Serological and molecular evidence of bluetongue in sheep and goats in Uttar Pradesh, India

Cross-sectional experimental study was conducted with the objective to estimate the seroprevalence on the basis of antibodies to VP7 protein of bluetongue virus by competitive enzyme linked immunosorbent assay (c-ELISA), to test the neutralizing ability of the antibody to reference strains of 4 BTV serotypes (BTV-1, 2, 10 and 23) by micro serum neutralization assay (m-SNT), to check the presence of BTV dsRNA and to isolate and characterize bluetongue virus (BTV). A total of 91 serum and 26 whole blood samples were obtained from sheep and goat. The study was conducted between September and November, 2012 when the culicoides midges’ activity is maximal. The animals were observed for clinical signs of BTV infection and serum samples were obtained from all animals for c-ELISA and m-SNT. Further, blood samples were collected from the c-ELISA positive animals and subjected to virus isolation and nested RT-PCR. Out of 91 animals tested, 26 (28.6%) were found to be seropositive by c-ELISA and one sheep showed neutralizing antibody against BTV-1 serotype at a titer of 1.2 log10. Multivalent logistic regression analysis of risk factors like age, sex, body condition and species of animals were considered during the serological study and found that species of animal had significant influence (χ2 = 17.111, P<0.05) in seropositivity of BTV. Goats showed more seropositivity to bluetongue as compared to sheep (OR = 0.233). Other risk factors had no significant influence (P>0.05) on seropositivity. It was worth enough to conclude that higher seroprevalence among goats indicated that goats would be the most important animals in the epidemiology of BTV with less clinical manifestation due to development of acquired immunity as the result of continuous exposure.Keywords: Bluetongue virus (BTV), competitive enzyme linked immunosorbent assay (c-ELISA), goat, seroprevalence, sheep, micro serum neutralization assay (m-SNT), nested reverse transcription polymerase chain reaction (RT-PCR)African Journal of Biotechnology Vol. 12(19), pp. 2699-270

Differential cytokine expression profile in peripheral blood mononuclear cells of endometritic buffaloes

The aim of present study was to elucidate the relative mRNA expression of certain cytokines, viz. interleukin- 1ß (IL-1ß), IL-8, tumor necrosis factor alpha (TNFa) and IL-4 in peripheral blood mononuclear cells (PBMCs) of endometritic buffaloes. Out of 15 buffaloes, 9 buffaloes were diagnosed positive for endometritis whereas 6 buffaloes having no symptoms of endometritis were considered as healthy control. The blood samples were collected from each animal during diagnosis of endometritis and PBMCs were separated from collected blood samples by density gradient centrifugation. Total RNA was extracted from the cultured PBMCs isolates, reverse transcribed to synthesize the cDNA and amplified in real time PCR system. The relative fold of expression was estimated using 2-∆∆Ct method. An up-regulation in all 3 pro-inflammatory cytokines (IL-1ß, IL-8 and TNFa) mRNA, ranging from 1.92 to 3.27 fold was observed in buffaloes with endometritis as compared to healthy. However, no alteration was detected in anti-inflammatory cytokine, IL-4 expression. Thus, it is concluded that certain cytokines expressed differentially in PBMCs in buffaloes with endometritis, that is first ever information in this species. Nevertheless, further research in respect to threshold level of each target cytokine in peripheral circulation involving more number of animals is required to establish these potential marker gene(s) for diagnosis of endometritis and for monitoring of new therapeutic approaches

Dodatak supranutritivnih doza cinkova sulfata i bakterija Bacillus firmus ubijenih toplinom u obrok rano odbijenih svinja: utjecaj na rast, funkciju neutrofila i upalne citokine

The objective of the study was to investigate the effect of supplementation of a supranutritional dose of zinc sulfate (ZnSO4) and Bacillus firmus derived bio-response modifier (BRM) on growth, blood neutrophil functions, pro-inflammatory and anti-inflammatory cytokine responses in early weanling piglets. In total, 45 piglets (age of 19.25 ± 0.84 days) were randomly divided into five groups: I (basal diet only), II (basal diet supplemented with ZnSO4), III (basal diet supplemented with BRM), IV (basal diet supplemented with ZnSO4 plus BRM) and V (basal diet without weaning from dam). The production of myeloperoxidase (MPO) and superoxide anion (O2-), and the concentration of transforming growth factor-β1 (TGF-β1) were markedly reduced, whereas the concentrations of intercellular adhesion molecule 1 (ICAM1), and monocyte chemoattractant protein 1 (MCP1) were significantly (P0.05) but a marked increase (P0,05), ali kod prasadi iz skupine II. i III. je 14. i 7. dana opaženo znakovito povišenje (P<0,05) ICAM1, MCP1, te sniženje koncentracije TGF-β1. U prasadi iz skupine IV dodavanje BRM i ZnSO4 u osnovnom obroku poboljšalo je MPO (2. dan) i O2- (7. dan), bez znakovitih promjena u rastu i koncentraciji citokina. Na kraju, zaključeno je da dodavanje kombinacije BRM i ZnSO4 u obroku potiče urođenu imunost prasadi što nije slučaj kada se ZnSO4 ili BRM dodaju zasebno. Rezultati ovog istraživanja pomoći će u formuliranju učinkovitog upravljanja hranidbom kod rano odbijene prasadi u krdu svinja

Molecular characterisation and nucleotide sequence analysis of canine parvovirus strains in vaccines in India

Canine parvovirus 2 (CPV‑2) is one of the most important viruses that causes haemorrhagic gastroenteritis and myocarditis of dogs worldwide. The picture has been complicated further due to the emergence of new mutants of CPV, namely: CPV‑2a, CPV‑2b and CPV‑2c. In this study, the molecular characterisation of strains present in the CPV vaccines available on the Indian market was performed using polymerase chain reaction and DNA sequencing. The VP1/VP2 genes of two vaccine strains and a field strain (Bhopal) were sequenced and the nucleotide and the deduced amino acid sequences were compared. The results indicated that the isolate belonged to CPV type 2b and the strains in the vaccines belonged to type CPV‑2. From the study, it is inferred that the CPV strain used in commercially available vaccine preparation differed from the strains present in CPV infection in dogs in Indi

Neutrophil Functions and Cytokines Expression Profile in Buffaloes with Impending Postpartum Reproductive Disorders

The study was conducted to correlate the periparturient immune status in terms of neutrophil functions and cytokine expression in peripheral blood mononuclear cell culture with impending postpartum reproductive disorders in buffaloes. Forty pregnant buffaloes were observed for occurrence of postpartum reproductive disorders (PRD), i.e., metritis, endometritis and delayed uterine involution etc., during one week prepartum to four weeks postpartum period. A representative number (n = 6) of buffaloes that did not develop any PRD were included in group I (healthy, control), while the animals which experienced PRD were assigned into group II (PRD, n = 8). The blood samples were collected at weekly interval from one week prepartum to four weeks postpartum period considering the day of calving as ‘d 0’. Differential leucocytes counts, superoxide and hydrogen peroxide production activity in isolated neutrophils and the mRNA expression profile of cytokines i.e., IL-2, IL-4 and IFN-γ in PBMC culture were studied in all the samples. A higher total leucocytes, neutrophil and band cells count along with impaired neutrophil functions i.e., lowered level of production of superoxide and hydrogen peroxide before parturition and during early postpartum period were observed in buffaloes developing PRD. Further, a lower expression of IL-2, IFN-γ and IL-4 mRNA in PBMC culture was observed at calving in buffaloes that subsequently developed PRD at later postpartum. Thus, suppression in neutrophil function and cytokine expression at prepartum to early postpartum period predisposes the buffaloes to develop postpartum reproductive disorders. Hence, monitoring of neutrophils function and cytokine expression profile would be effective to predict certain reproductive disorders at late pregnancy or immediately after parturition in buffaloes. In future, this may be a novel approach for determining suitable management and therapeutic decisions for prevention of commonly occurring reproductive disorders in farm animals

Concurrent testing of breeding bulls for bovine herpesvirus 1 infection (BHV-1) in India

In this study, sera from 65 breeding and 19 training bulls from Uttar Pradesh State in north India were tested for bovine herpesvirus 1 (BHV-1) antibodies by enzyme linked immunosorbent assay (ELISA) and virus neutralization test (VNT). The VNT test could detect 56 (86.15%) and 9 (47.37%) of the samples from breeding and training bulls as positive for BHV-1 antibodies whereas in ELISA 63 (96.92%) and 10 (52.63%) were found positive, respectively. Semen samples from the breeding bulls were simultaneously tested by the Taqman based real time PCR (qPCR). Of the 65 samples screened, only 40 (61.54%) were found to contain BHV-1 DNA indicating that all the seropositive bulls are not shedding the virus in semen. When the RT-PCR positive samples were subjected to virus isolation on Madin-Darby bovine kidney (MDBK) cells, no virus isolates could be obtained. The advantages of concomitant testing of serum and semen of breeding bulls and measures for control of BHV-1 infections in bull farms are discussed

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

Abstract 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) effect on seropositivity to bluetongue

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Not AvailableBluetongue (BT) disease poses a constant risk to the livestock population around the world. A better understanding of the risk factors will enable a more accurate prediction of the place and time of high-risk events. Mapping the disease epizootics over a period in a particular geographic area will identify the spatial distribution of disease occurrence. A Geographical Information System (GIS) based methodology to analyze the relationship between bluetongue epizootics and spatial–temporal patterns was used for the years 2000 to 2015 in sheep of Andhra Pradesh, India. Autocorrelation (ACF), partial autocorrelation (PACF), and cross-correlation (CCF) analyses were carried out to find the self-dependency between BT epizootics and their dependencies on environmental factors and livestock population. The association with climatic or remote sensing variables at different months lag, including wind speed, temperature, rainfall, relative humidity, normalized difference vegetation index (NDVI), normalized difference water index (NDWI), land surface temperature (LST), was also examined. The ACF & PACF of BT epizootics with its lag showed a significant positive autocorrelation with a month’s lag (r = 0.41). Cross-correlations between the environmental variables and BT epizootics indicated the significant positive correlations at 0, 1, and 2 month’s lag of rainfall, relative humidity, normalized difference water index (NDWI), and normalized difference vegetation index (NDVI). Spatial autocorrelation analysis estimated the univariate global Moran’s I value of 0.21. Meanwhile, the local Moran’s I value for the year 2000 (r = 0.32) showed a high degree of spatial autocorrelation. The spatial autocorrelation analysis revealed that the BT epizootics in sheep are having considerable spatial association among the outbreaks in nearby districts, and have to be taken care of while making any forecasting or disease prediction with other risk factors.Not Availabl