs in the framework of quality assurance systems for meat inspectors.

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.

Genotypic and phenotypic characterization of Staphylococcus aureus isolates from wild boars.

Eight Staphylococcus aureus isolates collected from 117 wild boars were characterized and compared to livestock isolates. They belonged to sequence types ST133, ST425, and the new type ST1643. The spa types were t1181, t6782, and the new types t6384, t6385, and t6386. Antimicrobial susceptibility testing and microarray-based genotyping confirmed the absence of important virulence/resistance genes.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.