Virological Characterization of Pigs with Erythema Multiforme

Erythema multiforme in pigs is an acute, self-limiting disease characterized by red skin areas and often associated with anorexia, fever and respiratory problems. The cause of the disease remains unknown. In a recent study, animals of a commercial breeding herd in Greece were examined, and all animals were found seropositive for porcine reproductive and respiratory syndrome virus (PRRSV). However, neither PRRSV and porcine circovirus type 2 (PCV2) viremia nor antibodies against Aujeszky’s disease virus, African swine fever virus and classical swine fever virus were detected. Here, an extended examination of these pigs was performed on a wide range of porcine viruses using highly sensitive polymerase chain reaction (PCR)-based methods. Affected skin of five animals revealed the presence of porcine lymphotropic herpesvirus-1 (PLHV-1) in all cases, PLHV-2 in one animal and PLHV-3 in four animals. However, neither porcine cytomegalovirus (PCMV) nor porcine circoviruses (PCV1, PCV2, PCV3 and PCV4) were detected. In blood samples, PLHV-1 was present in two animals and PLHV-2, PCV2 and PCV3 in one individual, with PCMV, PCV1 and PCV4 in none of the animals. In one animal, four viruses were found in the blood (PLHV-1, PLHV-2, PCV2 and PCV3). A PRRSV viremia was also not detected. All animals carried porcine endogenous retrovirus C (PERV-C) in their genome, but recombinant PERV-A/C was not detected. The results suggest that porcine viruses may be involved in erythema multiforme in these animals and that further studies are needed to assess the role of these pathogens in the disease

Fifth European Dirofilaria and Angiostrongylus Days (FiEDAD) 2016

Peer reviewe

Study of the experimental encephalomyocarditis in rats (rattus norvegicus) and mice (mus musculus)

The aim of the present study was to investigate the pathogenesis of encephalomyocarditis in rats and mice and specifically of the disease caused by two different strains, the Greek 424/90 and the Belgian B279/95, which were isolated from outbreaks that occurred in pig farms in Europe. For this purpose, rats and mice were experimentally infected and the histopathological lesions induced by the two strains in these animal species and the distribution of the viral antigen in the various organs were studied. The immunohistochemical examinations were performed using the avidin-biotin complex method. Furthermore, tissue samples were obtained for the purpose of isolation of the virus in cell cultures. In addition, the period of viral excretion in faeces, the production of neutralizing antibodies and the blood glucose levels were studied. The samples were obtained from 44 rats (Wistar rats) and 24 mice (BALB /c mice) aged 8 weeks, separated in six groups (A, B, C, D, E, F). Half of the animals of the groups A and C were inoculated oronasally with 104.5 TCID50 /ml (dose volume 0.5 ml) of the Greek strain 424/90 and half of the animals of the groups B and D were inoculated oronasally with 104.5 TCID50 /ml (dose volume 0.5 ml) of the Belgian strain B279/95. Each one of the rest of the animals of the same four groups (A, B, C and D) was placed in the same cage together with an inoculated animal. Groups E and F served as controls. Euthanasia of all animals was carried out in two phases: 11-22 (first phase) and 56-62 (second phase) days post infection. EMCV was isolated from faeces of rats on days 2 to 29 and from faeces of mice on days 2 to 15 post infection. In rats, encephalomyocarditis virus (EMCV) was also isolated from the heart, the thymus, the lungs, the liver, the pancreas, the kidneys, the spleen and the small intestine (Peyer’s patches) in the first phase and from the thymus and the small intestine (Peyer’s patches) in the second phase of the experiment. In mice, EMCV was isolated from the thymus, the lungs and the pancreas of mice in the first phase and from the thymus of only two animals in the second phase of the experiment. The main histopathological lesions observed both in rats and mice were interstitial pancreatitis, interstitial pneumonia, depletion and necrosis of lymphocytes of thymus and Peyer’s patches. In all cases the lesions were mild. Additionally, histopathological lesions in the heart were observed only in two mice that died. In rats, EMCV antigen was detected in the cardiac muscles cells, in the pancreatic acinar cells, in the epithelial cells of the liver and in macrophages of the lung, spleen and thymus in the first phase and in the cardiac muscles cells, in the pancreatic acinar cells and in macrophages of the thymus in the second phase of the experiment. In mice, EMCV antigen was detected in the pancreatic acinar cells and in macrophages of the lung and thymus in the first phase and in the pancreatic acinar cells and in macrophages of the thymus in the second phase of the experiment. The antigen was also detected in the cytoplasm of cardiac muscle cells of three mice. The titres of neutralizing antibodies reached the cut-off value in only a few animals. Blood glucose concentrations were always within normal limits. The two viral strains that we studied, cause inapparent infection in rats and mice and only occasionally deaths in mice and seem to be able to spread in a population of rodents by horizontal rat-to-rat and mouse-to-mouse transmission. Histopathological lesions of similar nature and intensity were observed in the same organs of both contact infected and experimentally infected animals. Moreover, a similar distribution of EMCV antigen in the various organs of contact infected compared to experimentally infected animals was observed.Σκοπός της παρούσας έρευνας ήταν η μελέτη της παθογένειας της εγκεφαλομυοκαρδίτιδας στους επίμυς και τους μυς και συγκεκριμένα της νόσου που προκαλούν δύο στελέχη του ιού, το Ελληνικό 424/90 και το Βελγικό Β279/95, τα οποία απομονώθηκαν από περιστατικά κλινικής νόσου σε χοίρους, σε περιοχές της Ευρώπης όπου η νόσος ενζωοτεί. Για το σκοπό αυτό πραγματοποιήθηκε πειραματική αναπαραγωγή της νόσου και μελετήθηκαν οι ιστοπαθολογικές αλλοιώσεις που προκαλούνται από τα συγκεκριμένα στελέχη του ιού σε αυτά τα είδη ζώων και η κατανομή του αντιγόνου του ιού στα διάφορα όργανα με τη χρήση της ανοσοϊστοχημικής μεθόδου του συμπλέγματος αβιδίνης-βιοτίνης. Επιπρόσθετα, ελήφθησαν δείγματα οργάνων για την απομόνωση του ιού σε κυτταροκαλλιέργειες. Επίσης, μελετήθηκαν η περίοδος απέκκρισης του ιού με τα κόπρανα, η παραγωγή εξουδετερωτικών αντισωμάτων και οι συγκεντρώσεις της γλυκόζης στο αίμα των πειραματόζωων. Το παθολογικό υλικό προήλθε από 44 επίμυς (Wistar rats) και 24 μυς (BALB/c mice) ηλικίας 8 εβδομάδων, που διαχωρίστηκαν σε έξι ομάδες (Α, Β, Γ, Δ, Ε, ΣΤ). Τα μισά πειραματόζωα των ομάδων Α και Γ μολύνθηκαν στοματορινικά με 0,5 ml εναιωρήματος 104.5 TCID50 /ml του Ελληνικού στελέχους 424/90 και τα μισά πειραματόζωα των ομάδων Β και Δ μολύνθηκαν στοματορινικά με 0,5 ml εναιωρήματος 104.5 TCID50 /ml του Βελγικού στελέχους Β279/95 του ιού. Καθένα από τα υπόλοιπα πειραματόζωα των τεσσάρων αυτών ομάδων (Α, Β, Γ και Δ) τοποθετήθηκε στον ίδιο κλωβό με ένα πειραματικά μολυσμένο πειραματόζωο. Οι ομάδες Ε και ΣΤ αποτέλεσαν τους μάρτυρες του πειραματισμού. Η ευθανασία των πειραματόζωων πραγματοποιήθηκε σε δύο φάσεις: πρώτη φάση 11-22 ημέρες μετά τη μόλυνση και δεύτερη φάση 56-62 ημέρες μετά τη μόλυνση. Ο ιός απομονώθηκε από τα κόπρανα των επίμυων και των μυών 2 έως 29 και 2 έως 15 ημέρες μετά τη μόλυνση αντίστοιχα. Στους επίμυς, ο ιός απομονώθηκε από την καρδιά, το θύμο αδένα, τους πνεύμονες, το ήπαρ, το πάγκρεας, τους νεφρούς, το σπλήνα και το λεπτό έντερο (πλάκες του Peyer) στην πρώτη φάση και από το θύμο αδένα και το λεπτό έντερο (πλάκες του Peyer) στη δεύτερη φάση του πειραματισμού. Στους μυς, ο ιός απομονώθηκε από το θύμο αδένα, τους πνεύμονες και το πάγκρεας στην πρώτη φάση και από το θύμο αδένα δύο πειραματόζωων στη δεύτερη φάση του πειραματισμού. Τόσο στους επίμυς όσο και στους μυς, οι κύριες ιστοπαθολογικές αλλοιώσεις που διαπιστώθηκαν ήταν διάμεση παγκρεατίτιδα, διάμεση πνευμονία και νέκρωση των λεμφοκυττάρων και μείωση του αριθμού τους στο θύμου αδένα και τις πλάκες του Peyer. Σε όλες τις περιπτώσεις οι αλλοιώσεις ήταν ήπιες. Επιπρόσθετα, ιστοπαθολογικές αλλοιώσεις στην καρδιά παρατηρήθηκαν σε δύο μόνο μυς οι οποίοι πέθαναν. Στους επίμυς, το αντιγόνο του ιού, στην πρώτη φάση, διαπιστώθηκε ανοσοϊστοχημικά στα καρδιακά μυϊκά κύτταρα, στα αδενικά κύτταρα της εξωκρινούς μοίρας του παγκρέατος, στα επιθηλιακά κύτταρα του ήπατος και στα μακροφάγα του πνεύμονα, του σπλήνα και του θύμου αδένα, και στη δεύτερη φάση, στα καρδιακά μυϊκά κύτταρα, στα αδενικά κύτταρα της εξωκρινούς μοίρας του παγκρέατος και στα μακροφάγα του θύμου αδένα. Στους μυς, το αντιγόνο του ιού διαπιστώθηκε ανοσοϊστοχημικά στα αδενικά κύτταρα της εξωκρινούς μοίρας του παγκρέατος και στα μακροφάγα του πνεύμονα και του θύμου αδένα, κατά την πρώτη φάση του πειραματισμού και στα αδενικά κύτταρα της εξωκρινούς μοίρας του παγκρέατος και στα μακροφάγα του θύμου αδένα, κατά τη δεύτερη φάση του πειραματισμού. Σε τρεις μόνο περιπτώσεις το αντιγόνο του ιού διαπιστώθηκε ανοσοϊστοχημικά στα καρδιακά μυϊκά κύτταρα μυών. Ο τίτλος των εξουδετερωτικών αντισωμάτων έφτασε το κρίσιμο όριο διάκρισης σε πολύ μικρό αριθμό πειραματόζωων. Οι συγκεντρώσεις της γλυκόζης στο αίμα όλων των πειραματόζωων διατηρήθηκαν πάντα σε φυσιολογικά επίπεδα

Effect of Bladder Injection of OnabotulinumtoxinA on the Central Expression of Genes Associated with the Control of the Lower Urinary Tract: A Study in Normal Rats

To investigate a possible central mechanism of action of Botulinum toxin A (BoNT/A) following injection in the bladder, complementary to the acknowledged peripheral bladder effect, we studied changes in the expression of neuropeptides and receptors involved in lower urinary tract function in the spinal cord (SC) and dorsal root ganglia (DRG) of normal rats following BoNT/A bladder injection. Thirty-six Sprague-Dawley rats, divided into three groups of n = 12, received bladder injections of 2U or 5U OnabotulinumtoxinA (BOTOX®), or saline. Six animals from each group were sacrificed on days 7 and 14. Expression of Tachykinin 1 (Tac1), capsaicin receptor (TRPV1), neuropeptide Y (NPY), proenkephalin (PENK) and muscarinic receptors M1, M2, M3, was evaluated in the bladder, L6-S1 DRG, and SC segments using real-time PCR and Western blotting. Real-time PCR revealed increased expression of NPY in all tissues except for SC, and increased TRPV1 and PENK expression in DRG and SC, whereas expression of Tac1, M1 and M2 was decreased. Less significant changes were noted in protein levels. These findings suggest that bladder injections of OnabotulinumtoxinA may be followed by changes in the expression of sensory, sympathetic and cholinergic bladder function regulators at the DRG/SC level

Τhe Effect of Opioid Administration on Cytologic and Histopathologic Diagnosis of Canine Cutaneous Mast Cell Tumors Treated by Surgical Excision

Mast cell tumor (MCT) is a frequent cutaneous tumor in dogs, with a variable biological behavior. Studies correlate cytologic and histopathologic features of MCTs with their biological behavior, prognosis, and response to treatment. The use of preoperative opioids is common in canine patients undergoing surgical removal of these tumors. Certain opioids can induce or downregulate mast cell degranulation and influence cancer progression. The aim of the present study was to investigate whether the administration of morphine or butorphanol during surgical excision of canine cutaneous MCTs affects their cytologic and histopathologic appearance, thus influencing cytologic and histopathologic grading. This was a prospective, blinded, randomized, cohort clinical study. Forty-five dogs with cutaneous MCTs were randomly allocated into three groups according to preanaesthetic medication: dexmedetomidine combined with morphine (group M) or butorphanol (group B) or normal saline (group C). Cytologic specimens and histopathologic samples were obtained both prior to and after surgery. Samples were graded according to Kiupel’s and Patnaik’s systems, examined immunohistochemically for Ki-67 protein (Ki-67) and c-kit proto-oncogene product (KIT) expression, and histochemically for argyrophilic nucleolar organizing regions (AgNORs). Based on both Kiupel’s and Patnaik’s systems, no statistically significant differences were noted concerning the number of cases with grading discrepancies in grades allocated prior to versus after surgery among the groups. The same applied for cytological grading and immunohistochemical and histochemical evaluation. It seems that administration of morphine or butorphanol as part of the preanesthetic medication for surgical removal of canine cutaneous mast cell tumors does not influence histopathologic and cytologic grading of MCTs

First evidence of the European wildcat (Felis silvestris silvestris) as definitive host of Angiostrongylus chabaudi

Angiostrongylus chabaudi (Strongylida, Angiostrongylidae) is a parasitic nematode described for the first time last century from the pulmonary arteries of six European wildcats (Felis silvestris silvestris) in central Italy. Since then, this parasite remained practically unknown until recently, when immature A. chabaudi have been reported from one wildcat in Germany and two domestic cats (Felis silvestris catus) in Italy. The present report describes the first record of A. chabaudi in Greece and, most importantly, the first known case of patent infection by A. chabaudi. The necropsy of a road-killed F. s. silvestris found near the lake Kerkini, in the municipality of Serres (Macedonia, Greece), revealed the presence of nematodes of both sexes in the right ventricle and the pulmonary artery of the heart. All parasites were mature adults and numerous eggs were present in the uteruses of females. The morphological characteristics of the parasites were consistent with those of A. chabaudi. Moreover, Angiostrongylus-like first stage larvae (L1) were present in the faeces of the animal that was negative for any other cardio-pulmonary parasite. Genetic examination of adult parasites and L1 confirmed the morphological identification as A. chabaudi. Histopathological examination of the lungs showed severe, multifocal to coalescing, chronic, interstitial granulomatous pneumonia due to the presence of adult parasites, larvae and eggs. These findings demonstrate for the first unequivocal time that this nematode reproduces in the European wildcat which should be ultimately considered a definitive host of A. chabaudi. Finally, the L1 of A. chabaudi are described here for the first time, opening new prospects for further studies on this neglected parasite

Regurgitations in a Lamb with Acute Coenurosis-A case Report

  Coenurosis is a disease of the central nervous system in sheep, caused by Coenurus cerebralis, the larval stage of Multiceps multiceps, which inhabits the small intestine of Canidae. A case of regurgitations in a 2.5 month old lamb with acute coenurosis is being reported. The lamb was presented with a sudden onset of ataxia and re-gurgitations for 10 days. The post-mortem examination revealed 4 immature C. cerebralis cysts between 0.5 and 1.5 cm in diameter located in the brainstem and cerebellum, and histopathological examination revealed multifocal pyogranuloma-tous meningoencephalitis, so a diagnosis of acute coenurosis was established. Thus, acute coenurosis should be included in the differential diagnosis of regurgi-tations in lambs

Erythema multiforme associated with respiratory disease in a commercial breeding pig herd

This study describes an erythema multiforme (EM) in breeding sows, after their mixing in the group housing system. Sows at 30-35 days of gestation showed red and raised skin areas, depression, anorexia, fever, respiratory problems, and increased return to estrus. Blood and nasal samples from diseased sows were examined by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay for respiratory pathogens. Hematological and biochemical analyses were performed on the blood samples. From diseased sows, vaginal swabs for microbiological examinations and samples at slaughterhouse for gross and microscopic examinations were collected. Samples from the complete gestation and lactation feed were examined for mycotoxins. All sampled sows were seropositive for Actinobacillus pleuropneumoniae (App) and porcine reproductive and respiratory syndrome virus (PRRSV). No viremia for PRRSV and porcine circovirus type 2 were detected. All nasal samples were positive for Streptococcus suis, one for Swine Influenza Virus and one for App, Hemophilus parasuis, and S. suis. In all vaginal swabs, Escherichia coli and Streptococcus spp. were detected. Diseased sows had moderate leukocytosis, mild anemia, and thrombocytopenia. No mycotoxins were detected in feed. Histopathological examination revealed increased vascularization of the superficial and middle dermis. EM was likely due to illness caused by viral and bacterial infections. This study suggests that stress caused by the sows' mixing might have triggered the problem