The induction of a protective immunity against Jembrana disease in cattle by vaccination with inactivated tissue-derived virus antigens

The ability to induce a protective immunity against Jembrana disease, an acute lentivirus disease of Bali cattle (Bos javanicus) present in Indonesia, was investigated. A protective immune response was induced in cattle by vaccination with virus-containing plasma and spleen tissue derived from acutely affected cattle. The virus was inactivated with Triton X-100 and emulsified in either incomplete Freund's adjuvant or a mineral oil adjuvant (MOA). The vaccination procedure suppressed the duration and severity of the disease but did not completely prevent the development of disease in animals challenged with 100 infectious doses of virus.     

Evidence for immunosuppression associated with Jembrana disease virus infection of cattle.

Jembrana disease virus (JDV) is a newly recognised bovine lentivirus causing an acute disease syndrome in Bali cattle (Bos javanicus) in Indonesia. We evaluated the effect of JDV infection on the antibody response to chicken ovalbumin (cOVA) and Brucella abortus Strain 19 in Bali cattle. In infected cattle the IgG and IgM response to cOVA was suppressed and delayed and the IgG response to B. abortus Strain 19 was delayed. The results indicate that the humoral immune response is suppressed and delayed in JDV infected cattle.     

Evidence for a viral aetiology of Jembrana disease in Bali cattle.

Jembrana disease is an acute infectious disease of Bali cattle (Bos javanicus) in Indonesia. High titres of the infectious agent are present in plasma during the febrile phase of the disease. The size of the agent determined by membrane filtration was between 50 and 100 nm, indicating it is a virus and not a rickettsia as previously proposed. Spherical virus-like particles of 75 to 130 nm diameter with a smooth membrane and frequently with an eccentric nucleoid were detected by electron microscopy in plasma from infected animals. The virus replicated in mononuclear cell cultures of peripheral blood origin but not in other cell cultures. The virus and the associated disease have characteristics consistent with viruses in the family Retroviridae.     

Scanning electron microscopy of vascular corrosion casts and histologic examination of pulmonary microvasculature in dogs with dirofilariosis

OBJECTIVE: To characterize structural changes in pulmonary vessels of dogs with dirofilariosis. ANIMALS: 8 dogs with dirofilariosis and 2 unaffected control dogs. PROCEDURE: Pulmonary artery pressure was measured in affected dogs, and dogs then were euthanatized. Scanning electron microscopy was used to examine vascular corrosion casts of pulmonary vasculature. Tissue sections of pulmonary vasculature were evaluated by use of histologic examination. RESULTS: Pulmonary artery pressure was higher in dogs with severely affected pulmonary vessels. In tissue sections, dilatation, as well as lesions in the tunica intima and proliferative lesions resulting in constriction or obstruction, were frequently observed in branches of the pulmonary artery. Numerous dilated bronchial arteries were observed around affected pulmonary arteries. Hyperplastic venous sphincters were observed in small pulmonary veins and venules. In corrosion casts, affected pulmonary lobar arteries had dilatation, pruning, abnormal tapering, constriction, and obstruction. In small arteries and arterioles, surface structures representing aneurisms and edema were seen. Bronchial arteries were well developed and extremely dilated, and they formed numerous anastomoses with pulmonary arteries at all levels, from the pulmonary trunk to peripheral vessels. Capillaries in the lungs were dilated with little structural change. Small pulmonary veins and venules had irregular annular constrictions that were caused by hyperplastic smooth muscle cells of venous sphincters. CONCLUSIONS AND CLINICAL RELEVANCE: Scanning electron microscopy of microvascular casts delineated links between the bronchial and pulmonary circulations in dogs with dirofilariosis. Results of scanning electron microscopy provided a structural explanation for the development of pulmonary circulatory disturbances and pulmonary hypertension in dogs affected by dirofilariosis.     

RADIOGRAPHY REVIEW: THE VASCULAR AND BRONCHIAL PATTERNS OF PULMONARY DISEASE

Pulmonary blood vessels account for the majority of radiographically visible structures in normal lung. The vascular pattern of pulmonary disease is characterized by changes in vessel size, shape, contour. Changes in size can be classified as hypo vasularity (decreased size) or hyper vascularity (increased size). The pulmonary arteries and veins may be affected separately or simultaneously. In normal lung, the bronchial structures peripheral to the hilar area are not seen clearly. The bronchial pattern of pulmonary disease is characterized by alteractions in bronchial wall thickness and dinsity, or in bronchial lumen diameter. Mixtures of the four lung patterns, i.e. , alveolar, interstitial, bronchial, and vascular, occur frequently. The lung radiographic pattern in an individual animal depends not only on the disease process present, but also on the stage of the disease during which they are radiographed.     

Microfilarial granulomas in the spleens of wild-caught cynomolgus monkeys (Macaca fascicularis)

Splenic nodules from 38 cynomolgus monkeys (Macaca fascicularis) which were captured in Malaysia and Indonesia were studied histologically. The lesions were characterized by well-circumscribed focal fibrosis, accumulation of eosinophils and histiocytes, hemorrhage or hemosiderosis, and loss of normal splenic architecture. Small arteries in the lesion frequently had intimal thickening and narrowing of the lumen in addition to the presence of microfilariae. Microfilariae were also seen in the extravascular area of the lesion, and were occasionally engulfed by multinucleated giant cells. The splenic lesion was thought to have been initiated by incomplete infarction caused by intimal thickening and microfilarial occupation of the small arteries.     

Flow cytometric analysis of lymphocyte subset kinetics in Bali cattle experimentally infected with Jembrana disease virus

Jembrana disease virus (JDV) is an unusual bovine lentivirus that causes an acute and sometimes fatal disease after a short incubation period in Bali cattle (Bos javanicus). The pathological changes occur primarily in lymphoid tissues, which feature proliferating lymphoblastoid-like cells predominantly throughout parafollicular (T-cell) areas, and atrophy of follicles (B-cell) areas. Five Bali cattle were experimentally infected with JDV and all developed typical clinical signs of Jembrana disease characterised by a transient febrile response, enlargement of superficial lymph nodes and a significant leukopenia. Flow cytometric analysis of PBMC during the acute (febrile) disease phase showed that the reduced number of lymphocytes was due to a significant decrease in both the proportion and absolute numbers of CD4(+) T cells, but not CD8(+) T-cells or CD21(+) B-cells. At the end of the febrile phase, total numbers of both CD8(+) T-cells and CD21(+) B-cells increased significantly, while CD4(+) T-cell numbers remained below normal values, resulting in a significantly reduced CD4(+):CD8(+) ratio. We speculate that the persistent depletion of CD4(+) T cells following JDV infection, through lack of CD4(+) T cell help to B cells, may explain the lack of production of JDV-specific antibodies for several weeks after recovery despite an increase in CD21(+) B cell numbers. Further, our previous data showing that IgG(+) plasma cells are targets for JDV infection, correlated with our current data demonstrating an increase in CD8(+) T cell numbers, supports the suggestion that anti-viral cytotoxic T cell or other cell-mediated immune responses may be critical in the recovery process, although this remains to be formally demonstrated for JDV.     

IBARAKI DISEASE AND ITS RELATIONSHIP TO BLUETONGUE

Ibaraki disease, an epizootic disease of cattle in Japan resembling bluetongue, is characterized by fever and lesions affecting the mucous membranes, the skin, the musculature and vascular system. Degeneration of striated muscular tissue is observed in the oesophagus, larynx, pharynx, tongue and the skeletal muscles. Oedema and haemorrhage are marked in the mouth, lips, abomasum, around the coronets, etc., and are occasionally followed by degeneration of the epithelium leading to erosions or ulcerations. Severe lesions affecting the oesophageal and laryngopharyngeal musculature cause difficulty in swallowing which in turn produces dehydration and emaciation, and occasionally the aspiration pneumonia, which constitute the major causes of death of affected animals. These clinical and pathological findings indicate the similarity of the disease to bluetongue in sheep and cattle. Ibaraki disease was first recognised in Japan in 1959 and 1960. Seasonally its occurrence is limited to late summer and autumn, and geographically to the central and western parts of Japan, roughly south of 37 degrees north latitude. It is absent from the higher altitudes. The seasonal and geographical incidence suggests the possibility of an arthropod vector; but direct evidence for such a vector is still lacking. Serological data suggest the presence of Ibaraki virus on Bali Island in Indonesia and in Taiwan. The disease can be transmitted serially in calves by the intravenous inoculation of blood obtained at the height of a febrile reaction. Ibaraki virus can be isolated in bovine cell cultures from both natural and experimentally produced cases of the disease. The virus multiplies and induces cytopathic effects in primary cultures of bovine, sheep and hamster lung origin, and L cells; but it does not grow in primary cultures of horse and swine kidney nor in HeLa cell cultures. The virus is readily passaged serially in 4 to 5-day-old eggs by yolk-sac inoculation and incubation at 33.5 degrees C. It multiplies in the brains of mice of any age after incracerebral inoculation but younger mice give a better viral growth and develop encephalitis. No evidence has been obtained that rabbits and guinea pigs are susceptible to Ibaraki virus...     

Seroprevalence of antibodies to <Emphasis Type="Italic">Neospora caninum</Emphasis> in <Emphasis Type="Italic">Bos javanicus</Emphasis> (‘Bali cattle’) from Indonesia

A cross-sectional survey was performed to obtain first information on the presence of Neospora caninum infection in Bos javanicus (‘Bali cattle’), the predominant beef cattle in the Eastern Islands of Indonesia. Serum samples were collected from 438 Bali cattle of two age classes (<2 years, >2 years) and both genders at three slaughterhouses in the Bali island, and examined for N. caninum-specific antibodies using native NcSRS2 (p38 antigen) as an ELISA antigen. The estimated overall seroprevalence of antibodies was 5.5% (95% CI: 3.5–8.0%). The seroprevalence was not significantly associated with age class or gender of the animals. The results give first serological evidence for the presence of natural N. caninum infection in Bos javanicus and indicate its occurrence in Indonesia.     

Histologic examination of selected areas of canine pulmonary arteries

Selected areas of pulmonary arteries from 18 healthy mixed-breed dogs were examined using histologic staining techniques. Smooth muscle cell, collagen, and elastin content of the tunica intima and tunica media were assessed. Fifteen dogs had abnormalities of tunica intima or tunica media in at least one arterial section examined. Of all arterial sections examined, 40% had histologic changes of the tunica intima or tunica media, and 42% of these vascular lesions were in the main pulmonary artery. The most commonly occurring pathologic change was loss of smooth muscle cells and elastin of the tunica media and replacement by collagen. This lesion is similar to cystic medionecrosis of the aorta. Seemingly, a high frequency of spontaneous vascular lesions exist in pulmonary arteries of young dogs.     

Comparative investigation of tissue alterations and distribution of BVD-viral antigen in cattle with early onset versus late onset mucosal disease

Tissue alterations and distribution of BVDV antigen were examined in nine cattle with early onset and five cattle with late onset mucosal disease (MD) to evaluate the possibility to differentiate between the two disease entities. MD was induced by inoculation of persistently viremic cattle with different strains of cytopathogenic BVDV. Animals which developed early onset MD became moribund approximately 2 weeks post-inoculation (pi); animals with late onset MD 42-115 days pi. All animals were euthanized and necropsied when moribund. Macroscopic lesions were found in the upper and lower digestive tract of cattle with early and late onset MD. In cattle with late onset MD, lesions in the oral cavity were generally milder and in the intestinal tract they were not only associated with GALT, but frequently affected the mucosa outside. Histologically, the abrupt changes between hyperplastic and atrophic areas of mucosa were striking in the cattle with late onset MD. This corresponded with the multifocal distribution of areas of mucosa in which intense staining for BVD-virus antigen could be demonstrated. In both courses of MD, a severe depletion of Peyer's patches was noted, but only in late onset MD, there was a complete loss of architecture. The most distinctive difference was the presence of vascular lesions which were observed in all five cattle with late onset MD, but in none of the animals with early onset MD. The vasculopathy was characterized by segmental necrosis of vascular walls and lymphohistiocytic perivasculitis in arterioles and small arteries in the submucosa of the intestine.     

Development of disabled, replication-defective gene transfer vectors from the Jembrana disease virus, a new infectious agent of cattle

Jembrana disease virus (JDV) is a newly isolated and characterised bovine lentivirus. It causes an acute disease in Bali cattle (Bos javanicus), which can be readily transmitted to susceptible cattle with 17% mortality. There is as yet no treatment or preventive vaccine. We have developed a gene transfer vector system based on JDV that has three components. The first of the components is a bicistronic transfer vector plasmid that was constructed to contain cis-sequences from the JDV genome, including 5'- and 3'-long terminal repeats (LTRs), 0.4kb of truncated gag and 1.1kb of 3'-env, a multiple cloning site to accommodate the gene(s) of interest for transfer, and an internal ribosome entry site plus the neomycin phosphotransferase (Neo) gene cassette for antibiotic selection. The second element is a packaging plasmid that contains trans-sequences, including gag, pol, vif, tat and rev, but without the env and packaging signals. The third is a plasmid encoding the G glycoprotein of vesicular stomatitis virus (VSV-G) to supply the vector an envelope for pseudotyping. Cotransfection of 293T cells with these three plasmid components produced VSV-G pseudotyped, disabled, replication defective, bicistronic JDV vectors encoding the green fluorescent protein (EGFP) and the Neo resistance selection maker simultaneously with a titre range of (0.4-1.2)x10(6)CFU/ml. Transduction of several replicating primary and transformed cells from cattle, primate and human sources and importantly growth-arrested cells with the JDV vectors showed high efficiency of EGFP gene transfer at 35-75%, which was stable and the expression of EGFP was long term. Furthermore, these JDV vectors were designed to suit the inclusion and expression of genes corresponding to JDV specific proteins, such as gag or env, for the development of vaccines for Jembrana disease. This strategy should also be applicable to other bovine diseases as well. The design and construction of the JDV vector system should facilitate the study of the lentivirology and pathogenesis of the diseases associated with JDV or other bovine virus infections. To our knowledge, this is the first such vector system developed from a cattle virus.     

Localization of potential binding sites for the edema disease verotoxin (VT2e) in pigs.

The purpose of this study was to identify organs and cells to which the edema disease verotoxin (VT2e) could bind in pigs. Frozen 4-5 microns thick sections of organs usually affected in edema disease (colon, spinal cord, cerebellum and eyelid) and organs not usually affected (liver, ileum) from two 5- to 6-week-old weaned pigs were permeabilized with acetone, then exposed to VT2e. Unbound VT2e was removed by washing and bound VT2e was detected by immunohistochemistry. In the eyelid, double-label immunofluorescence was used to identify the cells to which VT2e bound. VT2e was shown to bind to all six organs that were examined. The toxin bound to arteries in all organs, to veins in all organs except the liver, and to enterocytes in the ileal crypts. Double labelling of eyelid with monoclonal antibodies specific for von Willebrand factor or alpha-smooth actin and VT2e showed that the toxin bound to endothelial and vascular smooth muscle cells. The binding of VT2e to endothelium is consistent with findings for other verotoxins but binding to vascular smooth muscle has not been reported for other verotoxins. It is concluded that i) factors other than the presence of receptors for VT2e influence the development of lesions in edema disease, and ii) smooth muscle necrosis, which is characteristic of the vascular lesions in edema disease, may be due to a direct action of toxin on smooth muscle cells.     

Indigenous Bali cattle is most suitable for sustainable small farming in Indonesia

Livestock husbandry is essential for Indonesia. This study reviews cattle characteristics and husbandry methods in the country with special interest in describing the importance of indigenous breeds of cattle. As a conclusion, the Bali cattle ought to be considered the most suitable indigenous cattle breed for the low-input, high stress production system still practised by millions of families in Indonesia.     

High incidence of polyarteritis nodosa in the brains of culled sows

A total of 307 brains of purebred sows obtained from an abattoir were retrospectively examined. These sows were culled with reasons of reproductive failure, urogenital infections, or locomotor problems. The most common macroscopic lesions were cavitations or lacunae in the basal nuclei (9.1%, 28/307) and coarse and thickened leptomeninges with marked vessels (3.9%, 12/307). The most frequent microscopic lesion was polyarteritis nodosa (21.2%, 65/307), which was found in all 40 brains with the above-mentioned gross lesions and in all 25 brains with microscopic cerebral infarcts or cavitations. The affected arteries of polyarteritis nodosa were distributed primarily in the cerebral leptomeninges, basal nuclei, and internal and external capsules. Histopathologically, a characteristic change of the affected arteries was transmural fibrinoid necrosis with severe infiltration of mixed inflammatory cells; narrowing or occlusion of the lumen. The inflammatory cells were chiefly composed of lymphocytes, macrophages, and plasma cells, with a few eosinophils and occasional multinucleated giant cells. Polyarteritis nodosa was found at a high percentage in the brains from culled sows. It may result in cerebral ischemia, infarcts, and hemorrhage, and possibly play a role in the necessity for culling due to locomotor problems.     

Equine orbital angiomatosis

Angiomatosis refers to a vascular tumour consisting of several vascular, blood‐filled cavities with a lining of well‐differentiated endothelial cells. This condition is seen in very young cattle as a malformative process and has been documented in other animals as well. In horses, this disease has only rarely been described, with no reported lesions being found within the equine orbit. This report details the clinical findings, surgical exploration, histopathology, and long‐term follow‐up of an aged horse with orbital angiomatosis.     

Note on Onchocerca armillata in the Sudanese camel (C. dromedarius). A histological and anatomo-pathological approach

Aortic onchocercosis due to O. armillata was diagnosed in 45 (41%) out of 109 Sudanese camels. Although the thoracic aorta was regularly affected, involvement of other vessels such as the abdominal aorta, brachiocephalic, pulmonary and costocervical arteries was not excluded. Gross examination of the affected blood vessels revealed tortuous tunnels of parasitic tracks readily visible in the intimal surface of the vessels. Nodular lesions were more common on the adventitia. Microscopically, the lesions showed irregulary elevated intimal surfaces. The underlying tunica media contained a varying number of encapsulated and partially mineralized parasitic fragments. The inflammatory response varied in intensity and location and consisted of eosinophils, lymphocytes, macrophages and occasional giant cells.     

ANGIOGRAPHIC EVALUATION OF BOVINE FOOT ABNORMALITIES

Angiography of normal and diseased feet in cattle was carried out. In osteomyelitis of distal phalanx there was a narrowing of the digital artery with diminished vascular supply to the affected digit. An area of bone necrosis was found to be avascular. With overgrown hooves, narrowing of both the medial and lateral digital arteries with ill-defined laminar vessels was seen. The vascular pattern in cattle with cork screw claws was characterized by a dilated tortuous digital artery and a crowded arterial network resulting in increased vascular supply in the affected claw. Abrupt stricture of common digital artery, a crowded vascular network in the area of the middle phalanx and filling defects of terminal arches were demonstrated in animals with exostosis     

Sheep pox: experimental studies with a west african isolate

Under conditions of a maximum security laboratory, four cross-bred sheep were inoculated intradermally only or intradermally and intratracheally with a West African isolate of sheep pox virus. All sheep had increased temperature and depression by the fourth or fifth day after infection. Nasal and lacrimal discharge and coughing occurred in all sheep but were more severe in sheep receiving the virus via the tracheal route. From the fifth day after infection, numerous papular erythematous skin lesions developed at the inoculation sites. These were 3-7 mm in diameter and gradually became nodular. Some of these lesions healed and others coalesced to form tumorlike masses. In one sheep, euthanized 14 days after intradermal and intratracheal inoculation, nodular lesions were found in the skin around the eyes, nostrils, oral and perianal regions, the mucosa of the rumen and throughout the lungs. Histologically, skin nodules were characterized by ischemic necrosis, vasculitis, microvesicualtion, eosinophilic cytoplasmic inclusions in the dermal epithelial cells and vacuolar nuclear degeneration. The pulmonary lesion was that of proliferative alveolitis with occasional cytoplasmic inclusions in the alveolar cells and macrophages. Ultrastructurally, large cuboidal virus particles were found both in the skin lesion and inoculated tissue cultures. The sheep pox virus structure was easily distinguished from contagious ecthyma virus, a parapoxvirus which causes sporadic disease in Canada. Serum neutralizing antibodies developed in all the sheep by 14 days postinfection.

The clinical and pathological characteristics of experimental sheep pox produced with this West African isolate were similar to those caused by Neethling virus of lumpy skin disease in cattle.