The production of peste des petits ruminants hyperimmune sera in rabbits and their application in virus diagnosis

Hyperimmune sera were produced by serial inoculation of rabbits with Vero cell-adapted, sucrose gradient-purified Nigerian peste des petits ruminants virus (PPRV) isolate. Two antisera produced, neutralized the homologous PPRV but not the heterologous rinderpest Kabette "O" virus. The antisera gave strong precipitin lines with purified PPRV antigens and were used to detect PPRV and rinderpest virus antigens from ante-mortem secretions and post-mortem tissue homogenates from PPR and rinderpest virus infected goats and cattle by the agar gel precipitation tests (AGPT). The hyperimmune sera gave good titration curves with both purified Nigerian goat and the United Arab Emirate wildlife PPRV isolates in the indirect enzyme linked immunosorbent assay (ELISA). Results of indirect ELISA showed that although there were some cross reactions with the rinderpest, canine-distemper and measles viruses, at 1:100 dilution, the antisera would give a positive signal with only the homologous PPR virus.     

Improved isolation of rinderpest virus in transformed bovine T lymphoblast cell lines.

Bovine T lymphoblast cell lines transformed by the protozoan Theileria parva were compared with bovine kidney (BK) and Vero cells for their ability to isolate various strains of rinderpest virus from tissues and infected secretions. All of the strains of rinderpest virus that were tested, including attenuated cell-culture, caprinised and lapinised vaccines, and both mild and virulent pathogenic strains, readily induced syncytial cytopathic effect (cpe) in T lymphoblasts. The cpe could often be detected within one day of inoculation of lymphoblasts, whereas it took three to 14 days to appear in Vero and BK cells. Using lymphoblasts it was possible to reisolate rinderpest virus from nine of 42 swabs collected from three cattle experimentally infected with an isolate from a recent outbreak of mild disease whereas the same swabs yielded only one reisolate on BK cells. It was also possible using the lymphoblasts to detect infectious virus in the ocular, nasal and oral secretions of goats and rabbits infected with caprinised and lapinised virus, respectively. Peste des petits ruminants virus appeared to grow as rapidly as rinderpest virus in the lymphoblasts whereas canine distemper virus readily induced cpe on first passage but less readily on subsequent passage. Measles virus induced relatively little cpe when inoculated into lymphoblasts and did not appear to passage in these cells. The lymphoblasts are easy to maintain in culture and since they rapidly recovered 11 isolates from 37 diagnostic samples could prove useful in laboratories carrying out rinderpest diagnosis.     

Preliminary observations on rinderpest in pregnant cattle

A Kabete 'O' strain of rinderpest virus enhanced in virulence was inoculated subcutaneously into four cows which were between six and eight months pregnant. All the cows developed clinical signs of rinderpest from the third day after inoculation and shed high titres of virus in their ocular and vaginal secretions during the course of the clinical disease. Three of the cows died of rinderpest on the third day after the onset of fever but no virus was isolated from their fetuses which were examined post mortem. The fourth cow showed complete clinical and virological recovery by the eighth day after the onset of fever and aborted an eight-and-a-half-month-old fetus on the 12th day after it recovered. Rinderpest virus was demonstrated in a wide range of the aborted fetal tissues. Virus was also detected in the maternal vaginal discharges up to 24 hours after abortion. The only gross pathological change observed was a severe necrotising placentitis.     

Detection of rinderpest virus antigens in vitro and in vivo by direct immunofluorescence

Cell-culture attenuated and virulent strains of rinderpest virus (RV) were inoculated on to bovine kidney cell cultures. A direct immunofluorescent antibody test detected RV antigens in cell cultures within one to three days after inoculation whereas RV cytopathic effects usually took three to nine days to develop. Cells containing RV antigens were also detected in impression smears and frozen sections of tissues collected from RV infected animals at post mortem examination, and in smears of lymph node biopsies taken from cattle with clinical rinderpest. These techniques may offer additional methods for rapid diagnosis of rinderpest.     

Rinderpest Antibody Detected in Sheep and Goats Before an Outbreak of Rinderpest Reported in Cattle in Northern Tanzania

In January 1997, serum samples from 1346 adult sheep and goats were tested by a competitive ELISA to determine the prevalence of rinderpest in the northern zone of Tanzania. Seroconversion rates of 20%, 13%, 9%, 7% and 3% in sheep and goats were recorded in Ngorongoro, Monduli, Hai, Arumeru and Simanjiro districts, respectively. The low profile and insidious nature of the rinderpest virus involved caused very mild disease in cattle in some of these area. The mild signs associated with this outbreak of rinderpest resulted in difficulty in its diagnosis. In these circumstances, the presence of rinderpest antibody in sheep and goats served as a valuable and effective indicator of the rinderpest outbreak in cattle.     

Development and stability of rinderpest virus antigens in cattle tears and lymph nodes

Summary Diffusible rinderpest virus antigens were demonstrated in increasing quantities in ocular and lymph node biopsies from rinderpest-infected cattle using agar gel immunodiffusion (AGID) and counterimmunoelectrophoresis (CIEP) tests. Positive samples were detected from the second day of pyrexia to two days after death. The antigens in ocular secretions and lymph nodes were thermolabile being destroyed within five minutes at 56°C and within two weeks at 4°C.

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Resumen Se demostró un incremento de antígenos difusibles del virus de rinderpest, en biopsias oculares y linfáticas tomadas de ganado inoculado experimentalmente. Las pruebas utilizadas fueron la inmunodifusión en agar gelatina y la contrainmunoelectroforesis. Se detectaron muestras positivas desde el segundo día de pirexia, hasta dos días antes de la muerte. Los antígenos en las secreciones oculares y nódulos linfáticos fueron termolábiles, siendo destruidos a los cinco minutos a 56°C y en dos semanas a 4°C.

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Résumé Les tests d’immuno-diffusion en gélose (IDG) et de contre-immunoélectrophorèse (CIE) ont permis de mettre en évidence des quantités croissantes d’antigènes diffusibles du virus de la peste bovine dans les prélèvements de larmes et de ganglions lymphatiques. Des résultats positifs ont été enregistrés à partir du deuxième jour de la fièvre jusqu’au 2e jour après la mort. Les antigènes des secrétions oculaires et des ganglions lymphatiques sont thermolabiles: destruction en cinq minutes à 56°C et en deux semaines à 4°C.

     

Serological studies with peste des petits ruminants and rinderpest viruses in Nigeria

One hundred and ninety-five goat and 67 sheep sera collected from various parts of southern Nigeria were screened for neutralising antibodies to both the peste des petits ruminants (PPR) and rinderpest viruses. Neutralising antibodies against both viruses were found in the sheep and goat sera examined. Parallel titration of samples which neutralised both viruses indicated a primary infection with the PPR virus (PPRV). However, some samples which failed to neutralise PPRV neutralised the rinderpest virus (RV) indicating RV activity in sheep and goats in Nigeria. These findings are discussed in relation to the diagnosis of PPRV infection and the recent reappearance of bovine rinderpest in Nigeria.     

Serological studies with the virus of peste des petits ruminants in Nigeria.

It was possible to distinguish separate serological responses when experimental goats were inoculated with either rinderpest virus or peste des petits ruminants virus. Examination of field samples established that peste des petits ruminants occur commonly in Nigerian sheep and goats although some villages have escaped recent infection. There was no evidence of infection with rinderpest in these animals.     

The detection of antibody against peste des petits ruminants virus in Sheep,Goats, Cattle and Buffaloes

Monoclonal antibody-based competitive ELISA (C-ELISA) has been used for the specific measurement of antibodies to peste des petits ruminants (PPR) viruses in sheep, goats, cattle and Buffalo. Serum samples from sheep (n = 232), goats (n = 428), cattle (n = 43), buffalo (n = 89) were tested. The animals had not been vaccinated against rinderpest or PPR. Findings suggested that the sero-positive cases were significantly higher in sheep (51.29%) than in goats (39.02%) (P = 0.002). The overall sero-prevalence of PPRV in small ruminants was 43.33%. The PPR antibodies seroprevalence was 67.42% in buffalo and 41.86% in cattle which was significantly higher in buffalo (P = 0.005). The overall sero-prevalence of PPRV in large ruminants was 59.09%. Cattle and buffalo sera showed a high prevalence of antibody against PPR virus which may explain the difficulty experienced in achieving high post-vaccination immunity levels against rinderpest. Because antibodies against PPR virus are both cross-neutralizing and cross-protective against rinderpest virus, further vaccination in the presence of antibodies against PPR virus may be a waste of national resources. It was also suggested that antibodies to PPR virus could prevent an immune response to the rinderpest vaccine. This paper presents serological evidence for the transmission of PPR virus from sheep and goats to cattle and buffalo and highlights the need to include PPR serology in the sero-monitoring programme to give a better indication of national herd immunity of sheep and goats against PPR.     

Antibody seroprevalences against peste des petits ruminants (PPR) virus in camels, cattle, goats and sheep in Ethiopia

A questionnaire-survey data indicated that 26% of 276 farmers reported the presence of respiratory disease in their herds in 2001. The incidence was perceived as "high" in small ruminants and camels, but as "low" in cattle. Simultaneously, 2815 serum samples from camels (n=628), cattle (n=910), goats (n=442) and sheep (n=835) were tested. The peste des petits ruminants (PPR) antibody seroprevalence was 3% in camels, 9% in cattle, 9% in goats and 13% in sheep. The highest locality-specific seroprevalences were: camels 10%, cattle 16%, goats 22% and sheep 23%. The animals had not been vaccinated against rinderpest or PPR. Antibody seroprevalences detected in camels, cattle, goats and sheep confirmed natural transmission of PPR virus under field conditions.     

Experimental infection of equine herpesvirus 9 in dogs

Equine herpesvirus 9 (EHV-9), a new neurotropic equine herpesvirus, was inoculated intranasally at 107 plaque-forming units in five dogs to assess its pathogenicity. Dogs showed weight loss, pyrexia, anorexia, and neurologic signs on the fourth day. The EHV-9 virus was recovered from the examined brains. Histologically, dogs had a fulminant nonsuppurative encephalitis characterized by severe neuronal degeneration and loss, with intranuclear inclusions, slight glial reactions, perivascular cuffing, and multifocal hemorrhage. The olfactory bulb and the frontal and temporal lobes were predominantly affected. Immunohistochemistry revealed reactivity for EHV-9 antigen in neurons. All dogs had mild bronchopneumonia and various degrees of lymphoid necrosis. These findings indicate that dogs are fully susceptible to EHV-9 and that EHV-9 can cause fulminant encephalitis with high mortality in dogs, as in gazelles and goats.     

Evidence of identification of peste des petits ruminants from goats in Egypt

Peste des petits ruminants virus was isolated from young goats in an outbreak of the disease for the first time in Egypt. Affected goats showed symptoms simulating rinderpest, disease in cattle. The mortality rate was about 30%, and morbidity reached 90%. History, clinical symptoms, postmortem lesions, and diagnosis were discussed.     

Use of the caprinised strain of rinderpest virus for potency testing an attenuated cell culture rinderpest vaccine

The caprinised strain of rinderpest virus was inoculated into goats to produce a challenge stock. These goats were kept with control animals (goats, sheep, calves). In this trial the caprinised strain was shown to have a mild pathogenicity for goats and it spread to one of two contact goats but not from goats to other species. The caprinised strain was then tested on cattle where a febrile reaction was observed. The caprinised strain also did not spread between cattle. The cattle vaccinated with a freeze-dried vaccine produced from the attenuated Kabete RBKO strain on bovine kidney cells were then challenged with the caprinised strain with good results.     

Reaction of goats to infection with infectious bovine rhinotracheitis virus

Intranasal exposure of goats to infectious bovine rhinotracheitis virus resulted in mild respiratory disease and virus reisolation from nasal secretions. No disease was produced in goats exposed to the same virus by the genital or ocular routes. There was serological evidence of contact transmission of infection from infected goats to cattle. Virus recrudescence was not detected in goats treated with dexamethasone two months after virus inoculation.     

Etiology of the stomatitis pneumoenteritis complex in Nigerian dwarf goats.

The causative agent of stomatitis pneumoenteritis complex was isolated in domesticated goats and Vero cell culture. It was identified immunologically and morphologically as identical with the "Peste des Petits Ruminants" virus. There were cross reactions between stomatitis pneumoenteritis complex virus isolate and rinderpest virus by immunodiffusion and complement fixation tests but no cross neutralization. Goats recovered from stomatitis pneumoenteritis complex were protected against a challenge with rinderpest virus that was lethal to control goats. Ultrastructural morphology revealed intracytoplasmic and intranuclear inclusions made up of random arrays of fibrillar strands. Pleomorphic particles budded from the plasma membrane of infected cells and enveloped virions were seen extracellularly. Specific ferritin tagging was demonstrated in the stomatitis pneumoenteritis complex virus infected cells treated with homologous and peste des petits ruminants viral antibody systems but little, if any, tagging in the heterologous rinderpest system.     

Experimental infection of lactating goats with Leptospira interrogans serovars pomona and hardjo

Pathogenesis of Leptospira interrogans serovars pomona and hardjo was evaluated in 14 lactating goats. Although mild clinical signs of leptospiral infection characterized by pyrexia and reduction in milk yield appeared in some animals, a consistent clinical pattern was not observed in the inoculated animals. The pomona serovar was isolated from the kidney of 1 of the 4 goats inoculated with serovar pomona. The hardjo serovar (strain UI 750) was isolated in the rabbit serum-supplemented bovine albumin polysorbate-80 liquid medium only from the mammary gland of 1 of 4 goats at 13 days after inoculation with serovar hardjo. The positive culture was detected after an 8-month incubation period.     

The isolation of peste des petits ruminants virus from Nigerian sheep and goats.

Four isolates of peste des petits ruminants virus were obtained from sick Nigerian sheep and goats. One was identical antigenically with the prototype Senegalese strain. A cross relationship was found between peste des petits ruminants virus and rinderpest virus based on neutralisation in vitro.     

Observations on the pathogenicity for sheep and goats and the transmissibility of the strain of virus isolated during the rinderpest outbreak in Sri Lanka in 1987

The pathogenicity for sheep and goats of the virus strain that caused acute rinderpest in cattle and domestic buffalo in Sri Lanka after an interval of over 40 years has been examined. The results show that it can cause overt clinical disease in goats, but only mild or unapparent infection in sheep. The disease was transmitted from infected sheep to in-contact susceptible sheep and calves.     

A recent outbreak of rinderpest in East Africa

Rinderpest was brought under control in Kenya in 1976 but in April 1986 an outbreak of the disease occurred in cattle in Western Kenya, five kilometres from the Kenya-Uganda border. This was the first confirmed field outbreak of the disease in Kenya after a lull of over 10 years. Clinical disease was confined to unvaccinated zebu calves aged six to eight months from which rinderpest virus was isolated. High titres of antibodies to rinderpest virus were demonstrated in sera collected from sheep and goats that were grazing together with the affected cattle herds; there was, however, no evidence of clinical disease in these small ruminants and wildlife species in the affected area. The disease outbreak was rapidly stamped out by quarantine and vaccination.     

A Comparison of the Viruses of Infectious Bovine Rhinotracheitis (IBR), Infectious Pustular Vulvovaginitis (IPV), and Rinderpest: Part I. Studies of Antigenic Relationships

The viruses of infectious bovine rhinotracheitis (IBR), infectious pustular vulvovaginitis (IPV), and rinderpest were compared by specific methods. The results further confirmed that IBR and IPV are caused by agents with common antigens. No antigenic relationship was found between these viruses and rinderpest virus, which confirms earlier work.