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Toxoplasma gondii and Chlamydophila abortus are the 2 most common infectious causes of ovine abortion worldwide. These obligate intracellular pathogens are associated with severe placentitis leading to abortion or stillbirth in pregnant ewes, and resulting in significant economic losses. The objectives of the current study were the development, validation, and application of a duplex real-time polymerase chain reaction (PCR) assay capable of quantifying the burden of infection by T. gondii and C. abortus in material submitted for diagnostic purposes. The validation was carried out using samples from ewes experimentally infected with these organisms. Based on the numbers of genome copies detected, an arbitrary cutoff level was established to correlate with significant pathological changes sufficient to give rise to abortion. When the PCR assay was applied to samples from 66 Irish farms with naturally occurring outbreaks of ovine abortion, toxoplasmosis and enzootic abortion of ewes (EAE) accounted for 14% and 20% of the farms, respectively, while on 6% of the farms, there was evidence of dual infection. When standard diagnostic techniques including histopathological examination, serological analysis, chlamydial antigen detection, and bacteriological culture, were used on samples from the same farms, toxoplasmosis was diagnosed in 17% of farms, and EAE in 12%; dual infection was diagnosed on 3% of the farms. In general, good agreement was found between the PCR and the standard methods. The duplex real-time PCR assay developed in this study has proved to be a very sensitive and rapid tool that might provide a valuable addition to the methods currently available for routine diagnosis of ovine abortions.

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Enzootic abortion of ewes (EAE) caused by Chlamydophila abortus is an important disease resulting in significant lamb loss in most sheep producing countries. Ewes are considered to be naturally infected with C. abortus via the oral-nasal route and may become persistent carriers, shedding during subsequent oestrous cycles and at lambing. The aim of this study was to monitor the clinical outcomes, pathological changes and shedding of C. abortus in 18 periparturient orally infected sheep for two breeding seasons. In the first season, C. abortus was detected by real-time PCR (rt-PCR) in 13/18 conjunctival swabs at oestrus. Three out of the 15 pregnant ewes gave birth to 1 live and 1 dead lamb, and 2 of them aborted. Following parturition/abortion, C. abortus was detected in 12/15 vaginal swabs and in all the collected foetal membranes. However, only those membranes containing high copy numbers of the bacterium displayed the EAE typical lesions. In the second season, none of the 13 pregnant ewes aborted, and 5 of them gave birth to dead or weak lambs. C. abortus was not detected in conjunctival or vaginal swabs at oestrus or parturition. The bacterium was detected at low levels in 36% of the foetal membranes, but with no evidence of histopathological lesions. These results indicate that C. abortus can be detected in a large proportion of animals during the first pregnancy after oral infection. However, this proportion is reduced at the subsequent breeding season, confirming the occurrence of a chronic low level persistent infection in post-abortion/lambing ewes.

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Mammary tumors of all types are rare in herbivores, and there is a particular paucity of reports in sheep. The present report describes a case of mammary carcinoma in a 6-year-old uniparous ewe. The tumor was present for at least 18 months, during which time the ewe remained clinically well. At postmortem examination, the mass was found to be multilobulated with occasional cysts. Histologically, the lobules consisted of tubules lined by cuboidal to low columnar epithelium with loss of polarity and moderate anisokaryosis within a moderately extensive fibrous stroma. It was classified as a low-grade carcinoma. The histologic classification and lack of evidence of invasion correlated well with the slow clinical progression.

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A 39-yr-old, acyclic, uniparous, female white rhinoceros with a history of recurrent vaginal bleeding was euthanized following a period of respiratory distress and ill-thrift. The rhinoceros' uterus had previously been evaluated by ultrasound and diffuse endometrial hyperplasia and two benign uterine leiomyomas had been diagnosed. At necropsy examination, a large, infiltrative, metastatic uterine adenocarcinoma was found multifocally throughout the uterus, scattered within the peritoneal cavity, on the diaphragm, the splenic capsule, the pleural surface of the lung and mesenteric lymph nodes. A large volume (100 L) of ascites fluid was present in the abdominal and pleural cavities.

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A neuropathologic survey was conducted on mink brains from the 5 licensed mink farms in Ireland. The survey was part of a transmissible spongiform encephalopathy surveillance study. Aleutian disease (AD) was present on 4 of the 5 farms (80%). Neuropathologic features of nonsuppurative meningoencephalitis were common in mink from the 4 affected farms but were absent in the mink from the fifth farm, which was free of AD. The meningoencephalitis was characterized by infiltrates of lymphocytes and plasma cells, which were present in meninges, perivascular spaces, and the brain parenchyma. Fibrinoid necrotizing arteritis was seen in 11 mink brains, all of which were obtained from a single farm. Aleutian mink disease virus (AMDV) sequences for the capsid protein VP2 were obtained from brain samples from all affected farms. Although containing previously unreported amino acid residues, similarities with European and North American isolates were observed in the hypervariable regions within VP2, suggesting Irish AMDV is related to those isolates. The predicted amino acid residues, suspected of conferring pathogenicity at certain positions of the VP2 sequence, were present in the viral nucleic acid sequences.

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This study characterizes T- and B-lymphocyte responses in the peripheral blood and lesional skin of dogs with immunomodulatory-responsive lymphocytic-plasmacytic pododermatitis (ImR-LPP), a term previously proposed to denote a subpopulation of dogs with idiopathic pododermatitis. T-cell (CD3+, CD4+ and CD8+ ) and B-cell (CD21+) counts were significantly increased in both the epidermis and dermis of lesional ImR-LPP skin compared with that in pedal skin from healthy controls. CD3+ , CD4+, CD8+ and CD21+ cells were commonly observed in perivascular sites in the superficial dermis, periadnexally, beneath the dermal-epidermal (DE) junction and in the epidermis of lesional ImR-LPP skin. The CD8+ /CD3+ T-cell ratio in peripheral blood was significantly increased in the ImR-LPP group (0.42 versus 0.35 in controls). Serum IgA, IgG and IgM concentrations were all significantly elevated in affected dogs. Lymphocyte stimulation indices in ImR-LPP dogs were comparable with control levels except for a lower response to ionomycin (6.0 versus 11.1). Dogs with ImR-LPP had a higher incidence and mean (semi-quantitative) score for IgA, IgG and IgM deposits in the epidermis, and a significantly increased incidence of dermal IgA+, IgG+ and IgM+ mononuclear inflammatory cells. The results indicate that upregulated T- and B-lymphocyte responses may contribute to the pathogenesis of the skin lesions observed in dogs with ImR-LPP.

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The term immunomodulatory-responsive lymphocytic-plasmacytic pododermatitis (ImR-LPP) has previously been proposed to denote a sub-population of dogs with idiopathic pododermatitis. The objective of this study was to investigate dendritic cell (DC) and MHC class II antigen expression in lesional skin of dogs with ImR-LPP (n=47). Median epidermal CD1c(+) cell counts were 37.8 and 12.5 mm(-1) in ImR-LPP dogs and healthy controls (n=27), respectively (P<0.01), while the corresponding dermal cell counts were 180.9 and 45.0 mm(-2), respectively (P<0.01). Intra-epidermal clusters of DCs were observed in 18/47 dogs with ImR-LPP. Median epidermal MHC class II(+) cell counts were 32.5 and 10.5 mm(-1) in ImR-LPP dogs and healthy controls, respectively (P<0.01), while the corresponding dermal cell counts were 216.9 and 46.9 mm(-2), respectively (P<0.01). Dermal MHC class II(+) staining was primarily associated with DCs (47/47 dogs), mononuclear inflammatory cells (45/47), fibroblast-like cells (19/47) and vascular endothelium (14/47). The DC hyperplasia and increased MHC class II expression in lesional ImR-LPP skin are consistent with enhanced antigen presentation, and suggest that both parameters may contribute to the pathogenesis of ImR-LPP through the priming and activation of CD4(+) T cells. Equally, it is possible that the enhanced DC numbers observed in this study may contribute to the immunoregulation of steady-state pathology in lesional ImR-LPP skin through additional expanded, although as yet unresolved, mechanisms.

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Pododermatitis is a common inflammatory skin disease of dogs. As pedal lesions are reported in many canine dermatoses, a methodical series of diagnostic tests is required to establish the underlying aetiology. However, laboratory/ancillary investigations may prove unrewarding, prompting a diagnosis of idiopathic disease. Various hypotheses have been proposed to explain the pathogenesis of idiopathic pododermatitis including pedal conformation, trauma, immunosuppression, bacterial infection, furunculosis and dermal granuloma formation. Idiopathic pododermatitis accounts for 0.5% of all dermatology referrals to the authors' clinic. A sub-group within this population is characterised histopathologically by epidermal hyperplasia, hyperkeratosis, spongiosis, dermal oedema and perivascular aggregates of lymphocytes and plasma cells. The term lymphocytic-plasmacytic pododermatitis (LPP) has previously been proposed to reflect the histological appearance of such lesions. Affected dogs, although systemically well, characteristically have pruritus, erythema, swelling, pain and alopecia of the feet. Although non-responsive to antimicrobial therapy, antiparasitic agents and elimination diets, these dogs typically respond well to immunomodulatory therapy.

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Stomatitis in sheep caused by orf virus can be confused with lesions of more economically significant diseases, including foot-and-mouth disease, but there is no published account of the sequential development of oral orf lesions in the sheep. This report describes the clinical appearance of such lesions during a natural outbreak of the disease in young lambs. Lesions were seen on the gingiva, the tongue and the dental pad/hard palate, and progressed from small erythematous papules to larger, often coalescing papules that in some cases were ulcerated. Resolution started within seven days and was complete within 22 days. The lambs continued to suck and thrive throughout the infection. Lesions at all stages were proliferative, providing a major differentiating factor between orf and other causes of stomatitis in sheep.

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The term immunomodulatory-responsive lymphocytic-plasmacytic pododermatitis (ImR-LPP) has previously been proposed to denote a subpopulation of dogs with idiopathic pododermatitis. The objective of this study was to quantify the expression of mRNA encoding Th(1)-like [interferon (IFN)-gamma, interleukin (IL)-2 and IL-12], Th(2)-like [IL-4 and IL-6] and immunomodulatory cytokines [IL-10 and transforming growth factor (TGF)-beta] in lesional ImR-LPP, nonlesional ImR-LPP and healthy control pedal skin. Gene transcripts were quantified using TaqMan real-time reverse transcriptase-polymerase chain reaction assays. The skin of dogs with ImR-LPP had significant overexpression of IL-6 mRNA (P < 0.05) and significant underexpression of IL-12 mRNA (P < 0.01) compared to healthy controls. In addition, lesional ImR-LPP skin had significantly higher levels of IL-10 transcripts compared to healthy control pedal skin (P < 0.05). Although not attaining significance (P = 0.07), a trend towards reduced TGF-beta mRNA expression in lesional ImR-LPP skin was also evident. There were no significant differences in the levels of IFN-gamma or IL-2 mRNA transcripts among the three skin sample sources. IL-4 mRNA was detected in only one lesional sample. These results suggest that the pathogenesis of ImR-LPP may be associated with a T-cell-mediated inflammatory response characterized by impaired Th(1)-like, but enhanced Th(2)-like cytokine expression.

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Freshly excised rat, ovine and bovine ileal Peyer's patch (PP) and non-Peyer's patch tissues (NPP) were mounted in modified horizontal polyethylene diffusion chambers with a range of window areas. Rat tissue was initially used to establish that barrier function and histology were maintained for up to 60 min. Horse-radish peroxidase (HRP) fluxes and S. Typhimurium adherence and invasion were significantly higher in rat PP over NPP. Particle uptake was shown to be a rapid, energy-, time-, and size-dependent process, occurring more readily in PP than NPP tissue in each species. In a kinetic analysis, particles were localized initially in the follicle-associated epithelium and then in the dome region. For NPP uptake, particles were initially localized to villous epithelium, and were then detected in the crypts and lamina propria. Electrophysiological parameters including pharmacologically-stimulated inward short-circuit current responses were determined in isolated PP and NPP from each species mounted under identical conditions in Ussing chambers. In conclusion, comparative functional and histological characteristics of PP from several species were demonstrated in horizontal diffusion chambers. Horizontal diffusion chambers are therefore a useful in vitro model in which a range of functions including transport of particulate formulations by PP may be examined.

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A 6-year-old, intact female, Labrador Retriever/Terrier cross was presented to the University Veterinary Hospital, University College Dublin with a 3-week history of therapy-resistant cervical pain and intermittent fever. Physical examination findings included marked cervical pain resulting in neck extension and vocalization. Examination of the CSF revealed mild pleocytosis (total nucleated cells = 0.009 x 10(9)/L, reference interval <0.005 x 10(9)/L). Cytocentrifuged preparations of the CSF were of low cellularity, containing predominantly macrophages and occasional small lymphocytes. Several small- to medium-sized fragments of a slightly granular, amorphous, eosinophilic substance were observed. The majority of mononuclear cells were located within this material, in small groups of 3-13 cells. The amorphous foamy material stained positive with Luxol fast blue, suggestive of myelin-like material. The dog was euthanized and postmortem examination revealed intervertebral disk protrusion between C2 and C3. Hematoxylin- and Luxol fast blue-stained histopathologic sections of brain and spinal cord revealed only mild hemorrhage. The extracellular material in the CSF of this dog may have been caused by myelin degeneration or leakage of phospholipids from damaged cells. Because no histologic evidence of demyelination was observed with the disk extrusion, the myelin-like material in this case was thought to be the product of phospholipid breakdown from damaged cellular membranes. Three cases of dogs with spinal cord disease and myelin-like material in the CSF have been reported previously. The clinical significance of this finding is still unknown.

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: Infection with Angiostrongylus vasorum was diagnosed at necropsy on a dog that died from acute pulmonary haemorrhage, and on recovery of L1 larvae by Baermann examination of faeces from two dogs, one of which had abdominal pain and retroperitoneal haemorrhage, while the other had right-sided heart failure due to cor pulmonale. The presenting signs included syncope (one dog), exercise intolerance (two dogs), cough (two dogs), abdominal pain (one dog) and depression (one dog). One-stage prothrombin time and activated partial thromboplastin time were prolonged in two dogs, buccal mucosal bleeding time was prolonged in one dog and globulin was elevated in all three dogs. Two dogs were treated with fenbendazole and recovered.