Evaluation of three serological tests for the diagnosis of Brucella suis in dogs using Bayesian latent class analysis.

Brucella suis infection of dogs is an emerging issue worldwide requiring specific management to address zoonotic risks and animal welfare concerns. Diagnosis in dogs is routinely based on serological testing, but to date these tests have only been validated for use in production animal species and humans. This study aimed to assess the diagnostic performance of three commonly used serological tests in dogs. Canine sera (n = 989) were tested with the Rose Bengal rapid plate agglutination test (RBRPT), the complement fixation test (CFT) and a competitive ELISA (C-ELISA). Diagnostic test performance was evaluated using a three test, two population Bayesian latent class analysis accounting for conditional dependence between the three tests. Positive and negative predictive values (PPV, NPV) were calculated for a range of expected prevalence estimates for the individual tests and test combinations interpreted in series and parallel. The RBRPT showed the highest individual Se of 0.902 (95 % posterior credible interval [PCI] 0.759-0.978) and the CFT the highest individual diagnostic specificity (Sp) of 0.914 (95 % PCI 0.886-0.946). The C-ELISA had marginally the best overall diagnostic performance (Youden's index = 0.807). The CFT and the C-ELISA interpreted in parallel returned the highest combined Se and Sp (0.988 and 0.885, respectively). All tests returned NPVs of > 0.982 in 2-8 % prevalence settings. Series interpretation of the three-test combination as well as the two-test combinations of the RBRPT and the C-ELISA and the CFT and the C-ELISA produced a PPV of 0.502 when the estimated prevalence was 8 %. While all tests are suitable for the detection of B. suis antibodies in dogs, they should not be interpreted in isolation as their diagnostic value is dependent on the pre-test probability of the disease. As such they are useful tools for the diagnosis of B. suis in dogs when exposure, history and clinical presentation indicate a risk of brucellosis.

Automatic offline-capable smartphone paper-based microfluidic device for efficient biomarker detection of Alzheimer's disease.

BACKGROUND: Alzheimer's disease (AD) is a prevalent neurodegenerative disease with no effective treatment. Efficient and rapid detection plays a crucial role in mitigating and managing AD progression. Deep learning-assisted smartphone-based microfluidic paper analysis devices (µPADs) offer the advantages of low cost, good sensitivity, and rapid detection, providing a strategic pathway to address large-scale disease screening in resource-limited areas. However, existing smartphone-based detection platforms usually rely on large devices or cloud servers for data transfer and processing. Additionally, the implementation of automated colorimetric enzyme-linked immunoassay (c-ELISA) on µPADs can further facilitate the realization of smartphone µPADs platforms for efficient disease detection. RESULTS: This paper introduces a new deep learning-assisted offline smartphone platform for early AD screening, offering rapid disease detection in low-resource areas. The proposed platform features a simple mechanical rotating structure controlled by a smartphone, enabling fully automated c-ELISA on µPADs. Our platform successfully applied sandwich c-ELISA for detecting the ß-amyloid peptide 1-42 (Aß 1-42, a crucial AD biomarker) and demonstrated its efficacy in 38 artificial plasma samples (healthy: 19, unhealthy: 19, N = 6). Moreover, we employed the YOLOv5 deep learning model and achieved an impressive 97 % accuracy on a dataset of 1824 images, which is 10.16 % higher than the traditional method of curve-fitting results. The trained YOLOv5 model was seamlessly integrated into the smartphone using the NCNN (Tencent's Neural Network Inference Framework), enabling deep learning-assisted offline detection. A user-friendly smartphone application was developed to control the entire process, realizing a streamlined "samples in, answers out" approach. SIGNIFICANCE: This deep learning-assisted, low-cost, user-friendly, highly stable, and rapid-response automated offline smartphone-based detection platform represents a good advancement in point-of-care testing (POCT). Moreover, our platform provides a feasible approach for efficient AD detection by examining the level of Aß 1-42, particularly in areas with low resources and limited communication infrastructure.

Seroprevalence and Risk Factors Related to Bovine Brucellosis in Continental Ecuador.

Bovine brucellosis is a worldwide zoonotic contagious disease. According to World Animal Health Information System reports Ecuador has presented an increasing number of bovine brucellosis outbreaks in the continental territory over the past years (756 in 2018 versus 964 in 2021), generating economic losses for producers and causing a risk to public health. A cross-sectional study was conducted to investigate the seroprevalence of bovine brucellosis and associated risk or protective factors between May and June 2018. This stratified random study was implemented in 290 cattle herds located in the 23 provinces of continental Ecuador, which represents a total of 3737 cows aged 24 months or older. A competitive ELISA was used to detect Brucella antibodies. Simultaneously, an epidemiological survey was implemented to assess the brucellosis risk or protective factors. The apparent prevalence of bovine brucellosis at the herd level was 21.3% (95% CI: 16.8-26.6) and 6.2% (95% CI: 5.5-7) at the animal level. Univariate and multivariate logistic regression analyses were performed to determine the relationship between the potential factors associated with the presence of bovine brucellosis. The risk factors identified after multivariate analysis were a surface in ha per herd > 70 ha (OR = 2.73; 95% CI: 1.18-6.32) and the number of parturitions per animal (two or more with OR ≥ 1.8 and p-value ≤ 0.047). On the contrary, the protective factors were the region (farms located in the eastern region) and the absence of reported clinical signs. In addition, in herds where extensive production predominates, farmers have a low level of knowledge, and the farm biosecurity level is low. These results can guide the authorities in managing the risk factors identified, understanding the current epidemiological situation in Ecuador, improving the bovine brucellosis control program and food safety, as well as increase the one-health approach.

Serological and molecular surveillance of West Nile virus in domesticated mammals of peninsular Malaysia.

West Nile virus is a mosquito-borne neurotropic pathogen with a wide host range that constitutes a significant risk to public and animal health. There is limited information on WNV infection in domesticated mammals in Malaysia; however, current reports indicate infections in birds, macaques, bats and pigs from Malaysia. In this study, 203 serum samples from cattle, goats, and horses were tested for the presence of anti-WNV IgG using a competitive enzyme-linked immunosorbent assay (c-ELISA). Additionally, using one-step RT-PCR, nasopharyngeal swabs were analyzed for WNV RNA from all 203 animals in this study. The WNV seroprevalence was 32.53% (27/83) at 95% CI (0.2342-0.4319) in cattle, 48.27% (14/29) at 95% CI (0.3139-0.6557) in goats and 53.84% (49/91) at 95% CI (0.4366-0.6373) in horses. Cross-reactive JEV antibodies were detected in two cattle and 34 horses. None of the cattle or goats tested positive for WNV RT-PCR. Seven horses were positive for WNV RT-PCR, a molecular prevalence of 7.69% (7/91) at 95% CI (0.0353-0.1528). This is the first reported detection of WNV in domesticated mammals of Malaysia, a significant addition to the growing evidence that WNV is being transmitted from vectors to susceptible hosts in Malaysia.

Seroprevalence and associated risk factors of peste des petits ruminants among ovine and caprine in selected districts of Afar region, Ethiopia.

BACKGROUND: A Peste des petits ruminant is an acute, highly contagious and economically important transboundary viral disease of small ruminants. Despite the fact that food and agriculture organization and world organization for animal health plan to eradicate the disease by 2030, some studies indicated an increasing seropositivity of PPR infection in sheep and goats in Ethiopia. A cross-sectional study was employed to estimate the seroprevalence of PPR and to assess risk factors during the study period, February to April, 2020. Following purposive selection of the study districts, simple random sampling technique was employed to select individual animal during sample collection. A total of 384 serum samples were collected from apparently healthy sheep and goats. Competitive Enzyme Linked Immunosorbent Assay was used to detect the presence of antibodies against PPR at national veterinary institute. Descriptive statistics, Pearson's chi-square (X2) and logistic regression analysis were used is this study. RESULTS: The overall animal level seroprevalence of PPR virus was found to be 60.15% (n = 231/384) and species level prevalence rate was found to be 38.18% (n = 42) in sheep and 68.98% (n = 189) in goats in the study areas. Among the associated risk factors considered; species, sex, age and herd sizes were significantly associated (P < 0.05) with the disease occurrence. Among the associated risk factors considered in this study, species, sex, age and herd size were found to be statistically associated with the seropositivity of PPR infection. CONCLUSION: The present study finding revealed that a higher seroprevalence of PPR virus infection and this confirms peste des petits ruminant virus is circulating in Afar region. Further studies should be carried out on the entire region to determine PPR seroprevalence and to develop appropriate control and eradication strategies of PPR disease.

Evaluation of diagnostic performance of H-based blocking ELISA for specific detection of peste des petits ruminants in domestic sheep, goats, cattle and camels.

INTRODUCTION: Peste des petits ruminants virus (PPRV) causes a highly devastating disease of sheep and goats, peste des petits ruminants (PPR), which is targeted for global control and eradication by 2030. The serological diagnostic tool kits for accurate diagnosis of PPR have inherent strengths and weaknesses that require parallel validation and optimization across animal species. Thus, the objective of this study was to evaluate diagnostic performance of haemagglutinin based PPR blocking ELISA (HPPR- b-ELISA), that was developed by Africa Union Pan African Veterinary Vaccine Center for specific detection of anti- PPRV antibodies. METHODS: In preliminarily investigation, diagnostic performance of the HPPR-b-ELISA®, commercial PPR competition ELISA (c-ELISA) and virus neutralization test (VNT) were compared for the detection of anti-PPRV antibodies in goats, sheep, cattle and camels. RESULTS: The sensitivity and specificity of HPPR- b-ELISA® were 79.55 and 99.74%, respectively, compared to c-ELISA. The HPPR- b-ELISA® was in perfect agreement (κ = 0.86) with the c-ELISA in all sera collected from goats, sheep and cattle. There was almost perfect agreement between the species of goats (κ = 0.82) and sheep (κ = 0.98), while the agreement was substantial in cattle (κ = 0.78) and no agreement was observed in camels (κ = 0.00). Similarly, the sensitivity and specificity of the HPPR b-ELISA were 80 and 96.36%, respectively compared to VNT with almost perfect agreement in goats (κ = 0.83) and sheep (κ = 0.89), moderate in cattle (κ = 0.50) and none in camels (κ = 0.00). CONCLUSION: Our study revealed that HPPR- b-ELISA is a suitable and valid method that can alternatively be used for screening and monitoring of PPR in sheep, goats and cattle except for camels.

Seroprevalence of Bluetongue Virus Antibodies in Ovine in Maji District of West Omo Zone, Southwest Ethiopia.

Background: Bluetongue (BT) disease is an arthropod-transmitted viral disease of domestic and wild ruminant species caused by Bluetongue virus (BTV). It is of most importance in sheep and endemic primarily in the tropical and subtropical regions where vectors (Culicoides species) are present. Materials and Methods: A cross-sectional study was conducted in July-November 2019 to examine the seroprevalence of BTV infection in ovine in Maji district of West Omo zone. Serum samples were examined for the presence of specific antibodies of BTV using competitive enzyme-linked immunosorbent assay (c-ELISA) test. The collected data was coded and analyzed using STATA version 13 software. Associations between sero-prevalence and its risk factors were tested in a Chi-square analysis and with a P<0.05 were considered as statistically significant. Results: The individual animal prevalence was revealed as 39.23% (153/390). Herd size prevalence was: small size herd (37.42%; 61/163), medium size herd (32.35%; 55/170), and large size herd (64.91%; 37/57). Species-based prevalence showed ovine (38.00%; 141/371) and caprine (63.15%; 12/19). Age-based prevalence revealed adult (39.26%; 150/382) and young (37.5%; 3/8). The cumulative sex prevalence for both ovine and caprine was male (37.95%; 52/137) and female (39.92%; 101/253). Conclusion: The current prevalence of BTV antibodies in the area was found to be high. Lack of application of bluetongue disease control mechanisms like vaccination for the animals is a key factors for the high prevalence of the disease in the areas besides the existence of the vectors.

Peste des petits ruminants virus antibodies in domestic large ruminants in Bangladesh.

INTRODUCTION: Peste des petits ruminants (PPR) is an important transboundary animal disease of small ruminants which causes serious damage to the livelihood and food security of millions of small-scale farmers. PPR is endemic in goats in Bangladesh since 1993. The aim of this study was to determine the seroprevalence of PPR in sheep, cattle, and buffaloes in Bangladesh. METHODOLOGY: A total of 434 blood samples from sheep (n = 100), cattle (n = 190) and buffalo (n = 144) were collected aseptically. Sera were separated and antibody titer was determined using a commercially available c-ELISA kit. RESULTS: The overall seroprevalence was 16% and 3.68% in sheep and cattle, respectively, while buffaloes had a considerably higher seroprevalence of 42.36%. The study suggests that buffaloes are more prone to the PPR virus (PPRV) infection and cattle. CONCLUSIONS: This study provides serological evidence of PPRV infection in cattle and buffaloes. These results may warrant further studies to find out the role of large ruminants in transmitting PPRV infection to small ruminants and vice versa and inclusion of all domestic and wild ruminants for regular surveillance program.

Seroprevalence of Brucellosis in goats in some selected areas of Bangladesh.

OBJECTIVE: This study aimed to explore the seroprevalence of Brucella spp. in goats in some selected areas of Bangladesh. MATERIALS AND METHODS: This study was conducted in different goat-populated regions of Bangladesh from July 2017 to June 2018. A total of 208 serum samples were randomly collected from goats in Jashore (n = 50), Jhenidah (n = 22), Tangail (n = 40), Savar (n = 46), Thakurgaon (n = 18), and Bandarban (n = 32) areas. The samples were subjected to determine the presence of antibodies against Brucella spp. by rose bengal plate test (RBPT) and competitive enzyme-linked immunosorbent assay (c-ELISA). RESULTS: Overall, the seroprevalence of Brucellosis in goats was 4.33% (n = 9/208) by RBPT and 2.40% (n = 5/208) by c-ELISA. The seroprevalence of brucellosis on the basis of RBPT was 6% (buck: 0%, doe: 6%) in Jashore, 4.5% (buck: 0%, doe: 4.5%) in Jhenidah, 2.5% (buck: 0%, doe: 2.5%) in Tangail, 4.35% (buck: 0%, doe: 4.35%) in Savar, 6.25% (buck: 0%, doe: 6.25%) in Bandarban, and 5.56% (buck: 0%, doe: 5.56%) in Thakurgaon. On the other hand, the seroprevalence of brucellosis by c-ELISA was 4% (buck: 0%, doe: 4%) in Jashore, 4.5% (buck: 0%, doe: 4.5%) in Jhenidah, 3.13% (buck: 0%, doe: 3.13%) in Bandarban, and 5.56% (buck: 0%, doe: 5.56%) in Thakurgaon. Brucellosis was more prevalent (p > 0.001) in does aging 3-4 years. CONCLUSION: Goats from different areas of Bangladesh are caring antibodies against Brucella organisms. Further bacteriological investigations are necessary.

Exposure to Zoonotic West Nile Virus in Long-Tailed Macaques and Bats in Peninsular Malaysia.

The role of wildlife such as wild birds, macaques, and bats in the spreading and maintenance of deadly zoonotic pathogens in nature have been well documented in many parts of the world. One such pathogen is the mosquitoes borne virus, namely the West Nile Virus (WNV). Previous research has shown that 1:7 and 1:6 Malaysian wild birds are WNV antibody and RNA positive, respectively, and bats in North America may not be susceptible to the WNV infection. This study was conducted to determine the status of WNV in Malaysian macaques and bats found in mangrove forests and caves, respectively. Archive sera and oropharyngeal swabs from long-tailed macaques were subjected to the antibody detection using WNV competitive enzyme-linked immunosorbent assay (c-ELISA) and WNV RNA using RT-PCR, respectively, while the archive oropharyngeal and rectal swabs from bats were subjected to RT-PCR without serological analysis due to the unavailability of serum samples. The analysis revealed a WNV seropositivity of 29.63% (24/81) and none of the macaques were positive for WNV RNA. Meanwhile, 12.2% (5/41) of the bats from Pteropodidae, Emballonuridae, and Rhinolophidae families tested positive for WNV RNA. Here, we show a high WNV antibody prevalence in macaques and a moderate WNV RNA in various Malaysian bat species, suggesting that WNV circulates through Malaysian wild animals and Malaysian bat species may be susceptible to the WNV infection.

Prevalence and Distribution of Avian Influenza Viruses in Domestic Ducks at the Waterfowl-Chicken Interface in Wetlands.

Ducks are a natural reservoir of influenza A viruses (IAVs) and can act as a reassortment vessel. Wetlands, such as Hakaluki and Tanguar haor in Bangladesh, have unique ecosystems including domestic duck (Anas platyrhynchos domesticus) rearing, especially household and free-range ducks. A cross-sectional study was, therefore, conducted to explore avian influenza status and its distribution and risk factors in the wetland areas. During the three consecutive winters of 2015-2017, specifically in December of these years, we collected a total of 947 samples including blood, oropharyngeal and cloacal swabs from domestic ducks (free-range duck (n = 312 samples) and household ducks (n = 635 samples) in wetlands. We screened serum samples using a nucleoprotein competitive enzyme-linked immunosorbent assay (c-ELISA) to estimate seroprevalence of IAV antibodies and swab samples by real-time reverse transcriptase polymerase chain reaction (rRT-PCR) to detect IA viral M gene. Eleven (11) M gene positive samples were subjected to sequencing and phylogenetic analysis. Serological and viral prevalence rates of IAVs were 63.8% (95% CI: 60.6-66.8) and 10.7% (8.8-12.8), respectively. Serological and viral RNA prevalence rates were 51.8% (95% CI: 47.2-56.4) and 10.2% (7.6-13.3) in Hakaluki haor, 75.6% (71.5-79.4) and 11.1% (8.5-14.3) in Tanguar haor, 66.3% (62.5-69.9) and 11.2% (8.8-13.9) in household ducks and 58.7% (52.9-64.2) and 9.6% (6.5-13.4) in free-range ducks, respectively. The risk factors identified for higher odds of AI seropositive ducks were location (OR = 2.9, 95% CI: 2.2-3.8, p < 0.001; Tanguar haor vs. Hakaluki haor), duck-rearing system (OR = 1.4, 1.1-1.8, household vs. free-range), farmer's education status (OR = 1.5, 1.2-2.0, p < 0.05 illiterate vs. literate) and contact type (OR = 3.0, 2.1-4.3, p < 0.001; contact with chicken vs. no contact with chicken). The risk factors identified for higher odds of AI RNA positive ducks were farmer's education status (OR = 1.5, 1.0-2.3, p < 0.05 for illiterate vs literate), contact type (OR = 2.7, 1.7-4.2, p < 0.001; ducks having contact with chicken vs. ducks having contact with waterfowl). The phylogenetic analysis of 11 partial M gene sequences suggested that the M gene sequences detected in free-range duck were very similar to each other and were closely related to the M gene sequences of previously reported highly pathogenic avian influenza (HPAI) and low pathogenic avian influenza (LPAI) subtypes in waterfowl in Bangladesh and Southeast Asian countries. Results of the current study will help provide significant information for future surveillance programs and model IAV infection to predict the spread of the viruses among migratory waterfowl, free-range ducks and domestic poultry in Bangladesh.

Serological Evidence of West Nile Virus in Wild Birds in Bangladesh.

West Nile Virus (WNV) is a vector-borne zoonotic disease maintained in a sylvatic cycle involving mosquito vectors and birds. To detect WNV and other flavivirus infections in wild resident and migratory birds, we tested 184 samples from 19 identified species within nine families collected during 2012-2016 from four districts in Bangladesh. We tested serum samples for the immunoglobulin G (IgG) antibody against WNV using competitive Enzyme-Linked Immunosorbent Assay (c-ELISA), whereas tracheal and cloacal swabs were subjected to consensus Polymerase Chain Reaction (c-PCR) for the detection of the flavivirus RNA. Overall, we detected 11.9% (n = 22; 95% CI: 0.07-0.16) samples were seropositive, including 15.9% in the migratory wild birds and 10.7% in the resident wild birds. The migratory wild Tufted duck showed 28.5% seropositivity, whereas the resident wild house crows showed 12.5% seropositivity. None of the swab samples was positive for flavivirus RNA infection (0%, n = 184; 95% CI: 0-0.019). These study findings recommend continued surveillance for early detection and to better understand the epidemiology of WNV and other flavivirus circulation in both birds and mosquitoes in Bangladesh.

Serological Evidence of Avian Influenza in Captive Wild Birds in a Zoo and Two Safari Parks in Bangladesh.

Avian influenza (AI) is endemic and frequently causes seasonal outbreaks in winter in Bangladesh due to high pathogenic avian influenza (HPAI) H5N1 and low pathogenic avian influenza (LPAI) H9N2. Among avian influenza A viruses (AIV), H5, H7, and H9 subtypes have the most zoonotic potential. Captive birds in zoos and safari parks are used for educational, recreational, breeding, and conservational purposes in Bangladesh. To screen for AIV in captive birds to assess potential public health threats, we conducted a cross-sectional study in two safari parks and one zoo in Bangladesh for four months, from November to December 2013 and from January to February 2014. We collected blood samples, oropharyngeal, and cloacal swabs from 228 birds. We tested serum samples for AIV antibodies using competitive enzyme-linked immunosorbent assay (c-ELISA) and AIV sero-subtype H5, H7, and H9 using hemagglutination inhibition (HI) test. Swab samples were tested for the presence of avian influenza viral RNA using real-time reverse transcription-polymerase chain reaction (rRT-PCR). Across all the samples, AIV antibody prevalence was 9.7% (95% CI: 6.1-14.2, n = 228) and AIV HA subtype H5, H7 and H9 sero-prevalence was 0% (95% CI: 0-1.6, n = 228), 0% (95% CI: 0-1.6, n = 228) and 6.6% (95% CI: 3.72-10.6, n = 228), respectively. No AI viral RNA (M-gene) was detected in any swab sample (0%, 95% CI: 0-1.6, n = 228). Birds in the Safari park at Cox's Bazar had a higher prevalence in both AIV antibody prevalence (13.5%) and AIV H9 sero-prevalence (9.6%) than any of the other sites, although the difference was not statistically significant. Among eight species of birds, Emu (Dromaius novaehollandiae) had the highest sero-positivity for both AIV antibody prevalence (26.1%) and AIV H9 prevalence (17.4%) followed by Golden pheasant (Chrysolophus pictus) with AIV antibody prevalence of 18.2% and AIV H9 prevalence of 11.4%. Our results highlight the presence of AI antibodies indicating low pathogenic AIV mingling in captive birds in zoos and safari parks in Bangladesh. Continuous programmed surveillance is therefore recommended to help better understand the diversity of AIVs and provide a clear picture of AI in captive wild birds, enabling interventions to reduce the risk of AIV transmission to humans.

Evidence of West Nile virus infection in migratory and resident wild birds in west coast of peninsular Malaysia.

West Nile virus (WNV) is a zoonotic mosquito-borne flavivirus that is harbored and amplified by wild birds via the enzootic transmission cycle. Wide range of hosts are found to be susceptible to WNV infection including mammals, amphibians and reptiles across the world. Several studies have demonstrated that WNV was present in the Malaysian Orang Asli and captive birds. However, no data are available on the WNV prevalence in wild birds found in Malaysia. Therefore this study was conducted to determine the serological and molecular prevalence of WNV in wild birds in selected areas in the West Coast of Peninsular Malaysia. Two types of wild birds were screened, namely migratory and resident birds in order to explore any possibility of WNV transmission from the migratory birds to the resident birds. Thus, a cross-sectional study was conducted at the migratory birds sanctuary located in Kuala Gula, Perak and Kapar, Selangor by catching 163 migratory birds, and 97 resident birds from Kuala Gula and Parit Buntar, Perak at different time between 2016 and 2017 (Total, n = 260). Blood and oropharyngeal swabs were collected for serological and molecular analysis, respectively. Serum were screened for WNV antibodies using a commercial competitive ELISA (c-ELISA) (ID Screen® West Nile Competition Multi-species ELISA, ID VET, Montpellier, France) and cross-reactivity towards Japanese Encephalitis virus (JEV) was also carried out using the JEV-double antigen sandwich (DAS) ELISA. Oropharyngeal swabs were subjected to one-step RT-PCR to detect WNV RNA, in which positive reactions were subsequently sequenced. WNV seropositive rate of 18.71% (29/155) at 95% CI (0.131 to 0.260) and molecular prevalence of 15.2% (16/105) at 95% CI (0.092 to 0.239) were demonstrated in migratory and resident wild birds found in West Coast Malaysia. Phylogenetic analyses of the 16 WNV isolates found in this study revealed that the local strains have 99% similarity to the strains from South Africa and were clustered under lineage 2. Evidence of WNV infection in resident and migratory birds were demonstrated in this study. As a summary, intervention between migratory birds, resident birds and mosquitoes might cause the introduction and maintenance of WNV in Malaysia, however the assumption could be further proven by studying the infection dynamics in the mosquitoes present in the studied areas.

Evaluation of newly developed 'six recombinant secretary proteins based 'cocktail ELISA' and 'whole cell lysate' based 'indigenous ELISA' and tissue microscopy' with 'Gold standard' histo-pathology for the diagnosis of Johne's disease in slaughtered goats and buffaloes.

Three screening tests {(newly developed, six recombinant secretory proteins based 'cocktail ELISA', in-house robust 'indigenous ELISA' based on semi-purified protoplasmic antigens and tissue microscopy were evaluated with 'Gold standard', histo-pathology for the diagnosis of Johne's disease in goats and buffaloes. Serum and tissues {mesenteric lymph nodes and intestines) were driven from farmer's goats (n = 77) and buffaloes (n = 40) slaughtered for harvesting meat and farm goats (n = 77), died and necropsied. Twenty seven (35%) goats and 23 (57.5%) buffaloes were positive in all the four tests. Of 134 tissues screened by histo-pathology, 79.8% MLN and 76.8%, intestines, were positive for MAP infection. In tissue microscopy, 55.2 and 52.3%, goats and buffaloes were positive, respectively. Of 117 sera screened by i_ELISA, 58.4 and 70.0%, goats and buffaloes were positive, respectively. Whereas, c_ELISA detected 55.8 and 62.5%, goats and buffaloes, positives, respectively. Twelve tissues (70.5%) of goats necropsied were positive, both in tissue microscopy and histo-pathology. Most significant gross findings were serous atrophy of the fat and mild to moderate, diffuse thickening of terminal ileum, especially at ileo-caecal junction with or without transverse / longitudinal corrugations. In histo-pathology grade III and IV lesions were significantly low as compared to grade I and II. Of the four tests used for screening 268 samples, histo-pathology was most sensitive (78.3%), followed by i_ELISA (62.3%), c_ELISA (58.9%) and tissue microscopy (58.9%). Between two ELISA tests, c_ELISA using six recombinants secretory proteins, had higher specificity as compared to i_ELISA.

Rift Valley fever virus: a serological survey in Libyan ruminants.

A serological survey was carried out in Libya to investigate the circulation of Rift Valley fever virus (RVFV) among domestic ruminants. A total of 857 serum samples were collected from year 2015 to 2016 in eleven provinces of Libya belonging to five branches of the country. Samples were tested for RVFV antibodies using a competitive Enzyme-Linked Immunosorbent Assay (c-ELISA). Antibodies specific for RVFV were not detected in any of the 857 samples. However, a statistical analysis was carried out to assess the maximum expected number of infected animals and the maximum expected prevalence of RVFV among Libyan ruminants' populations according to the sampled population. The overall maximum expected prevalence was estimated to be 1.8% for cattle and 0.4% for small ruminants. Results seem to exclude the circulation of RVFV, however, a surveillance plan should be implemented in areas at risk of RVFV introduction.

Serological Prevalence of Contagious Bovine Pleuropneumonia in Niger in 2017.

Contagious bovine pleuropneumonia (CBPP) is a highly contagious disease of cattle caused by Mycoplasma mycoides subsp. mycoides Biotype Small Colony (MmmSC). The disease currently occurs in most of sub-Saharan Africa and where it is endemic and a major constraint for improving pastoral productivity. Following the persistence of this scourge, and in order to control this disease, a serological survey was conducted to determine the prevalence of CBPP in Niger. In fact, 1,590 sera were collected following a stratified sampling plan based on the risk factor of dissemination of CBPP. The analysis were performed at the Central Livestock Laboratory using the c-Elisa test. The results obtained show a wide distribution of the disease with an overall prevalence of 4.15% at individual level. The highest prevalences were recorded in the South-East regions [Zinder (7.5%), Diffa (7.5%)] and the West part [Tahoua (6.9%)]. The prevalence at the commune level was about 36.55%, which was relatively high. The prevalence at strata level was 36.55% (95% PI 0.2428-0.4882). The expected prevalences did not match those found. The results of this serological survey are considered the reference situation (T0) of CBPP in Niger with the PRAPS project, and allowed to the country to redefine control policies for better control of the disease at national and sub-regional level.

Bluetongue disease in small ruminants in south western Ethiopia: cross-sectional sero-epidemiological study.

OBJECTIVE: The status of bluetongue disease, vectors for transmission of the disease and the serotypes involved are not clearly known in Ethiopia. This sero-epidemiological study was conducted to determine the seroprevalence and associated risk factors of bluetongue in small ruminants of South Western Ethiopia. RESULT: 422 serum samples were screened for the presence of bluetongue virus (BTV) specific antibodies using competitive enzyme-linked immunosorbent assay (c-ELISA) and 30.6% (129/422) (confidence interval CI 26.2-35%) of the sheep and goat serum samples were found positive. Multivariate analysis of several risk factors like age, sex, altitude, body condition and species of animals were studied and it was observed that species of animals, age and altitude had significant influence (P < 0.05) on seropositivity to BTV. Goats showed more seropositivity to bluetongue as compared to sheep [AOR = 2.4, 95% CI (1.5-3.9), P = 0.001], adult animals were more seropositive [AOR = 3.1, 95% CI (1.9-5.1), P = 0.001] than other age groups and animals at the lowland [AOR = 3.1, 95% CI (1.5-6.4), P = 0.002] showed more seropositivity to bluetongue than midland and high land. Sex and body condition of the animals had no statistically significant (P > 0.05) effect on seropositivity to bluetongue.

Evaluation of three serological tests for diagnosis of bovine brucellosis in smallholder farms in Pakistan by estimating sensitivity and specificity using Bayesian latent class analysis.

The present study aimed to estimate the herd-level sensitivity (Se) and specificity (Sp) of three commonly used serological tests in naturally-infected cattle and buffalo in smallholder farms in Pakistan. Between February and June 2015, a cross-sectional study was carried out in five districts of Punjab (Kasur, Okara, Pakpattan, Jhelum, and Bhakkar) and two districts of Sindh (Badin and Thatta). Serum samples from mixed farms of cattle (n=441) and buffalo (n=621) were collected and tested using the Rose Bengal test (RBT), indirect ELISA (I-ELISA) and competitive ELISA (C-ELISA). A Bayesian latent class analysis (LCA) approach was used to estimate the Se and Sp of these three serological tests and the true herd-level prevalence in each district. The model was fitted under the assumption of conditional independence between three tests and also conditional dependence by including covariances between the two ELISAs. In addition, the model was fitted using three different shapes of beta distributions to incorporate prior information in the model. The test with the highest Se was the C-ELISA, with a range from 76.3% (95% PCI (Posterior Credibility Interval), 62.6-88.2%) to 81.4% (95% PCI, 68.2-92.8%). The RBT was found to have the highest Sp (99.1-99.4%) of the tests. The highest estimated herd-level prevalence, 45% (95% PCI, 32-59%), was found in Jhelum district and the lowest in Thatta district, 1.1% (95% PCI 0.04-6.0%). The results of this study identified some discrepancy between the published literature on the level of Se of these tests, especially for RBT. It appears that RBT has lower Se and higher Sp when used in the field conditions of the present study. Consequently, it is recommended that none of the evaluated tests should be performed in isolation for the diagnosis of bovine brucellosis in the field conditions of Pakistan, but the use of tests in combination, with RBT and C-ELISA used in parallel returning optimal Se and Sp, is warranted.

Development of Competitive Enzyme-Linked Immunosorbent Assays for Antibody Detection Based on Bluetongue Virus Monoclonal Antibodies.

Bluetongue is a ruminant infectious disease that is characterized by hyperpyrexia, leukocyte decrease and mucosal ulcerative inflammation changes. In this study, three segments of Bluetongue virus (BTV)-8 VP2 protein (BTV-8A, 8B, and 8C) were cloned into pET-28a (+) and pMAl-c5X vectors and expressed in Escherichia coli BL21 (DE3) as histidine (His)-tagged (His-8A/8B/8C) and maltose-binding protein (MBP)-tagged (MBP-8A/8B/8C) fusion proteins. After purification, His-8A/8B/8C were used to immunize BALB/mice and MBP-8A/8B/8C were used to screen for monoclonal antibody (mAb)-secreting hybridomas. Two mAbs (8B-5D4 and 8B-3G11) that could recognize BTV-8 were obtained. Two competitive enzyme-linked immunosorbent assays with good specificity and sensitivity using mAbs 8B-5D4 or 8B-3G11 as competitive antibody were established. With the joint reaction of these methods, serum samples containing anti-BTV-7 or anti-BTV-8 antibody could be screened out, suggesting that these methods would be useful for the diagnosis and epidemiological studies of BTV-8.