Yersinia species infection of lambs and cull cows at an abattoir

The rectal contents of 330 cull cows and 66 lambs were sampled over the 1984-1985 killing season at a New Zealand freezing works for the presence of Yersinia species. Samples from the cows revealed one isolation of Y. pseudotuberculosis and one isolation of Y. intermedia. Samples from the lambs gave thirteen isolates of Y. enterocolitica, three of Y. inter-media, two of Y. pseudotuberculosis and one of Y. frederiksenii. Two of the isolates of Y. enterocolitica were shown to be serotype 0:3. Although no parallel studies were carried out on young cattle or adult sheep, these findings are consistent with the hypothesis that Yersinia infection primarily involves young animals. Thus there exists the potential for transmission of the above species of Yersinia from young animals to man which needs further detailed study.     

The isolation of Yersinia sp. from feral and farmed deer faeces

Faecal samples from clinically normal farmed red deer, wapiti, fallow deer; and feral red deer and white tail deer were examined for members of the genus Yersinia. From 922 samples 176 strains of Y.enterocolitica, 56 strains of Y.frederiksenii, 29 strains of Y.kristensenii, eight strains of Y.intermedia, and seven strains of Y.pseudotuberculosis were isolated. High isolation rates of Yersinia sp. were recorded from some farms. Two herds had isolation rates of 33.3% and 36.8%. Sixteen strains of Yersinia sp. in addition to strains of Y.psuedotuberculosis were found to be Hela cell invasive. The majority of these strains were confined to a single herd and represented Y.enterocolitica biotypes I, II and III, Y.intermedia, Y. fredericksenii, and Y.kristensenii.     

A cohort study of Yersinia infection in goats

OBJECTIVE: To determine the temporal pattern of Yersinia infections in three goat flocks and examine the influence of management and seasonal factors on the incidence of those infections over a 1-year period. METHODS: A longitudinal study involving monthly culture of faeces for Yersinia spp. from age groups of randomly selected goats on three farms in the Manawatu region of New Zealand. RESULTS: The incidence of excretion of potentially pathogenic Yersinia (Yersinia pseudotuberculosis and Y enterocolitica biotypes 2, 3 and 5) peaked in winter and fell in summer. In contrast, environmental Yersinia (Y enterocolitica biotype 1A, Y frederiksenii, Y intermedia and Y rohdei) showed no clear pattern of seasonal variation. Pathogenic Yersinia were more prevalent in young animals than in adults, while environmental Yersinia were more prevalent in adults. The same type was isolated from the same animal in two or more successive months in about 20 to 25% of cases, and in the remaining cases there was a gap of at least one month between successive isolations, with many animals yielding a particular type on only a single occasion. A notable difference was that with the potentially pathogenic types, no animal had more than one period of time when it was found to be excreting a particular type, suggesting that immunity develops following exposure. In contrast, it was common for environmental types to be isolated from the same animal throughout the study period. Two goats were suspected to have developed clinical yersiniosis but all remaining infected animals showed no clinical signs of infection. CONCLUSIONS: Asymptomatic Yersinia carriage was common in goats in New Zealand, with a clear seasonal and age group pattern of infection with potentially pathogenic types. There was evidence that immunity developed to potentially pathogenic types. This is the first time that Y rohdei has been isolated from goats.     

The role of wild birds and the environment in the epidemiology of Yersiniae in New Zealand

A survey was carried out to determine the prevalence of Yersiniae in wild passerines in the lower half of the North island of New Zealand over a period of 12 months. Samples of soil, water and foliage were also collected. Out of a total of 1370 avian samples, only two strains of Y. pseudotuberculosis were isolated and a total of 98 strains of environmental yersiniae were identified, including Y. enterocolitica biotype 1a, Y. frederiksenii, Y. kristensenii and Y. intermedia. No strains of Y. pseudotuberculosis were isolated from 1032 non-avian samples collected, which included 100 samples taken from wild mammals. From the non-avian samples, 51 strains of environmental Yersiniae were identified, of which the relative prevalence of Yersinia enterocolitica, biotype 1a, Y. frederiksenii, Y. kristensenii and Y. intermedia was similar to that in the rural passerines. The prevalence of Yersiniae in soil samples was greater in rural areas than in urban areas of the survey region. In both rural and urban passerine populations, the prevalence of Yersiniae was greater in the winter and early summer than at other times of the year.     

The activity of chosen bacteriophages on Yersinia enterocolitica strains

The aim of the present study was to evaluate the lytic activity of three bacteriophages on Yersinia enterocolitica strains isolated from humans and pigs. The Y. enterocolitica strains tested belonged to 0:3, 0:9 and 0:2 serogroups. The ZD5 phage was obtained from a water sample, but remaining phages were obtained from the lysogenic Y. frederiksenii 7291 and Y. enterocolitica 8684 strains. All the Y. enterocolitica strains tested which belonged to 0:9 serogroup did not show any susceptibility to the bacteriophages used. The bacteriophages tested showed different lytic activity on the Y. enterocolitica 0:3 strains investigated. The phage susceptibility of Y. enterocolitica 0:3 strains revealed 9 different phage patterns. ZD5 phage showed the highest lytic activity, because it produced confluent lysis of the most Y. enterocolitica 0:3 strains tested. The Y. enterocolitica 0:2 strains isolated from pigs showed the similar phage susceptibility. The Y. kristensenii and Y. pseudotuberculosis strains tested were not sensitive to the bacteriophages used.     

The relationship of plasmids from environmental Yersinia isolates and the virulence plasmid of enteropathogenic Yersinia strains

The human pathogenic strains of Yersinia harbour a conserved plasmid carrying the Yop virulon. The virulence plasmid of Yersinia enterocolitica strains belonging to the serogroups O:3 and O:9 were used as probes to detect homologous sequences in plasmids of "avirulent" Yersinia strains. "Avirulent" Yersinia strains (Y. enterocolitica biogroup 1A, Y. intermedia, Y. kristensenii and Y. frederiksenii) lack the virulence plasmid. They are widely distributed in the environment and can frequently be isolated from clinical samples. Hybridisation experiments revealed a number of common genetic elements of the virulence plasmid and the plasmids of "avirulent" Yersinia strains. These elements were identified as genes involved in plasmid replication, as an endonuclease gene and as mobile genetic elements. However, none of the plasmid encoded virulence genes was present in the plasmids of "avirulent" Yersinia strains. The frequent occurrence and the possible etiological relevance of "avirulent" isolates will be discussed.     

Faecal bacteria of wild ruminants and the alpine marmot

Faecal samples from 60 red deer (Cervus elaphus), 13 roe deer (Capreolus capreolus), 7 chamois (Rupicapra rupicapra), 41 alpine marmot (Marmota marmota) and soils mixed with deer faeces from the Stelvio National Park were examined for Campylobacter sp. and Salmonella sp. with negative results. The same material, especially deer faeces, was a habitat highly suitable for Yersinia sp.: Y. enterocolitica (two biotypes) was isolated twice, Y. kristensenii (two serotypes) was isolated 19 times, Y. frederiksenii and Y. intermedia were isolated once. Antibiotic-resistant Escherichia coli were isolated from 16 specimens from wild ruminants, one from marmot and two from feeding places.     

Enterocolitis in cattle associated with Yersinia pseudotuberculosis infection

A syndrome in cattle of diarrhoea and death associated with enteric Yersinia pseudotuberculosis infection is described. Outbreaks occurred during winter and early spring in adult cattle grazing pastures waterlogged by recent flooding or persistent heavy rain. Antibiotic therapy was effective early in the course of the syndrome. At necropsy there was severe acute enterocolitis, and bacteria consistent with Y. pseudotuberculosis were observed in the lesions. This organism could usually be isolated from the intestines of affected animals but was recovered less often from other organs. Representative isolates were identified as Y. pseudotuberculosis serotype III. The association of this syndrome with waterlogged pastures and low temperatures suggests that these conditions favour transmission of Y. pseudotuberculosis infection in cattle. The role of Y. pseudotuberculosis as primary pathogen requires confirmation.     

Yersiniosis in deer from the Otago-Southland region of New Zealand

Samples from 350 routine deer cases submitted to Invermay Animal Health Laboratory for diagnosis during 1979-1982 were examined specifically for the presence of Yersinia sp. An analysis of 57 cases of yersiniosis due to Yersinia pseudotuberculosis is made and two cases due to Yersinia enterocolitica are described. The occurrence of cases appeared strongly correlated with periods of stress, predominantly in winter when cool wet conditions and lack of grazing combined to precipitate the disease. Animals up to one year old were most commonly affected (64% of cases). Of the 61 strains of Yersinia pseudotuberculosis isolated 35 were of Serotype I, 17 of Serotype II and nine of Serotype III. All strains of Yersinia pseudotuberculosis and the strains of Yersinia enterocolitica isolated from the two cases described were Hela cell invasive and gave a positive autoagglutination test for virulence.     

Diagnosis o Yersinia enterocolitica infections: a review on classical identification techniques and new molecular biological methods

Only three of the eleven species of the genus Yersinia are associated with disease. Y. pestis is the causative agent of plague, Y. pseudotuberculosis and several pathogenic bio/serovars of the species Y. enterocolitica cause yersiniosis. New Y. enterocolitica subspecies with diagnostic relevance have been proposed allowing the differentiation of European and American isolates. The ISO-standard (ISO 102739) summarizes the knowledge gained from enrichment and isolation of Y. enterocolitica from food and feed samples. The final biochemical identification must be carried out by classical tube testing, as commercially available test-systems are not sensitive and specific. For the assessment of the presumptive pathogenicity of a Y. enterocolitica isolate empiric virulence markers can be replaced by PCR assays targeting plasmoidal or chromosomal genes. Their evaluation in terms of routine diagnostic procedures is still missing. The definite identification of Y. enterocolitica isolates can also be achieved by sequencing the 16S rRNA gene. Immunoblot based on plasmoidal encoded Yersinia proteins enables the serological determination of animal and human infections. The development of simple, sensitive and specific rapid identification systems applicable for the direct and indirect diagnosis for veterinary use is a challenge for the future.     

Occurrence of virulence markers in species of Yersinia isolated from animals in Nigeria

Fourteen strains of Yersinia species isolated from apparently healthy pigs and cattle in Nigeria were screened for four virulence markers using six test systems. These were two in vitro assays, namely, calcium dependency and autoagglutination, both at 37 degrees C, the Serény test in guinea-pigs and the detection of heat-stable enterotoxin (ST) by the rabbit ileal loop test, the ligated intestine test in pigs and the infant mouse system. Seven of the 14 strains of Yersinia were positive for one or more of these tests. Six of nine strains of Y. enterocolitica and one of four Y. intermedia were positive in one or more tests. The only strain of Y. frederiksenii isolated was negative in all six test systems. All three strains of Y. enterocolitica, serotype 0:8 and the only serotype 0:3 isolated were positive in one or more tests. However, only two of five strains of Y. enterocolitica serotype 0:12, 26, the most frequently encountered, were positive. A good correlation was observed between test results of calcium dependency, autoagglutination and Serény assays. The results indicate that cattle and pigs have the potential to transmit virulent strains of Y. enterocolitica to human beings in Nigeria.     

Faecal survey of deer for Yersinia pseudotuberculosis and Salmonella sp

The results of a national survey of faeces from clinically normal deer for Salmonella sp. and Yersinia pseudotuberculosis are reported. Five isolates of Y. pseudotuberculosis and none of Salmonella sp. were made from 3810 faeces representing 122 farms.     

Isolation of pathogenic yersiniae from wild animals in Bulgaria

Pathogenic Yersinia strains were isolated between December 1998 and April 1999 from 37 wild animals: rabbit (Lepus europeus), boar (Sus scrofa scrofa), asiatic jackal (Canis aureus), red fox (Vulpes vulpes), mouflon (Ovis musimon), european river otter (Lutra lutra), beech marten (Martes foina), polecat (Musleta putorius) and wild cat (Felis silvestris). It was established that among the wild animals Y. enterocolitica strains of serotype 0:3 predominated, accompanied by Y. pseudotuberculosis strains of serotype 0:3. In one sample from asiatic jackal and one sample from rabbit, Y. enterocolitica serotype 0:8 was isolated. Yersinia enterocolitica and Y. pseudotuberculosis strains were isolated from tonsils and tongues as well as from the viscera--lung, liver, heart, spleen, kidney and lymph nodes, mainly in young animals (1-2 years of age). The results showed that wild animals are a possible natural reservoir for pathogenic Y. enterocolitica and Y. pseudotuberculosis and are included in the epidemiological chain of yersinioses.     

Epidemiological study of Yersinia enterocolitica in swine herds in Québec

The objectives of this study were the identification of the different contamination sources of Yersinia enterocolitica, as well as the determination of the prevalence and the distribution of the different genotypes in swine herds. The owners of 20 farms, located in the Richelieu-Yamaska region, agreed to participate in the study. Each farm was visited a minimum of 5 times between May and October 1997, and, at each visit, 20 environmental and 10 fecal samples were collected. Yersinia enterocolitica isolates were identified, serotyped, and submitted to a genetic characterization by pulsed-field gel electrophoresis. The correlation coefficient (0.61) between prevalence in environment and in feces was significant (P = 0.004). Among the 153 positive samples, 93.5% belonged to serotype 0:3. The comparison of PFGE profiles revealed that all environmental Y. enterocolitica isolates had a profile identical to that of isolates recovered in feces from the corresponding farms. Also, when the genetic profiles of isolates recovered from feces collected at the first visit were compared with the profiles of isolates obtained from the subsequent visits, the same profile was observed on every farm. We concluded that environment does not represent the main source of contamination of swine by Y. enterocolitica and that, in most instances, the same strain persists in a barn from one production lot to another.     

Yersinia enterocolitica in sheep - a high frequency of biotype 1A

Background

Pigs are regarded as the main reservoir for human pathogenic Yersinia enterocolitica, which is dominated by bioserotype 4/O:3. Other animals, including sheep, have occasionally been reported as carriers of pathogenic strains of Y. enterocolitica. To our knowledge, this is the first study performed in the Nordic countries in which the presence of Y. enterocolitica in sheep is investigated.

Methods

Tonsils and faecal samples collected from sheep slaughtered on the island Gotland (Sweden) from September 2010 through January 2011 were analysed for presence of Y. enterocolitica. In an attempt to maximize recovery, several cultural strategies were applied. Various non-selective media were used and different temperatures and durations of the enrichment were applied before subculturing on Cefsulodin Irgasan Novobiocin (CIN) agar. Presumptive Y. enterocolitica colonies were subjected to urease, API 20E and agglutination test. Yersinia enterocolitica isolates were biotyped, serotyped, and tested for pathogenicity using a TaqMan PCR directed towards the ail-gene that is associated with human pathogenic strains of Y. enterocolitica.

Results

The samples collected from 99 sheep yielded 567 presumptive Y. enterocolitica colonies. Eighty urease positive isolates, from 35 sheep, were identified as Y. enterocolitica by API 20E. Thirty-four of 35 further subtyped Y. enterocolitica isolates, all from faecal samples, belonged to biotype 1A serotype O:5, O:6. O:13,7 and O:10. One strain was Yersinia mollaretii serotype O:62. No human pathogenic strains of Y. enterocolitica were found in the investigated sheep. Other species identified were Y. kristensenii (n = 4), Y. frederiksenii/intermedia (n = 3), Providencia rettgeri (n = 2), Serratia marcescens (n = 1) and Raoultella ornithinolytica (n = 1).

Conclusions

This study does not support the hypothesis that sheep play an important role in transmission of the known human pathogenic Y. enterocolitica in the studied geographical region. However, because there are studies indicating that some strains of Y. enterocolitica biotype 1A may cause disease in humans, the relative importance of sheep as carriers of human pathogenic strains of Y. enterocolitica remains unclear. Tonsils do not appear to be favourable sites for Y. enterocolitica biotype 1A in sheep.     

Suppression of heat-stable enterotoxin production by Yersinia spp. in milk

One of 16 raw milk isolates of Yersinia enterocolitica and Y. intermedia produced heat-stable enterotoxin (ST) in milk at 25 degrees C but not at 4 degrees C after 21 days of incubation. A catabolite repression of ST synthesis by the lactose-fermenting strain of Y. enterocolitica was observed when 4.6% lactose was added to trypticase soy broth. However, the lactose-fermenting strain was killed by acid produced by lactose fermentation in milk and did not produce ST in milk with the pH adjusted to neutrality. This study suggested that lactose and fat in milk are not the fundamental inhibitors of ST synthesis by Y. enterocolitica and that repression of ST synthesis may be related to other components.     

Prevalence of anti-Yersinia antibodies in European brown hares in North-Rhine Westphalia, Germany

Yersinia (Y.) pseudotuberculosis infections may lead to significant lethality in European brown hare (Lepus europaeus, Pallas) populations especially during the cold and wet seasons. In recent decades, also Y. enterocolitica was isolated from hares found dead. Consequently, a Western-blot technique proved to be valuable for the detection of antibodies against all pathogenic Yersinia isolates was applied to monitor the prevalence of antibodies in hare populations in North-Rhine Westphalia, Germany. A total of 89.6% of the 230 animals tested was seropositive. Further investigations should be performed to elucidate the role of subclinical yersiniosis in the decline of European brown hare populations in Germany.     

Development of an ELISA procedure to detect swine carriers of pathogenic Yersinia enterocolitica

An enzyme immunoassay (ELISA) was developed to detect antibodies in pigs against the lipopolysaccharidic antigen of the three serotypes of Yersinia enterocolitica mostly associated with human infections. Recent epidemiological evidence has demonstrated that pigs and pork are important sources of yersiniosis in humans. The purpose of this study was to clarify the use of an ELISA to detect swine carriers of this enteroinvasive bacteria by examining seroconversion and tissue distribution of Y. enterocolitica following experimental infection and then screening pigs at a slaughterhouse by bacterial culture and ELISA. It was observed that seroconversion occurred in animals experimentally inoculated with Y. enterocolitica but not with other enterobacteria. It was also found that 27% of swine at a slaughterhouse carried the bacterium in their tonsils and/or intestinal tract, whereas 66% showed serological evidence of previous infection. About 6% of swine at slaughter were culture-positive, but seronegative. Although, similar numbers of swine showed serological evidence of previous infection by each of the three Y. enterocolitica serotypes tested, virtually all culture isolates belonged to serotype O:3. This ELISA appears as a valuable control tool that can be used, in conjunction with culture, to identify pigs or herds infected by strains of Y. enterocolitica associated with human infections.     

Analysis of the presence of ail, ystA and ystB genes in Yersinia enterocolitica strains isolated from aborting sows and aborted fetuses

Forty-five Yersinia enterocolitica strains isolated from aborted fetuses and placentas and from vaginal and rectal swabs of aborting sows were subjected to serotyping, biochemical typing and polymerase chain reaction multiplex analyses to detect the presence of the ail, yst A and ystB genes. The isolates were recovered from the internal organs (tonsil, lung, liver, spleen, kidney, mesentheric lymph nodes, small intestine and rectal intestine) of 18 (18.6%) of 97 aborted fetuses examined, two (8%) of 25 aborted placentas and 27 (15.8%) of 172 examined aborting sows. Serotyping of Y. enterocolitica revealed that only six (13.3%) of the examined isolates belonged to serotype O:3, with a considerable number of isolates (31.1%) having serotype O:5, while biochemical studies showed that as many as 40 of the 45 strains belonged to biotype 1A. As expected, the Y. enterocolitica strains of bioserotype 4/O:3 contained ail and ystA genes, while strains of biotype 1A contained only the ystB gene.     

Prevalence of Salmonella, Yersinia and Campylobacter spp. in feral raccoons (Procyon lotor) and masked palm civets (Paguma larvata) in Japan

To estimate the public and animal health risk that alien species pose, the prevalence of Salmonella, Yersinia, and Campylobacter spp. in feral raccoons (Procyon lotor, n=459) and masked palm civets (Paguma larvata, n=153), which are abundant alien species in Japan, was investigated in urban and suburban areas of Japan. Salmonella enterica was detected from 29 samples [26 raccoons, 5.7%, 95% confidence interval (CI) 7.8-3.5%; three masked palm civets, 2.0%, 95% CI 4.2-0%]. Many of the isolates belonged to serovars that are commonly isolated from human gastroenteritis patients (e.g. S. Infantis, S. Typhimurium, and S. Thompson). The antimicrobial susceptibility test showed that 26.9 % of the isolates from raccoons were resistant to at least one antimicrobial agent, whereas none of the isolates from masked palm civets were resistant. Yersinia sp. was detected from 193 samples (177 raccoons, 38.6%, 95% CI 43.0-34.1%; 16 masked palm civets, 10.5%, 95% CI 15.3-5.6%). All virulent Yersinia strains belonged to Yersinia pseudotuberculosis, which was isolated from seven (1.5%, 95% CI 2.6-0.4%) raccoons and six (3.9%, 95% CI 7.0-0.8%) masked palm civets. According to the detection of virulence factors, all the Y. pseudotuberculosis isolates belonged to the Far Eastern systemic pathogenicity type. Campylobacter spp. was detected from 17 samples (six raccoons, 1.3%, 95% CI 2.3-0.3%; 11 masked palm civets, 7.2%, 95% CI 11.3-3.1%). Among these, three isolates from raccoons were identified as C. jejuni. These results showed that these pathogens can be transmitted by human activities, other wild animals, and the environment to feral raccoons and masked palm civets, and vice versa. As these animals have omnivorous behaviour and a wide range of habitats, they can play an important role in the transmission of the enteric pathogens.