Foodborne pathogenic bacteria in wild European hedgehogs (Erinaceus europaeus).

BACKGROUND: European hedgehogs (Erinaceus europaeus) are widely distributed across Europe. They may play an important role by spreading zoonotic bacteria in the environment and to humans and animals. The aim of our work was to study the prevalence and characteristics of the most important foodborne bacterial pathogens in wild hedgehogs. RESULTS: Faecal samples from 148 hospitalised wild hedgehogs originating from the Helsinki region in southern Finland were studied. Foodborne pathogens were detected in 60% of the hedgehogs by PCR. Listeria (26%) and STEC (26%) were the most common foodborne pathogens. Salmonella, Yersinia, and Campylobacter were detected in 18%, 16%, and 7% of hedgehogs, respectively. Salmonella and Yersinia were highly susceptible to the tested antimicrobials. Salmonella Enteritidis and Listeria monocytogenes 2a were the most common types found in hedgehogs. All S. Enteritidis belonged to one sequence type (ST11), forming four clusters of closely related isolates. L. monocytogenes was genetically more diverse than Salmonella, belonging to 11 STs. C. jejuni ST45 and ST677, Y. pseudotuberculosis O:1 of ST9 and ST42, and Y. enterocolitica O:9 of ST139 were also found. CONCLUSIONS: Our study shows that wild European hedgehogs should be considered an important source of foodborne pathogens, and appropriate hygiene measures after any contact with hedgehogs and strict biosecurity around farms are therefore important.

Pathogenic profile and antimicrobial resistance of Escherichia coli, Escherichia marmotae and Escherichia ruysiae detected from hunted wild boars in Sardinia (Italy).

The objective of this study was to evaluate the occurrence of E. coli in hunted wild boars in Sardinia (Italy) and to further characterize the isolates with Whole Genome Sequencing to assess the genetic relatedness and the presence of virulence and antimicrobial resistance (AMR) genes. Samples were taken from 66 wild boars between 2020 and 2022 slaughtered in five hunting houses. A total of 181 samples were tested, including 66 samples from mesenteric lymph nodes, 66 samples from colon content and 49 samples from carcass surface. Isolates referable to Escherichia species were detected in all of the wild boars sampled. On a selection of 61 isolates, sequencing was conducted and antimicrobial susceptibility was tested. Among these, three isolates were confirmed to be two Escherichia marmotae (cryptic clade V) and one Escherichia ruysiae (cryptic clade III). E. coli pathotypes identified were UPEC (13 %), ExPEC-UPEC (5.6 %) and ETEC (3.7 %). Moreover, 3/6 E. marmotae isolates had typical ExPEC genes. Genetic similarity was observed in isolates collected from animals slaughtered in the same hunting house; this suggests epidemiological links deriving from the presence of animals infected with closely related strains or the result of cross-contamination. Antimicrobial resistance genes were detected in three non-pathogenic E. coli isolates: one isolate had sul2, tet(B), aph(6)-ld and aph(3″)-lb resistance genes and two had the fosA7 gene. This study confirmed that wild boars can act as reservoirs and spreaders of pathogenic Escherichia species and it provides information for future comparative genomic analysis in wildlife. Although isolates showed a limited resistome, the detection of resistance in non-pathogenic isolates underlines the need to monitor antimicrobial resistance in the wild boar population. To the best of our knowledge, this is the first detection of E. mamotae and E. ruysiae isolates in wild boars in Italy and the presence of this pathogen in wildlife and livestock need to be investigated further.

Enteropathogenic Yersinia with Public Health Relevance Found in Dogs and Cats in Finland.

Yersiniosis is a common zoonotic enteric disease among humans, which has been linked to pigs and contaminated food, especially pork. The epidemiology of yersiniosis is still obscure, and studies on yersiniosis in pets are very scarce. In this study, we performed pheno- and genotypic characterisation of 50 Yersinia strains isolated from pets in Finland between 2012 and 2023. Y. enterocolitica 4/O:3/ST135, the most common type in human yersiniosis, was also the most common type (68%) found in clinical faecal samples in our study. Also, human pathogenic Y. enterocolitica 2/O:9/ST139 and Y. pseudotuberculosis O:1/ST9 and O:1/ST42 strains carrying all essential pathogenic genes were identified. Three Y. enterocolitica 4/O:3/ST9 strains were multi-drug-resistant and two of them were highly related, showing one allelic difference (AD) with core genome multi-locus sequence typing. Non-pathogenic, genotypically highly diverse Y. enterocolitica 1A strains, showing more than 1000 ADs and missing the essential virulence genes, were also recognised in dogs and cats. Our study demonstrates that pets can excrete human pathogenic Yersinia in their faeces and may serve as an infection source for human yersiniosis, especially in families with small children in close contact with their pets.

Prevalence and genotype diversity of Campylobacter jejuni in hunted reared pheasants (Phasianus colchicus) in Finland.

Campylobacter spp., especially C. jejuni, is the most common zoonotic pathogen in humans worldwide. In Nordic countries, the prevalence of C. jejuni in broilers, which is an important reservoir of human infections, is generally low. Thus, other sources of domestically acquired infections besides chicken meat need to be considered. Game birds are known to carry a variety of zoonotic agents, including Campylobacter. The aim of this study was to investigate Campylobacter spp. carriage in a flock of reared pheasants at hunting in two successive samplings to better understand the dynamics of Campylobacter infections in pheasants. Overall, 72% of the intestinal samples were positive for Campylobacter spp. by direct culture on mCCDA. C. jejuni was the only species identified. The isolates were genotyped using whole genome sequencing (WGS), multilocus sequence typing (MLST), and ad hoc whole genome MLST (wgMLST). Two distinct C. jejuni clones were identified among the 18 isolates studied, representing MLST sequence types (STs) ST-45 and ST-699. The ST-45 isolates were closely related to previous human clinical isolates using core genome MLST (cgMLST). In contrast, the ST-699 isolates forming the dominant clone in the latter sampling were quite distinct from previously described cgMLST profiles from different hosts and sources worldwide. In conclusion, the intestine of reared pheasants is commonly colonized by C. jejuni and may carry genotypes relevant to infections in livestock and humans. Hygienic measures are needed to limit the spread of infection in reared flocks. Especially farmers and hunters having direct contact with pheasant offal need to be aware of the associated zoonosis risk to protect themselves and their working dogs alike. Biosecurity measures to improve the safety and reduce the zoonosis risk associated with pheasant farming should be further investigated.

Antimicrobial Resistance of Campylobacter coli Isolated from Caecal Samples of Fattening Pigs at Slaughter.

Pigs are known as the main Campylobacter coli reservoirs. Campylobacteriosis, the most commonly reported gastrointestinal disease in humans, is mainly caused by the consumption of poultry meat, and little is known about the role of pork. Pigs are often associated with C. coli, including antimicrobial-resistant isolates. Therefore, the entire pork production chain must be considered as an important source of antimicrobial-resistant C. coli. This study aimed to determine the antimicrobial resistance of Campylobacter spp. isolated from caecal samples of fattening pigs at the Estonian slaughterhouse level over a five-year period. The proportion of Campylobacter-positive caecal samples was 52%. All Campylobacter isolates were identified as C. coli. A high proportion of the isolates were resistant to most of the studied antimicrobials. The resistance to streptomycin, tetracycline, ciprofloxacin and nalidixic acid was 74.8%, 54.4%, 34.4% and 31.9%, respectively. In addition, a high proportion (15.1%) of the isolates were multidrug-resistant and, in total, 93.3% were resistant to at least one antimicrobial.

Traceability, virulence and antimicrobial resistance of Yersinia enterocolitica in two industrial cheese-making plants.

Between 2018 and 2019, 309 environmental and food samples were collected from two industrial cheese-making plants located in Sardinia, in order to investigate Y. enterocolitica presence and to characterize the isolates. Y. enterocolitica isolates were further compared with isolates detected during a previous investigation from sheep and goat raw milk samples. Y. enterocolitica was detected in 7.4 % of the samples and the prevalence was higher, even if not significantly (P > 0.05) higher in non-food contact surface samples (10.2 %) than in food contact surface samples (3.8 %). The highest prevalence was detected in floor samples (13.5 %), followed by drain samples (7.2 %), which might serve as main harborage sites for further contamination. Y. enterocolitica was also detected in food contact surfaces, namely shelves of the Ricotta cooling room and packaging room, one cheese cutting machine surface and one raw milk filter sample. The biotype 1A isolates identified in this study were classified into six different serotypes. Additionally, a bioserotype 2/O:5,27 isolate was identified in one goat milk sample. All 1A isolates possessed the virulence genes invA and ystB while the 2/O:5,27 isolate showed the presence of ail, ystA, invA and yadA genes, thus confirming a pathogenic potential. The isolates showed intrinsic resistance to amoxicillin-clavulanic acid, ticarcillin and cefoxitin due to the presence of the blaA gene. Whole genome sequencing allowed to identify seven different sequence types among the 1A isolates, thus showing a high genetic diversity. The same Y. enterocolitica sequence type (ST3) was detected from three different areas of the same cheese-making plant, indicating a possible transfer of the microorganism along the processing lines. Y. enterocolitica contamination in cheese-making plants can pose a risk to human health. Preventive measures include the hygienic design of the plant layout and equipment, in association with proper cleaning and disinfection programmes.

Microbiological quality and safety of vacuum-packaged white-tailed deer meat stored at 4 °C.

Vacuum packaging is widely used for extending the shelf life of commercial fresh meat products. It also ensures product hygiene during distribution and storage. However, very little information exists concerning the effects of vacuum packaging on the shelf life of deer meat. One of our aims was to evaluate how storage under vacuum at 4 °C affects the microbial quality and safety of white-tailed deer (Odocoileus virginianus) meat cuts. This was assessed in a longitudinal study based on sensory analyses and measurements of (1) mesophilic aerobic bacteria (MAB), (2) lactic acid bacteria (LAB), (3) enterobacteria (EB), (4) and Escherichia coli (EC) counts, and the presence of foodborne pathogens (Campylobacter, Salmonella, stx-harbouring E. coli (STEC), Yersinia and Listeria). Microbiomes were additionally investigated by 16S rRNA gene amplicon sequencing at the time of spoilage. In total, 50 vacuum-packaged meat cuts from the carcasses of 10 wild white-tailed deer harvested in southern Finland in December 2018 were analysed. A significant (p < 0.001) drop in the odour and appearance scores and a significant increase in MAB (p < 0.001) and LAB (p = 0.001) counts of the vacuum-packaged meat cuts were observed after 3 weeks of storage at 4 °C. A very strong correlation (rs = 0.9444, p < 0.001) between the MAB and LAB counts were found during the 5-week sampling period. Clear spoilage changes, manifested as sour off-odours (odour scores ≤2) and pale colour, were detected in the meat cuts spoilt after 3-week storage. High (≥8 log10 cfu/g) MAB and LAB counts were also detected. According to the 16S rRNA gene amplicon analyses, Lactobacillus was the dominant bacterial genus in these samples, demonstrating that LAB can cause rapid spoilage of vacuum-packaged deer meat cuts stored at 4 °C. The rest of the samples were spoilt after four or five weeks of storage, and a vast number of bacterial genera were identified in them. Listeria and STEC were detected by PCR in 50 % and 18 % of the meat cut samples, respectively, which may indicate a public health problem. Our results demonstrate that it is very challenging to ensure the quality and safety of vacuum-packaged deer meat stored at 4 °C, and freezing is therefore recommended to prolong the shelf life.

Hunted Wild Boars in Sardinia: Prevalence, Antimicrobial Resistance and Genomic Analysis of Salmonella and Yersinia enterocolitica.

The objective of this investigation was to evaluate Salmonella and Yersinia enterocolitica prevalence in wild boars hunted in Sardinia and further characterize the isolates and analyse antimicrobial resistance (AMR) patterns. In order to assess slaughtering hygiene, an evaluation of carcasses microbial contamination was also carried out. Between 2020 and 2022, samples were collected from 66 wild boars hunted during two hunting seasons from the area of two provinces in northern and central Sardinia (Italy). Samples collected included colon content samples, mesenteric lymph nodes samples and carcass surface samples. Salmonella and Y. enterocolitica detection was conducted on each sample; also, on carcass surface samples, total aerobic mesophilic count and Enterobacteriaceae count were evaluated. On Salmonella and Y. enterocolitica isolates, antimicrobial susceptibility was tested and whole genome sequencing was applied. Salmonella was identified in the colon content samples of 3/66 (4.5%) wild boars; isolates were S. enterica subs. salamae, S. ser. elomrane and S. enterica subs. enterica. Y. enterocolitica was detected from 20/66 (30.3%) wild boars: in 18/66 (27.3%) colon contents, in 3/66 (4.5%) mesenteric lymph nodes and in 3/49 (6.1%) carcass surface samples. In all, 24 Y. enterocolitica isolates were analysed and 20 different sequence types were detected, with the most common being ST860. Regarding AMR, no resistance was detected in Salmonella isolates, while expected resistance towards ß-lactams (blaA gene) and streptogramin (vatF gene) was observed in Y. enterocolitica isolates (91.7% and 4.2%, respectively). The low presence of AMR is probably due to the low anthropic impact in the wild areas. Regarding the surface contamination of carcasses, values (mean ± standard deviation log10 CFU/cm2) were 2.46 ± 0.97 for ACC and 1.07 ± 1.18 for Enterobacteriaceae. The results of our study confirm that wild boars can serve as reservoirs and spreaders of Salmonella and Y. enterocolitica; the finding of Y. enterocolitica presence on carcass surface highlights how meat may become superficially contaminated, especially considering that contamination is linked to the conditions related to the hunting, handling and processing of game animals.

Characterisation of Listeria monocytogenes Isolates from Hunted Game and Game Meat from Finland.

Listeria monocytogenes is an important foodborne zoonotic bacterium. It is a heterogeneous species that can be classified into lineages, serogroups, clonal complexes, and sequence types. Only scarce information exists on the properties of L. monocytogenes from game and game meat. We characterised 75 L. monocytogenes isolates from various game sources found in Finland between 2012 and 2020. The genetic diversity, presence of virulence and antimicrobial genes were studied with whole genome sequencing. Most (89%) of the isolates belonged to phylogenetic lineage (Lin) II and serogroup (SG) IIa. SGs IVb (8%) and IIb (3%) of Lin I were sporadically identified. In total, 18 clonal complexes and 21 sequence types (STs) were obtained. The most frequent STs were ST451 (21%), ST585 (12%) and ST37 (11%) found in different sample types between 2012 and 2020. We observed 10 clusters, formed by closely related isolates with 0-10 allelic differences. Most (79%) of the virulence genes were found in all of the L. monocytogenes isolates. Only fosX and lin were found out of 46 antimicrobial resistance genes. Our results demonstrate that potentially virulent and antimicrobial-sensitive L. monocytogenes isolates associated with human listeriosis are commonly found in hunted game and game meat in Finland.

Prevalence and Persistence of Multidrug-Resistant Yersinia enterocolitica 4/O:3 in Tonsils of Slaughter Pigs from Different Housing Systems in Croatia.

Yersinia enterocolitica is one of the priority biological hazards in pork inspection. Persistence of the pathogen, including strains resistant to antimicrobials, should be evaluated in pigs from different housing systems for risk ranking of farms. In this 2019 study, tonsils were collected from 234 pigs, of which 69 (29.5%) were fattened on 3 big integrated farms, 130 (55.5%) on 10 medium-sized farms, and 35 (15%) on 13 small family farms. In addition, 92 pork cuts and minced meat samples from the same farms were tested for the presence of Y. enterocolitica using the culture method. Phenotypic and genetic characteristics of the isolates were compared with previously collected isolates from 2014. The overall prevalence of Y. enterocolitica in pig tonsils was 43% [95% CI 36.7−49.7]. In pigs from big integrated, medium-sized, and small family farms, the prevalence was 29%, 52%, and 40%, respectively. All retail samples of portioned and minced pork tested negative for pathogenic Y. enterocolitica, likely due to high hygienic standards in slaughterhouses/cutting meat or low sensitivity of culture methods in these matrices. The highest recovery rate of the pathogen from tonsils was found when alkali-treated PSB and CIN agar were combined. The biosecurity category of integrated and medium farms did not affect the differences in prevalence of Y. enterocolitica (p > 0.05), in contrast to family farms. Pathogenic ail-positive Y. enterocolitica biotype 4 serotype O:3 persisted in the tonsils of pigs regardless of the type of farm, slaughterhouse, and year of isolation 2014 and 2019. PFGE typing revealed the high genetic concordance (80.6 to 100%) of all the Y. enterocolitica 4/O:3 isolates. A statistically significant higher prevalence of multidrug-resistant Y. enterocolitica 4/O:3 isolates was detected in the tonsils of pigs from big integrated farms compared to the other farm types (p < 0.05), with predominant and increasing resistance to nalidixic acid, chloramphenicol, and streptomycin. This study demonstrated multidrug resistance of the pathogen in pigs likely due to more antimicrobial pressure on big farms, with intriguing resistance to some clinically relevant antimicrobials used in the treatment of yersiniosis in humans.

Prudent Antimicrobial Use Is Essential to Prevent the Emergence of Antimicrobial Resistance in Yersinia enterocolitica 4/O:3 Strains in Pigs.

Yersinia enterocolitica is a psychrotrophic zoonotic foodborne pathogen. Pigs are considered the main reservoir of Y. enterocolitica 4/O:3, which is the most commonly isolated bioserotype in many European countries. Consuming pork contaminated with Y. enterocolitica can be a health threat, and antimicrobial-resistant strains may complicate the treatment of the most severe forms of yersiniosis. We analyzed the antimicrobial resistance of 1,016 pathogenic porcine Y. enterocolitica 4/O:3 strains originating from Belgium, Estonia, Finland, Germany, Italy, Latvia, Russia, Spain, and the United Kingdom. Based on available reports, we also compared antimicrobial sales for food production animals in these countries, excluding Russia. Antimicrobial resistance profiles were determined using a broth microdilution method with VetMIC plates for 13 antimicrobial agents: ampicillin, cefotaxime, ceftiofur (CTF), chloramphenicol (CHL), ciprofloxacin, florfenicol, gentamicin, kanamycin, nalidixic acid (NAL), streptomycin (STR), sulfamethoxazole (SME), tetracycline (TET), and trimethoprim (TMP). The antimicrobial resistance of Y. enterocolitica 4/O:3 strains varied widely between the countries. Strains resistant to antimicrobial agents other than ampicillin were rare in Estonia, Finland, Latvia, and Russia, with prevalence of 0.7, 0.4, 0, and 8.3%, respectively. The highest prevalence of antimicrobial resistance was found in Spanish and Italian strains, with 98 and 61% of the strains being resistant to at least two antimicrobial agents, respectively. Resistance to at least four antimicrobial agents was found in 34% of Spanish, 19% of Italian, and 7.1% of English strains. Antimicrobial resistance was more common in countries where the total sales of antimicrobials for food production animals are high and orally administered medications are common. Our results indicate that antimicrobials should be used responsibly to treat infections, and parenteral medications should be preferred to orally administered mass medications.

Osteomyelitis in a slaughter turkey flock caused by Yersinia pseudotuberculosis sequence type ST42.

A Yersinia pseudotuberculosis outbreak was diagnosed in a male turkey flock in Finland. Y. pseudotuberculosis is a quite rare zoonotic bacterium, which typically causes enteritis in humans and sudden death in animals. In this study, osteomyelitis was diagnosed in small, lame, 11- to 12-wk-old male turkeys. Lameness and slower growth among the turkeys was observed on the farm. During pathological examination, multiple lesions were found in the metaphyseal and physeal areas of the femurs, tibiotarsi, and tarsometatarsi, with multifocal to coalescing mixed heterophilic/granulomatous necrotizing osteomyelitis. Y. pseudotuberculosis was isolated from the femoral and tibiotarsal bones or from the joints of six lame turkeys sent for necropsy. The isolation required homogenizing of lesion tissue in phosphate-mannitol-peptone broth, which was cultured directly - and, if needed, after cold enrichment - on selective cefsulodin-irgasan-novobiocin agar. Whole-genome sequencing was used for identification and typing. All isolates belonged to bio/serotype 1/O:1a and sequence type ST42 (Achtman scheme), which is commonly reported in both human and animal Y. pseudotuberculosis infections in Europe. The isolates from all six turkeys showed only one to two allele differences in the core genome comparison, indicating a common source of infection. All asymptomatic turkeys were slaughtered at the age of 17 weeks. Whole and partial carcass condemnation rates at the slaughterhouse were high, but no macroscopic changes in the skeletal system were found, showing that food chain information is essential. This study confirms earlier findings that Y. pseudotuberculosis can cause osteomyelitis in fattening turkeys, leading to lameness. Food chain information is essential for slaughterhouse operations, to protect the workers and emphasize good working hygiene during slaughter.

Prevalence and Serotype Diversity of Salmonella enterica in the Estonian Meat Production Chain in 2016-2020.

BACKGROUND: Salmonella enterica represents a considerable public concern worldwide, with farm animals often recognised as an important reservoir. This study gives an overview of the prevalence and serotype diversity of Salmonella over a 5-year period in the meat production chain in Estonia. Data on human salmonellosis over the same period are provided. METHODS: Salmonella surveillance data from 2016 to 2020 were analysed. RESULTS: The prevalence of Salmonella at the farm level was 27.7%, 3.3% and 0.1% for fattening pigs, cattle and poultry, respectively. S. Derby was the most prevalent serotype at the farm level for fattening pigs and S. Dublin for cattle. The top three serotypes isolated at the slaughterhouse and meat cutting levels were S. Derby, monophasic S. Typhimurium and S. Typhimurium with proportions of 64.7%, 9.4% and 7.0%, respectively. These serotypes were the top five most common Salmonella serotypes responsible for human infections in Estonia. S. Enteritidis is the main cause (46.9%) of human salmonellosis cases in Estonia, but in recent years, Enteritidis has not been detected at the slaughterhouse or meat cutting level. CONCLUSION: In recent years, monophasic S. Typhimurium has become epidemiologically more important in Estonia, with the second-highest cause in human cases and third-highest among the most prevalent serotypes of Salmonella enterica in the meat chain.

Bacterial quality and safety of raw beef: A comparison between Finland and Nigeria.

Beef can easily be contaminated with bacteria during the meat production chain. In this work, we studied the contamination levels of mesophilic aerobic bacteria (MAB) and thermotolerant coliform bacteria (TCB) on raw beef surfaces from small shops in Helsinki, Finland and meat markets in Benin City, Nigeria. We also investigated the prevalence of Salmonella, Campylobacter, Yersinia, Shiga toxin-producing Escherichia coli (STEC), Listeria, and cephalosporin-resistant E. coli (CREC). In total, one hundred unpacked raw beef samples from Finland and Nigeria were collected in 2019. The median MAB and TCB counts were significantly (P < 0.001) higher on beef from Nigeria than from Finland. The median MAB and TCB counts in Nigeria were 7.5 and 4.0 log10 cfu/cm2, respectively, and 6.5 and 2.8 log10 cfu/cm2 in Finland, respectively. Most (94%) Nigerian samples were unacceptable according to limits set by the EU. Beef samples from meat markets in Benin City were significantly (P < 0.05) more frequently contaminated with Salmonella, STEC, and CREC than beef samples from small shops in Helsinki. Salmonella, STEC, and CREC were isolated from 30, 36, and 96% of Nigerian samples, respectively, and from <2, 12, and 2% of Finnish samples, respectively. Our study demonstrates a significant difference between the bacterial contaminations of raw beef in Nigeria and Finland, along with a possible misuse of cephalosporins in animal husbandry in Nigeria.

Hunted game birds - Carriers of foodborne pathogens.

Game birds may carry zoonotic bacteria in their intestines and transmit them to hunters through bird handling or through the handling and consumption of contaminated meat. In this study, the prevalence of foodborne bacteria was screened from game bird faeces and mallard breast meat using PCR. The sampling occurred in southern Finland from August to December during the hunting season. Isolates were characterized by multi-locus sequence typing. Mesophilic aerobic bacteria and Escherichia coli counts were used to assess the microbial contamination of mallard meat. In total, 100 woodpigeon (Columba palumbus), 101 pheasants (Phasianus colchicus), 110 mallards (Anas platyrhynchos), and 30 teals (Anas crecca) were screened during the hunting season. Additionally, 100 mallard breast meat samples were collected. Campylobacter and Listeria were commonly detected in the faeces and Listeria on mallard meat. L. monocytogenes of sequence types associated with human listeriosis were frequently found in game bird faeces and on mallard meat. Good hygiene during game bird handling, storing the game bird meat frozen, and proper heat treatment are important measures to minimize the health risk for hunters and consumers.

Foodborne Zoonoses Common in Hunted Wild Boars.

The northern European wild boar population has increased during the last decade. Highest wild boar numbers in Finland have been reported in the southeastern part near the Russian border. Wild boars may be infected with several human and animal pathogens. In this study, we investigated the presence of important foodborne pathogens in wild boars hunted in 2016 in Finland using serology, PCR and culturing. Seroprevalence of Salmonella (38%) and Yersinia (56%) infections was high in wild boars. Antibodies to hepatitis E virus, Toxoplasma gondii and Brucella were found in 18%, 9% and 9% of the wild boars, respectively. Trichinella antibodies were detected in 1% of the animals. We recorded no differences in the seroprevalence between males and females. However, Yersinia and T. gondii antibodies were detected significantly more often in adults than in young individuals. Listeria monocytogenes (48%) and stx-positive Escherichia coli (33%) determinants were frequently detected in the visceral organs (spleen and kidneys) by PCR. Yersinia pseudotuberculosis O:1 and L. monocytogenes 2a and 4b were identified by culturing from the PCR-positive samples. Brucella suis biovar 2 was isolated from visceral organs. No African swine fever, classical swine fever or Aujeszky's disease were detected in the wild boars. Our study shows that wild boars are important reservoirs of foodborne pathogens.

Two copies of the ail gene found in Yersinia enterocolitica and Yersinia kristensenii.

Yersinia enterocolitica is the most common Yersinia species causing foodborne infections in humans. Pathogenic strains carry the chromosomal ail gene, which is essential for bacterial attachment to and invasion into host cells and for serum resistance. This gene is commonly amplified in several PCR assays detecting pathogenic Y. enterocolitica in food samples and discriminating pathogenic isolates from non-pathogenic ones. We have isolated several non-pathogenic ail-positive Yersinia strains from various sources in Finland. For this study, we selected 16 ail-positive Yersinia strains, which were phenotypically and genotypically characterised. Eleven strains were confirmed to belong to Y. enterocolitica and five strains to Yersinia kristensenii using whole-genome alignment, Parsnp and the SNP phylogenetic tree. All Y. enterocolitica strains belonged to non-pathogenic biotype 1A. We found two copies of the ail gene (ail1 and ail2) in all five Y. kristensenii strains and in one Y. enterocolitica biotype 1A strain. All 16 Yersinia strains carried the ail1 gene consisting of three different sequence patterns (A6-A8), which were highly similar with the ail gene found in high-pathogenic Y. enterocolitica biotype 1B strains (A2). The Ail protein encoded by the ail1 gene was highly conserved compared to the Ail protein encoded by the ail2 gene. Multiple sequence alignment of the ail gene and Ail protein were conducted with MAFF. In total, 10 ail sequence variations have been identified, of which 8 conserved ones belonged to the ail1 gene. According to our results, the detection of ail alone is not sufficient to predict the pathogenicity of Yersinia isolates.

Atypical Hemolytic Listeria innocua Isolates Are Virulent, albeit Less than Listeria monocytogenes.

Listeria innocua is considered a nonpathogenic Listeria species. Natural atypical hemolytic L. innocua isolates have been reported but have not been characterized in detail. Here, we report the genomic and functional characterization of representative isolates from the two known natural hemolytic L. innocua clades. Whole-genome sequencing confirmed the presence of Listeria pathogenicity islands (LIPI) characteristic of Listeria monocytogenes species. Functional assays showed that LIPI-1 and inlA genes are transcribed, and the corresponding gene products are expressed and functional. Using in vitro and in vivo assays, we show that atypical hemolytic L. innocua is virulent, can actively cross the intestinal epithelium, and spreads systemically to the liver and spleen, albeit to a lesser degree than the reference L. monocytogenes EGDe strain. Although human exposure to hemolytic L. innocua is likely rare, these findings are important for food safety and public health. The presence of virulence traits in some L. innocua clades supports the existence of a common virulent ancestor of L. monocytogenes and L. innocua.

Assessment of the feasibility of serological monitoring and on-farm information about health status for the future meat inspection of fattening pigs.

Current macroscopic meat inspection cannot detect the most common pork-borne pathogens (Salmonella spp., Yersinia enterocolitica and Toxoplasma gondii). Furthermore, food chain information (FCI) may not provide sufficient data for visual-only inspection, which is supposed to be the common way of inspection of pigs in the European Union. Our observational study aimed to evaluate the serological monitoring and the clinical evaluation of on-farm health status of pigs and assess the feasibility of these data as part of the FCI in meat inspection. We studied the serological status of Salmonella spp., Yersinia spp. and T. gondii in pigs during the fattening period. Additionally, we evaluated the association between on-farm health status and meat inspection findings. On 57 indoor fattening pig farms in Finland, we collected blood samples (mean of 20 pigs/farm) and assessed the on-farm health (coughing, tail biting, lameness) at the end of the fattening period. We visited 34 of these farms also at the beginning of the fattening for sampling and on-farm health evaluation of the same pigs. Meat inspection results were obtained after slaughter for all 57 farms. Salmonella seroprevalence was low at the end of the fattening period: it was 17.6%, 10.6% or 1.9%, with the cut-off values of OD15% (recommended by the test manufacturer), OD20% (used by Danish monitoring programme) and OD40% (used by German monitoring programme), respectively. The overall seroprevalence of Salmonella spp. and Yersinia spp. increased significantly (P < 0.001) during the fattening period (from 8.1% to 17.2% and from 30.3% to 72.3%, respectively), while the seroprevalence of T. gondii remained low (<1%). The within-farm seroprevalences of Salmonella spp. and Yersinia spp. differed significantly between the farms and this farm-level serological data could be used as FCI for risk-based decisions to improve food safety. Such potentially feasible decisions could include additional carcass testing, carcass decontamination, carcass processing, slaughtering arrangements and improved biosecurity measures at the farm. However, risk mitigation targets and procedures must be carefully adjusted for each pathogen regarding also economic aspects. Tail biting observed on farm was associated with partial carcass condemnations and arthritis at slaughter. This information could be included in the FCI and used when making decisions regarding meat inspection procedure: visual-only or additional inspections.