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The aim of the study was to investigate variations in lengths and lesions in claws of culled sows and to evaluate their association. All four feet of 185 sows from three Greek farrow-to-finish farms (Farm A: 57 sows, Farm B: 64 sows, Farm C: 64 sows) were examined for lesions and their lengths were measured. All claw lengths were lower in sows of farm C compared to those from sows of B and A. Claw lengths in sows of farm B were lower compared to those from A for all lateral toes of front feet and for all medial and four out of three lateral toes of rear feet. Sum of length measurements of the main toes of the front feet (SLF) associated with lesions on sole, white line and heel of front feet, while sum of length measurements of the main toes of the rear feet (SLR) associated with all lesions of the rear feet. The lengths of the main toes were correlated with the length of dew claws on front and rear feet. Overall, sows’ claw lesion severity and claw lengths may differ between farms and frequency of lesions is higher in longer claws.

Background Urinary tract infections (UTI) of sows characterized by cystitis, which may progress to ureteritis and pyelonephritis, can affect their productivity, longevity and welfare. In this study, we determined the prevalence of UTI by histopathology and bacteriology. Furthermore, we investigated possible associations between histologically confirmed cystitis and the results of urinalysis and urine cultures in culled sows from three farrow-to-finish herds in Greece. Materials and methods One hundred eighty-five routinely culled sows were included in the study. Their urinary bladder was collected from abattoirs and subjected to histopathology. Furthermore, urinalysis and urine cultures were performed on urine samples aseptically collected from the bladders. Results Histologically confirmed cystitis was evident in 85/185 (45.94%) culled sows. Among those, 44 (51.76%) suffered from acute and 41 (48.24%) from chronic inflammation. The majority of positive urine cultures were due to colonization of the urinary tract with E.coli, which accounted for 55.81% of the total cases, followed by Staphylococcus spp. which accounted for 18.60% of detected infections. Evidence of cystitis was associated with bacteriuria and sows with bacteriuria were 2.30 (P = 0.03, 95% CI: 1.10–4.83) times more likely to have histologically confirmed cystitis compared to sows with negative urine cultures. Bacteriuria was associated with proteinuria (P P P P P P P P = 0.006, 1.39–7.69) times less likely for a sow to have chronic or acute cystitis compared to absence of cystitis. Conclusion In the studied population, UTI affected almost one out of two culled sows. Bacteriuria, which was more common among sows with UTI than those without, was mainly ascribed to members of the intestinal and environmental bacteria. Proteinuria and the existence of urine sediment which were associated with UTI, could be used as proxy traits for UTI in live sows.

Enzootic nasal adenocarcinoma (ENA) is a contagious neoplasm of sheep and goats, associated with the oncogenic retroviruses enzootic nasal tumor virus (ENTV) 1 and 2, respectively. It appears to be common in countries with substantial small ruminant-production. ENA diagnosis in goats is based on autopsy and histopathology, and there is no real-time PCR method available for ENTV-2 detection. Here, a novel one-tube real-time RT-PCR (RT-qPCR) method for the detection and quantification of ENTV-2 in nasal swabs is presented. The method targets the env gene/U3 region. For the design of ENTV-2-specific oligonucleotides, molecular characterization of seven Greek ENTV-2 strains was performed. Phylogenetic analysis revealed three distinct phylogenetic clades of ENTV-2 that correlate with the country of sample collection. Evaluation of the analytical performance of the RT-qPCR revealed an amplification efficiency of 92.8% and a linear range of quantification between 2 × 108 and 2 × 102 RNA transcripts. Analysis of nasal swabs from 23 histopathologically confirmed, naturally occurring ENA cases via RT-qPCR yielded positive results. Moreover, modification of the method for use in a real-time PCR (qPCR) assay enables detection of proviral DNA in tumor specimens. Both methods are highly specific and can be used for the confirmation of ENA-suspected cases. Future applications could include ante-mortem diagnosis, verification of the ENTV-2-free status in animal trade, disease surveillance, and control programs.

Ένα κατοικίδιο κουνέλι 6.5 eτών, αρσeνικό, ακέραιο προσκομίστηκe μe γeνικeυμένη αποφολιδωτική δeρματίτιδα, κατάπτωση και ανορeξία. Οι eργαστηριακές eξeτάσeις του δέρματος ήταν αρνητικές για eξωπαράσιτα, βακτήρια και μύκητeς, eνώ στην ακτινογραφία του θώρακος παρατηρήθηκe eυμeγέθης μάζα στο μeσοπνeυμόνιο. Η γeνική eξέταση αίματος και οι βιοχημικές eξeτάσeις δeν eμφάνιζαν αποκλίσeις από τις φυσιολογικές τιμές. Υπήρξαν ισχυρές υποψίeς για θύμωμα και οι θeραπeυτικές eπιλογές συζητήθηκαν, αλλά ο ιδιοκτήτης αρνήθηκe. Η υποστηρικτική και συμπτωματική θeραπeία αποδeίχθηκαν αναποτeλeσματικές, γeγονός το οποίο, και σe συνδυασμό μe την σοβαρή eπιδeίνωση των συστηματικών συμπτωμάτων καθιστούσαν αναγκαία την eυθανασία του ζώου. Κατά τη νeκροψία παρατηρήθηκe μάζα λeυκόφαιου χρώματος στο πρόσθιο και κοιλιακό τμήμα του μeσοπνeυμόνιου, καθώς και ωχρό και διογκωμένο ήπαρ. Οι ιστοπαθαλογικές eξeτάσeις eπιβeβαίωσαν την ύπαρξη θυμώματος και έθeσαν διάγνωση παρανeοπλασματικής αποφολιδωτικής δeρματίτιδας λόγω θυμώματος, σe συνδυασμό μe κίρρωση του ήπατος.

Inoculation of embryonated chicken eggs is the standard method for the titration of infectious Bluetongue virus (BTV). Here, six RNA extraction methods coupled with optimised dsRNA denaturation and real-time RT-PCR were evaluated for the quantitation of BTV in blood samples from experimentally infected sheep and results were correlated to infectious virus titres. An exogenous dsRNA internal control (IC) from the closely related Epizootic hemorrhagic disease virus (EHDV) was used to assess the efficiency of BTV genome extraction, dsRNA denaturation, RT, and PCR amplification. Recovery rates of IC and BTV dsRNA copies from extracted blood samples were highly correlated. Adjustment of BTV concentrations according to the IC recovery reduced variation in sample analyses among the different extraction methods and improved the accuracy of BTV quantitation. The EID(50)/ml titre, determined in blood samples from sheep infected experimentally with BTV-1 or BTV-9, correlated highly with the assessed concentration of BTV dsRNA copies. However, this correlation was consistent only during the first 28 days post-infection. The optimised extraction methods and quantitative RT-PCR could be useful for experimental studies of BTV transmission, pathogenesis and vaccine efficacy, or adapted further for the detection and quantitation of EHDV, African horse sickness virus and other dsRNA viruses.

In 2002, postweaning multisystemic wasting syndrome (PMWS) was diagnosed in a European female wild boar (Sus scrofa), based on the detection of porcine circovirus 2 (PCV2) DNA in various organs, including the uterus, and on histopathologic lesions. This is the first detection of PCV2 DNA in the uterus of a wild boar. Three years later (2005), a wild boar6-8 mo of age was found moribund. It presented wasting and dyspnea and finally died. PCV2 DNA was detected in tissue samples, and histopathologic lesions consistent with PMWS were observed. Both wild boars were from neighboring hunting areas in central Greece. Two PCV2 strains from the wild boars were genetically characterized and compared to other reported PCV2 sequences from wild boars and domestic pigs. The PCV-2 sequences from the wild boars in this study were closely related to each other and were grouped with two isolates from wild boars from Hungary. The phylogenetic analysis revealed that the virus might be transmitted between hunting areas. In addition, PCV2 may spread from domestic pigs to wild boars and vice versa.

International audience; During 2014, an outbreak of Bluetongue virus (BTV) infections attributed to serotype 4 occurred in Greece and spread to south-eastern Europe. In the present article, the clinical and epidemiological data of 15 sheep flocks and 5 dairy cattle herds affected in Greece are described. In sheep, the most frequent clinical signs observed were fever, hyporexia, and edema of the face. A number of clinically affected sheep had chronic laminitis resulting in chronic lameness. Confirmation of suspect clinical cases was performed using BTV-specific real-time RT-PCR, and serotype 4-specific RT-PCR. The average morbidity of bluetongue in the sheep flocks was estimated to be 15.3 % (95 % C.I. 6.8-23.8 %) and the average mortality and case fatality were 4.5 % (95 % C.I. 1.5-7.6 %) and 32.0 % (95 % C.I. 18.1-42.9 %), respectively. The BTV seroprevalence and the ratio of clinical manifestations-to-infections determined in seven of these flocks, were on average 36.5 % (95 % C.I. 15.7-57.3 %) and 24.6 % (95 % C.I. 12.8-36.3 %). BTV ratio of clinical manifestations-to-infections was higher in the imported western European sheep breeds examined compared to the local ones. In dairy cattle, the average herd prevalence of viremia was 48.8 % (95 % C.I. 15.3-82.4 %) and none had signs associated with bluetongue. The results of this study indicate that the 2014 Greek BTV-4 has significant impact on the health status and the viability of sheep in affected flocks but does not cause clinical signs in cattle, despite the high prevalence of viremia.

An imported 2.5-yr-old female green iguana (Iguana iguana rhinolopa) kept in Greece was presented with a history of anorexia and allotriophagy of 1 mo duration. Upon clinical examination, it was cachectic and had severe abdominal distension and fibrous osteodystrophy. Despite treatment, it died a month later. On necropsy, massive accumulations of threadlike nematode parasites were observed in the large intestine, identified as Ozolaimus megatyphlon, a member of the Oxyuridae family of Pharyngodonidae, a usually nonpathogenic intestinal parasite of iguanas. To the authors' knowledge, its presence has not been reported previously in Europe, although one pathogenic infection has been reported previously in Japan. The animal was presumably infected before importation. Although death was attributed to the heavy parasitic overload, the poor diet and terrarium hygiene, and absence of an anthelminthic regime further contributed to the deterioration of the animal's condition. Recognition of this condition, which may be recently introduced or underdiagnosed, may help improve medical and trade standards concerning this species in practice.

Rats (n=40) aged 8 weeks were infected, either by oronasal inoculation or by contact, with one of two different myocardial strains of encephalomyocarditis virus (EMCV), namely, the Greek strain 424/90 and the Belgian strain B279/95. The animals were killed at 11-62 days post-infection (dpi) and samples of brain, heart, pancreas, kidney, Peyer's patches, spleen, lung and thymus were processed for virological, histopathological and immunohistochemical evaluation. This experimental infection was inapparent, but virus was isolated from faeces and several organs of all animals. The main histopathological changes were focal interstitial pancreatitis, degeneration and necrosis of pancreatic acinar cells, depletion of thymus and Peyer's patches, and interstitial pneumonia. EMCV antigen was detected in the cytoplasm of cardiac muscle cells, pancreatic acinar cells and hepatic epithelial cells, and in macrophages of the spleen, lung and thymus. In the heart (the target organ of EMCV in pigs), the presence of EMCV in cardiac muscle cells without lesions lends support to the hypothesis that the rat is a natural reservoir host species of EMCV. The persistence of virus in the macrophages of the thymus may represent a mechanism of perpetuation and reactivation, under immunosuppressive conditions, of the infection.

Summary Histopathological and immunohistochemical examinations were performed on the brain and spinal cord of 37 goats from two Greek herds in which scrapie had been reported. Of the 37 animals, 18 were from a herd consisting only of goats and 19 were from a herd of goats mixed with sheep. The goats studied were grouped on the basis of the presence or absence of clinical signs. Distinctive lesions and PrP sc (PrP, prion protein) deposition were found in the central nervous system (CNS) of eight clinically affected animals and six symptomless animals. The lesion profile and PrP sc distribution varied both between and within groups, variation being particularly pronounced in the symptomless goats. The results concerning the latter group suggested a poor correlation between the intensity of lesions, the amount of PrP sc in the CNS, and the manifestation of clinical signs. Immunohistochemical examination revealed 10 different PrP sc types, four of which are reported for the first time in goats. All scrapie-affected animals carried the VV 21 II 142 HH 143 RR 154 genotype, with the exception of two goats that carried the HR 143 dimorphism and had detectable PrP sc deposits. The results suggest that the histopathological and immunohistochemical profile of the natural disease in goats is influenced by the PrP genotype and age of the animals but may not be directly associated with the presence or otherwise of clinical signs.

From 1999 to mid-2003, 97 European brown hares (Lepus europaeus) found dead throughout Greece were examined by necropsy, histopathology, and reverse-transcription polymerase chain reaction (RT-PCR) for the presence of European brown hare syndrome (EBHS) and EBHS virus (EBHSV), respectively. Hare losses were sporadic, starting in the cold season and lasting for many months (December to May). The most prominent gross lesions were observed in the liver and included swelling and discoloration; congestion and hemorrhages were present mainly in lungs and tracheal mucosa. Necropsy findings were suggestive of EBHS, which was confirmed by histopathology and RT-PCR. This study documents, for the first time, EBHS in Greece.

Canine sclerosing encapsulating peritonitis is a rarely reported condition. A 10-year-old male German shepherd dog cross was presented with a history of ascites, vomiting, soft faeces, anorexia and depression. Gathering of the intestinal loops in the middle portion of the abdomen was detected by radiography and ultrasonography. Cytological examination of Giemsa-stained smears from the popliteal lymph nodes revealed Leishmania species. The results of culture of serosanguineous fluid obtained by abdominocentesis were negative for bacteria and fungi. Laparotomy revealed a sac of fibrous tissue encasing most of the intestinal loops and numerous adhesions extending between them. Histologically, an uneven, diffusely thickened, visceral peritoneal membrane was found. A diagnosis of idiopathic sclerosing encapsulating peritonitis was made. The dog was euthanased because the intestinal wall was torn at many sites during dissection of the membrane.

Six piglets aged 20 days were inoculated oranasally with 5 ml of a suspension (106 TCID50/ml) of a Greek myocardial strain of encephalomyocarditis virus (EMCV). The animals either died (n=2) or were killed for examination on days 1,2 or 3 post-inoculation (pi). EMCV was isolated from virtually all organs examined (heart, tonsils, palatine glands, pancreas, spleen, small intestine and mesenteric lymph nodes). Histopathologically, interstitial myocarditis, necrosis of cardiac muscle cells and Purkinje fibres, and necrotizing tonsillitis were detected in all inoculated piglets. Focal interstitial pancreatitis, necrosis of pancreatic acinar cells and Langerhans islet cells, and necrosis of germinal centre lymphocytes of the lymph nodes and Peyer's patches were detected in two piglets that died or were killed on day 3 pi. Immunohistochemically, viral antigen was detected in epithelial cells of all organs examined, including the tonsils, palatine glands, pancreatic interlobular ducts and small intestine. This suggests that EMCV is epitheliotropic, in addition to its known myocardial tropism. The frequent presence of intracytoplasmic EMCV in macrophages of the tonsils and spleen supports the hypothesis that macrophages play a role in viral replication and dissemination in the body.

There appears to be a lack of information concerning responses of mules to natural infection or experimental inoculation with equine infectious anemia virus (EIAV). In the present study EIAV was isolated from mules, for the first time, and its pathogenicity in naturally infected and experimentally inoculated animals was investigated. Two naturally infected (A and B) and three EIAV free mules (C, D and E) were used for this purpose. Mule A developed clinical signs, whereas mule B remained asymptomatic until the end of the study. Mules C and D were each inoculated with 10 ml of blood from mule A and developed signs of the disease; they were euthanatized or died at day 22 and 25 post-inoculation, respectively. Mule E served as a negative control. The virus was isolated from the plasma samples of mules with clinical signs of the disease (A, C and D), but not from the asymptomatic mule B. Both proviral DNA and viral RNA were amplified from blood and tissues of the infected animals by nested polymerase chain reaction (nPCR). Antibodies were not detected in the two experimentally infected mules until their natural death or euthanasia. Clinicopathological and laboratory findings showed that, in mules, EIAV produced clinical signs similar to those observed in horses and ponies. Nested PCR proved to be a rapid, sensitive and specific diagnostic method for the detection of EIAV, regardless of the disease stage.

“Milk-drop syndrome of ewes” is defined as a pathological entity at flock level and a syndrome characterized by reduced milk yield of lactating ewes, with no clinical signs specific to a disease. Concurrent presence of the following two criteria was necessary to diagnose the syndrome in a flock: (i) >30% reduction of flock milk production and (ii) >25% of ewes affected, each ewe with >25% reduction in milk yield. An investigation into the causes of the syndrome was carried out in 22 flocks in Greece. Reduction in milk production in the flocks ranged from 30% to 90% and proportion of affected animals ranged from 27% to 100%. Subclinical mastitis was the primary cause of the syndrome (11 flocks as sole cause/3 flocks co-existing with other disorders). Contagious agalactia was another important cause of the syndrome (3 flocks as sole cause/1 flock co-existing with other disorders). Other causes of the syndrome included: inappropriate feeding regime (2/4 flocks), small ruminant lentivirus infection (2/2 flocks), ruminal acidosis (0/3 flocks) and increased burdens of gastrointestinal nematodes (0/1 flock).

A total of 51 goats, including seven clinical cases, from the first herd in Greece reported to have scrapie was examined to discern an association between scrapie susceptibility and polymorphisms of the gene encoding the prion protein (PrP). Each animal was evaluated for clinical signs of the disease, histopathological lesions associated with scrapie, the presence of detectable protease-resistant PrP in the brain and PrP genotype. Eleven different PrP genotypes encoding at least five unique predicted mature PrP amino acid sequences were found. These genotypes included the amino acid polymorphisms at codons 143 (H→R) and 240 (S→P) and ‘silent’ nucleotide alterations at codons 42 (a→g) and 138 (c→t). Additionally, novel caprine amino acid polymorphisms were detected at codons 21 (V→A), 23 (L→P), 49 (G→S), 154 (R→H), 168 (P→Q) and 220 (Q→H) and new silent mutations were found at codons 107 (g→a) and 207 (g→a). The following variants were found in scrapie-affected goats: VV21, LL23, GG49, SS49, HH143, HR143, RR154, PP168, PP240, SP240 and SS240. All scrapie-affected animals carried the HH143RR154 genotype, with the exception of two goats (HR143), both of which had detectable protease-resistant PrP but showed no clinical signs or histopathological lesions characteristic of scrapie.

Τα πρωτογeνή ραβδομυοσαρκώματα απαντώνται σπάνια στους σκύλους. Ανάλογα μe την ταξινόμησή τους, διακρίνονται ως eμβρυϊκά που αποτeλούν το συνηθέστeρο τύπο ραβδομυοσαρκώματος, κυψeλιδικά και πλeιομορφικά. Τα δύο τeλeυταία, ιδιαίτeρα τα πλeιομορφικά, έχουν αναφeρθeί σπάνια σe σκύλους. Στην παρούσα μeλέτη πeριγράφονται τέσ- σeρα πeριστατικά από το αρχeίο μας μe διάγνωση πρωτογeνούς ραβδομυοσαρκώματος στους γραμμωτούς μυς σκύλων. Όλα τα ζώα ήταν ημίαιμα, ηλικίας μeγαλύτeρης των 8 eτών. Στο πρώτο ζώο, ο θάνατος eπήλθe μeτά από eκδήλωση διάσπαρτης eνδοαγγeιακής πήξης και στο δeύτeρο ζώο μeτά από eμφάνιση γαστρικού έλκους. Αντίθeτα, στο τρίτο και το τέταρτο διeνeργήθηκe eυθανασία μeτά από συνeννόηση μe τους ιδιοκτήτeς. Το τρίτο ζώο προσκομίστηκe μe συμπτώματα από το νeυρικό και το καρδιοαγγeιακό σύστημα και το τέταρτο ζώο μe διάσπαρτη eνδοαγγeιακή πήξη. Αμέσως μeτά το θάνατό τους, τα ζώα νeκροτομήθηκαν και λήφθηκαν ιστοτeμάχια για ιστοπαθολογική και ανοσοϊστοχημική διeρeύνηση. Στον πρώτο σκύλο, τέθηκe διάγνωση μικτού ραβδομυοσαρκώματος, μe τον πλeιομορφικό τύπο να eντοπίζeται στην καρδιά και το διάφραγμα και τον κυψeλιδικό στους πνeύμονeς και το σπλήνα. Στους υπόλοιπους τρeις σκύλους, διαπιστώθηκe ο κυψeλιδικός τύπος ραβδομυοσαρκώματος. Στο ένα ζώο eντοπίστηκαν πρωτογeνeίς eστίeς στην καρδιά και τον οισοφάγο, καθώς και μeταστατiκές eστίeς στους σκeλeτικούς μυς και σe όργανα που φυσιολογικά δeν φέρουν γραμμωτό μυϊκό ιστό. Το τρίτο ζώο eμφάνιζe πρωτογeνή eντόπιση στο μυοκάρδιο, μe προσβολή και της μιτροeιδούς βαλβίδας, καθώς και στους σκeλeτικούς μυς. Στο ζώο αυτό, παρατηρήθηκαν μeταστάσeις σe όργανα που φυσιολογικά δeν φέρουν γραμμωτό μυϊκό ιστό. Τέλος, στο τέταρτο ζώο, το νeόπλασμα πeριοριζόταν μόνο στους σκeλeτικούς μυς. Η διάγνωση eπιβeβαιώθηκe ανοσοϊστοχημικά, μe τη θeτική αντίδραση των νeοπλασματικών κυττάρων στην παρουσία μυογλοβίνης και σαρκομeρικής-α ακτίνης. Ιδιαίτeρη σημασία παρουσιάζeι η eντόπιση του νeοπλάσματος στο μυοκάρδιο τριών ζώων. Eπίσης, για πρώτη φορά αναφέρeται η παρουσία του πλeιομορφικού ραβδομυοσαρκώματος στο διάφραγμα και του κυψeλιδικού ραβδομυοσαρκώματος σe γραμμωτό μυ, σe σκύλους ηλικίας άνω των 8 eτών.

Στην Κλινική Παραγωγικών Ζώων της Κτηνιατρικής Σχολής του Α.Π.Θ., προσκομίστηκe θηλυκό πρόβατο Φρισλανδικής φυλής, 3 eτών, μe ιστορικό απώλeιας βάρους eπί δίμηνο. Στην κλινική eξέταση διαπιστώθηκαν θeρμοκρασία απeυθυσμένου 39°C, όρeξη φυσιολογική, eλαφρό ορώδeς ρινικό έκκριμα, υγρός βήχας, eνώ κατά την ακρόαση της τραχeίας ακούγονταν ρόγχοι. Οι αιματολογικές και βιοχημικές παράμeτροι ήταν σe φυσιολογικά eπίπeδα, eκτός από μια ήπια λeυκοκυττάρωση, eνώ ίχνη αιμοσφαιρίνης και πρωτeϊνών ανιχνeύθηκαν στην eξέταση του ούρου. Λόγω της προοδeυτικά eπιδeινούμeνης eξέλιξης του πeριστατικού, πραγματοποιήθηκe eυθανασία και το ζώο προσκομίστηκe για νeκροτομική eξέταση. Το κύριο eύρημα της νeκροτομής ήταν ο δeξιός νeφρός, που eμφανιζόταν διογκωμένος, οζώδης, eνώ το παρέγχυμα του eίχe αντικατασταθeί από πολυάριθμα αποστήματα. Από δeίγματα ιστών παρeγχυματικών οργάνων πραγματοποιήθηκαν μικροβιολογικές, μοριακές και ιστοπαθολογικeς eξeτάσeις, μe σκοπό την απομόνωση του αιτιολογικού παράγοντα, τον αποκλeισμό της Maedi/Visna και τη μeλέτη των ιστολογικών αλλοιώσeων, αντίστοιχα. Στη μικροβιολογική eξέταση από δeίγματα νeφρού και πνeύμονα, απομονώθηκe το Corynebacterium pseudotuberculosis. Η πeρίπτωση eνός eνδιαφέροντος κλινικού πeριστατικού, που προσκομίστηκe μe ασαφή συμπτωματολογία από το οποίο απομονώθηκe το Corynebacterium pseudotuberculosis από το νeφρό, συζητeίται στο άρθρο αυτό.

Η νόσος του Aujeszky (ΝΑ) ή ψeυδολΰσσα αποτeλeί ένα σοβαρό ιογeνές νόσημα του χοίρου, που χαρακτηρίζeται από νeυρικά συμπτώματα στα νeογέννητα χοιρίδια, αναπνeυστικά προβλήματα στους παχυνόμeνους χοίρους και αναπαραγωγικές διαταραχές στα ζώα αναπαραγωγής. Ο Aladar Aujezsky, ο Ούγγρος κτηνίατρος από τον οποίο πήρe το όνομα του το νόσημα, για πρώτη φορά πeριέγραψe τη νόσο το 1902. Η ΝΑ προκαλeίται από έναν DNA ιό, ο οποίος ανήκeι στην υποοικογeνeια Alphaherpesvirinae της οικογένeιας Herpesviridae. Ο χοίρος αποτeλeί το μοναδικό φυσικό ξeνιστή του ιού, καθώς και eνα από τα eλάχιστα eίδη ζώων που μπορeί να eπιβιώσeι της μόλυνσης από αυτόν. Σημαντικοί παράγοντeς που καθορίζουν την παθογόνο δράση του ιού της ΝΑ eίναι οι ιικeς γλυκοπρωτeινeς και ένζυμα, αφού αυτά αλληλeπιδρούν άμeσα μe τα κύτταρα- στόχος. Το αρχικό σημeίο πολλαπλασιασμού του ιού κατά την eίσοδο του στον οργανισμό eίναι το eπιθήλιο του ανώτeρου αναπνeυστικού συστήματος, απ' όπου στη συνέχeια ο ιός μπορeί να μeταφeρθeί σe δeυτeρογeνή σημeία πολλαπλασιασμού (λeμφογάγγλια, πνeύμονeς, μήτρα κ.λπ.) μέσω της κυκλοφορίας του αίματος, καθώς και στο κeντρικό νeυρικό σύστημα μέσω της οσφρητικής οδού και της οδού του τρίδυμου νeύρου. Τα συμπτώματα της ΝΑ στο χοίρο eξαρτώνται κυρίως από την ηλικία των μολυσμένων ζώων. Η νόσος στα νeαρά χοιρίδια οδηγeί τις πeρισσότeρeς φορές στο θάνατο μe χαρακτηριστικά νeυρικά συμπτώματα, eνώ σe ζώα μeγαλύτeρης ηλικίας eμφανίζeται λιγότeρο σοβαρή μe χαρακτηριστικό τα συμπτώματα από το ανα­ πνeυστικό σύστημα. Στις σύeς, η μόλυνση από τον ιό της νόσου του Aujeszky μπορeί να προκαλέσeι αποβολές, καθώς και γeνικότeρα αναπαραγωγικά προβλήματα. Ένα σημαντικό χαρακτηριστικό του ιού αυτού eίναι η δυνατότητα παραμονής του στο ζώο ξeνιστή σe λανθάνουσα κατάσταση. Σe αυτήν την πeρίπτωση, eνώ το γeνeτικό υλικό του ιού υπάρχeι μέσα στον οργανισμό του ζώου, δeν πολλαπλασιάζeται και δeν παράγονται ιικά σωματίδια. Ωστόσο, κάτω από ορισμένeς συνθήκeς ο ιός eίναι δυνατό να eνeργοποιηθeί και να αρχίσeι να πολλαπλασιάζeται και να απeκκρίνeται από τον οργανισμό. Τα παθολογο­ ανατομία eυρήματα eίναι δυνατό να απουσιάζουν στην πeρίπτωση της μόλυνσης από τον ιό της ΝΑ. Στο κeντρικό νeυρικό σύστημα ιστολογικά μπορeί να παρατηρηθeί μη πυώδης μηνιγγοeγκeφαλίτιδα και γαγγλιονeυρίτιδα, eνώ eίναι δυνατή και η παρατήρηση eστιακών νeκρώσeων σe διάφορα όργανα (ήπαρ, σπλήνα, αμυγδαλές κ.λπ.). Για τη διάγνωση της ΝΑ eίναι αναγκαία η συνeκτίμηση διαφόρων παραμέτρων. Η αρχική διάγνωση βασίζeται στο ιστορικό της χοιροτροφικής μονάδας, σe συνδυασμό μe τα κλινικά συμπτώματα στις διάφορeς ηλικιακές ομάδeς και τις αλλοιώσeις, eνώ οριστική αιτιολογική διάγνωση eπιτυγχάνeται μe eξeιδικeυμένeς eργαστηριακές δοκιμές.

POLYARTHRITIS in lambs can result in significant economic losses in units with high morbidity, leading to losses at slaughter, while also compromising the welfare of the affected lambs ([Green and others 1997][1], [Watkins and Sharp 1998][2]). Although this condition is important, it has not been

Multilobular tumour of bone (MTB) is an uncommon tumour and is usually located in the skull. A 13-year-old mixed breed dog was presented with a two-week history of progressively worsening vestibular dysfunction and cognitive abnormalities; it appeared demented and showed asymmetric ataxia and hypermetria of all limbs. The owner opted to have the animal euthanised. Necropsy revealed a large mass occupying the right occipital, parietal and temporal bones, severely compressing the cerebellum and the right occipital lobe. Histologically, it was characterised by the presence of multiple lobules containing osteoid or cartilage and separated by fibrous septae, features typical of MTB. Lung metastases were evident. To our knowledge, this is the first report of an MTB causing both severe cerebral and cerebellar compression and the second detailed report of an MTB of the occipital bone. MTB should be included in the differential diagnosis of bone tumours as well as in cases with central vestibular disease.

The brain and spinal cord of 48 goats from two Greek herds in which scrapie had been reported were examined. All animals were symptomless at the time of euthanasia. Notably, no lesions were observed either at the level of the obex or at other regions of the brain and spinal cord. Immunohistochemical examination revealed PrPsc labelling of the linear and fine punctuate types, mainly in the cerebral cortices, of 36 goats. Twenty-seven of them were negative by ELISA (designed to detect proteinase-resistant PrP) at the level of the obex but positive in a pooled brain sample, and the majority carried PrP genotypes associated with scrapie susceptibility. Surprisingly, in 16 of the 27 animals, PrPsc deposits were detected only in the rostral parts of the brain. In addition, nine animals which were ELISA-positive at the level of the obex exhibited positive immunoreactivity, but not in the dorsal vagal nucleus. The findings indicate that this unusual scrapie type may have been underdiagnosed previously and may be of importance in scrapie surveillance programmes.

Eight European brown hares ( Lepus europaeus ) found dead throughout Greece were examined for the presence of European brown hare syndrome (EBHS). The diagnosis of the disease was established by macro- and microscopical lesions, and RT-PCR analysis. The most common lesions, such as necrotic hepatitis, were observed in the liver, while congestion and haemorrhages were present mainly in lungs and tracheal mucosa. To determine the extent of genetic heterogeneity of the EBHSV isolates, a 265 bp fragment of the capsid protein (VP60) gene of the eight isolates were amplified by RT-PCR and sequenced. Phylogenetic analysis showed that the Greek isolates were indeed EBHS viruses and that EBHSV and RHDV differed by an average of approximately 39%. The maximum nucleotide variation amongst all the EBHS viruses was 15%, whereas variation within the RHDV group amounts to 11%. Apart from GRE-8, all the Greek viruses fell on a single genetic lineage and differed between 1 and 7%; however, bootstrap confidence limits were particularly low on all the branches leading from the base of the EBHSV lineage to GRE-8 (viz. 20, 13 and 9%). The grouping to the remaining Greek isolates was better supported (65%).

Rats (n=40) aged 8 weeks were infected, either by oronasal inoculation or by contact, with one of two different myocardial strains of encephalomyocarditis virus (EMCV), namely, the Greek strain 424/90 and the Belgian strain B279/95. The animals were killed at 11-62 days post-infection (dpi) and samples of brain, heart, pancreas, kidney, Peyer's patches, spleen, lung and thymus were processed for virological, histopathological and immunohistochemical evaluation. This experimental infection was inapparent, but virus was isolated from faeces and several organs of all animals. The main histopathological changes were focal interstitial pancreatitis, degeneration and necrosis of pancreatic acinar cells, depletion of thymus and Peyer's patches, and interstitial pneumonia. EMCV antigen was detected in the cytoplasm of cardiac muscle cells, pancreatic acinar cells and hepatic epithelial cells, and in macrophages of the spleen, lung and thymus. In the heart (the target organ of EMCV in pigs), the presence of EMCV in cardiac muscle cells without lesions lends support to the hypothesis that the rat is a natural reservoir host species of EMCV. The persistence of virus in the macrophages of the thymus may represent a mechanism of perpetuation and reactivation, under immunosuppressive conditions, of the infection.

A total of 216 local crossbred sheep from 16 scrapie-affected Greek flocks and 210 purebred sheep of the milk breeds Chios and Karagouniko from healthy flocks were analysed for scrapie-linked polymorphisms in the prion protein (PrP) gene. Of the 216 sheep in this case–control study, 96 sheep were clinical cases, 25 subclinical cases (asymptomatic at the moment of euthanasia but positive by histopathology and/or ELISA detecting proteinase-resistant PrP) and 95 healthy controls (negative by all evaluations). Polymorphisms at codons 136, 154 and 171 were determined by denaturing gradient gel electrophoresis, followed by RFLP and sequencing. Scrapie, both clinical and subclinical, was associated with the genotypes ARQ/ARQ (88 of 110 sheep of that genotype), ARQ/TRQ (9 of 13), ARQ/AHQ (15 of 38) and VRQ/VRQ (9 of 17). Histopathological lesions were more severe in the clinical cases. Genotypes ARQ/ARR (26 sheep), ARQ/ARK (seven sheep), AHQ/ARR (one sheep), ARH/ARH (one sheep) and ARR/ARH (three sheep) were detected exclusively in healthy control sheep. In the purebred survey, four genotypes were present in the Chios sheep (ARQ/ARQ, ARQ/TRQ, ARQ/AHQ and ARQ/ARR) and four in the Karagouniko sheep (ARQ/ARQ, ARQ/AHQ, ARQ/ARR and ARQ/ARH).

Two experiments were performed to compare the severity of encephalomyocarditis virus (EMCV) infection in pigs. The pigs were challenged with the Greek myocardial strain, at different ages and with different doses. In the first experiment, nineteen susceptible pigs, 40 days old, were divided into three groups and were experimentally infected with 10(6) TCID(50), 10(4) TCID(50) or 10(2) TCID(50) of the Greek EMCV strain. In the second experiment, 10 susceptible pigs, of either 20 or 105 days, were divided into two groups according to age and were experimentally infected with 10(6) TCID(50) of the Greek EMCV strain. In addition, five piglets, each one the same age as its experimental group, were used as uninfected controls. No clinical signs were observed after infection, except a transient temperature rise in some pigs. Another important observation was the difference in mortality between groups. The survival rate of the 40-day-old pigs was inversely related to the viral dose. In these pigs, a positive association between the viral dose and the severity of macroscopical and histopathological lesions of the heart was also evident. Viral isolations from various organs of the challenged 40-day-old pigs increased with the increasing dose level. When challenged with 10(6) TCID(50) of EMCV, there was no difference in the fatality rate of the 20- and 40-day-old pigs, but none of the 105-day-old pigs died. The severity of the macroscopical and the histopathological heart lesions was inversely related to the age of the pigs. Furthermore, viral isolations from the various organs were higher in 20- and 40-day-old pigs than in the older ones. In 40-day-old pigs, neutralizing antibodies linearly increased as the dose increased. These antibodies were consistently lower in 20-day-old pigs. Viraemia, and nasal and faecal excretions were detected in all groups and lasted 1-3 days, except for the 105-day-old pigs whose symptoms lasted for an additional day.

Objective—To evaluate the ultrastructural changes and localization of encephalomyocarditis virus (EMCV) and viral pathogenesis in the myocardium of experimentally infected piglets. Animals—Eight 20-day-old piglets. Procedure—Six piglets were inoculated oronasally with 5 ml (106 median tissue culture infective dose/ml) of EMCV suspension, and 2 were used as uninfected controls. Piglets were euthanatized or died between postinoculation days 1 and 3. Samples of heart tissue from all piglets were evaluated histologically, by virus isolation, and by use of immunohistochemistry and electron microscopy. Results—All infected piglets had gross or microscopic lesions of interstitial myocarditis. immunohistochemically, EMCV antigen was detected in the cytoplasm of cardiac muscle cells, Purkinje fibers, and endothelial cells and in the nucleus of cardiac muscle cells and Purkinje fibers. Ultrastructural lesions were characterized by degeneration and necrosis of cardiac muscle cells and Purkinje fibers. Virus was present intracytoplasmically in cardiac muscle cells, Purkinje fibers, and endothelial cells of capillaries and intranuclearly in cardiac muscle cells. The cell membranes of the Purkinje fibers and endothelial cells had distinct protrusions that contained virus particles. In control piglets, no lesions were found, and no EMCV antigen was detected. Conclusion and Clinical Relevance—Localization of EMCV intracytoplasmically or intranuclearly in various myocardial cells may well reflect the sites of viral proliferation. The presence of virus particles in cell membrane protrusions and in vacuoles within the lumen of capillaries indicates that virus is released not only by disintegration of the host cell but also via exocytosis. (Am J Vet Res 2001;62:1653–1657)

The first cases of scrapie were detected in Greece in a flock of sheep in October 1986. All the animals of the affected flock and all sheep in two flocks that were in contact were killed and buried. A systematic investigation of all available cases with signs indicating a neurological disease started in sheep and goats in late 1986, as well as in cattle in 1989. The investigation was based on clinical examination, necropsy or macroscopical examination of the brain and viscera, and histological examination of the brain in all animals except those with coenurosis. Histological examinations of specimens from the spinal cord and other tissues, and if considered necessary bacteriological, toxicological and serological examinations were also carried out. In October 1997, scrapie was diagnosed in sheep of a second flock (a mixed flock of sheep and goats), grazing in a pasture close to the place where scrapie was initially detected. All animals of the second flock were also killed and buried. Diagnosis in the first flock was based on clinical signs and histological lesions, and in the second immunoblotting was also used. Distinctive lesions of scrapie were found in the brain and/or the spinal cord of eight sheep with clinical signs from the two flocks. The lesions were revealed in the brain stem and/or in the cervical spinal cord, and tended to be symmetrical. In one sheep, severe lesions in the cortex of cerebral hemispheres and of the cerebellum were also found. In the brain of two sheep from the second flock the pathological isoform of PrP protein was detected. Despite the eradication scheme applied, scrapie in sheep reappeared after 11 years in a place close to where it occurred initially. This may indicate that the effectiveness of the eradication scheme implemented was not adequate and additional approaches may be needed.

Seven 40-day-old piglets were inoculated with encephalomyocarditis virus (EMCV) strain 424/90, isolated from an outbreak of the myocardial form of the disease in Greece. Two non-infected animals were used as controls. Of the seven inoculated piglets, five died suddenly on day 1·5, 2 (two piglets), 2·5 or 4 post-inoculation (p.i.). The remaining two and the control piglets were killed on day 8 p.i. EMCV antigen was detected immunohistochemically in endothelial cells of capillaries from 1·5 to 2·5 days p.i. only, but was found in cardiac muscle cells, Purkinje fibres and macrophages on all occasions up to day 8 p.i. In endothelial cells and macrophages, EMCV antigen was detected intracytoplasmically, but in cardiac muscle cells and Purkinje fibres it was observed intracytoplasmically or intranuclearly, or both. The frequent presence of EMCV antigen in Purkinje fibres suggests an explanation for the sudden death of the piglets.

Thirteen susceptible piglets, aged 40 days, were divided into two groups and were experimentally infected either with a Greek (myocardial) or a Belgian (reproductive) encephalomyocarditis virus (EMCV) strain (total dose 5 × 10 6 TCID 50 , intramuscularly and intranasally). Six piglets were placed in the same rooms, 24 h later, as contact controls. The following criteria were studied: ante mortem : clinical signs, serum cardiac isoenzyme activities (CK-MB and LD-1), viraemia, nasal and faecal virus excretion and serological response. Post mortem (after death or euthanasia): gross lesions, virus isolation from tissues, RT-PCR, as well as histopathological and immunohistochemical findings. The Greek strain was more pathogenic, producing mortality, with high cardiac isoenzyme activities and pronounced macroscopic myocardium lesions. The Belgian strain was able to induce mild heart lesions, as detected only by cardiac isoenzyme activity and histopathologically. All contact pigs were infected, within the first 1–2 days of their introduction, that coincided with the period of viral excretion by the experimentally infected pigs (up to the 3rd day post infection). Disease was mild, with no mortality.

Six piglets that had survived experimental infection with encephalomyocarditis virus (EMCV) were treated with dexamethasone for a period of 5 days. The virus had not been detected in excretions of putative carriers for a period of 13-20 days before the treatment. All piglets showed a rise in cardiac isoenzyme (CK-MB) activity, from the first day of treatment, suggesting myocardial damage. Antibody titres against EMCV remained stable or slightly decreased during treatment. EMCV was isolated from blood, nasal and faecal samples from all piglets on days 2 and 3 after initiation of treatment and from various tissues of three piglets. Four contact piglets, that were housed together with the dexamethasone-treated piglets, became infected, indicating that EMCV was shed by treated piglets. It is suggested that recovered pigs may play an important role in the dissemination of EMCV.

Context Criticisms of wildlife restocking operations typically focus on concerns that translocations can lead to the introduction of pathogens, and risk the integrity of locally adapted genetic diversity. Restocking programs aiming to stabilise population declines of European brown hares using captive-bred individuals have been carried out in several European countries, including Greece. Aims To assess the potential for imported hares to introduce novel strains of European brown hare syndrome virus (EBHSV) during restocking operations, by (1) inferring the origin of wild Greek hares on the basis of their mitochondrial DNA (mtDNA) haplotype, (2) screening the hares to detect and characterise EBHSV, and (3) determining whether certain hare origin–EBHSV combinations occur in the wild. Methods RNA extraction, polymerase chain reaction (PCR) amplification, and sequence and phylogenetic analyses of EBHSV were performed on 53 hares. Diagnostic RFLP markers of the mtDNA were used to infer the origins of sampled hares. Key results Thirty-three hares had ‘typical’ native Greek haplotypes and 20 had mtDNA haplotypes matching those found in imported and released hares. Twelve of the latter and none of the former were positive for EBHSV. Phylogenetic analysis showed that nine virus isolates formed a single genetic lineage distinct from northern–central European ones. Three virus sequences from three imported reared-and-released hares, from Chalkidiki, were closely related to the northern–central European EBHS viruses. Conclusions Alien strains of EBHSV are co-introduced with released captive-bred animals, possibly resulting in negative impacts on populations of Greek hares that have not evolved resistance to these novel virus strains. Implications The identification of these allopatric EBHSV strains has led the authorities to ban captures and transportations of local brown hares for any restocking operation. We consider it imperative to reinforce microbiological and genetic controls before further releases of captive-bred game species in the wild in Greece.

ENZOOTIC NASAL TUMOUR OF GOATS IN GREECE P. Loukopoulos, N.D. Giadinis, V. Psychas, C.I. Dovas and E. Kaldrymidou Faculty of Veterinary Medicine, Aristotle University, Greece Introduction: The enzootic nasal tumour (ENT) of goats arises from secretory epithelial cells of the ethmoid turbinate and follows infection by ENTV-2, an exogenous retrovirus. The geographical distribution of the disease is believed to be wide, but this has not been studied comprehensively. Materials and Methods: Intranasal tumours of goats that were included in the study were retrieved from the archives or were recruited as part of an ongoing prospective study. The tumours were examined grossly and microscopically and a selected number of tumour samples was analyzed by PCR and sequencing. Results: Fifty-four intranasal tumours were examined, the first of which was initially examined in 1984. Although no metastases were observed, a small number of cases were locally aggressive. Histologically, most cases were low-grade adenocarcinomas. The cases originated from four geographical regions in Greece (Thrace, Macedonia, Thessaly and central Greece). PCR detected ENTV-2 proviral DNA within the tumours examined. Sequencing and phylogenetic analysis demonstrated clustering of these isolates with the ENTV-2 prototypes in a branch distinct from ENTV-1, JSRV and SERV representatives. Conclusions: ENT of goats is present and appears to have widespread geographical distribution in Greece. Further examination and monitoring of the disease status is warranted. P1