Toxoplasmosis in sheep III. Further evaluation of the ability of a live Toxoplasma gondii vaccine to prevent lamb losses and reduce congenital infection following experimental oral challenge

The aim of this study was to compare the ability of a live incomplete strain (Strain 48) and a live complete strain (Strain 89) of Toxoplasma gondii to protect against abortion and congenital infection following an oral challenge of T. gondii oocysts. Sixty-nine two-tooth ewes were immunised pre-tupping with live Strain 48 of T. gondii tachyzoites and seventy ewes were immunised with Strain 89. Eighty-two serologically negative ewes served as controls. At mid-pregnancy half of the ewes were challenged orally with T. gondii oocysts (2×10⁵/ewe).

The ewes vaccinated with Strain 48 were significantly (p<0.05) protected against the effects of experimental challenge and the rate of congenital infection was also significantly (p<0.15) reduced. The ewes vaccinated with Strain 89 were also significantly (p<0.05) protected.

The serological response to challenge as measured by both the Dye test and the Indirect Haemagglutination test varied considerably between the two vaccinated groups.     

Long-Term Study of Toxoplasma gondii Infection in a Swedish Sheep Flock

Lundén, A., A. Näsholm and A. Uggla: Long term study of Toxoplasma gondii infection in a Swedish sheep flock. Acta vet. scand. 1994,35,273-281.– The infection rate of Toxoplasma gondii was studied during 6 years in a sheep flock in central Sweden. The flock consisted of 165-249 breeding ewes of which 20-35% were lambs less than 1 year old. Most ewes were slaughtered when 5 years old. The sheep were kept indoors from end of September to early May. Lambing took place in March and April. Individual serum samples were collected twice a year, once just before turning the sheep out to pasture in the spring, and again after housing in the autumn. Sera were analysed by ELISA for antibodies to T. gondii. The seroprevalence varied between 10% and 45% during the 6 years of observation. Seroconversion was detected predominantly at the autumn sampling, indicating that in most cases infection was acquired at pasture. Subclinical effects of T. gondii infection on lamb weight, litter size, total litter weight and ewe weight were also studied. Lambs born to chronically infected ewes were lighter at birth than those of uninfected ewes, but this disparity was no longer evident at weaning.     

Vaccination of sheep with a live incomplete strain (S48) of Toxoplasma gondii and their immunity to challenge when pregnant

Sixty-four ewes were vaccinated with tachyzoites of an incomplete strain (S48) of Toxoplasma gondii grown either in the peritoneal cavity of mice (group 1) or vero cell culture (group 2) and 30 ewes (group 3) were not vaccinated. All the ewes were mated 77 days later and challenged orally with 2000 sporulated oocysts at 89 to 90 days of gestation. Ten additional unvaccinated (group 4) and 10 vaccinated (group 5) control ewes were not challenged. The unvaccinated ewes developed a characteristic febrile response to challenge while in the vaccinated ewes the fever commenced earlier but was less severe and of shorter duration. After challenge, the antibody titres against T gondii rose rapidly to high values in the vaccinated ewes while the ewes in group 3 responded more slowly. Only eight of the 45 fetuses/lambs (17.8 per cent) from group 3 were viable compared with 72.3 per cent of those in group 1 and 80.8 per cent of those in group 2. Gestation in the unvaccinated challenged ewes was shortened and the mean birthweight of their single, viable offspring was significantly lower than the weight of single lambs from the vaccinated (groups 1 and 2) and control ewes (groups 4 and 5). Examination of precolostral sera showed that almost two-thirds of the lambs from the vaccinated ewes were infected in utero. The 20 control ewes appeared clinically normal at all times and lambed normally. The two vaccine preparations were equally effective.     

Majority of T. gondii seropositive chickens (Gallus domesticus) in Central Ethiopia carries the infective parasite

Background

The prevalence of Toxoplasma gondii in free range chickens is a good indicator of the prevalence of T. gondii oocysts in the environment. The aim of this study was to isolate T. gondii parasites from heart and brain of seropositive free range (FR) chickens.

Findings

Isolation of T. gondii from pooled heart and brain of 41 direct agglutination test (DAT) positive (≥1:40) free range chickens (Gallus domesticus) was carried out by bioassay in mice. T. gondii specific antibodies in mice were assayed by DAT and microscopy was employed for detection and enumeration of brain tissue cysts. Overall, bioassay was positive in 29 (70.7%) chicken samples. T. gondii tissue cysts were isolated from 59% (24/41) of bioassayed chickens: from 2 of 7 chickens with a titer of 1: ≤ 60, 2 of 5 with titer 1: 180, 6 of 8 with titer 1: 540, 10 of 15 with titer 1: 1620, 1 of 2 with titer 1: 6000, 2 of 3 with titer 1:18000, 1 of 1 with titer 1:54000. None of the isolates was pathogenic for mice. Tissue cysts were detected from 61% of seropositive mice (DAT ≥ 1:40). Generally, tissue cyst counts per brain of mouse were low (mean: 132.7 ± 84.4; range: 47–352).

Conclusions

Majority of T. gondii seropositive chickens (Gallus domesticus) in Central. Ethiopia carries the infective parasite. Tissues from the free range chicken might be a source infection for animals and humans.     

Trial of a novel experimental Toxoplasma iscom vaccine in pregnant sheep

Fifteen vaccinated ewes (group 1) and 13 unvaccinated ewes (group 2) were each challenged orally with 2000 sporulated Toxoplasma gondii oocysts at 91 (+/- 1) days' gestation. Another four pregnant ewes acted as unvaccinated unchallenged controls. Lamb mortality in group 1 was 36.4% after a mean gestation of 141 days while in group 2 it was 64.7% after a mean gestation of 131.5 days. These differences were not statistically significant. However, substantially more specific antibody was detected in precolostral sera from live lambs from vaccinated ewes than in live lambs from unvaccinated ewes in group 2.     

Increased Toxoplasma gondii positivity relative to age in 125 Scottish sheep flocks; evidence of frequent acquired infection

Toxoplasma gondii seroprevalence was determined in 3333 sheep sera from 125 distinct sheep flocks in Scotland, with the majority of flocks being represented by 27 samples, which were collected between July 2006 and August 2008. The selected farms give a representative sample of 14 400 sheep holdings identified in the Scottish Government census data from 2004. Overall T. gondii seroprevalence, at individual sheep level, was determined to be 56.6%; each flock tested, had at least a single positive animal and in four flocks all ewes tested positive. The seroprevalence of sheep increased from 37.7% in one year old stock to 73.8% in ewes that were older than six years, showing that acquired infections during the life of the animals is frequent and that environmental contamination by T. gondii oocysts must be significant. The median within-flock seroprevalence varied significantly across Scotland, with the lowest seroprevalence of 42.3% in the South and the highest seroprevalence of 69.2% in the far North of Scotland and the Scottish Islands, while the central part of Scotland had a seroprevalence of 57.7%. This distribution disequilibrium may be due to the spread and survival of oocysts on pasture and lambing areas. A questionnaire accompanying sampling of flocks identified farms that used Toxovax^®, a commercial vaccine that protects sheep from abortion due to T. gondii infection. Only 24.7% of farmers used the vaccine and the vaccine did not significantly affect the within flock seroprevalence for T. gondii. The implications for food safety and human infection are discussed.     

Neospora caninum infection as a cause of reproductive failure in a sheep flock

Neospora caninum has been detected only sporadically in cases of ovine abortion, and it has therefore traditionally been considered as an unimportant parasite in small ruminants. This study was carried out with the aim of identifying the pathogen causing serious reproductive problems on a commercial sheep farm. Sera from all rams and ewes tested negative for antibodies against Border disease virus, Schmallenberg virus and Coxiella burnetii, and infections by these agents were therefore ruled out. Nevertheless, seropositivity to N. caninum and/or Toxoplasma gondii was detected, although the seroprevalence was higher in the case of N. caninum. The percentage of lambings and the number of lambs per dam were significantly lower in ewes that were seropositive to N. caninum while no effect on these parameters was detected in ewes that were seropositive to T. gondii. There was also no evidence of infection by T. gondii in the foetal/lamb tissues analyzed by PCR and/or immunohistopathological techniques. On the contrary, the DNA of N. caninum was detected in 13 out of 14 foetuses/lambs descendant from dams seropositive to this parasite. Characteristic lesions caused by N. caninum and/or its antigen were also detected. Genotyping of the N. caninum DNA revealed only two closely related microsatellite multilocus genotypes. The results clearly demonstrate that infection by N. caninum was the cause of the low reproductive performance of this sheep flock.     

A field evaluation of two vaccines against Mycoplasma hyopneumoniae infection in pigs

Background

A field trial was carried out with two Mycoplasma hyopneumoniae vaccines in order to investigate the benefit of vaccination under field conditions in modern Danish pig production facilities with pigs being positive for M. hyopneumoniae. The M. hyopneumoniae infection of the herd was confirmed through blood samples that were positive for antibodies against M. hyopneumoniae combined with gross lesions of the lungs related to M. hyopneumoniae at slaughter and detection of M. hyopneumoniae by polymerace chain reaction in these lesions.

Results

A total of 2,256 pigs from two herds were randomly divided into three groups. Group 1 received 2 mL ThoroVAX^®VET, Group 2 received 1 mL Ingelvac^®MycoFLEX, and Group 3 was a non-vaccinated control group. The vaccination was performed by a person who was not involved in the rest of the trial and vaccination status thereby blinded to the evaluators.The prevalence of lung lesions related to M. hyopneumoniae were significantly lower for pigs vaccinated with ThoroVAX^®VET but not for pigs vaccinated with Ingelvac^®MycoFLEX^®, when compared to non-vaccinated pigs. There was no significant effect of vaccination on growth rate, antibiotic consumption or mortality.

Conclusion

This trial demonstrated that vaccination with Thoro^®VAX VET was effective in reducing the prevalence of lung lesion in pig units infected with M. hyopneumoniae.     

The role of rpoS,hmp, and ssrAB in Salmonella enterica Gallinarum and evaluation of a triple-deletion mutant as a live vaccine candidate in Lohmann layer chickens

Salmonella enterica Gallinarum (SG) causes fowl typhoid (FT), a septicemic disease in avian species. We constructed deletion mutants lacking the stress sigma factor RpoS, the nitric oxide (NO)-detoxifying flavohemoglobin Hmp, and the SsrA/SsrB regulator to confirm the functions of these factors in SG. All gene products were fully functional in wild-type (WT) SG whereas mutants harboring single mutations or a combination of rpoS, hmp, and ssrAB mutations showed hypersusceptibility to H₂O₂, loss of NO metabolism, and absence of Salmonella pathogenicity island (SPI)-2 expression, respectively. A triple-deletion mutant, SGΔ3 (SGΔrpoSΔhmpΔssrAB), was evaluated for attenuated virulence and protection efficacy in two-week-old Lohmann layer chickens. The SGΔ3 mutant did not cause any mortality after inoculation with either 1 × 10⁶ or 1 × 10⁸ colony-forming units (CFUs) of bacteria. Significantly lower numbers of salmonellae were recovered from the liver and spleen of chickens inoculated with the SGΔ3 mutant compared to chickens inoculated with WT SG. Vaccination with the SGΔ3 mutant conferred complete protection against challenge with virulent SG on the chickens comparable to the group vaccinated with a conventional vaccine strain, SG9R. Overall, these results indicate that SGΔ3 could be a promising candidate for a live Salmonella vaccine against FT.     

Western blot analysis of the IgG response of sheep vaccinated with S48 Toxoplasma gondii (Toxovax)

The IgG antibody responses of sheep vaccinated by the subcutaneous injection of live tachyzoites of ‘incomplete’ strain S48 toxoplasma (Toxovax) were analysed by Western blotting. Antibodies corresponding to a range of tachyzoite antigens (13 to 48 kD) were detected, but the response was dominated by antibody recognising a 30 to 32 kD band. Unvaccinated ewes challenged orally with oocysts of the ‘complete’ M3 toxoplasma strain had a more complex IgG response that recognised antigens in six dominant bands of similar intensity as those in sheep vaccinated with S48 tachyzoites and then challenged with M3 oocysts. No differences were detected between the antigenic structures of the S48 tachyzoites and RH strain tachyzoites when the antigens were probed with immune ovine sera. Many of the anitgens of the S48 tachyzoites that were recognised had molecular weights similar to those of antigens that have been identified in other strains of toxoplasma.     

The incidence of caseous lymphadenitis in Alberta sheep and assessment of impact by vaccination with commercial and experimental vaccines.

In Alberta, caseous lymphadenitis (CLA) is one of the leading causes of lamb and mutton carcass condemnation. In this study, serologic results confirmed a high (50-94%) incidence of exposure to Corynebacterium pseudotuberculosis, the causative agent of CLA, in mature, unvaccinated sheep in southern Alberta. To assess the efficacy and impact of vaccination with 2 commercial (Glanvac-6 and Case-Vac) and 1 experimental (WC+ MDP-GDP) CLA vaccines, a series of 3 field trials in 3249 ewes and lambs was conducted in affected flocks from 1992-1996. Efficacy was assessed from the serological response to vaccination, prevalence and size of injection site reactions by treatment, and the incidence of CLA abscesses. Overall, agglutinating antibody titres to C. pseudotuberculosis in lambs vaccinated with WC+MDP-GDP and Case-Vac remained significantly elevated above nonvaccinated control lambs for the 12 mo period after the initial vaccination. Lambs vaccinated with the WC/MDP-GDP maintained higher titres (P < 0.06) than those vaccinated with Case-Vac for the period from 6 to 12 mo after vaccination. Agglutinating antibody titres for lambs vaccinated with Glanvac did not differ from those of controls at any point during the 12 mo period after vaccination. The number of injection site reactions was elevated in lambs vaccinated with Glanvac as compared to those vaccinated with WC+MDP-GDP but the size of injection site reactions did not significantly differ. Sheep vaccinated with WC+ MDP-GDP also had a reduced incidence of putative CLA abscesses, although confirmation of the presence of C. pseudotuberculosis was only successful in a small number of instances.     

Titration of Toxoplasma gondii oocysts in non-pregnant sheep and the effects of subsequent challenge during pregnancy

Fifty-nine ewes, seronegative to Toxoplasma gondii, were allocated to four groups which received 2000, 200, 20 or no M1 strain toxoplasma oocysts 56 days before mating. Fifty-one of them subsequently became pregnant and were challenged with 10,000 oocysts between 78 and 83 days of gestation. Infection with 2000 oocysts induced a pyrexia, seroconversion and protective immunity in all the recipient animals. Ewes that received either 20 or no oocysts before pregnancy were susceptible to subsequent challenge and severe fetal mortality occurred. In this study 200 oocysts was the threshold value for the induction of toxoplasma infection in sheep, although not all the ewes that seroconverted to this dose were protected against further challenge.     

Protection from parainfluenza-3 virus and persistence of infectious bovine rhinotracheitis virus in sheep vaccinated with a modified live IBR-PI-3 vaccine.

Ewes (N = 7) and their lambs (N = 12) were vaccinated with a commercial modified live infectious bovine rhinotracheitis-parainfluenza type 3 virus vaccine. Both the vaccinated ewes and lambs and a group of unvaccinated ewes (N = 8) and their lambs (N = 13) were subsequently challenged with virulent parainfluenza type 3 virus. Although absolute immunity to infection and clinical response was not conferred, the clinical response was less severe in vaccinated lambs. Vaccinated animals also shed parainfluenza type 3 virus in nasal secretions for a shorter time than nonvaccinated animals. Some vaccinated lambs developed a persistent infectious bovine rhinotracheitis virus infection that was recrudesced by treatment with dexamethasone. It was concluded that vaccination was of benefit in reducing the severity of infection with parainfluenza type 3 virus. However, the inclusion of infectious bovine rhinotracheitis virus in a vaccine for sheep respiratory tract disease is highly questionable as it might increase the risk factor associated with vaccination. The consequences of the persistence of infectious bovine rhinotracheitis virus are now known.     

Pathogenesis of reproductive failure induced by Trypanosoma vivax in experimentally infected pregnant ewes

The present study was aimed at investigating the effect of experimental infection by Trypanosoma vivax in different stages of pregnancy, determining the pathogenesis of reproductive failure, and confirming transplacental transmission. We used 12 pregnant ewes distributed into four experimental groups: G1, was formed by three ewes infected with T. vivax in the first third of pregnancy (30 days); G2 comprised three infected ewes in the final third of pregnancy (100 days); G3 and G4 were composed of three non-infected ewes with the same gestational period, respectively. Each ewe of G1 and G2 was inoculated with 1.25 × 10⁵ tripomastigotes. Clinical examination, determination of parasitemia, serum biochemistry (albumin, total protein, glucose, cholesterol, and urea), packed cell volume (PCV), serum progesterone, and pathological examination were performed. Placenta, amniotic fluid, blood and tissues from the fetuses and stillbirths were submitted to PCR. Two ewes of G1 (Ewe 1 and 3) presented severe infection and died in the 34^th and 35^th days post-infection (dpi), respectively; but both fetuses were recovered during necropsy. In G2, Ewe 5 aborted two fetuses on the 130^th day (30 dpi) of pregnancy; and Ewe 6 aborted one fetus in the 140^th day (40 dpi) of gestation. Ewes 2 and 4 delivered two weak lambs that died five days after birth. Factors possibly involved with the reproductive failure included high parasitemia, fever, low PCV, body score, serum glucose, total protein, cholesterol, and progesterone. Hepatitis, pericarditis, and encephalitis were observed in the aborted fetuses. The presence of T. vivax DNA in the placenta, amniotic fluid, blood, and tissues from the fetuses confirms the transplacental transmission of the parasite. Histological lesion in the fetuses and placenta also suggest the involvement of the parasite in the etiopathogenesis of reproductive failure in ewes.     

Preslaughter diet management in sheep and goats: effects on physiological responses and microbial loads on skin and carcass

Sixteen crossbred buck goats (Kiko x Spanish; BW = 32.8 kg) and wether sheep (Dorset x Suffolk; BW = 39.9 kg) were used to determine the effect of preslaughter diet and feed deprivation time (FDT) on physiological responses and microbial loads on skin and carcasses. Experimental animals were fed either a concentrate (CD) or a hay diet (HD) for 4 d and then deprived of feed for either 12-h or 24-h before slaughter. Blood samples were collected for plasma cortisol and blood metabolite analyses. Longisimus muscle (LM) pH was measured. Skin and carcass swabs were obtained to assess microbial loads. Plasma creatine kinase activity (863.9 and 571.7 ± 95.21 IU) and non-esterified fatty acid concentrations (1,056.1 and 589.8 ± 105.01 mEq/L) were different (P < 0.05) between sheep and goats. Species and diet treatments had significant effects on the ultimate pH of LM. Pre-holding total coliform (TCC) and aerobic plate counts (APC) of skin were significantly different between species. Goats had lower (P < 0.05) TCC (2.1 vs. 3.0 log₁₀ CFU/cm²) and APC (8.2 vs. 8.5 log₁₀ CFU/cm²) counts in the skin compared to sheep. Preslaughter skin E. coli counts and TCC were different (P < 0.05) between species. Goats had lower (P < 0.05) counts of E. coli (2.2 vs. 2.9 log₁₀ CFU/cm²) and TCC (2.3 vs. 3.0 log₁₀ CFU/cm²) in the skin compared with those in sheep. Diet, species, and FDT had no effect (P > 0.05) on E. coli and TCC in carcass swab samples. The APC of carcass swab samples were only affected (P < 0.05) by the FDT. The results indicated that preslaughter dietary management had no significant changes on hormone and blood metabolite concentrations and sheep might be more prone for fecal contamination than goats in the holding pens at abattoir.     

Seroprevalence of Toxoplasma gondii infection in Norwegian dairy goats

Background

Toxoplasma gondii is a major problem for the sheep industry as it may cause reproduction problems. The importance of T. gondii in Norwegian goat herds is uncertain, but outbreaks of toxoplasmosis in dairy goat farms have been recorded. The aim of this study was to describe the prevalence of T. gondii infection in Norwegian dairy goats by using serology.

Findings

Goat serum originally collected as part of two nationwide surveillance and control programmes between 2002 and 2008 were examined for T. gondii antibodies by using direct agglutination test. In total, 55 of 73 herds (75%) had one or more serologically positive animals, while 377 of 2188 (17%) of the individual samples tested positive for T. gondii antibodies.

Conclusions

This is the first prevalence study of T. gondii infection in Norwegian goats. The results show that Norwegian goat herds are commonly exposed to T. gondii. Nevertheless, the majority of goat herds have a low prevalence of antibody positive animals, which make them vulnerable to infections with T. gondii during the gestation period.     

Difference in virulence between Staphylococcus aureus isolates causing gangrenous mastitis versus subclinical mastitis in a dairy sheep flock

Staphylococcus aureus mastitis in dairy sheep ranges from subclinical mastitis to lethal gangrenous mastitis. Neither the S. aureus virulence factors nor the host-factors or the epidemiological events contributing to the different outcomes are known. In a field study in a dairy sheep farm over 21 months, 16 natural isolates of S. aureus were collected from six subclinical mastitis cases, one lethal gangrenous mastitis case, nasal carriage from eight ewes and one isolate from ambient air in the milking room. A genomic comparison of two strains, one responsible for subclinical mastitis and one for lethal gangrenous mastitis, was performed using multi-strain DNA microarrays. Multiple typing techniques (pulsed-field-gel-electrophoresis, multiple-locus variable-number, single-nucleotide polymorphisms, randomly amplified polymorphic DNA, spa typing and sas typing) were used to characterise the remaining isolates and to follow the persistence of the gangrenous isolate in ewes’ nares. Our results showed that the two strains were genetically closely related and they shared 3 615 identical predicted open reading frames. However, the gangrenous mastitis isolate carried variant versions of several genes (sdrD, clfA-B, sasA, sasB, sasD, sasI and splE) and was missing fibrinogen binding protein B (fnbB) and a prophage. The typing results showed that this gangrenous strain emerged after the initial subclinical mastitis screening, but then persisted in the flock in the nares of four ewes. Although we cannot dismiss the role of host susceptibility in the clinical events in this flock, our data support the hypothesis that S. aureus populations had evolved in the sheep flock and that S. aureus genetic variations could have contributed to enhanced virulence.     

Lambs immunized with an inactivated variant of Anaplasma phagocytophilum

Background

Anaplasma phagocytophilum (formerly Ehrlichia phagocytophila) is an obligate intracellular bacterium causing the disease tick-borne fever (TBF) in domestic ruminants. An effective vaccine against the infection has been demanded for livestock by sheep farmers and veterinary practitioners for years.

Findings

In the present study, we immunized lambs with an inactivated suspension of 1 × 10⁸ killed A. phagocytophilum organisms mixed with adjuvant (Montanide ISA 61VG; Seppic). Twelve 9-months-old lambs of the Norwegian White Sheep breed were used. A full two-dose series of immunization was given subcutaneously to six lambs with a 4 week interval between injections. One month after the last immunization, all lambs were challenged with the homologous viable variant of A. phagocytophilum. After challenge, all lambs showed clinical responses for several days, although the immunized lambs reacted with an anamnestic response, i.e. significant reduction in infection rate and a significantly higher antibody titer.

Conclusion

Immunization with inactivated A. phagocytophilum did not protect lambs TBF.     

Distribution of cysts and tachyczoites in calves and pregnant cows inoculated with toxoplasma gondii oocysts

Sixteen calves and 6 cows were each inoculated with 100 000 infective oocytes of the GT-1 strain of Toxoplasma gondii. Cattle were necropsied between 3 and 287 days post-inoculation (DPI) and their tissues were inoculated into mice or fed to Toxoplasma-free cats for the detection of Toxoplasma in bovine tissues. Ten to 10 000-fold more T. gondii were recovered from small intestine and mesenteric lymph nodes of calves at 3 and 6 DPI than from lungs and liver, and the number of T. gondii in bovine tissues was reduced 1000-fold between 6 and 8 DPI. By using the pepsin digestion technique, or feeding tissues to Toxoplasma-free cats, it was demonstrated that T. gondii encysted in bovine tissues as early as 11 DPI and persisted as late as 287 DPI. More Toxoplasma gondii cysts occurred in livers than in any other bovine tissue. Of the 6 cows inoculated at 95–155 days after breeding, 5 delivered normal calves and T. gondii was isolated from only one of these calves. One cow was barren. Toxoplasma gondii was not isolated either by mouse inoculation or by feeding cats tissues from 2 cows killed 132 and 190 DPI. Toxoplasma gondii was not isolated in mice inoculated with tissues of cows killed 98 and 109 DPI, but cats fed on bovine tissues shed T. gondii oocysts. The organism, however, was isolated in mice inoculated with the mesenteric lymph nodes of 1 of the 2 cows killed 162 and 168 DPI, and from the small intestine of the other. Cats fed tissues of these cows later shed T. gondii oocysts.     

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.