Antibody seroprevalences against Peste des Petits Ruminants (PPR) virus in sheep and goats in Sudan

Counter immnuo-electrophoresis (CIEP) and Competitive ELISA (C-ELISA) tests were employed for seroprevalence of Peste des Petits Ruminants (PPR) infection in Sudan. The result of both tests showed high prevalence of PPRV antibodies in sheep and goats sera collected from six different regions of Sudan. Of the 519 serum samples examined for the presence of PPRV antibodies 307(59.15%) were positive by CIEP while 263(50.67%) were positive by C-ELISA. CIEP technique was shown to be more sensitive than C-ELISA technique for detection of PPRV antibodies (Kappa statistics 0.259). C-ELISA allowed rapid, simple, specific, sensitive and differential sero-diagnosis of PPRV and RPV in sheep, goats and cattle. CIEP is, unlike competitive ELISA, is group-specific test and can not differentiate between PPR and RP infections. Despite its low specificity CIEP can be a useful indicative screening test for PPRV antibodies in flocks that neither been vaccinated nor otherwise exposed to PPR or RP virus. Results obtained suggest that CIEP, like the HI test, could be a useful screening test where it is not possible to use C-ELISA.     

Current situation of Peste des petits ruminants (PPR) in the Sudan

The current situation of PPR in Sudan was investigated. A total of 61 tissue samples were collected from various PPR suspected outbreaks in sheep in Sudan during 2008. Collected tissue samples were tested for PPR antigen using IcELISA, PPR antigen was detected in 26 out of 61 samples (42.6%). Highest antigen detection rate was in specimens collected from western Sudan. A total of 1198 serum samples were collected from sheep (n = 500), camels (n = 392), and goats (n = 306) from different areas in Sudan (Khartoum, Gezira, Tambool, River Nile, Kordofan, White Nile, Blue Nile, Gedarif, Kassala, Halfa ElGadida, Port Sudan). Collected sera were examined for PPR antibodies using cELISA, a total of 336 (67.2%) sheep, 170 (55.6%) goat and 1 (0.3%) camel samples were found to be positive.     

Peste des petits ruminants infection in domestic ruminants in Sudan

The existence of peste des petits ruminants (PPR) in domestic ruminants and camels in Sudan during 2008–2012 was investigated. Lung tissues and serum samples were randomly collected from sheep, goats, cattle, and camels at different areas of Sudan. A total of 12,384 serum samples were collected from clinically healthy 7413 sheep, 1988 camels, 1501 cattle, 1459 goats, and 23 gazelles at different areas in the Sudan. They were examined for PPR antibodies using competitive ELISA (cELISA). The overall detected seroprevalence of PPR in tested sera was 49.4%; seroprevalence values within species were 67.1, 48.2, 25.8, 2.1, and 21.7% in sheep, goat, cattle, camels, and gazelles, respectively. The highest seroprevalence (68.1%) was observed in sera collected from Darfur states, then the central states (54.3%). A total of 1276 lung tissue samples (623 sheep, 324 cattle, 220 camels, and 109 goats) were collected. The majority of lung samples were collected from clinically healthy animals that showed lesions on PM in slaughterhouses (95%) and during PPR outbreaks; samples were tested for PPR antigen using immunocapture ELISA (IcELISA). PPR antigen was detected in 233 out of the 1276 tested samples (18.3%). Positive results were observed in samples collected from clinically healthy and diseased animals. The observed prevalence values in each species were 33.6, 21.1, 15.4, and 12.3% in camel, goat, sheep, and cattle, respectively. PPR antigen was detected in samples from different areas; however, the highest prevalence (63.9%) was found in samples collected from the eastern states, then Khartoum state (28%). Trials for virus isolation were done in different cell cultures. Out of 30 IcELISA-positive samples inoculated in primary bovine and ovine kidney cells, Vero cells, the PPR virus was successfully isolated from 15 (eight sheep, five camels, and two goats) samples in the three cell culture types. Using RT-PCR, PPRV nucleic acid was detected in all 25 IcELISA-positive tested samples.     

Prevalence of peste des petits ruminants among sheep and goats in India

This study measured the clinical prevalence of peste des petits ruminants (PPR) among sheep and goats in India between 2003 and 2009 by analyzing clinical samples from suspected cases of PPR that were submitted to the Rinderpest and Allied Disease Laboratory, Division of Virology, IVRI, Mukteswar for PPR diagnosis. PPR outbreaks were confirmed by detecting PPR virus (PPRV)-specific antigen in the clinical samples. Clinical samples (blood, nasal swabs, spleen, lymph node, kidney, liver, intestine, and pooled tissue materials) were taken from a total of 592 sheep and 912 goats in different states of India and screened for the presence of PPRV antigen using a monoclonal antibody-based sandwich ELISA kit. A total of 20, 38, and 11 laboratory-confirmed PPR outbreaks occurred among sheep, goat, and combined sheep and goat populations, respectively. Our findings provide evidence of widespread PPR endemicity in India. The underlying reasons could be variations in husbandry practices in different geographical regions, agro-climatic conditions, and livestock migration. Furthermore, decrease in the number of PPR outbreaks over time might be due to the effectiveness of current live PPR vaccines and timely vaccination of target species. Vaccination against PPR has been practiced in India since 2002 to control this disease.     

Prevalence and distribution of Peste des petits ruminants virus antibodies in various districts of Tanzania

Despite the widespread prevalence of infection with Peste des petits ruminants virus (PPRV) in goats and sheep industry in Asia and sub-Saharan Africa, there have been few, if any, structured population-based studies examining the epidemiology of this infection in Tanzania. In this study, we investigated the seroprevalence, and risk factors, of Peste des petitis ruminants(PPR) in sheep and goat flocks from seven different geographical administration authorities (Ngorongoro, Monduli, Longido, Karatu, Mbulu, Siha and Simanjiro) located in Northern Tanzania. Serum samples from 657 and 892 sheep and goats, respectively, corresponding to 91 sheep/goat flocks and 43 villages were collected. Competitive enzyme linked immunosorbent assay (c-ELISA) was used to detect the presence of antibodies in the serum against PPRV. Chi-square analysis and multivariable logistic regression model were used to identify risk factors for PPRV seropositivity. Findings suggested that the sero-positive cases were significantly higher in goats than in sheep (49.5% versus 39.8%; P = 0.002). The overall seroprevalence of PPRV infection in small ruminants was 45.8%. Highest seroprevalence (42.6–88.02%) was observed in Mbulu, Siha, Longido, Ngorongoro districts, while antibodies less than 40% to none were found in serum from Monduli, Karatu and Simanjiro, respectively. These findings confirm natural transmission of PPRV under field condition for the first time in Tanzania. Results may be correlated with variations in the sheep and goat husbandry practices within different geographic localities, the uncontrolled movement of animals, the levels of natural immunity and the sharing of grazing field amongst agro and pastoralists.     

Seroprevalence of Peste des petits ruminants in cattle and buffaloes from Southern Peninsular India

This study describes seroprevalence of Peste des petits ruminants (PPR) in cattle and buffaloes carried out during the period 2009–2010 using the randomly collected serum samples from different parts of Southern peninsular India. The report presents the results of PPR virus (PPRV)—specific antibodies in situations where either the subclinical or inapparent or non-lethal infection was there in cattle and buffaloes. A total of 2,548 serum samples [cattle = 1,158, buffaloes = 1,001, sheep = 303 and goat = 86] were collected and screened for PPRV antibodies by using a PPR monoclonal antibody-based competitive ELISA kit. Analysis of 2,159 serum samples indicates an overall 4.58% prevalence of PPRV antibody in cattle and buffaloes. The presence of PPRV-specific antibodies demonstrates that cattle and buffaloes are exposed to PPR infection naturally, and the transmission mode may be direct or indirect. Further, it implies the importance of bovines as subclinical hosts for the virus besides widespread presence of the disease in sheep and goats in the country.     

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.     

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.     

Experimental infection of alpine goats with a Moroccan strain of peste des petits ruminants virus (PPRV)

Peste des petits ruminants virus (PPRV) recently caused a serious outbreak of disease in Moroccan sheep and goats. Alpine goats were highly susceptible to PPRV with mortality rates approaching 100%, as opposed to local breeds of sheep which were less susceptible to the disease. The relative susceptibility of alpine goats was investigated through an experimental infection study with the Moroccan strain of PPRV. Severe clinical signs were observed in the alpine goats with virus being excreted through ocular, nasal and oral routes. No difference in the severity of the disease in goats was observed with different inoculation routes and transmission of the virus by direct contact was confirmed. This study confirmed the susceptibility of the alpine goat to PPRV infection and describes a challenge protocol that effectively and consistently reproduced severe clinical signs of PPR in experimentally infected goats.     

小反刍兽疫活疫苗的安全性评价试验

2007年小反刍兽疫(PPR)在我国西藏首次暴发,在西藏和新疆部分地区使用PPR Nigeria 75/1疫苗株制造的疫苗进行免疫接种。为明确疫苗的安全性,中国兽医药品监察所国家牛瘟参考实验室对其安全性能进行了系统评价。健康易感山羊、绵羊及怀孕山羊、怀孕绵羊按不同剂量接种疫苗后,均未观察到异常临床反应;怀孕母羊所产羔羊数量与对照组无明显差异。疫苗对小白鼠、豚鼠的非特异性安全试验表明,所有接种动物均健活。结果表明该疫苗安全性良好,可在田间大规模使用。     

Development of a monoclonal antibody based competitive-ELISA for detection and titration of antibodies to peste des petits ruminants (PPR) virus

Peste des petits ruminants (PPR) is an acute febrile, viral, disease of small ruminants with great economic importance. A competitive-ELISA (c-ELISA) test was developed for detection of antibodies to PPR virus in the sera samples of goats and sheep. The test uses monoclonal antibody to a neutralizing epitope of haemagglutinin protein of the virus. Based on the distribution of known negative sera samples (n=933) in respect of PPR virus antibodies in the test, a cut-off value was set as 38%. This value was the result of mean of negative population added with two times the standard deviations. A total of 1668 sera samples from goat and sheep and 32 sera from cattle were screened by c-ELISA and virus neutralization test (VNT). Efficacy of c-ELISA compared very well with VNT having high relative specificity (98.4%) and sensitivity (92.4%). The sensitivity of c-ELISA for PPR sero-surveillance could further be increased (95.4%), if the target population is non-vaccinated. c-ELISA test correlated well with VNT (r=0.845) for end-point titration of PPR virus antibody in 64 goat sera samples. It could clearly separate infected population from uninfected in field sera. Using c-ELISA test paired sera samples from 13 goats provided a clear diagnosis of PPR virus infection. Furthermore, antibodies to PPR virus could be successfully detected during 1 year after vaccination in four goats inoculated with an experimental PPR vaccine. Findings suggest that the c-ELISA test developed can easily replace VNT for sero-surveillance, sero-monitoring, diagnosis from paired sera samples and end-point titration of PPR virus antibodies.     

小反刍兽疫免疫荧光诊断技术研究

为了建立小反刍兽疫(PPR)荧光抗体诊断方法,研究选用小反刍兽疫病毒(PPRV)Nigeria75/1减毒疫苗株制备抗原,免疫试验山羊,制备小反刍兽疫高免血清,用硫酸铵盐析法粗提免疫球蛋白(IgG),再应用提取的IgG制备异硫氰酸荧光素标记抗体。结果表明:所制备的荧光抗体对试验用Vero细胞组织抗原和犬瘟热病毒(CDV)抗原无反应性,对小反刍兽疫病毒感染细胞的检出敏感性为1×106稀释度。     

Evidence of mixed infection of peste des petits ruminants virus and bluetongue virus in a flock of goats as confirmed by detection of antigen, antibody and nucleic acid of both the viruses

A mixed infection with peste des petits ruminants virus (PPRV) and bluetongue virus (BTV) occurred in goats which exhibited symptoms characteristic of PPR. A number of samples were collected from ailing or dead goats for labrotory diagnosis. Antibody to BTV and PPRV was detected in sera samples by competitive ELISA. No PPRV antigen was detected in tissue samples like lung and spleen, however, presence of PPRV antigen in some sera samples was confirmed by sandwich ELISA. All the blood samples collected from the ailing animals were found positive for BTV antigen by a sandwich ELISA. BTV- and PPRV nucleic acids were amplified from the pooled blood and tissue samples respectively by RT-PCR assays. The identity of the amplicons was confirmed by cloning and sequencing. All these tests confirm that the goats were infected with PPRV and BTV simultaneously. Isolation of viruses from the clinical samples is underway.     

Evidence of peste des petits ruminants virus (PPRV) infection in Sindh Ibex (<Emphasis Type="Italic">Capra aegagrus blythi</Emphasis>) in Pakistan as confirmed by detection of antigen and antibody

An outbreak resulting in mortality in Sindh Ibex (Capra aegagrus blythi) was investigated. There was a history of about 36 deaths (both young and adult) during the period of 1 month. Disease appeared in a generalized form, affecting the respiratory and digestive systems. Major lesions were respiratory distress, pustules on and in the mouth, ocular–nasal discharges, and severe diarrhea. The most significant lesion was the oculonasal discharges and diarrhea. Deaths were mainly due to blindness, anorexia, diarrhea, and respiratory arrest. Both adult (mortality = 21) and young (mortality = 15) animals were affected with the disease. Peste des petits ruminants virus (PPRV) antigen was detected in the spleen, lung, lymph node, and swab samples by immunocapture enzyme-linked immunosorbent assay. Spleen and lung samples were also tested and found positive for the presence of F-gene of PPRV by polymerase chain reaction. Thirteen of 20 serum samples from nearby sheep and goats were found positive for antibodies to PPRV. The disease threatened the huge population of ibex in the wild life park, which was spread over a large area, but vaccination of the domestic population of sheep and goats in the surrounding villages appeared to control the disease.     

Prevalence and mortality rate of peste des petitis ruminant (ppr): possible association with abortion in goat

Present study was designed to investigate the prevalence and mortality (%) caused by Peste des Petitis Ruminant (PPR) and its possible association with abortion in goat flocks at different areas of Pakistan. A total of 140 animals were samples in the population of 650 which was having 185 deaths (Mortality rate = 28 %) from three different regions of the country. There were 58 abortions in the 140 pregnant goats of above said population One hundred & ten (110) serum samples from diseased, recovered and apparently healthy animals were tested for the presence of PPR antibodies by competitive ELISA (c ELISA). Eighty-four (84) animals were positive for PPR antibodies whereas in apparently healthy adult goats in the same flock, no PPR antibodies were detected. Twenty-four (24) tissue samples collected from the dead animals and six samples from aborted fetus were tested for the presence of PPR antigen by Immuno-capture ELISA (Ic ELISA). Nineteen (19) out of thirty (30) organ samples mainly from lung, spleen, lymph node were found positive for PPR antigen but negative from lungs of aborted fetus. There was a high rate of abortions (28–45 %) in each of the outbreak and it was highest in the outbreak of Golra Sharif, Islamabad (No. = 21 in total population of 100). As the serum samples from the aborted dams were found positive for PPR antibodies so the study provides the possible association of mortality and prevalence of PPR disease with high rate of abortions in goat.     

Monoclonal antibody-based competitive ELISA for simultaneous detection of rinderpest virus and peste des petits ruminants virus antibodies

An experimental competitive enzyme-linked immunosorbent assay (morbillivirus cELISA) using a recombinant N antigen (rRPV N) expressed in a baculovirus and a ruminant morbillivirus (RPV and PPRV)-specific monoclonal antibody (P-13A9) was developed for simultaneous detection of rinderpest virus (RPV) and peste des petits ruminants virus (PPRV) antibodies and its diagnostic performance was evaluated. A set of known reference antisera against RPV and PPRV belonging to different lineages, experimental sera from cattle vaccinated for a RPV of Asian lineage, and field sera from cattle and sheep/goat populations known to be positive (West Africa) and negative (Korea) for RPV and PPRV were used for the evaluation. Morbillivirus cELISA results on the panel of experimental RPV and PPRV antisera showed high correlation (r=0.97) between the whole virus and the rRPV N antigens, suggesting that the rRPV N contains a ruminant morbillivirus-specific antigenic determinant recognized by the P-13A9 and it may be suitable as an ELISA antigen in place of the whole virus. Morbillivirus cELISA detected anti-RPV and anti-PPRV antibodies in all reference RPV and PPRV antisera containing VN titers >/=1:8, suggesting that the assay can simultaneously detect antibodies against RPV and PPRV. Anti-RPV antibody was detected by morbillivirus cELISA in vaccinated cattle as early as the VNT and continued to be detectable by both the cELISA and the VNT until termination of the study. When applied to field samples from Africa, morbillivirus cELISA showed good agreement with a RP cELISA kit (kappa value of 0.86) in bovine sera and with a peste des petits ruminant cELISA kit (kappa value of 0.81) in caprine/ovine sera. Usefulness of morbillivirus cELISA using the rRPV N protein was discussed.