The morphology and morphogenesis of jaagsiekte retrovirus (JSRV)

Jaagsiekte retrovirus ( JSRV ) was recently shown to be the aetiological agent of jaasiekte (ovine pulmonary adenomatosis). The morphogenesis of JSRV was studied in jaagsiekte tumour tissue. Intracytoplasmic particles, often associated with centrioles, were found in tumour cells. JSRV budded from tumour cells with a complete core which appeared to mature during the budding process. Extracellular particles were found in the alveolar lumen. Immature extracellular particles were rare. Mature extracellular JSRV was membrane-bound and had a slightly eccentric nucleoid with an electron-dense perinucleoidal space. In negatively stained preparations of JSRV the envelope was covered with spikes. JSRV is morphologically distinct from all known retroviruses.     

Enzootic nasal adenocarcinoma of sheep in Canada

A survey of veterinary diagnostic laboratories revealed that intranasal tumors occur in sheep in most provinces of Canada. Tumors were diagnosed in 44 sheep of several breeds including Polled Dorset, Suffolk, Cheviot, Rambouillet and various crossbreeds. Twenty-seven percent of tumors occurred in sheep that were less than two years old. Most tumors were sporadic but 33% of cases occurred in six related flocks, indicating that this disease can be an enzootic problem. The clinical signs were persistent serous, mucous or mucopurulent nasal discharge and stridor. Affected sheep progressively developed anorexia, dyspnea and mouth breathing and most died from effects of asphyxia and inanition within 90 days of the onset of clinical signs. Tumors originated unilaterally or occasionally bilaterally in the olfactory mucosa of the ethmoid turbinates. They were expansive and sometimes locally invasive but metastases were not found. Histologically, the tumors were classified as adenomas or, more frequently, adenocarcinomas. The etiology was not established but retrovirus like particles were observed in tumor tissue from one affected sheep.     

A stereological study on calculation of volume values regarding lumbosacral segments of quails

In the study, total segment, grey matter and white matter volume values of lumbosacral segment of spinal cord in quails (Coturnix coturnix japonica) were stereologically examined. Regardless of sex, six adult quails were used as material. After the materials were perfused, they were dissected and their spinal cords were uncovered. Tissue samples were taken from each lumbosacral segment of spinal cord. After the tissue fixation process, 6–7 sections having a 5‐µm thickness were separated in every 50th section as from the first section, where the tissue was seen, among tissue samples of each segment. These segments were stained with haematoxylin–eosin staining. They were photographed via a microscope. By using the Cavalieri's principle, the volume values of both the total segment, grey matter (GM) and white matter (WM) in each lumbosacral segment were separately calculated. All these calculations were performed by using SHTEREOM 1.5 program. As a result, total volume, grey matter and white matter volume values of lumbosacral segments and white matter/total volume, grey matter/total volume, and grey matter/white matter volume fractions were obtained.     

Detection of foreign DNA in transgenic chicken embryos using the polymerase chain reaction

Chicken primordial germ cells were infected with a defective retrovirus containing the Escherichia coli lacZ gene and injected into the heart of stage 15 embryos. DNA samples were isolated from various tissues of the injected embryos at different stages of development and were examined for the presence of the lacZ gene using the polymerase chain reaction. Integration of the retrovirus DNA was demonstrated with a 32P-labelled oligonucleotide in five-, 10- and 18-day embryos. This quick procedure provides an opportunity for the early detection of foreign DNA in small numbers of transfected cells and is a valuable tool in the detection of transgenic animals.     

Detection and quantitation of a type D retrovirus gag protein in ovine pulmonary carcinoma (sheep pulmonary adenomatosis) by means of a competition radioimmunoassay

A heterologous competition radioimmunoassay (RIA) which consisted of 125I-labeled langur retrovirus major gag protein and goat anti-squirrel monkey retrovirus serum was used to detect a type D retrovirus-associated antigen in tumor cell homogenates, lung fluid, and cell culture supernatant fluids of naturally occurring and experimentally-induced ovine pulmonary carcinoma (OPC, sheep pulmonary adenomatosis). In this assay, there was no cross reactivity between structural proteins of the type D retrovirus and an ovine lentivirus, which frequently co-infects OPC-affected sheep. The sensitivity of the assay was similar to an immunoblotting assay using antiserum to Mason-Pfizer monkey virus major gag protein which had been used previously to detect the OPC retrovirus antigen in tumor homogenates and lung fluids of OPC-affected sheep. All unconcentrated samples of lung fluid collected from five sheep with naturally occurring OPC or six sheep with experimentally induced OPC competed in the competition RIA. The competition RIA titers of the type D retrovirus antigen in lung fluids of lambs with induced OPC were relatively higher than the titers of this antigen in the naturally occurring OPC cases. The competition RIA detected the retrovirus antigen associated with OPC in the culture fluids of four out of five primary lung cultures from OPC sheep tested between 1 and 56 days after culture initiation. Because this RIA is appropriate for the quantitation of OPC-associated antigen, it will provide a means for determination of the target cell type for OPC virus replication in vitro.     

A rapid method for detecting Trichinella spiralis larvae in pork using a monoclonal antibody-latex conjugate

A simple, non-microscopic method was developed for the direct detection of Trichinella spiralis in pork. Samples of meat were treated with pepsin for 2 h to liberate the larvae, and sonicates were made from the deposit and used in an antigen detection system. In this system, latex particles, sensitized with a parasite-specific monoclonal antibody, were utilized in a slide or tube agglutination test. The efficiencies of these immunological methods were compared with that of trichinelloscopy in the examination of 102 pork specimens, 35 of which contained varying numbers of larvae after enzymatic digestion of 2 g of the meat. Trichinelloscopy detected 77.1% of these positive samples, but this was lower (55.5 and 41.7%) for those samples with a low larval burden (less than 50 or less than 10 larvae g-1 tissue, respectively). Better results (88.6, 76.5 and 60.0%) were obtained with the slide latex agglutination technique. The efficiency of detection was further improved (100%) using the more sensitive tube agglutination method. However, this was based on a small number (20) of samples examined and the limit of detection of this technique was subsequently found to be three larvae in total. No false-positives were seen with the latex agglutination methods from a total of 67 uninfected specimens examined.     

The demonstration by interference tests of an infective agent in fetuses from ewes inoculated with Border disease tissue.

Fetuses and placental tissues were taken from pregnant ewes at intervals varying between eight and 21 days after inoculation with tissue suspensions from cases of Border disease. Virus isolation procedures involving the detection of a cytopathic effect in tissue cultures with or without interference tests produced universally negative results but interference tests, using a plaque technique with the NADL strain of bovine virus diarrhoea virus as a challenge virus, detected the presence of an agent in tissues from six out of 10 fetuses. Inoculated ewes allowed to proceed to term showed a serological response characteristic of Border disease infection, as measured by four different tests. Although hairy shaker lambs were not seen, the occurence of abortion and stillbirth due to causes other than bacterial agents, was an indication that the Border disease agent was present. Electron microscopy of fetal fluids failed to detect viral particles.     

A quantitative study of the neural changes underlying pyloric stenosis in dogs

This study aimed to quantify the neural changes in congenital pyloric stenosis in dogs and to study the comparative anatomy between this condition in dogs and that in infantile hypertrophic pyloric stenosis. Eight specimens from the pylorus of dogs with pyloric stenosis and six control specimens were examined using conventional histology and immunohistochemistry for a range of neural antigens. The changes in the proportion of nerves immunoreactive for each antigen were quantified and analysed statistically. The morphology of the nerves in the diseased dogs was similar to that in controls. Only vasoactive intestinal peptide was reduced in expression in dogs (median proportion in control dogs 0.57, in diseased dogs 0.17; P = 0.065). This study demonstrates both morphological similarities and significant differences between closely related conditions in dogs, humans and other species.     

Astrovirus: a cause of an enteric disease in turkey poults

Virus particles of 30 nm diameter and star-shaped morphology were detected in intestinal contents of turkey poults and were identified as astroviruses. Seventy-six intestinal samples from 65 commercial turkey flocks between 6 and 35 days of age were evaluated for the presence of astroviruses by immune electron microscopy. Astroviruses were frequently detected in intestinal samples from poults that had enteritis and diarrhea of undetermined etiology. Astroviruses were geographically widespread and were present in poults from all six operations evaluated. Astroviruses were inoculated into specific-pathogen-free poults. Changes observed in the gastrointestinal tract were: dilatated ceca containing yellowish frothy contents, gaseous fluid in the intestinal tract, and loss of tone of the intestinal tract (gut thinness). Poults experimentally inoculated with astrovirus gained significantly less body weight and absorbed significantly less D-xylose than uninoculated controls.     

Necrosis of the nasal conchae in 12 horses

Twelve horses with a history of a unilateral malodorous nasal discharge were examined clinically, endoscopically and radiographically. The head of one of the horses was examined by computed tomography. Tissue samples were taken for microbiology from four of the horses and for histology from three. The conchal necrosis was unilateral in 11 of the horses and bilateral in the other, and no particular concha was affected more than the others. The affected tissue was removed transendoscopically with a polypectomy snare while the horses were sedated and standing. Mild disinfectants were used locally in six of the horses. After the removal of all the necrotic tissue, the affected concha healed without any complications in all 12 horses.     

Evidence for the expression and enzymatic activity of haem oxygenase-1 in the lungs of horses

The aim of this study was to characterise the expression of haem oxygenase-1 (HO-1) in healthy lung tissue from horses and to measure its activity. Samples of lung tissue were collected from six horses euthanased for reasons other than respiratory disease. HO-1 expression and activity were detected in type II alveolar epithelial cells, macrophages and neutrophils in all the samples examined. The activity was dependent on the presence of NADPH and inhibited quantitatively by the addition of increasing concentrations of a competitive inhibitor of HO-1, tin mesoporphyrin IX.     

Eradication of ovine pulmonary adenocarcinoma by motherless rearing of lambs

The principles of maedi-visna eradication programmes were applied to a field trial for the eradication of ovine pulmonary adenocarcinoma (OPA). In two maternal flocks the prevalence of gross and histological lesions in slaughtered animals was 18.3 per cent and 29.8 per cent, respectively. The lambing period was supervised for three consecutive years from 1999 to 2001, during which the lambs were taken away from their mothers at birth, deprived of maternal colostrum, and hand-reared away from other sheep. Over the three-year period, 322 hand-reared animals, mainly male lambs between 10 and 14 months old, were slaughtered; their lungs were examined grossly, 52.5 per cent of them were examined histologically, and 105 samples of caudal mediastinal lymph nodes were examined by PCR. No OPA tumours were detected in the slaughter specimens from the derived flock, but one lamb had histological lesions in one lung location; intrauterine transmission was ruled out in this case. No clinical OPA has subsequently been observed in the hand-reared flock. Bronchoalveolar lavage samples from the breeding stock were examined by PCR in order to rule out further subclinical cases of OPA. No Jaagsiekte retrovirus was detected in any of the 488 samples.     

Rumen papillomas in sheep

Out of a sample of 200 rumens from sheep slaughtered in Edinburgh, papillomas were found in 25. They occurred as fibro-papillomas, mostly along the pillar of muscle between the dorsal and ventral rumen, and were often multiple. No inclusion bodies were seen in the lesions or papilloma virus particles on electron microscopy. Homogenisation of papillomatous tissue followed by various methods of purification did not yield identifiable virus particles, and viral DNA was not detected. Immunoperoxidase staining showed a very small number of positive cells at or on the surface of 6 out of 10 lesions examined. Thus, it seems probable that virus particles are not found in large numbers in the rumen papillomas of sheep, unlike the situation in ovine skin warts, but are present in a few epithelial cells which are rapidly shed from the surface of the mucous membrane.     

Feline bronchial cytology: histologic/cytologic correlation in 22 cats

Trichrome-stained bronchial washings were obtained from 22 cats at necropsy and compared to lung tissue sections obtained at the same time. In three of the 22 cats, antemortem bronchial washings also were obtained. Sixteen of the cats were clinically ill and six were clinically normal and served as controls. On the basis of comparison of histologic sections and bronchial washings, patterns were detected in the washings indicative of general processes such as alveolar edema, chronic passive congestion, mucous metaplasia, and inflammation. Two specific etiologic agents, Histoplasma capsulatum and Toxoplasma gondii, each were seen in one washing.     

Caprine arthritis-encephalitis: clinical features and presence of antibody in selected goat populations

Features of caprine arthritis-encephalitis, a retrovirus disease of domestic goats, were studied in 60 goats over a 10-year period. The rate of progression and the severity of the disease process were highly variable within and among animals, but the most salient features were chronically swollen joints and bursae, lameness, weight loss, poor coat, mineralization of soft tissue, and death. Of 1,160 goat serum samples from 24 states tested by the immunodiffusion technique, 81% were positive for antibody to caprine arthritis-encephalitis virus antigens.     

A survey for torovirus in New Zealand cats with protruding nictitating membranes

Faecal samples from 51 cats were examined by negative stain electron microscopy to investigate a possible association between the syndrome of bilateral protrusion of the nictitating membranes and torovirus infection. Although toroviruses were not observed in any sample, other viruses or virus-like particles were observed in faeces from one of 15 cats with the syndrome, six of 21 cats which had experienced prior episodes of the syndrome and four of 15 healthy control cats. The findings of this study do not support the hypothesis that the syndrome is associated with torovirus infection.     

Effects of Ureaplasma diversum on bovine oviductal explants: quantitative measurement using a calmodulin assay.

Calmodulin (CAM) acts as an intracellular regulator of calcium, an important mediator of many cell processes. We used the CAM assay and electron microscopy to investigate the effects of Ureaplasma diversum on bovine oviductal explants obtained aseptically from slaughtered cows. A stock suspension of U. diversum (treated specimens) and sterile broth (controls) was added to replicates of cultured explants and incubated at 38 degrees C in an atmosphere of 5.5% CO2 for 48 hours. Explants were examined for ciliary activity, extracellular CAM loss, and for histological and ultrastructural changes. Explants and their culture media were examined for changes in CAM concentration. All experiments were replicated three times. In addition, U. diversum, medium and broth were assayed for CAM content. The concentrations of CAM in explants and media changed significantly (p < 0.05) in samples which were inoculated with U. diversum when compared to controls. The controls and infected specimens did not differ histologically or ultrastructurally, but U. diversum was seen to be closely associated with infected explant tissue. In view of this close affinity it is assumed the loss of CAM from the oviductal cells was causally related, but this was not proven. The failure to show cell membrane injury on light and electron microscopic examination was probably related to the short duration of the experiment and may only point out the sensitivity of the CAM assay in detecting early cell membrane injury. Compromise in characteristics of the medium to support both, the viability of oviductal cells and U. diversum limited the experimental time to 48 hours.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pheasant ataxia: a new condition in pheasant poults

Between 1995 and 1997 a neurological condition in pheasant poults from 24 sites in England and Scotland was investigated. Affected birds showed varying degrees of ataxia and incoordinated movements and, in severe cases, recumbency, but generally remained alert with their heads held upright. The condition characteristically affected poults from seven weeks of age and the incidence on any one site was low. No significant bacteria were isolated consistently from brain tissue. The condition was characterised histologically by a non-suppurative meningoencephalitis, in which lesions were found predominantly in the cerebellum in 61 of 81 samples examined (75.3 per cent). A non-suppurative myelitis was recorded in 16 of 20 spinal cords examined. No lesions were recorded in peripheral neural tissue and lesions were rare in other tissues. The condition appeared not to have been recorded previously in pheasants. A viral aetiology was suspected but Newcastle disease virus was not involved.