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Complete genome sequences of bovine viral diarrhoea virus types 1 and 2 (BVDV-1 and 2) deposited in the GenBank were submitted to bioinformatic analysis using a recombination-detecting software. The results indicate that recombination events are not rare in the case of BVDV, which frequently causes immunotolerance and, consequently, persistent infection in calves. The lack of specific immunity provides an ideal possibility for multiple infections by antigenically related but genetically different BVDV strains, and hence recombinations may occur. Among the 62 BVDV-1 genomes five recombinants and their possible parent strains, while among the 50 BVDV-2 genomes one simple recombinant and its parent strains were identified, which were supported by extremely strong probability values (P values varying between 1.26 × 10–4 and 1.58 × 10–310). Besides the newly identified recombinants, recombination events described previously were confirmed, but in some of these cases former information was completed with new data, or different parent(s) were suggested by the programme (RDP 4.46 BETA) used in this study.

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Claudins are integral membrane proteins of the tight junction structures expressed by epithelial and endothelial cells. The present study has evaluated the expression of claudin-4 in 10 normal canine hepatoid glands and in 67 hepatoid glands with hyperplastic and neoplastic lesions. The lesions studied included normal hepatoid glands (n = 10), nodular hyperplasias (n = 10), adenomas (n = 12), epitheliomas (n = 15), differentiated carcinomas (n = 15) and anaplastic carcinomas (n = 15). There was an intensive expression of claudin-4 in normal canine hepatoid glands as well as in hyperplasias and adenomas. Claudin-4 was detected as a well-localised linear circumferential membranous staining pattern of epithelial cells (mature hepatoid cells) in normal hepatoid glands, perianal gland hyperplasias and adenomas. In nodular hyperplasia and adenoma, the reserve cells showed membrane positivity for the claudin-4 molecule. There was a weaker expression in hepatoid gland epitheliomas. In the epitheliomas, the basaloid reserve cells never expressed the claudin-4 molecule. The multiple small parts of epitheliomas in which the cells exhibited typical hepatoid features showed a well-localised linear circumferential membranous staining pattern for claudin-4. The numerical score for cellular expression of claudin-4 was higher in differentiated carcinomas than in epitheliomas, but moderately lower than in adenomas. The anaplastic, poorly differentiated hepatoid gland carcinomas showed an overexpression of claudin-4. These results suggest that low claudin-4 expression in epitheliomas is a molecular characteristic indicative of increasing cellular disorientation, detachment motility and invasion by tumour cells, and claudin-4 seems to be helpful in distinguishing undifferentiated carcinomas from differentiated carcinomas and epitheliomas of the hepatoid gland. In addition, claudin-4 can help distinguish epithelioma from differentiated carcinoma of the canine hepatoid gland.

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The authors describe a case of unilateral adenocarcinoma emerging from the Harderian gland, filling the right orbital cavity of a Florida Red-bellied Turtle ( Pseudemys nelsoni ). The tumour did not produce any metastasis but presented an expansive growth and led to the dislocation and protrusion of the right eyeball. Histopathological analysis revealed the presence of numerous mitotic figures in the cellular population that made up the tumour. The tumour cells completely filled the alveoli of the gland and had a nest-like structure. The authors also emphasise the importance of the differential diagnosis of this rare pathological change in turtles. Epithelial hyperplasia of the Harderian gland’s duct, observed in animals suffering from vitamin A deficiency, can also lead to an enlargement of the eyelid, but in these cases the change usually involves both eyelids symmetrically. This is the first description of a Harderian gland adenocarcinoma in a Florida Red-bellied Turtle.

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Claudin-5 is an endothelium-specific tight junction protein. The aim of the present study was to detect the expression pattern of this molecule in intact pancreatic tissues and in well-differentiated and poorly differentiated pancreatic acinar cell carcinomas from dogs by the use of cross-reactive humanised anticlaudin-5 antibody. The necropsy samples taken from dogs included 10 nonneoplastic pancreatic tissues, 10 well-differentiated pancreatic acinar cell carcinomas, 10 poorly differentiated pancreatic acinar cell carcinomas, 5 intrahepatic metastases of well-differentiated and 5 intrahepatic metastases of poorly differentiated acinar cell carcinomas. A strong lateral membrane claudin-5 positivity was detected in exocrine cells in all intact pancreas samples. The endocrine cells of the islets of Langerhans and the epithelial cells of the ducts were negative for claudin-5. The endothelial cells of vessels and lymphatic channels in the stroma of the intact pancreas showed strong membrane positivity for this claudin. All well-differentiated exocrine pancreas carcinomas and all poorly-differentiated pancreatic acinar cell carcinoma samples showed a diffuse loss of claudin-5 expression. The claudin-5-positive peritumoural vessels and lymphatic channels facilitated the detection of vascular invasion of the claudin-5-negative cancer cells. In liver metastasis samples, the pancreatic carcinomas were negative for claudin-5. It seems that the loss of expression of claudin-5 may lead to carcinogenesis in canine exocrine pancreatic cells.

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In this study, synchronous spontaneous, independent liver and gallbladder tumours were detected in a Bearded dragon (Pogona vitticeps). The multiple tumours consisted of intrahepatic cholangiocarcinoma as well as in situ adenocarcinoma and two adenomas of the gallbladder. The biliary epithelial cells and the cholangiocarcinoma showed membranous cross-immunoreactivity for claudin-7. The gallbladder epithelial cells, its adenoma and adenocarcinoma showed basolateral cross-reactivity for claudin-7. We think that the humanised anti-claudin-7 antibody is a good marker for the detection of different primary cholangiocellular and gallbladder tumours in Bearded dragons. The cholangiocytes, the cholangiocarcinoma, the endothelial cells of the liver and the epithelial cells and gallbladder tumours all showed claudin-5 cross-reactivity. The humanised anti-cytokeratin AE1–AE3 antibody showed cross-reactivity in the biliary epithelial cells, cholangiocarcinoma cells, epithelial cells and tumour cells of the gallbladder. It seems that this humanised antibody is a useful epithelial marker for the different neoplastic lesions of epithelial cells in reptiles. The humanised anti-α-smooth muscle actin (α-SMA) antibody showed intense cross-reactivity in the smooth muscle cells of the hepatic vessels and in the muscle layer of the gallbladder. The portal myofibroblasts, the endothelial cells of the sinusoids and the stromal cells of the cholangiocarcinoma and gallbladder tumours were positive for α-SMA. The antibovine anti-vimentin and humanised anti-Ki-67 antibodies did not show crossreactivity in the different samples from the Bearded dragon.

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Adenoviral nucleic acid was detected by polymerase chain reaction (PCR) in formalin-fixed paraffin-embedded tissue samples of a cat that had suffered from disseminated adenovirus infection. The identity of the amplified products from the hexon and DNA-dependent DNA polymerase genes was confirmed by DNA sequencing. The sequences were clearly distinguishable from corresponding hexon and polymerase sequences of other mastadenoviruses, including human adenoviruses. These results suggest the possible existence of a distinct feline adenovirus.

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Acta Veterinaria Hungarica
Authors: Elena Palade, Nóra Biró, Mihály Dobos-Kovács, Zoltán Demeter, Míra Mándoki, and Miklós Rusvai

From a total of 1819 great tits ( Parus major ) ringed in 2007 in Pilis Mountains, Hungary, 15 birds presented nodular proliferative lesions on different areas of the head and eyelids, suggesting a poxvirus infection. Three birds were submitted for analysis. The presence of avipoxvirus infection was confirmed by histopathology, electron microscopy (EM) and a polymerase chain reaction (PCR) based technique. Nucleotide sequence analysis of a 428 base pairs (bp) fragment of the viral 4b core protein gene revealed 100% identity between two of the Hungarian isolates (PM9 HUN, PM33 HUN) and two great tit poxvirus strains isolated in Norway in 1973 (GTV A256, GTV A311). The third Hungarian isolate (PM34 HUN) was more closely related to a different Norwegian isolate (GTVA310) than to the Hungarian isolates. The nucleotide sequence analysis of a shorter fragment of the viral 4b core protein (227 bp) gene revealed 100% identity between the Hungarian isolates, the same Norwegian isolates and a great tit poxvirus strain detected in Austria in 2007.

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Acta Veterinaria Hungarica
Authors: Csaba Jakab, Attila Szász, Janina Kulka, Ferenc Baska, Miklós Rusvai, Péter Gálfi, and Tibor Németh

This short report describes a case of tricuspid valvular metastasis of canine disseminated histiocytic sarcoma in a 9-year-old female Rottweiler. Immunohistochemically the malignant neoplastic cells gave a strong reaction for vimentin and lysozyme, and showed negativity for serotonin, CD3, CD79a and cytokeratin. The intratumoural microvessels were detected by immunohistochemistry using CD31 and claudin-5. This appears to be the first report of a valvular metastasis of canine malignant histiocytosis.

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Acta Veterinaria Hungarica
Authors: Zsuzsanna Tapaszti, Petra Forgách, Csaba Kővágó, László Békési, Tamás Bakonyi, and Miklós Rusvai

Microsporidiosis (nosema disease) of the European honeybee ( Apis mellifera L.) is present in bee colonies worldwide. Until recently, Nosema apis had been regarded as the causative agent of the disease, which may have many negative effects on the colony and cause heavy economic losses in apicultures. Another microsporidium species, Nosema ceranae , was reported to infest the Asian honeybee ( Apis ceranae ), but both honeybee species are susceptible to both microsporidia. In the European honeybee N. ceranae was first detected in Spain in the year 2006. As it is difficult to distinguish N. ceranae and N. apis morphologically, a rapid and accurate assay has been developed to differentiate N. apis and N. ceranae based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the partial large subunit ribosomal RNA. The assay was tested on 38 Nosema -infested bee samples, which were collected from geographically distant Hungarian bee colonies representing all regions of the country. Only one sample contained N. apis , and in the other 37 samples N. ceranae was detected, which indicates the dominance of N. ceranae in Hungarian apiaries. This is the first report on the presence of N. ceranae in Hungary.

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Acta Veterinaria Hungarica
Authors: Sanja Aleksić-Kovačević, József Özvegy, Nikola Krstić, Miklós Rusvai, Csaba Jakab, Zoran Stanimirović, and Zsolt Becskei

Water pollution is known to play an important role in the pathogenesis of plastron, carapace and skin diseases of turtles. In this study, a total of 150 European pond turtles (Emys orbicularis) of different age and both sexes, originating from natural habitats in Serbia, were examined for morphological changes of the skin, plastron, carapace and skeletal system. The turtles were taken out from their natural habitats in Lake Ludas, Lake Palic and Lake Tresetiste. After artificial hibernation, they were subjected to detailed examination, sampled and treated, and finally returned into their natural habitat. Biopsies from the skin and shell were subjected to histopathological examination and microbiological analysis. X-ray scanning was also performed to detect changes in the skeletal system. Macroscopic changes of the skin, most frequently degenerative, inflammatory or neoplastic diseases, were diagnosed in 49.33% of the turtles examined. Dermatitis of different origin and form was the most prominent histopathological finding (28.00%). In the plastron, inflammatory and degenerative processes were frequently found. Osteopathy and mechanical injuries were the dominant findings. Macroscopic changes of the plastron, carapace and skeletal system were diagnosed in 67.33% of the turtles examined. Using X-ray scanning, generalised osteopathy, anomalies and malformations of different aetiology were also diagnosed on the tail and legs. Microbiological examinations showed the presence of a variety of bacterial and fungal agents, either primary pathogens or potential polluters, which invaded the skin and shell, or were present in cloacal swab samples. Bacterial infection was diagnosed in 76.66% of the turtles, first of all in those with skin and shell necrosis. Mycoses were diagnosed in 33.33% of the animals.

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