Agata Bancerz-Kisiel

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.

Presence of ail and ystB genes in Yersinia enterocolitica biotype 1A isolates from game animals in Poland

The pathogenicity of Yersinia enterocolitica is associated with the presence of plasmid and chromosomal virulence genes. Strains belonging to biotype 1A do not possess pYV plasmids, often harbour the ystB gene and usually lack the ail gene, which is the main virulence marker for Y. enterocolitica. The simultaneous presence of ail and ystB is uncommon. In this study, 21/218 (9.6%) biotype 1A Y. enterocolitica isolates from rectal swabs of wild boar (Sus scrofa; n = 18), red deer (Cervus elaphus; n = 2) and roe deer (Capreolus capreolus; n = 1) in Poland harboured both ail and ystB genes.

Ultrasonographic fetometry formulas of inner chorionic cavity diameter and biparietal diameter for medium-sized dogs can be used in giant breeds

The aim of this study was to evaluate the suitability of the ultrasonographic fetometry method, involving inner chorionic cavity diameter (ICC) and biparietal diameter (BP) measurements, to predict the parturition date in giant breed dogs. Overall, 30 ICC and 24 BP measurements were taken on 24 giant breed bitches. The measured values were substituted into Luvoni and Grioni (2000) formulas for medium-sized bitches because formulas with ICC and BP to dogs with a body mass greater than 40 kg have not been defined. The accuracy of the parturition date predictions proved the method to be highly useful in the observed group of dogs. Prediction accuracy in the giants ranged between 54.16% (± 1 day, using BP) and 90% (± 2 days, using ICC), depending on the parameter measured and precision levels used. Numerically, the results obtained using ICC were better; however, no statistically significant differences between ICC and BP accuracy were found when comparing the effectiveness of the parturition date predictions. Regression lines based on the own fetometric measurements were highly convergent with the lines defined by Luvoni and Grioni (2000) formulas for medium-sized bitches. This outcome suggests a similar gestational development of fetuses in giant dogs and the possible use of Luvoni and Grioni (2000) formulas for medium-sized dogs with breeds weighing greater than 40 kg.

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.

Yersinia enterocolitica strains isolated from beavers (Castor fiber)

Pseudocloacal swabs and palatine tonsils from beavers have been examined for the Yersinia enterocolitica presence. Thirty-six samples from 24 beavers were collected and subjected to bacteriological examinations including sero- and biotypisation. Amplicons confirmed by PCR as Y. enterocolitica were sequenced. Positive samples originated from 4 out of the 24 beavers (16.7 %) and all the strains belonged to biotype 1A. The study suggested that Y. enterocolitica could be isolated from beavers, which may therefore be treated as a reservoir, a significant factor of water contamination and a vector of the Y. enterocolitica.

Monitoring of Yersinia enterocolitica strains from free-living animals using different methods.

The aim of the study was to monitor Y. enterocolitica strains from free-living animals captured during 2011-2014 hunting seasons in Poland using warm (ITC) and cold (PSB) enrichment and molecular examination. Over 1600 samples have been cultured. After ITC/PSB enrichment 237 strains presenting features characteristic for Y. enterocolitica were isolated. Molecular examination using multiplex PCR revealed 140 isolates from PSB and 78 from ITC. The concentration of pathogenic Yersinia in asymptomatic carriers is low and the PCR detection should be preceded by bacteriological examination.

Characterization of Yersinia enterocolitica strains potentially virulent for humans and animals in river water.

The aim of this study was to isolate and identify potentially pathogenic strains of Yersinia enterocolitica in water samples collected from the upstream section of the Drwęca River in Poland.

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.

Phylogenetic analysis of bovine papillomavirus E5 detected in equine sarcoids in Poland

The aim of the study was to analyse a part of the sequence of the E5 gene of bovine papillomaviruses (BPV) associated with equine sarcoids in Polish horses. Samples of 40 skin lesions obtained from 29 horses were collected for molecular examination. The PCR amplicons of BPV DNA were detected in 38 specimens. After phylogenetic analysis 37 specimens were recognized as BPV-1 and one as BPV-2. Phylogenetic analysis has allowed the classification of the amplicons into two phylogenetic groups (A1,) and four separate isolates (2, 10, 16, 17).

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.