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.     

Antibodies against certain bluetongue and epizootic haemorrhagic disease viral serotypes in Indonesian ruminants.

The orbiviruses contain several important viruses of livestock including bluetongue (BT) and epizootic haemorrhagic disease of deer (EHD) which share some group antigens. Preliminary screening of sera for antibodies to orbiviruses by the agar gel immunodiffusion (AGID) test has previously revealed widespread infections with the BT group in Indonesia. However serum neutralization (SN) tests give a more accurate estimate of exposure to each serotype in the BT and EHD groups, and in this study were applied to sera that had reacted previously in the AGID test. Five different serotypes of BT and one serotype of EHD virus were studied. Reactors to BT serotype 20 were the most prevalent, followed by EHD type 5 and BT types 21, 12, 1 and 17. Antibodies against BT serotype 20 were present in cattle, buffaloes, goats and sheep, but were most common in buffaloes. Buffaloes showed the highest exposure to the BT serotypes tested. Antibody to EHD type 5 occurred most frequently in cattle. Antibodies against all BT and EHD serotypes tested were found in buffaloes and cattle while goats had antibodies against BT types 20, 21 and EHD type 5 and sheep had antibodies only against BT type 20.     

Evaluation of bluetongue virus diagnostic tests in free-ranging bighorn sheep

Five bluetongue virus (BTV) diagnostic tests were evaluated for use in free-ranging bighorn sheep. We sampled one bighorn sheep population four times between 1989 and 1995. The tests evaluated included virus isolation (VI), polymerase-chain reaction (PCR), serum neutralization (SN), agar-gel immunodiffusion (AGID), and competitive enzyme-linked immunosorbent assay (c-ELISA). The c-ELISA, AGID and SN tests had high levels of agreement in determining serogroup exposure in bighorn sheep. We used maximum-likelihood algorithms to estimate the parameters of each diagnostic test used. Although the c-ELISA and AGID had high sensitivity and specificity, the SN had perfect specificity but lower apparent sensitivity. Due to the potential of cross-reactions among multiple serotypes, results of the SN must be interpreted with caution when assessing serotype exposure in an area where multiple serotypes are endemic. The PCR assay delineated convalescent antibody titers from more-recent infections, and consequently, was pivotal in distinguishing a different exposure pattern between the bighorn sheep and cattle in an adjacent herd. Based on an increasing seroprevalence (50% to 100%), BTV circulated through this bighorn sheep population between 1989 and 1993. This increase in seroprevalence coincided with a bighorn die-off due to BTV infection in June, 1991. An adjacent cattle herd was sampled in 1995 for comparison. The bighorn sheep and adjacent cattle had different patterns of exposure to BTV between 1994 and 1995. There was no evidence that BTV circulated through the bighorn sheep population from 1994 to 1995. In 1995, seroprevalence to BTV decreased to 72%, none of yearling bighorn was seropositive, and all of the 39 bighorn sheep were PCR-negative. In contrast, all adult cattle were seropositive to BTV by c-ELISA and SN, and 4 of the calves were seropositive; 11 of the 24 cattle were PCR-positive, including all five calves. Overall, the pattern of temporal herd immunity in the bighorn sheep appeared to follow a classic epidemic curve, with the appearance and subsequent disappearance of herd immunity coinciding with the 1991 die-off in this population. As low levels of herd immunity and high proportions of susceptible animals are key factors in the development of epidemics, this population of bighorn sheep may be at increased risk for a BTV epidemic in the future.     

Assessment of serological response of young and adult sheep to conjunctival vaccination with Rev-1 vaccine by fluorescence polarization assay (FPA) and other serological tests for B. melitensis

The serological response of young and adult sheep vaccinated conjunctivally with Rev-1 vaccine was assessed by fluorescence polarization assay (FPA), Rose Bengal test (RBT), complement fixation test (CFT), modified Rose Bengal test (m-RBT), indirect ELISA (i-ELISA) and competitive ELISA (c-ELISA), at different post vaccination intervals. One hundred and thirty six adult sheep and 64 lambs were used in the study. The vaccinated animals were bled prior to vaccination (0 day) and thereafter at 21st, 42nd, 35th, 63rd, 91st, 125th, 159th, and 223rd and 330th day post vaccination. The majority of animals (young and adult) showed positive reaction by FPA, RBT, CFT, m-RBT and c-ELISA 21 days post vaccination, whereas by i-ELISA at 42 days. All tests perform equal when animals vaccinated as young are tested 125 days (4 months) post vaccination. In case of animals vaccinated at adulthood, FPA, RBT, CFT and c-ELISA perform equal if the animals are tested 223 days (approximately 8 months) post vaccination. I-ELISA and m-RBT show low specificity if ewes vaccinated at adulthood are tested 330 days (11 months) post vaccination. If control of brucellosis in sheep is based on conjunctivally vaccination of lambs with Rev-1, the vaccinated animals can be tested by any test used for diagnosis of B.melitensis infection accurately at least 4 months post vaccination. If brucellosis control is based on mass vaccination the use of m-RBT and i-ELISA is not recommended for testing adult animals at least for 330 days (11 months) post vaccination due to tests low specificity. Further research is needed so the appropriate cut-offs to be established for FPA, c-ELISA or i-ELISA to become valuable tools for the eradication of Brucella spp. infection in small ruminants in areas where vaccination is practiced.     

Early detection of maedi-visna (ovine progressive pneumonia) virus seroconversion in field sheep samples.

The aim of this work was to investigate whether an enzyme-linked immunosorbent assay (ELISA) was useful for early detection of maedi-visna virus (MVV) infection in sheep under field conditions. An ELISA based on p25 recombinant protein and a gp46 synthetic peptide was used. Sequentially obtained serum samples (n = 1,941) were studied for 4 years. ELISA results were compared with those of the agar gel immunodiffusion (AGID) test, and results of both tests were compared with a reference result established using consensus scores for at least 2 of 3 serologic techniques (AGID, ELISA, and western blotting, which was used to resolve result discrepancies between the other 2 techniques). A total of 247 discrepancies were observed between ELISA and AGID. Of these, 131 were due to an earlier detection of 120 sera by the ELISA and 11 sera by AGID. The remaining discrepancies (116) were due to the presence of false reactions in both tests. Fewer false-negative results were found by ELISA than with AGID (6 vs. 69 sera, respectively), whereas the number of false-positive results was virtually the same for ELISA and AGID (21 vs. 20, respectively). In relation to the reference result, ELISA sensitivity and specificity were 97.8% and 98.2%, respectively, whereas values for AGID were 76.3% and 98.3%, respectively. The agreement between ELISA and the reference result was higher than that between AGID and the reference result (K value: 0.96 and 0.77, respectively). A variation in the ELISA signal (based on optical density) was observed during the study period, suggesting different antibody levels throughout the animal's life. The ELISA was useful for detecting MVV-infected sheep in field conditions and has potential for use in control and eradication programs.     

Bluetongue disease in Swiss sheep breeds: clinical signs after experimental infection with bluetongue virus serotype 8

Clinical disease of bluetongue (BT) in sheep may differ depending on breed, age and immunity of infected sheep and may also vary between serotype and strain of BT virus (BTV). Since there are no data available on the susceptibility of Swiss sheep breeds for BT, we performed experimental infection of the 4 most common Swiss sheep breeds and the highly susceptible Poll Dorset sheep with the BTV serotype 8 (BTV-8) circulating in Northern Europe since 2006. Clinical signs were assessed regarding severity, localisation, progression and time point of their appearance. The results clearly show that the Swiss sheep breeds investigated were susceptible to BTV-8 infection. They developed moderate, BT-characteristic symptoms, which were similar to those observed in Poll Dorset sheep. Regardless of breed, the majority of infected animals showed fever, swelling of the head as well as erosions of the mouth and subcutaneous haemorrhages.     

Specificity of absorbed ELISA and agar gel immuno-diffusion tests for paratuberculosis in goats with observations about use of these tests in infected goats

OBJECTIVE: To determine the specificity of serological tests that are currently used in veterinary diagnostic laboratories in Australia for detection of Mycobacterium avium subsp paratuberculosis infection in goats. DESIGN: A laboratory study. PROCEDURE: Four tests were studied, comprising AGID with M. a. paratuberculosis antigen derived from cattle isolates of caprine or bovine origin, the EMAI caprine Johne's disease absorbed ELISA and the CSL PARACHEK Johne's absorbed EIA. The specificities of AGID and ELISA for paratuberculosis (Johne's disease) were estimated after examining a panel of 1000 serum samples collected from goats in Western Australia, a region free of paratuberculosis. In addition a comparison was made of test performance in a small number of paratuberculous goats from New South Wales using sera from two archival collections. RESULTS: The specificity of the AGID tests was 100% while the specificities of the two absorbed ELISA were 99.7 to 99.8% at appropriate positive-negative cut-offs. Based on testing the small sample of sera from infected goats, the absorbed ELISA tests detected about twice as many goats with Johne's disease as the AGID. Each test detected paratuberculous animals regardless of whether infection was caused by cattle or sheep strains of M. a. paratuberculosis. CONCLUSIONS: Both ELISA and AGID tests for paratuberculosis have high specificity and can be used in a market assurance program without risk of generating large proportions of false positive test results. However, the results suggested the ELISA is more sensitive for detection of infected goats and should be used in preference to the AGID. The two formats of ELISA evaluated in this study have similar characteristics and could be used in paratuberculosis control programs for the goat industries, but further data on sensitivity would increase confidence in their application.     

Seroprevalence of antibodies against bluetongue virus in animals imported to Poland from EU countries

The aim of this study was to determine the seroprevalence of BTV-specific antibodies in animals imported to Poland from EU countries after 15 June 2006. From 1 January 2007 to 22 January 2008, a total of 10719 samples of sera collected from cattle, goats and fallow deer were tested. Sera were screened using the highly sensitive and specific c-ELISA test and positive results were confirmed by the AGID assay. Out of 10719 sera, 30 (0.28% of the total number of samples) were found to be positive in both tests applied. All of 21 seropositive cattle specimens were imported to Poland from Germany whereas 9 seropositive fallow deer were of Dutch origin. In conclusion, it can be stated that because BTV situation in Europe is getting worse, implemented surveillance studies should be continued to monitor the actual BT status in Poland.     

Ovine progressive pneumonia (Maedi-Visna): an emerging respiratory disease of sheep in Ethiopia

A serological study was done to assess the role of Maedi-Visna (MV) infection in sheep from flocks with high respiratory tract disease morbidity in Ethiopia. Of 105 sheep examined from central Ethiopia 78 (74%) were positive for MV-infection. However, antibodies to the virus were not detected in 48 sheep and 70 goats from elsewhere in Ethiopia. The infection was detected in all breeds of sheep examined (Awassi, Hampshire, Corriedale, indigenous Menz breeds and their crosses) but with a significant breed difference (chi 2 = 20, p < 0.001) varying from 48% in imported Awassi sheep to 92% in the indigenous Menz sheep. This suggests that Menz sheep are more susceptible to infection, which may support the observation of a higher incidence of clinical disease in these sheep compared to exotic breeds and their crosses. It also supports recent studies indicating that MV is becoming one of the most important respiratory tract diseases in sheep in central Ethiopia. Our findings indicate that MV was introduced into Ethiopia via sheep imported into the central highlands and that it now constitutes an important emerging disease is discussed. Measures to control the disease are suggested.     

Pathobiochemical mechanisms involved in the control of the disease caused by Trypanosoma congolense in African grey duiker (Sylvicapra grimmia)

The course of Trypanosoma congolense infections in African grey duiker (Sylvicapra grimmia) and sheep and goats were studied. Several parameters suggested that the grey duiker was much more resistant to trypanosomosis than sheep and goats. They showed increases in weight during infection, had a much longer pre-patent period, and their peak parasitaemia levels were about 100-fold lower than those of sheep and goats. Parasites were no longer detected in grey duiker blood 35 days after infection. Anaemia, measured as drops in packed cell volume (PCV), haemoglobin (Hb) concentration and erythrocyte (RBC) counts were not observed in the grey duiker. In contrast, sheep and goats suffered severe weight losses and had continuously high parasitaemia levels. Sheep and goats developed progressively severe normocytic normochromic anaemia and leucopenia from day 14 post-infection onwards.Serum levels of total protein, globulin and albumin of grey duiker did not change significantly throughout the course of infection, while the levels of total serum protein, globulin and gamma-globulin exhibited significant increases from day 21 post-infection onwards in sheep and goats, with peak values recorded on 28 and 35 days post-infection in sheep and goats, respectively. There were inconsistent variations in albumin levels in sheep and goats throughout the course of infection.There were no significant changes in erythrocyte activities of AST and ALT, while there were transient but significant elevations of ALP level on day 35, and GGT levels between 14 and 35 days post-infection in grey duiker. Conversely, the levels of all the enzymes were progressively depressed, especially from 14 to 49 days post-infection.In vitro erythrocyte peroxidation remained relatively unchanged throughout the period of the experiment in the grey duiker, except for slight but significant increase on day 42 post-infection. However, in vitro erythrocyte peroxidation increased significantly by between 100 and 300% of pre-infection levels from 14th to 42nd day p.i. both in sheep and goats, before returning to pre-infection levels after 14 days of treatment.Haematological values, serum and erythrocyte indices studied returned to near pre-infection levels 14 days after treatment with Berenil((R)).It is concluded that the grey duiker is inherently trypanotolerant. This is shown by its ability to control parasitaemia, suffer less severe anaemia, and to a relative degree resist pathobiochemical derangements of some serum and erythrocyte metabolites and enzymes, as well as reduction of infection-induced erythrocyte lipid peroxidase damage than sheep and goats.     

Comparison of a maedi-visna virus CA-TM fusion protein ELISA with other assays for detecting sheep infected with North American ovine lentivirus strains.

A maedi-visna virus CA-TM fusion protein ELISA (MVV ELISA) was evaluated for the detection of antibody in sheep infected with North American ovine lentivirus (OvLV). The results of the MVV ELISA were compared with other assays for OvLV antibody and with viral infection in an intensively studied group of 38 sheep with a high prevalence of OvLV infection and disease. The sensitivity, specificity, and concordance of assays for OvLV antibody (MVV ELISA, indirect ELISA, Western blot, and AGID), virus (virus isolation, PCR, antigen ELISA), and OvLV-induced disease in each animal were compared with OvLV infection status as defined by a positive result in two or more of the assays. Five sheep met the criteria for absence of OvLV infection. The sensitivity of the MVV ELISA in detecting OvLV infected sheep was 64%, whereas the sensitivity of the other three tests for antibody ranged from 85 to 94%. All the antibody assays were 100% specific in this group of animals. Of the assays for virus, the PCR test had the highest sensitivity and the best concordance with OvLV infection, but it also had the lowest specificity of any of the virus or antibody assays. Among the antibody tests, the concordance of the MVV ELISA compared most favorably with the AGID test for detecting OvLV-infected sheep. Analysis of serum samples from 28 lambs experimentally-infected with one of three North American strains of OvLV suggested that there were no significant strain differences detectable by antibody assay. Twenty virus-inoculated lambs were positive by both the MVV ELISA and the AGID test, five lambs were MVV ELISA negative and AGID test positive, and three lambs were MVV ELISA positive and AGID test negative. No pre-inoculation samples were positive by either assay. In a longitudinal study involving seven lambs, antibodies to OvLV were detected by AGID 3-5 weeks post-inoculation, but were not detected by MVV ELISA until 5-10 weeks post-inoculation. Among 128 naturally and experimentally-infected sheep that were seropositive in the AGID test, the overall sensitivity of the MVV ELISA was higher in the naturally infected sheep (84%) than in the experimentally infected sheep (69%). The data indicated that the MVV ELISA represents a less sensitive, but specific alternative for the detection of OvLV antibodies.     

Comparison of competitive ELISA, indirect ELISA and standard AGID tests for detecting blue-tongue virus antibodies in cattle and sheep

The performances of a competitive enzyme-linked immunosorbent assay (ELISA) using a group specific monoclonal antibody against bluetongue virus, an indirect ELISA and the standard agar gel immunodiffusion (AGID) test were compared in the detection of serum antibody against bluetongue virus. Test sera consisted of 1300 bovine, 530 ovine and 160 carpine samples from bluetongue-free areas of Canada, 605 bovine and ovine field samples from the USA and Barbados and 464 samples from 79 cattle and sheep experimentally infected with 19 South African and five USA serotypes of bluetongue virus. The diagnostic specificity of the competitive ELISA, as determined for the bluetongue virus-free cattle sera was superior (99.92 per cent) to that of the indirect ELISA (99.85 per cent) and the AGID (99.0 per cent). The specificities of the competitive ELISA for sheep (99.63 per cent) and goats (100.0 per cent) sera were also higher than those of the AGID test. The performance of the ELISA tests was similar whether a gamma-ray-irradiated (2.0 Mrad) or a non-irradiated bluetongue virus antigen preparation was used. The competitive ELISA results for bovine field sera from endemic areas demonstrated a relatively low level of agreement (92.04 per cent) with AGID test results, with 9.7 per cent false negatives. The possible presence in these sera of antibody to cross-reacting antigens or to other orbiviruses, eg, epizootic haemorrhagic disease virus, which react in the AGID but not in the competitive ELISA may account for this lack of agreement.(ABSTRACT TRUNCATED AT 250 WORDS)     

Bluetongue and related viruses in Nigeria: Experimental infection of West African dwarf sheep with Nigeria strains of the viruses of epizootic haemorrhagic disease of deer and bluetongue

West African dwarf sheep were inoculated by the subcutaneous route with epizootic haemorrhagic disease of deer (EHD) virus or bluetongue (BT) virus. No clinical disease was observed following primary EHD or BT infection, or subsequent challenge with either homologous or heterologous virus. However, viraemia was detected in non-immune sheep exposed to BT virus, but not in BT- or EHD-immune sheep challenged with either virus, or in non-immune sheep exposed to primary EHD virus infection. Complement fixing antibodies developed against both EHD and BT viruses following the primary infection with either virus, or subsequent challenge with homologous or heterologous virus. Following a primary infection, virus-neutralising (VN) antibodies developed only against the inoculated virus, while the detection of VN antibodied to both viruses followed the challenge of an EHD- or BT-immuned sheep with either the homologous or heterologous virus. These findings further support previous reports of a relationship between EHD and BT viruses. between EHD and BT viruses.     

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.     

Detection of contagious ovine ecthyma (orf) and risk factors for infection in small ruminants in Iran

The primary cause of contagious ecthyma is the orf virus, the parapoxvirus prototype. It is a viral problem observed in goat and sheep flocks in Iran, causing economic loss. Orf is a zoonosis with little epidemiological investigation present in Iran. The current research aims at determining the status of this virus, and a PCR was used as a confirmatory instrument. We sampled 668 goats and sheep and various breeding systems. Besides, the orf prevalence was studied, and vaccination efficacy was determined. Moreover, the potential risk factors surveyed for infection with ecthyma were identified. Samples were taken from goat and sheep flocks in the present cross-sectional research, and PCR was used for testing orf DNA. A checklist including animals’ general information was completed. Data were analyzed using univariate tests (chi-square and t-tests) and multivariable binary logistic regression analysis. Three hundred one (45%) goats and sheep detected orf DNA. The age of 70% of positive cases was below one month. Ecthyma infection was significantly higher in imported breeds (87.3%) than indigenous (39.3%). Ninety-six percent of infected goats and sheep in the present work were not vaccinated against ecthyma. The high prevalence of the orf virus was confirmed among goat and sheep flocks in Iran. It is necessary to train ranchers regarding sanitary actions, quarantine, and application of orf vaccination plans.     

Evaluation of three serological tests for diagnosis of Maedi-Visna virus infection using latent class analysis

Maedi-Visna virus (MVV) infection in sheep is present in several European countries, including Norway. The current Norwegian surveillance and control programme for MVV infection uses three serological tests: an agar gel immunodiffusion test (AGID) and two commercially available indirect ELISAs (Institut Pourquier, P-ELISA and HYPHEN BioMed, H-ELISA). From 18 flocks with suspected or confirmed MVV infection, sera from naturally infected sheep were obtained, and sensitivity (Se) and specificity (Sp) of the three tests were estimated in absence of a perfect reference test using latent class models in a Bayesian analysis. The AGID had higher Sp (95% posterior credibility interval (PCI) [98.4; 99.9]) than either ELISA (95% PCI: P-ELISA, [95.1; 99.0]; H-ELISA, [91.4; 96.6]), but much lower Se (95% PCI: AGID, [41.4; 59.8]; P-ELISA, [92.7; 100.0]; H-ELISA, [90.9; 99.4]). Currently the P-ELISA is used for screening and positive samples are subsequently confirmed by a setup using all three tests in a serial reading. The Se and Sp of the serial interpretations with and without the H-ELISA were estimated. The results suggested that the H-ELISA could be dropped as a confirmatory test as the Se of the three test serial reading was reduced significantly without adding a significant improvement of the Sp compared to the serial reading of the P-ELISA and AGID alone. However, the perceived cost of false positives versus false negatives will influence this decision. Estimates of the predictive values for the tests and combinations suggested that the P-ELISA is a good choice of screening, but confirmatory tests are needed to achieve acceptable levels of positive predictive values.     

An enzyme-linked immunosorbent assay for detection of antibodies to maedi-visna virus in sheep. II. Comparison to conventional agar gel immunodiffusion test.

A study was conducted to compare the indirect enzyme-linked immunosorbent-assay (i-ELISA) test using antigen prepared by a simple technique using sodium dodecyl sulfate (SDS) treatment to the conventional agar gel immunodiffusion test (AGID). Ten specific-pathogen-free (SPF) sheep were inoculated with maedi-visna virus (MVV) and serum antibody titers compared over a period of 14 weeks. All the sheep seroconverted by the i-ELISA compared to 90% by the AGID. The i-ELISA detected antibody at a mean of 2.6 weeks prior to the AGID. In both tests, fluctuations were observed in the serum antibody response of two sheep. The i-ELISA had a specificity of at least 98.8% and an increased relative sensitivity of 15.5% compared to the AGID, based on the analysis of sera from experimental sheep with MVV free status and sera from sheep from various sources. Of the sera from a seronegative flock which had been monitored with the AGID after a "test and remove" eradication program, 10.2% were positive by the i-ELISA. It was concluded that the AGID test may not be adequate to monitor samples for an eradication scheme.     

Epizootiologic study of bluetongue: virologic and serologic results

Heparinized blood and serum samples were obtained from 1,295 ruminants in herds or flocks with bluetongue virus (BTV) infection in 4 western states. Submissions were from herds or flocks with clinical bluetongue (BT), as well as from animals on premises with no history of BT disease. Insects, including Culicoides variipennis, were collected in areas enzootic for BT disease. Viral isolations were in 10-day-old embryonating chicken eggs that were then adapted to Vero cells for serotyping. Sera were tested from group-specific antibody to BTV by the micro agar gel precipitin (AGP) test. Viral isolations were from cattle (81), sheep (122), goats (9), antelope (2), and C varipennis (5). There were 7 isolates of serotype 120, 114 of serotype 11, 42 of serotype 13, and 56 of serotype 17. In herds or flocks from which BTV was isolated, 51% of cattle, 56% of sheep, 21% of goats, and 52% of antelope had AGP antibodies. Virus was isolated from 43% of the cattle and 23% of the sheep that had no demonstrable evidence of AGP antibodies. Viral isolations were seasonal, occurring from August until December. Approximately 30% of the herds or flocks from which virus was isolated had more than one serotype of virus causing infection.     

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.     

Comparison of IS900 tissue PCR, bacterial culture, johnin and serological tests for diagnosis of naturally occurring paratuberculosis in goats

Comparative efficacy of an IS900 tissue PCR, bacterial culture, johnin, agar-gel immunodiffusion (AGID) and absorbed-ELISA tests was investigated in 43 goats naturally infected with paratuberculosis. On histological examination, tissue sections from all animals showed typical granulomatous inflammatory changes. The lesions were classified as multibacillary (MB) (n=30), which had diffuse granulomatous lesions with abundant acid-fast bacilli (AFB), and paucibacillary (PB) (n=13), which had focal or multifocal granulomatous lesions with few AFB. The sensitivities of johnin test, tissue culture, faecal culture, tissue PCR, AGID and ELISA were 68% (17/25), 100% (30/30), 84.6% (22/26), 100% (30/30), 96.2% (25/26) and 100% (26/26) in MB goats, and 88.8 (8/9), 46.1% (8/13), 40% (4/10), 61.5% (8/13), 50% (5/10), and 70% (7/10) in PB goats, respectively. Except for the johnin test, which showed higher sensitivity in PB goats, all other tests displayed significantly higher sensitivities in MB goats. The results indicate the usefulness of tissue PCR, culture and serological tests in the diagnosis of clinically affected paratuberculous goats, especially with multibacillary pathology.