Long-term Study of Chlamydophilosis in Slovenia

Immune reactivity for Chlamydophila (C.) psittaci in Slovenia was monitored in parrots, canaries, finches and nine species of recently captured free-living birds (house sparrows, Eurasian goldfinches, tree sparrows, chaffinches, European greenfinches, European serines, Eurasian siskins, Eurasian linnets and Eurasian bullfinches) in the period from 1991 to 2001. In subsequent years, specific IgG antibodies were found using immunofluorescence in parrots (0.7– 53.6%), canaries (0.0–3.5%), finches (0.0–5.7%) and in captured free-living birds (33.3% of Eurasian goldfinches in 1994). An experimental infection with C. psittaci was performed in order to study clinical signs and pathological changes in canaries and finches. The C. psittaci strain used for experimental infection was isolated from a cockatiel (Nymphicus hollandicus). Chlamydial DNA was extracted from clinical material followed by RFLP-PCR analysis. Infection of canaries and finches was confirmed in organ smears by direct immunofluorescence and a modified Gimenez staining method. In addition, serological tests of indirect immunofluorescence and complement fixation were applied. However, in spite of positive immunological reaction there were no clinical signs three weeks after infection. The present study includes results of a serological survey of persons belonging to the most important risk groups (breeders, pet shopkeepers and veterinarians). The results of microimmunofluorescence to identify the presence of specific antibodies and correlation between appearance of infection in birds and important risk groups are presented. Out of 143 persons belonging to the high-risk group we found 10 (7%) persons who were immunologically positive. Testing of two successive samples was used to demonstrate an increase in IgG and IgA titres in human sera. However, IgM, which is indicative of acute infection, could not be detected.     

Fish Health Management in Slovenia

Veterinary Research Communications -     

Enzootic nasal adenocarcinoma of sheep in Slovenia

SUMMARY Clinicopathological and electron microscopical findings of eight cases of enzootic nasal adenocarcinoma of sheep, diagnosed solely in one big flock in Slovenia between years 2001 and 2003 are described. All affected sheep were female, their mean age was 4.5 +/- 1.5 years and they either belonged to the Istrian pramenka breed (five sheep) or were crossbreeds (three sheep). Tumours that arose from the ethmoid area of the nasal cavity were unilateral in six cases (75%) and bilateral in two cases (25%). All tumours were classified as adenocarcinomas by histopathological examination and they displayed either a combination of tubular and papillary growth or less often solely tubular proliferation. No metastases were detected in regional lymph nodes, brain or other organs. Electron microscopical studies performed on the reprocessed paraffin-embedded tissues revealed the presence of the virus-like particles with an average diameter between 70 and 90 nm.     

Long-term case study of myelodysplastic syndrome in a dog

A 10-year-old, female shih tzu was diagnosed as having myelodysplastic syndrome (MDS) based on the presence of a nonregenerative anemia, dysplastic changes in the three hematopoietic cell lines, a normal to hypercellular bone marrow, and less than 30% blast cells of all nucleated cells in the bone marrow. Low-dose aclarubicin, a differentiation-induction therapy for MDS and atypical leukemias in humans, was administered. Hematological improvement was observed, and the dog lived for 809 days after the first presentation.     

Phylogenetic analysis of small ruminant lentiviruses detected in Slovenia

Small ruminant lentiviruses (SRLV), which belong to the Retroviridae family, infect goats and sheep worldwide. The aim of this study was to characterize the SRLV strains circulating in Slovenia, by phylogenetic analysis of two genomic regions, 1.8 kb gag-pol fragment and 1.2 kb pol fragment. The results of our study revealed that Slovenian SRLV strains are highly heterogeneous, with ovine strains belonging to genotype A and caprine strains to genotypes A and B. The closest relatives of sheep virus sequences from two flocks that clustered together (SLO 35, 36) were found to be in subtype A5. A cluster composed of four sheep virus sequences (SLO 31) was clearly divergent from all other subtypes in group A and could not be assigned to any of them. The virus sequences from one goat flock belonged solely to subtype B1, whereas virus sequences from more than one genotype were found to circulate within the other two goat flocks, belonging to subtype B1 (SLO 1 and SLO 37) and to genotype A (SLO 2 and 78–88 g). Two goat virus sequences (SLO 2) were found to belong to genotype A and could not be assigned to existing subtypes. One goat virus sequence (37–88 g) from flock 37 was clearly different from other sequences of this flock and was more closely related to genotype A sequences. We propose two new subtypes within genotype A, subtype A14 (SLO 2) and A15 (SLO 31).     

A serological survey of selected pathogens in wild boar in Slovenia

Serum samples collected from 178 shot wild boars (Sus scrofa) were tested for the presence of antibodies against classical swine fever virus, Aujeszky's disease virus (ADV), porcine reproductive and respiratory syndrome virus, porcine respiratory coronavirus (PRCV), transmissible gastroenteritis virus, swine influenza virus, porcine parvovirus (PPV), swine vesicular disease virus, Actinobacillus pleuropneumoniae (APP), Mycoplasma hyopneumoniae, Salmonella spp., Brucella spp. and Haemophilus parasuis (HPS) throughout Slovenia during the hunting season 2003/2004. The number of samples corresponds to 3% of the total hunting bag. By enzyme-linked immunosorbent assay (ELISA) antibodies against ADV were detected in 55 sera (31%), against PRCV in five sera (3%), PPV in 87 sera (49%), APP in 93 sera (52%), M. hyopneumoniae in 38 sera (21%), Salmonella spp. in 85 sera (47%) and HPS in 33 sera (18%).     

Steroid responsive meningitis-arteritis in dogs: Long-term study of 32 cases

Steroid responsive meningitis-arteritis in 32 cases was studied. Two forms were recognised: The typical form showing cervical rigidity and pain, fever and pleocytosis with polymorphonuclear cells in the cerebrospinal fluid (CSF); and the atypical form, a more protracted type characterised by additional neurological deficits and lack of typical CSF changes. Both groups responded to steroid application. Several ancillary laboratory procedures were applied in an attempt to improve the diagnosis. Electroencephalography was found to be helpful in detecting an inflammatory process. Measuring IgA levels in the serum and CSF was also another useful technique for detecting atypical cases. Long-term treatment with prednisolone was undertaken in 20 dogs. The treatment was monitored by rigorous control examinations of the blood and CSF at regular intervals. Twelve dogs were considered cured after this treatment. Thirteen dogs were euthanased; five of these in the acute stage of the disease and eight because of progressive neurological disease. In all these cases histopathological examination was performed. The data are compared to reports in the literature on meningitis and arteritis in dogs and the pathogenesis of the disease is briefly discussed.     

Long-term retrospective study of 52 horses with sinunasal cysts

REASONS FOR PERFORMING STUDY: The spectrum of clinical presentation, methods of diagnosis, management and prognosis in cases of sinunasal cysts (SNCs) requires more extensive study. Despite the extensive nature of SNCs, the prognosis for afflicted horses appears to have improved since earlier studies were conducted. OBJECTIVES: To evaluate prognosis and cosmetic outcome of surgical ablation in 52 cases of SNCs. METHODS: Fifty-two case records (1982-2005) of horses affected with SNCs were retrieved from the archives. Subject details, clinical signs, diagnostic techniques, surgical management and post surgical complications were extracted. Owners and referring veterinarians were contacted to assess the results of treatment. RESULTS: There was no biphasic age distribution. The major presenting signs in descending order of frequency were facial swelling, mucopurulent nasal discharge, nasal airflow obstruction and abnormal respiratory noise. Common endoscopic findings included narrowed nasal meati, a cyst in the nasal cavity and a cyst visible caudal to the nasal septum viewed from the contralateral unaffected nasal meatus. Typical radiological signs included a discrete mass in the sinunasal region, a diffuse increase in opacity over the sinunasal region, free fluid lines, nasal septal deviation, expansion of the ventral conchal sinus and distortion of dental apices. Cyst extirpation using an osteoplastic flap provided a successful outcome. Forty-five of 48 horses showed complete resolution of clinical signs after subtotal or total extirpation of the cyst wall. Follow-up information was not available for the other 3 horses. Thirty-nine of 45 horses had a good to fair cosmetic appearance, judged to be good in 26 horses, fair in 13 and poor in 6. Results confirmed that the prognosis for full recovery has improved since a previous report of 15 cases. CONCLUSIONS AND POTENTIAL RELEVANCE: Equine sinunasal cysts may arise in horses of any age; presenting signs should alert clinicians to their likely presence. A discrete well-circumscribed mass found during radiographic examination provides supporting evidence of a cyst. The vivid yellow, translucent, seromucoid fluid aspirated from cysts is characteristic of the condition.     

Molecular epidemiology of the rabies virus in Slovenia 1994-2010

A molecular epidemiology study was performed on a selection of 30 rabies-positive brain samples collected between 1994 and 2010 in Slovenia and originating from the red fox (n=19), badger (n=3), cattle (n=3), dog (n=2), cat (n=1), marten (n=1) and horse (n=1). Based on the comparison of 1092 and 672 nucleotide sequences of nucleoprotein (N) and partial glycoprotein (G) gene regions, a low genetic diversity of the circulating strains was detected, but both phylogenetic trees were consistent with the topology where partial nucleoprotein or glycoprotein genes were used. A high sequence identity in the N and G gene to rabies virus isolates from neighbouring countries was found. The Slovenian strains were clearly different from the vaccine strains SAD B19 and SAD Bern, which have been used in Slovenia since 1988.     

Prevalence of pigeon circovirus infections in feral pigeons in Ljubljana, Slovenia

Pigeon circovirus (PiCV) was detected by real-time PCR in cloacal swabs, pharyngeal swabs, and serum samples taken from 74 feral pigeons (Columba livia var. domestica) that were caught at various locations in the city of Ljubljana, Slovenia. PiCV infections were detected in the majority of the tested birds. The highest (74.3%) detection rate was observed in the cloacal swabs and the lowest (31.1%) in serum samples. PiCV DNA was more readily detected in the cloacal swabs, pharyngeal swabs, and serum samples of birds younger than 1 yr. Molecular analysis of partial open reading frame V1 sequences showed that PiCV strains detected in feral pigeons share high nucleotide and amino acid sequence identities with PiCV strains detected in ornamental, racing, meat, and feral pigeons.     

Long-term study of aerobic bacteria of the genital tract in stud dogs.

The aerobic bacterial flora of the genital tract was characterized in 15 stud dogs in an 18-month study. The dogs represented 4 breeds and were from 3 kennels. Bacterial samples from the prepuce and semen were collected every month, except in connection with matings, when they were collected weekly (464 samples). The dogs that were included all mated at least once during the study. The mean pregnancy rate, litter size, and pup mortality for the bitches with which they had mated were all within normal limits. The most frequent bacteria isolated from the prepuce and semen were Pasteurella multocida, beta-hemolytic streptococci, and Escherichia coli. There was a tendency for breeds to differ in frequency of the most common bacterial species. Bacterial culture yielded no aerobic growth in 14.2% of the preputial samples and 69.8% of the semen samples. Bacteria were transferred between dog and bitch at mating. In this study of healthy breeding dogs, neither the fertility of the dog nor that of the bitch was affected by the bacteria transferred.     

Long-term study of aerobic bacteria of the genital tract in breeding bitches.

The aerobic bacterial flora of the genital tract was characterized in 59 bitches in an 18-month study. The bitches represented 4 breeds and were from 3 kennels. Collection of vaginal swab specimens for bacterial culturing was performed every month, except during estrus when specimens were collected every week (n = 826). The capsule of the swab containing transport media was broken before specimen collection to moisten the tip, which helped to reduce the number of negative cultures. All bitches helped at least once during the study and, thus, had known reproductive functions. Pregnancy rates, litter sizes, and pup mortality were within normal limits. Pasteurella multocida, beta-hemolytic streptococci group G, and Escherichia coli were the most common bacteria isolated. Although these species generally were isolated from mixed cultures, pure cultures were obtained from 18% of the specimens. There was a tendency for the various breeds to differ in their vaginal bacterial flora. The flora also varied during the reproductive cycle. Pasteurella multocida was isolated significantly more often during proestrus, estrus, metestrus, and pregnancy, than during anestrus and the postpartum period, and beta-hemolytic streptococci were isolated significantly more often during proestrus than during estrus, pregnancy, or the postpartum period. Staphylococcus intermedius was almost exclusively found after parturition. Culture results were negative for only 5.2% of specimens cultured. On the basis of our findings, bacterial culturing of vaginal swab specimens from bitches without signs of genital disease is of little value.     

Detection of Vibrio parahaemolyticus in Mediterranean mussels (Mytilus galloprovincialis) in Slovenia

The aim of this study was to determine the prevalence of Vibrio parahaemolyticus in shellfish samples harvested along the Slovenian coast. Shellfish samples of Mediterranean mussels (Mytilus galloprovincialis) were collected along the Slovenian coast at four locations (Se?a, Piran, Strunjan and Debeli Rti?) between 2006 and 2008. Samples were examined and analysed for the presence of V. parahaemolyticus by conventional and molecular methods. The presence of Vibrio in the samples was examined by conventional methods on plate grown bacterial cells before and after enrichment in alkaline saline peptone water (ASPW). PCR methods were used for the detection of V. parahaemolyticus-specific toxR and tlh genes and of the virulence-associated tdh and trh genes. Out of 168 samples examined, 24 were positive for toxR and tlh genes by PCR from enrichment broth. Five out of 62 (8.1%), 4 out of 32 (12.5%) and 15 out of 74 (20.2%) samples were positive in 2006, 2007 and 2008, respectively. Colonies of V. parahaemolyticus were isolated from only one sample positive for V. parahaemolyticus by PCR.     

Long-term study of Cryptosporidium prevalence on a lowland farm in the United Kingdom

A longitudinal sample survey testing for Cryptosporidium in livestock and small wild mammals conducted over 6 years (1992-1997) on a lowland farm in Warwickshire, England, has shown the parasite to be endemic and persistently present in all mammalian categories. Faecal samples were taken throughout the year and oocysts concentrated by a formal ether sedimentation method for detection by immunofluorescence staining using a commercially available genus specific monoclonal antibody. Cryptosporidium parvum was identified by morphology and measurement of modified Ziehl-Neelsen stained oocysts. C. muris was rarely found in wild mammals and C. andersoni oocysts were never detected in livestock. Faecal samples from 3721 individuals gave cumulative 6-year prevalences for C. parvum as follows: bull beef, 3.6%; dairy cows, 3.5%; ewes, 6.4%; horses, 8.9%; calves (home bred), 52%; calves (bought-in) 23.2%; lambs, 12.9%; small wild mammals (rodents) living in and around farm buildings, 32.8%; small wild mammals (mainly rodents) living in areas of pasture, 29.9%. Animal categories with the highest prevalences also shed the highest average oocyst numbers per gram of faeces (ranging from 1.4 x 10(3) for bull beef to 1.1 x 10(5) for calves). Analysis of annual and seasonal data for each animal category revealed that patterns of infection were variable and sporadic; this means that short-term sampling was never likely to provide a true or representative picture. Seasonally combined data for adult livestock, young livestock and small wild mammals showed all three categories tended to have the highest Cryptosporidium prevalences in the autumn. Calves were separated from their dams within 24h of birth and yet showed high prevalence of infection in most years despite the low prevalence for the dairy herd. It is possible the coincidence of high autumn prevalence in mice with the main period for the rearing of calves contributed to the infection of the latter. The farming estate was used to teach students of agriculture and took pride in good land management and husbandry practices that produced well fed and healthy livestock. The data from this estate may represent, therefore, the baseline, the lowest possible levels to be expected, for Cryptosporidium infection and oocyst production on a lowland farm in the United Kingdom.