Inspection of chicken wings and legs for animal welfare monitoring using X-ray computed tomography, visual examination, and histopathology.

In broiler chickens, fractures of wings and legs are recorded at poultry slaughterhouses based on the time of occurrence. Prekilling (PRE) fractures occur before the death of animal, so the chicken was still able to experience pain and distress associated with the injury (an animal welfare issue). Postkilling (POST) fractures occur when the chickens are deceased and fully bled-out and consequently unable to feel pain (not an animal welfare issue). Current practice dictates that fractures are recognized visually and recorded by the animal welfare officers as mandated by European Union and/or national regulations. However, new potential monitoring solutions are desired since human inspection suffers from some significant limitations including subjectivism and fatigue. One possible solution in detecting injuries is X-ray computed tomography (CT) scanning and in this study we aim to evaluate the potential of CT scanning and visual inspection in detecting limb fractures and their causes. Eighty-three chicken wings and 60 chicken legs (n = 143) were collected from a single slaughterhouse and classified by an animal welfare officer as PRE, POST or healthy (HEAL). Samples were photographed and CT scanned at a veterinary hospital. The interpretation of CT scans along with photographs took place in 3 rounds (1. CT scans only, 2. CT scans + photographs, 3. photographs only) and was performed independently by 3 veterinarians. The consistency of the interpretation in 3 rounds was compared with the animal welfare officer's classification. Furthermore, selected samples were also analyzed by histopathological examination due to questionability of their classification (PRE/POST). In questionable samples, presence of hemorrhages was confirmed, thus they fit better as PRE. The highest consistency between raters was obtained in the 2nd round, indicating that interpretation accuracy was the highest when CT scans were combined with photographs. These results indicate that CT scanning in combination with visual inspection can be used in detecting limbs fracture and potentially applied as a tool to monitor animal welfare in poultry slaughterhouses in the future.

The Value of Current Ante Mortem Meat Inspection and Food Chain Information of Dairy Cows in Relation to Post Mortem Findings and the Protection of Public Health: A Case for a More Risk-Based Meat Inspection.

In this study, the contribution of the ante mortem (AM) inspection and the food chain information (FCI) to ensuring meat safety and public health was investigated, by evaluating the slaughterhouse findings of 223,600 slaughtered dairy cows in the Netherlands. The outcome of this study was that the ante mortem (AM) and post mortem (PM) inspections have a substantial overlap, and that with regard to food safety and public health in over 99% of cases the PM could even be omitted on the basis of the AM. In this study, the data provided by the dairy farmers on the current FCI forms contributed little to nothing with regard to the outcomes of AM and PM inspection. It is concluded that current meat inspection procedures need an update and a more risk-based approach needs to be adopted. Regarding this, the AM inspection of dairy cattle should remain, because it plays an important role in ensuring food safety (e.g., by preventing contamination of the slaughter line by excessively dirty animals, or animals with abscesses), monitoring animal welfare and in detecting some important notifiable diseases. The PM inspection, however, could in many cases be omitted, provided there is a strict AM inspection complemented by a vastly improved (automated) way of obtaining reliable FCI.

Small Contaminations on Broiler Carcasses Are More a Quality Matter than a Food Safety Issue.

Depending on the interpretation of the European Union (EU) regulations, even marginally visibly contaminated poultry carcasses could be rejected for human consumption due to food safety concerns. However, it is not clear if small contaminations actually increase the already present bacterial load of carcasses to such an extent that the risk for the consumers is seriously elevated. Therefore, the additional contribution to the total microbial load on carcasses by a small but still visible contamination with feces, grains from the crop, and drops of bile and grease from the slaughter line was determined using a Monte Carlo simulation. The bacterial counts (total aerobic plate count, Enterobacteriaceae, Escherichia coli, and Campylobacter spp.) were obtained from the literature and used as input for the Monte Carlo model with 50,000 iterations for each simulation. The Monte Carlo simulation revealed that the presence of minute spots of feces, bile, crop content, and slaughter line grease do not lead to a substantial increase of the already existing biological hazards present on the carcasses and should thus be considered a matter of quality rather than food safety.

Analysis of a Clostridium difficile PCR ribotype 078 100 kilobase island reveals the presence of a novel transposon, Tn6164.

BACKGROUND: Clostridium difficile is the main cause of antibiotic associated diarrhea. In the past decade, the number of C. difficile patients has increased dramatically, coinciding with the emergence of two PCR ribotypes 027 and 078. PCR ribotype 078 is also frequently found during C. difficile outbreaks in pigfarms. Previously, the genome of the PCR ribotype 078 strain M120, a human isolate, was described to contain a unique insert of 100 kilobases. RESULTS: Analysis of this insert revealed over 90 open reading frames, encoding proteins originating from transposons, phages and plasmids. The insert was shown to be a transposon (Tn6164), as evidenced by the presence of an excised and circularised molecule, containing the ligated 5'and 3'ends of the insert. Transfer of the element could not be shown through filter-mating experiments. Whole genome sequencing of PCR ribotype 078 strain 31618, isolated from a diarrheic piglet, showed that Tn6164 was not present in this strain. To test the prevalence of Tn6164, a collection of 231 Clostridium difficile PCR ribotype 078 isolates from human (n = 173) and porcine (n = 58) origin was tested for the presence of this element by PCR. The transposon was present in 9 human, tetracycline resistant isolates, originating from various countries in Europe, and none of the pig strains. Nine other strains, also tetracycline resistant human isolates, contained half of the transposon, suggesting multiple insertion steps yielding the full Tn6164. Other PCR ribotypes (n = 66) were all negative for the presence of the transposon. Multi locus variable tandem repeat analysis revealed genetic relatedness among transposon containing isolates. Although the element contained several potential antibiotic resistance genes, it did not yield a readily distinguishable phenotype. CONCLUSIONS: Tn6164 is a newly described transposon, occurring sporadically in C. difficile PCR ribotype 078 strains. Although no transfer of the element could be shown, we hypothesize that the element could serve as a reservoir of antibiotic resistance genes for other bacteria. Further research is needed to investigate the transfer capabilities of the element and to substantiate the possible role of Tn6164 as a source of antibiotic resistance genes for other gut pathogens.

Allergy among veterinary medicine students in The Netherlands.

BACKGROUND: Veterinary medicine students who practice with animals are potentially exposed to many occupational agents, yet sensitisation and allergic symptoms among this group have not been studied extensively. OBJECTIVE: The objective of this study was to estimate the prevalence of sensitisation and allergic symptoms in veterinary medicine students in association with study specialisation over time. METHODS: A questionnaire-based cross-sectional study was conducted. Blood was collected and tested for total and specific serum IgE for 16 different common and study-specific allergens using enzyme immunoassay. RESULTS: New development of self-reported allergic symptoms to various allergens occurred in 8.7%, of which 44% was deducted against animals. Handling farm animals was strongly associated with self-reported allergies to various allergens (OR=6.9, 95% CI 1.9 to 25) and animal allergens (OR=12, 95% CI 1.4 to 103). Sensitisation to at least one allergen occurred in 33.1%. Sensitisation prevalence tended to be elevated in later years of the equine study program. In contrast to self-reported allergies, the prevalence of sensitisation to any allergen decreased with prolonged study duration for those specialising in farm animal health (years 3-5: OR=0.5, 95% CI 0.3 to 1.1; year 6: OR=0.2, 95% CI 0.1 to 0.5). This was independent of whether people were raised on a farm, which is in itself a protective factor for allergy and sensitisation. CONCLUSION: This study provides evidence of an elevated prevalence of allergic symptoms with increasing years of veterinary study, suggesting that contact with animals, more specifically contact to farm animals, is a risk factor for the development of symptoms.

A One Health Perspective on the Human-Companion Animal Relationship with Emphasis on Zoonotic Aspects.

Over time the human-animal bond has been changed. For instance, the role of pets has changed from work animals (protecting houses, catching mice) to animals with a social function, giving companionship. Pets can be important for the physical and mental health of their owners but may also transmit zoonotic infections. The One Health initiative is a worldwide strategy for expanding collaborations in all aspects of health care for humans, animals, and the environment. However, in One Health communications the role of particularly dogs and cats is often underestimated. OBJECTIVE: Evaluation of positive and negative One Health issues of the human-companion animal relationship with a focus on zoonotic aspects of cats and dogs in industrialized countries. METHOD: Literature review. RESULTS: Pets undoubtedly have a positive effect on human health, while owners are increasing aware of pet's health and welfare. The changing attitude of humans with regard to pets and their environment can also lead to negative effects such as changes in feeding practices, extreme breeding, and behavioral problems, and anthropozoonoses. For the human, there may be a higher risk of the transmission of zoonotic infections due to trends such as sleeping with pets, allowing pets to lick the face or wounds, bite accidents, keeping exotic animals, the importation of rescue dogs, and soil contact. CONCLUSIONS: One Health issues need frequently re-evaluated as the close human-animal relationship with pet animals can totally differ compared to decennia ago. Because of the changed human-companion animal bond, recommendations regarding responsible pet-ownership, including normal hygienic practices, responsible breeding, feeding, housing, and mental and physical challenges conforming the biology of the animal are required. Education can be performed by vets and physicians as part of the One Health concept.

Rat bite fever.

Rat bite fever (RBF) is a bacterial zoonosis for which two causal bacterial species have been identified: Streptobacillis moniliformis and Spirillum minus. Haverhill fever (HF) is a form of S. moniliformis infection believed to develop after ingestion of contaminated food or water. Here the infectious agents, their host species, pathogenicity (virulence factors and host susceptibility), diagnostic methods, therapy, epidemiology, transmission and prevention are described. Special emphasis is given on information from the field of laboratory animal microbiology and suggestions for future research.

Arcobacter spp. possess two very short flagellins of which FlaA is essential for motility.

Like Campylobacter and Helicobacter spp., Arcobacter spp. possess two flagellin genes (flaA and flaB) located adjacent to each other. The aim of this study was to characterize the flagellin proteins of Arcobacter spp., because these proteins are known virulence factors in the Epsilonproteobacteria, to which these three species belong. With the exception of Arcobacter nitrofigilis, Arcobacter flagellins are almost half the size of those in other Epsilonproteobacteria. Arcobacter flagellin proteins lack a large part of the variable central region. The low homology observed among flagellins of different Arcobacter species indicates genetic heterology between the members of this genus. Unlike in other Epsilonproteobacteria, the transcription of flagellin genes is not regulated by sigma 28- or sigma 54-dependent promoters, which suggests that transcription must be regulated in a different way in Arcobacter spp. Mutational studies revealed that only FlaA is needed for the motility of Arcobacter spp. Quantitative PCR analysis showed that transcription of flaB is higher at 30 degrees C than at 37 degrees C. Mutation of flaB had no effect on motility or on flaA transcription while mutation of flaA abolished motility and increased the transcription of flaB. These results underline that the genus Arcobacter is an unusual taxon in the epsilon subdivision of the Proteobacteria.

The introduction of Arcobacter spp. in poultry slaughterhouses.

Despite the presence high levels of Arcobacter spp. on chicken carcasses, the source of arcobacter contamination in slaughterhouses still remains unclear. It has been hypothesised in the literature that Arcobacter species that contaminate carcasses originate in in-plant slaughterhouses and/or supply water. The present study aimed to determine the source of Arcobacter contamination in two poultry slaughterhouses in The Netherlands. Carcasses and intestinal tracts from 3 hen flocks and 2 broiler flocks were collected. Water draining off carcasses during processing in 2 slaughterhouses and supply water in one slaughterhouse were also taken. For one flock, cloacal swabs and faecal samples were taken on the farm before slaughtering. ERIC-PCR was applied to study the genetic diversity and relationship among the isolates. No Arcobacter spp. were found in the supply water but on almost all of the sampled carcasses and in carcass-draining-off water arcobacters were identified. Arcobacter spp. were detected in the gut systems of chickens, ranging from 20% to 85% in hens and 3.3% and 51% in broilers. Similar ERIC-PCR genotypes were detected in gut contents as well as on carcasses from the same flock. The present study demonstrated that Arcobacter spp. can be detected in chicken intestines at slaughter and could be brought in this way into slaughterhouses where the bacteria contaminate carcasses during processing.

Dogs as vectors of Streptobacillus moniliformis infection?

Rat bite fever is a bacterial zoonosis transmitted through the bite of rats. One of the two etiological agents that cause rat bite fever is Streptobacillus moniliformis. Rat bite fever is rare and very likely under diagnosed but occurs worldwide. Other animals, like dogs and cats that have mouthed a rat are often mentioned in the literature as potential risks for the attraction of rat bite fever. However, rat bite fever caused by the bite of a dog or cat has very seldom been documented. Therefore, to identify the possible risk for humans to become infected with S. moniliformis after having been bitten by a dog that has been in contact with rats, the presence of S. moniliformis in the mouth of these dogs was tested with molecular methods. Swabs taken from the mouth of 18 dogs with proven contacts with rats were tested for the presence of S. moniliformis DNA by PCR. An amplicon of the right size was obtained in 10 of the 18 dogs. Nucleotide sequencing of five amplicons of PCR positive samples demonstrated the presence of S. moniliformis DNA in the mouth of three dogs. A bite by these dogs therefore might infect humans with S. moniliformis and cause rat bite disease.

Zoonotic bacteria and parasites found in raw meat-based diets for cats and dogs.

Feeding raw meat-based diets (RMBDs) to companion animals has become increasingly popular. Since these diets may be contaminated with bacteria and parasites, they may pose a risk to both animal and human health. The purpose of this study was to test for the presence of zoonotic bacterial and parasitic pathogens in Dutch commercial RMBDs. We analysed 35 commercial frozen RMBDs from eight different brands. Escherichia coli serotype O157:H7 was isolated from eight products (23 per cent) and extended-spectrum beta-lactamases-producing E coli was found in 28 products (80 per cent). Listeria monocytogenes was present in 19 products (54 per cent), other Listeria species in 15 products (43 per cent) and Salmonella species in seven products (20 per cent). Concerning parasites, four products (11 per cent) contained Sarcocystis cruzi and another four (11 per cent) S tenella In two products (6 per cent) Toxoplasma gondii was found. The results of this study demonstrate the presence of potential zoonotic pathogens in frozen RMBDs that may be a possible source of bacterial infections in pet animals and if transmitted pose a risk for human beings. If non-frozen meat is fed, parasitic infections are also possible. Pet owners should therefore be informed about the risks associated with feeding their animals RMBDs.

Interaction of Arcobacter spp. with human and porcine intestinal epithelial cells.

Little is known about the pathogenic mechanisms or potential virulence factors of Arcobacter spp. The aim of the study described here was to obtain more insights in the pathogenicity mechanisms of Arcobacter spp. by testing their ability to adhere to, invade and induce interleukin-8 expression in human Caco-2 and porcine IPI-2I cell lines. Eight Arcobacter strains were tested. Four strains were obtained from a culture collection, and represent the four Arcobacter spp. known to be associated with animals and humans. The other four strains were field isolates from the amniotic fluid of sows and from newborn piglets. All eight Arcobacter strains were able to adhere to both cell lines, and induced interleukin-8 production as early as 2 h after a 1h incubation period. This production was still increased 6 h postinfection. Differences in the cell association of the eight strains were obvious, with A. cibarius showing the highest adhesion ability. Invasion of intestinal epithelial cells was only observed for A. cryaerophilus strains. No correlation between invasiveness or strong adhesion of the tested strains and the level of interleukin-8 induction was observed.

Farmers' knowledge and expectations of antimicrobial use and resistance are strongly related to usage in Dutch livestock sectors.

Comprehensive strategies to improve on-farm antimicrobial use (AMU) are needed to contain antimicrobial resistance (AMR). Little is known about farmers' motivating and enabling factors, and about their influence on AMU. In a cross-sectional online survey, Dutch dairy, veal and pig farmers (n=457) reported their on-farm AMU as "Defined Daily Dose Animal" per year (DDDAF) and completed a detailed questionnaire on their view, knowledge and behavior towards AMU and AMR. Exploratory factor analysis (EFA) on the questionnaire items identified four psychological factors labeled as 'referent beliefs', 'perceived risk', 'knowledge', and 'undesired attitude to regulations'. Linear regression was done to explore the relationship between the obtained factors and on-farm AMU across the three animal sectors. Dairy farmers showed the highest factor scores for 'knowledge' and the lowest for 'perceived risk'. 'Knowledge' scores were significantly and inversely related to AMU (P=0.0004). Borderline significant associations with AMU were found for 'perceived risk' and 'undesired attitude to regulations' (negative and positive relationships respectively). There were no apparent differences for these relationships between the three livestock sectors. Behavioral interventions in farmers such as educational campaigns or increased support by veterinarians could empower farmers with more prudent and rational practices, eventually reducing AMU in food animals.

Arcobacter, what is known and unknown about a potential foodborne zoonotic agent!

Since the introduction of the genus Arcobacter in 1991, the association of Arcobacter butzleri, Arcobacter cryaerophilus and Arcobacter skirrowii with humans and animals has been clearly established. These bacteria have been detected world wide in products of animal origin and in healthy animals as well as in surface water. A fourth species Arcobacter cibarius was recently discovered on chicken carcasses. Although evidence was found for the connection of Arcobacter spp. with human and animal illness, Arcobacter spp. can be pathogens, opportunistic pathogens and commensals. Their potential as zoonotic foodborne and waterborne agents, the routes of transmission and the pathogenic mechanisms of these bacteria are largely unknown. Production of toxins or other virulence factors has not been demonstrated but adhesive and/or invasive properties were apparent. Antibiotic resistance is present in Arcobacter strains to significant levels. The tools to genetically access Arcobacter-like transformation of strains, construction of mutants are not yet available. Nor have genes (i.e. potential virulence factors) been cloned, expressed and characterized in other host organisms. Therefore those interested in the microbiology of these organisms eagerly await publication of the complete nucleotide sequence of the Arcobacter genome. The abundant presence of four Arcobacter species in foods of animal origin and the recovery of these bacteria from surface and drinking water suggest an important role of these bacteria as foodborne or waterborne agent and possibly as zoonotic agent.