The importance of slaughterhouses in monitoring the occurrence of tail biting in pigs - Review.

Tail biting in pigs represents a very serious problem in modern pig farming, particularly with the intensification of the industry. It is considered a multifactorial syndrome and can be caused by various factors, leading to significant economic losses through reduced weight gain and partial or total condemnation of slaughtered carcasses due to secondary bacterial infections. The aim of this article is to summarise the current knowledge regarding tail biting in pigs, with a primary focus on the use of slaughterhouses for evaluating tail lesions and monitoring their prevalence. The introduction addresses the factors influencing the incidence of tail biting in pig farms and prevention strategies. Subsequent sections cover topics such as tail docking, the negative effects of pig tail biting, the advantages and drawbacks of examining tail lesions in slaughterhouses, and the methodical procedure for evaluating such lesions. Additionally, the article discusses the relationship between tail lesions and meat inspection findings, as well as the prevalence of tail lesions in various European countries.

Shiga Toxin-Producing Escherichia coli in Faecal Samples from Wild Ruminants.

Wildlife can harbour Shiga toxin-producing Escherichia coli (STEC). In the present study, STEC in faecal samples from red deer (n = 106) and roe deer (n = 95) were characterised. All isolates were non-O157 strains. In red deer, STEC were detected in 17.9% (n = 19) of the isolates, and the eae/stx2b virulence profile was detected in two isolates (10.5%). One STEC strain harboured stx1a (5.3%) and eighteen STEC strains harboured stx2 (94.7%). The most prevalent stx2 subtypes were stx2b (n = 12; 66.7%), stx2a (n = 3; 16.7%), and stx2g (n = 2; 11.1%). One isolate could not be subtyped (NS) with the applied primers (5.6%). The most widely identified serotypes were O146:H28 (n = 4; 21%), O146:HNM (n = 2; 10.5%), O103:H7 (n = 1; 5.3%), O103:H21 (n = 1; 5.3%), and O45:HNM (n = 1; 5.3%). In roe deer, STEC were detected in 16.8% (n = 16) of the isolates, and the eae/stx2b virulence profile was detected in one isolate (6.3%). Two STEC strains harboured stx1a (12.5%), one strain harboured stx1NS/stx2b (6.3%), and thirteen strains harboured stx2 (81.3%). The most common subtypes were stx2b (n = 8; 61.5%), stx2g (n = 2; 15.4%), non-typeable subtypes (NS) (n = 2; 15.4%), and stx2a (n = 1; 7.7%). Serotype O146:H28 (n = 5; 31.3%) was identified. The study demonstrated that the zoonotic potential of STEC strains isolated from wildlife faeces should be monitored in the context of the 'One Health' approach which links human health with animal and environmental health.

Seroprevalence and Molecular Epidemiology of Aleutian Disease in Various Countries during 1972-2021: A Review and Meta-Analysis.

Aleutian disease (AD) poses a serious threat to both free-ranging and farmed mink around the world. The disease is caused by the Aleutian mink disease virus (AMDV), which also poses a health risk for other members of the family Mustelidae, including wild mink, weasels, badgers and other animal species. This article analyses the seroprevalence of AMDV infections in mink and other species around the world, and reviews recent knowledge relating to the molecular epidemiology of the AMDV. Depending on the applied diagnostic technique and the country, the prevalence of anti-AMDV antibodies or AMDV DNA was established at 21.60-100.00% in farmed American mink, 0.00-93.30% in free-ranging American mink and 0.00-25.00% in European mink. Anti-AMDV antibodies or AMDV DNA were also detected in other free-living fur-bearing animals in Europe and Canada, where their prevalence was determined at 0.00-32.00% and 0.00-70.50%, respectively. This may indicate a potential threat to various animal species. AMDV strains are not clustered into genotypes based on the geographic origin, year of isolation or pathogenicity. The isolates that were identified on mink farms around the world originated from North America because American mink were introduced to Europe and Asia for breeding purposes and to restock natural populations.

Influence of Infectious Pancreatic Necrosis Virus and Yersinia ruckeri Co-Infection on a Non-Specific Immune System in Rainbow Trout (Oncorhynchus mykiss).

BACKGROUND: The IPNV is one of the most common viral pathogens of rainbow trout (Oncorhynchus mykiss), while Y. ruckeri infections are widespread among bacterial agents. The current study aimed to determine the influence of IPNV and Y. ruckeri co-infection on a non-specific immune response. METHODS: Two experiments were conducted. The first experiment determined the changes in non-specific immunity parameters upon the simultaneous occurrence of IPNV and Y. ruckeri infection. In the second experiment, infection with the IPNV was performed two weeks before Y. ruckeri infection. The level of total protein, gamma globulins, the activity of lysozyme and ceruloplasmin, as well as the metabolic activity and potential killing activity of phagocytes were measured: 0, 24 h, 72 h, 7 days, 14 days, and 21 days after co-infection. RESULTS: A differentiated effect on the parameters of the non-specific immune response was shown between single infections with the IPNV and Y. ruckeri as well as co-infection with these pathogens. CONCLUSIONS: The immune response in the course of a co-infection depended on the time between infections. IPNV infection causes lysozyme activity suppression, which may lead to secondary bacterial infections.

The Prevalence of Yersinia enterocolitica and Yersinia pseudotuberculosis in Small Wild Rodents in Poland.

Rodents are a large group of mammals that can be carriers of zoonotic pathogens such as Yersinia strains that cause yersiniosis. The prevalence of Yersinia enterocolitica and Yersinia pseudotuberculosis was determined in 214 small wild rodents from south-eastern Poland. Samples were analyzed by precultivation and PCR. Nine (4.2%) Y. enterocolitica and one (0.5%) Y. pseudotuberculosis isolates were received. Most of them (n = 5) were obtained from the common vole (Microtus arvalis). All Y. enterocolitica strains were classified as biotype (BT) 1A. A PCR analysis of virulence markers revealed that all Y. enterocolitica isolates contained the ystB gene and five isolates harbored a rare genetic combination of ail/ystB. Three of the four ail/ystB-positive isolates belonged to serotype O:5.27. The Y. pseudotuberculosis inv-positive isolate was classified as BT 1. A genetic analysis of Y. enterocolitica harboring the ystB gene revealed 100% similarity between the analyzed sequences and the sequences from diarrhea patients in India and the United Kingdom as well as high similarity with the sequences from different species of wild animals from Poland. The Y. pseudotuberculosis inv sequence was 100% identical to the sequence isolated from fully virulent clinical strain from France and Australia. The results of our study suggest that small wild rodents, especially voles and yellow-necked mice, may act as carriers of Yersinia strains. The high similarity of the tested gene sequences between our isolates and the isolates from other free-living animals indicates that small wild rodents can play a role in the epidemiology of yersiniosis and can shed Yersinia spp. into the environment.

Shiga toxin-producing Escherichia coli isolates from red deer (Cervus elaphus), roe deer (Capreolus capreolus) and fallow deer (Dama dama) in Poland.

Shiga toxin-producing Escherichia (E.) coli (STEC) pathogens are responsible for the outbreaks of serious diseases in humans, including haemolytic uraemic syndrome (HUS), bloody diarrhoea (BD) and diarrhoea (D), and they pose a significant public health concern. Wild ruminants are an important environmental reservoir of foodborne pathogens that can cause serious illnesses in humans and contaminate fresh products. There is a general scarcity of published data about wildlife as a reservoir of foodborne pathogens in Poland, which is why the potential epidemiological risk associated with red deer, roe deer and fallow deer as reservoirs of STEC/AE-STEC strains was evaluated in this study. The aim of the study was to investigate the prevalence of STEC strains in red deer (Cervus elaphus), roe deer (Capreolus capreolus) and fallow deer (Dama dama) populations in north-eastern Poland, and to evaluate the potential health risk associated with wild ruminants carrying STEC/AE-STEC strains. We examined 252 rectal swabs obtained from 134 roe deer (Capreolus capreolus), 97 red deer (Cervus elaphus) and 21 fallow deer (Dama dama) in north-eastern Poland. The samples were enriched in modified buffered peptone water. Polymerase chain reaction (PCR) assays were conducted to determine the virulence profile of stx1, stx2 and eae or aggR genes, to identify the subtypes of stx1 and stx2 genes, and to perform O and H serotyping. E. coli O157:H7 isolates were detected in the rectal swabs collected from 1/134 roe deer (0.75%) and 4/97 red deer (4.1%), and they were not detected in fallow deer (Dama dama). The remaining E. coli serogroups, namely O26, O103, O111 and O145 that belong to the "top five" non-O157 serogroups, were detected in 15/134 roe deer (11.19%), 18/97 red deer (18.56%) and 2/21 fallow deer (9.52%). STEC/AE-STEC strains were detected in 33 roe deer isolates (24.63%), 21 red deer isolates (21.65%) and 2 fallow deer isolates (9.52%). According to the most recent FAO/WHO report, stx2a and eae genes are the primary virulence traits associated with HUS, and these genes were identified in one roe deer isolate and one red deer isolate. Stx2 was the predominant stx gene, and it was detected in 78.79% of roe deer and in 71.43% of red deer isolates. The results of this study confirmed that red deer and roe deer in north-eastern Poland are carriers of STEC/AE-STEC strains that are potentially pathogenic for humans. This is the first report documenting the virulence of STEC/AE-STEC strains from wild ruminants in Poland.

Applicability of Thyroxine Measurements and Ultrasound Imaging in Evaluations of Thyroid Function in Turtles.

INTRODUCTION: The thyroid and parathyroid glands play a major role in maintaining physiological homeostasis in all vertebrates. Reptiles have plasma concentrations of thyroid hormones far lower than mammals. Low levels of these hormones in reptiles impede thyroid hormone detection with assays designed for the higher levels of mammals. The aim of this study was to explore teaming this with ultrasound imaging of the thyroid to appraise glandular function. MATERIAL AND METHODS: Thyroid function of four pond sliders was evaluated based on the results of T4 analyses and ultrasound. RESULTS: The concentrations of T4 varied considerably between the examined animals from <9 nmol/L to >167.3 nmol/L. Ultrasound examination revealed uniform echogenicity and a smooth outline of the thyroid gland in all animals. CONCLUSION: Monitoring of thyroid function based on T4 and electrolyte concentrations is helpful in assessing the health and living conditions of reptiles, which is important in veterinary practice but problematic. Ultrasound examinations are useful in diagnosing changes in gland structure, such as tumours and goitres, and a combination of both methods supports comprehensive assessments of the anatomy and function of the thyroid gland.

Pathogenic potential to humans of Shiga toxin-producing Escherichia coli isolated from wild boars in Poland.

The aim of the study was to investigate the presence of Shiga toxin-producing Escherichia coli (STEC) in the wild boar population of north-eastern Poland, and to evaluate the potential health risk associated with wild boars carrying STEC/AE-STEC strains. In Poland, the African Swine Fever (ASF) virus has been a growing problem in domestic pigs and wild boars, one of the main reservoirs of the virus, because of this hunters, veterinary practitioners and foresters thus face a greater risk of coming into contact with animals. Rectal swabs samples were obtained from 152 wild boars hunter-harvested in the 2017/2018 season (autumn-winter) in north-eastern Poland. The samples were enrichment in modified buffered peptone water. Polymerase chain reaction (PCR) assays were conducted to determine the virulence profile of stx1, stx2 and eae and aggR genes, identify subtypes of stx1 and stx2 genes, and perform O and H serotyping. STEC/AE-STEC virulence genes were detected in 43 isolates (28.29%): STEC in 17 isolates (11.18%) and AE-STEC in 26 isolates (17.11%), respectively. None of the tested isolates carried the aggR gene. The most dangerous AE-STEC virulence profile associated with HUS was found in 2 isolates (1.32%): stx1NS/stx2a/d/eae serotype ONT:H7 and stx2a/eae serotype O146:H7. Six of the 152 tested samples belonged to serogroup O157 (3.95%), including one AE-STEC isolate with virulence profile stx2g/eae and five EPEC isolates. The results of this study suggest that wild boars in north-eastern Poland can carry STEC/AE-STEC strains that are potentially pathogenic for humans. This is the first report documenting the virulence of STEC and AE-STEC isolates from wild boars in Poland.

Phenotypic and genotypic presence of the Yersinia virulence plasmid do not affect the production of enterotoxin YstA by Yersinia enterocolitica strains.

The aim of the study was to determine whether the presence of the Yersinia virulence plasmid could affect the production of enterotoxin YstA by Y. enterocolitica strains isolated from pigs which are the main source of infection for humans. The phenotypic features characteristic for the Yersinia virulence plasmid were detected on CRMOX agar in 8 out of 12 strains producing enterotoxin YstA, in 5 out of 12 doubtful strains, and in 11 out of 12 strains not producing YstA. Autoagglutination ability was detected in all 12 Y. enterocolitica strains that were positive in the suckling mice bioassay, in 11 doubtful strains and 10 negative strains. CRMOX+ colonies were generally ystA, myfA, virF and yadA positive, while CRMOX- colonies were only ystA and myfA positive. The amplicons of yadA were not detected in 2 (8.3%) out of 24 CRMOX+ and virF positive strains. The results of this study indicate that the presence of pYV does not affect the enterotoxin-producing ability of Y. enterocolitica strains.

The Most Important Virulence Markers of Yersinia enterocolitica and Their Role during Infection.

Yersinia enterocolitica is the causative agent of yersiniosis, a zoonotic disease of growing epidemiological importance with significant consequences for public health. This pathogenic species has been intensively studied for many years. Six biotypes (1A, 1B, 2, 3, 4, 5) and more than 70 serotypes of Y. enterocolitica have been identified to date. The biotypes of Y. enterocolitica are divided according to their pathogenic properties: the non-pathogenic biotype 1A, weakly pathogenic biotypes 2⁻5, and the highly pathogenic biotype 1B. Due to the complex pathogenesis of yersiniosis, further research is needed to expand our knowledge of the molecular mechanisms involved in the infection process and the clinical course of the disease. Many factors, both plasmid and chromosomal, significantly influence these processes. The aim of this study was to present the most important virulence markers of Y. enterocolitica and their role during infection.

The prevalence of Yersinia enterocolitica in game animals in Poland.

Natural reservoirs of Yersinia (Y.) enterocolitica comprise different animal species, but little is known about the role of wild animals in the epidemiology of yersiniosis. The aim of the study was to evaluate the prevalence of Y. enterocolitica among game animals in Poland. The bio-serotypes and the pathogenicity markers of the analyzed isolates were determined. The experimental material comprised rectal swabs from 857 free-living animals hunter-harvested over a period of 2 years (2013-2014) in hunting districts across Poland. The isolates from bacteriological studies were confirmed by PCR and bio-serotyped based on the results of biochemical and agglutination tests. In the group of the 218 analyzed isolates of Y. enterocolitica, 133 were derived from wild boars, 70 from red deer, 11 from roe deer and 4 from fallow deer, and they accounted for 61.0%, 32.1%, 5.1% and 1.8% of all isolates, respectively. Bio-serotyping assays revealed that 91.7% of the examined isolates belonged to biotype 1A (200/218). The remaining 18 isolates belonged to bio-serotypes 1B/NI (3/218, 1.4%), 1B/O:8 (1/218, 0.5%), 2/NI (6/218, 2.8%), 2/O:27 (1/218, 0.5%), 2/O:3 (1/218, 0.5%), 2/O:9 (2/218, 0.9%), 3/NI (2/218, 0.9%), 4/O:3 (1/218, 0.5%) and 4/O:9 (1/218, 0.5%). The ail gene, a suggestive virulence gene for Y. enterocolitica, has been found in 30 isolates from 20 wild boars, in 6 isolates from red deer, and in 1 isolate from roe deer. Our study demonstrated that Y. enterocolitica is frequently isolated from game animals in Poland, which poses a risk of spreading these infectious agents to other animal species and humans.

Characterisation of ail-positive Yersinia enterocolitica of different biotypes using HRMA.

Yersiniosis is one of the four most frequent foodborne zoonotic diseases in Europe, and Yersinia enterocolitica is the primary agent in human infections. The ail gene is an important chromosomal virulence marker of Y. enterocolitica which encodes Ail, a 17-kDa outer membrane protein that promotes attachment and invasion. In the present study, ail-positive Y. enterocolitica strains of different biotypes were examined using high resolution melting analysis (HRMA) and DNA sequencing. Genotype data relating to Y. enterocolitica strains isolated from different sources and belonging to different biotypes were compared. Applied method allowed efficient distinguishing of three genotypes and phylogenetic groups: 1A - included non-pathogenic Y. enterocolitica strains; 1B - consisted of highly pathogenic Y. enterocolitica strains and 2/4 - involved weakly pathogenic Y. enterocolitica strains. Amplicon genotyping based on HRMA supports rapid identification of ail SNPs correlated with biotype of examined Y. enterocolitica strains.

A study of single nucleotide polymorphism in the ystB gene of Yersinia enterocolitica strains isolated from various wild animal species.

INTRODUCTION AND OBJECTIVE: Y. enterocolitica is the causative agent of yersiniosis. The objective of the article was a study of single nucleotide polymorphism in the ystB gene of Y. enterocolitica strains isolated from various wild animal species. MATERIALS AND METHOD: High-resolution melting (HRM) analysis was applied to identify single nucleotide polymorphism (SNP) of ystB gene fragments of 88 Y. enterocolitica biotype 1A strains isolated from wild boar, roe deer, red deer and wild ducks. RESULTS: HRM analysis revealed 14 different melting profiles - 4 of them were defined as regular genotypes (G1, G2, G3, G4), whereas 10 as variations. 24 of the examined Y. enterocolitica strains were classified as G1, 18 strains as a G2, 21 strains as a G3, and 15 strains as a G4. Nucleotide sequences classified as G1 revealed 100% similarity with the Y. enterocolitica D88145.1 sequence (NCBI). Analysis of G2 revealed one point mutation - transition T111A. One mutation was also found in G3, but SNP was placed in a different gene region - transition G193A. Two SNPs - transitions G92C and T111A - were identified in G4. Direct sequencing of 10 variations revealed 5 new variants of the ystB nucleotide sequence: V1 - transition G129A (3 strains); V2 - transitions T111A and G193A (2 strains); V3 - transitions C118T and G193A (1 strain); V4 - transitions C141A and G193A (2 strains); and V5 characterized by 19 SNPs: G83A, T93A, A109G, G114T, C116T, A123G, T134C, T142G, T144C, A150C, G162A, T165G, T170G, T174A, T177G, G178A, A179G, A184G and G193A (2 strains). The predominant genotype in isolates from wild ducks was G1; in red deer G2; in wild boar G3; in roe deer G1 and G4. CONCLUSIONS: The proposed HRM method could be used to analyze Y. enterocolitica biotype 1A strains isolated from different sources, including humans.

Zearalenone and its metabolites in the tissues of female wild boars exposed per os to mycotoxins.

The study was performed on 18 clinically healthy female wild boars with initial body weight of 35 ± 2 kg. The animals were divided into two experimental groups (group I and group II) and one control group (group C) of 6 female wild boars per group. Group I animals were administered per os pure zearalenone (ZEN) at 150 µg/kg BW every two months for 7 subsequent days, whereas group II animals received feed naturally contaminated with ZEN at 50 µg/kg BW/day. Female wild boars were exposed to ZEN over a period of 1 year. Control group animals were fed a placebo. Tissue samples (dorsal muscles, left lobe of liver, left kidney, spleen, apical part of the cardiac muscle, cranial lobe of lung, left ovary, central part of the left horn of the uterus) were collected on the last day of the experiment within 3 min after slaughter. In group I, the highest ZEN levels were noted in the spleen (19.813 ng/g), cardiac muscle (18.105 ng/g) and kidneys (14.555 ng/g). In group II, the highest concentrations of ZEN were observed in muscle tissue (12.033 ng/g), uterus (10.821 ng/g) and kidneys (10.463 ng/g). The highest values of the carry-over factor were noted in the same tissues. In the examined female wild boars, per os exposure to natural sources of the parent substance or a combination of ZEN and its metabolites led to different concentrations of ZEN in the analyzed tissues. Zearalenone concentrations were compatible with CF values in both experimental groups.

Detection and characterisation of Yersinia enterocolitica strains in cold-stored carcasses of large game animals in Poland.

Yersinia enterocolitica is an important foodborne pathogen. The aim of the present study was to identify the bioserotypes and virulence markers of Y.enterocolitica strains isolated from three different anatomical regions of cold-stored carcasses of large game animals intended for human consumption. Y.enterocolitica strains were found in 12/20 (60%) of the roe deer carcasses examined, 7/16 (43.8%) of red deer carcasses and 11/20 (55%) of wild boar carcasses. Of the 52 Y.enterocolitica strains, 19 were isolated from the perineum, followed by 17 strains from the peritoneum of the longissimus dorsi muscle and 16 from the tonsils. Only one strain was isolated from warm culture. Bioserotype 1A/NI was the most commonly found and was detected in 29/52 isolates. All isolates contained amplicons corresponding to ystB gene fragments. The relatively high degree of carcass contamination with Y.enterocolitica is of concern due to the growing popularity of game meat with consumers.

Yersiniosis - a zoonotic foodborne disease of relevance to public health.

INTRODUCTION: Y. enterocolitica is the causative agent of yersiniosis - a foodborne zoonosis with substantial importance to public health. Y. enterocolitica is widespread in the environment and animal populations, posing a potential source of infection to humans. OBJECTIVE: Presentation of yersiniosis as a zoonotic foodborne disease of relevance to public health. State of knowledge. Swine play an important role as a reservoir of Y. enterocolitica and insufficiently thermally processed pork is the main source of infection to humans. The correlation between strains isolated from pigs and from clinical cases of human yersiniosis has been sufficiently proven. Yersiniosis usually appears with gastrointestinal disturbances in children, whereas in adults it manifests in a pseudo-appendix form. The extra-enteric form of yersiniosis is rare. Classical bacteriological methods used for classifying Y. enterocolitica as pathogenic does not take into account the new aspects of the pathogenesis of yersiniosis. The examples are biotype 1A strains, commonly regarded as non-pathogenic, although they are increasingly often isolated from clinical cases of yersiniosis. Molecular methods seem much more effective and accurate in the diagnostic. New diagnostic tools such as real-time PCR, allows not only qualitative examination, but also quantitative evaluation of genes expression level, or single nucleotide polymorphism detection. CONCLUSIONS: Yersiniosis is an important food-borne zoonosis with wide range of clinical symptoms. Considering the fact that pork is the main source of infection for humans, public information campaigns seems to be an important element of the preventive measures against Y. enterocolitica infections.

The use of the HRM method for identifying possible mutations in the ymoA gene region and evaluating their influence on the enterotoxic properties of Y. enterocolitica strains.

BACKGROUND: The yst gene that encodes the production of Yst enterotoxins is one of the most important and reliable virulence markers. Its ability to produce Yst has been demonstrated in pathogenic strains isolated from clinical cases of yersiniosis with diarrhea. However, not all yst positive strains produce enterotoxins. According to some authors, Yst production can be restored in a silent strain by ymoA mutation. In this study, the HRM method was applied to identify ymoA single nucleotide polymorphism with the aim of evaluating their influence on the enterotoxic properties of Y. enterocolitica strains. RESULTS: Two genotypes (A and G) of the examined nucleotide sequence and some variations were detected in the HRM analysis. A phylogenetic analysis of 10 genotype A nucleotide sequences revealed 100% similarity with the Yersinia enterocolitica subsp. enterocolitica 8081 genome NCBI Acc. No. AM286415. An analysis of 10 genotype G nucleotide sequences and 3 variations sequences revealed two point mutations in the examined region: transition A3387326G and insertion A in position 3387368. However, no mutations were observed in the coding region of any of the examined ymoA gene fragments. Genotype G was identified in nearly all Y. enterocolitica strains isolated from pigs. Only 4 nucleotide sequences were similar to AM286415 and did not feature point mutations. In case of human Y. enterocolitica strains 31 were classified as belonging to genotype A, the remaining 59 belonged to genotype G and were characterized by the presence of point mutations. CONCLUSIONS: No correlations were observed between enterotoxic properties and the presence of mutations in the ymoA gene region of Y. enterocolitica strains isolated from both humans and pigs.

Bioserotypes and virulence markers of Yersinia enterocolitica strains isolated from mallards (Anas platyrhynchos) and Pheasants (Phasianus colchicus).

Yersinia enterocolitica is the causative agent of yersiniosis in different animal species and in humans. Food contaminated with Y. enterocolitica is the main source of infection for humans, and swine plays a major role in the transmission of the disease. There are a limited number of reports of the prevalence of Y. enterocolitica in wild animals and birds. This study characterized virulence markers associated with Y. enterocolitica isolates recovered from mallards and pheasants. Y. enterocolitica strains were isolated from 5 (11.11%) of 45 mallards originating from a cold culture (peptone, sorbitol, and bile salts medium) belonging to biotype 1A. Serotyping showed that three of these five serotypes represented serotype O:8, one belonged to serotype O:5, and one did not agglutinate with any of the sera and was classified as nonidentified. Molecular analysis for virulence markers detected the ystB gene, which encodes an enterotoxin, in five isolates. Y. enterocolitica was not detected in any of the 16 examined pheasants.