BackgroundThe devastating viral disease of small ruminants namely Peste des petits ruminants (PPR) declared as target for “Global Eradication” in 2015 by the Food and Agriculture Organization (FAO) and the World Organization for Animal Health (OIE). For a successful eradication campaign, molecular diagnostic tools are preferred for their specificity, efficacy and robustness to compliment prophylactic measures and surveillance methods. However, molecular tools have a few limitations including, costly equipment, multi-step template preparation protocols, target amplification and analysis that restrict their use to the sophisticated laboratory settings. As reverse transcription-loop mediated isothermal amplification assay (RT-LAMP) has such an intrinsic potential for point of care diagnosis, this study focused on the genetic detection of causative PPR virus (PPRV) in field conditions. It involves the use of a sample buffer that can precipitate out virus envelope and capsid proteins through ammonium sulphate precipitation and exposes viral RNA, present in the clinical sample, to the LAMP reaction mixture.ResultsThe test was evaluated using 11 PPRV cultures, and a total of 46 nasal swabs (n = 32 collected in the field outbreaks, n = 14 collected from experimentally inoculated animals). The RT-LAMP was compared with the reverse transcription-PCR (RT-PCR) and real-time quantitative RT-PCR (RT-qPCR) for its relative specificity, sensitivity and robustness. RT-LAMP detected PPRV in all PPRV cultures in or less than 30 min. Its detection limit was of 0.0001TCID50 (tissue culture infective dose-50) per ml with 10-fold higher sensitivity than that of RT-PCR. In 59.4% of the field samples, RT-LAMP detected PPRV within 35–55 min. The analytical sensitivity and specificity of the RT-LAMP were equivalent to that of the RT-qPCR. The time of detection of PPRV decreased by at least forty minutes or 3–4 h in case of in the RT-LAMP as compared with the RT-qPCR and the RT-PCR, respectively.ConclusionsThe sensitive and specific RT-LAMP test developed in this study targeting a small fragment of the N gene of PPRV is a rapid, reliable and applicable molecular diagnostic test of choice under the field conditions. RT-LAMP requiring minimal training offers a very useful tool for PPR diagnosis especially during the “Global PPR Eradication Campaign”.Electronic supplementary materialThe online version of this article (doi:10.1186/s12917-016-0940-0) contains supplementary material, which is available to authorized users.
A study was conducted to evaluate three different probiotics, using drinking water supplemented with protexin, biovet and yoghurt. The day old broiler chicks were randomly divided into 12 separate floor pens each comprising 25 birds and three pens (replicates) per treatment group following completely randomised design. At 28 and 39 days of age body weight (BW) and feed to gain ratio (FCR) were determined. At the end of experiment, nine birds per treatment were sacrificed to evaluate carcass characteristics, abdominal fat contents and the internal organs. Blood haemato-biochemical parameters were determined. Haemagglutination inhibition antibody titres against Newcastle disease virus and lymphoid organs weight/body weight ratio were also determined. The BW of birds given probiotics was significantly greater than control (without probiotics) at both 28 and 39 days of age. Similarly, better FCR was observed in birds those given drinking water with probiotics. There was less mortality recorded with probiotics treatments. Differences in carcass characteristics, organs weight, meat composition and haematological values among all the treatments were non-significant. However, abdominal fat contents reduced significantly in probiotics supplemented groups as compared to control and cholesterol contents were reduced significantly supplemented groups as compared to control at both 21 and 39 days of age. Feeding of probiotics did positively affect the immune system within the parameters measured. It may be concluded that performance, blood chemistry, immunity against disease and economic efficiency in broilers could be maintained when supplementing any probiotic incorporated in broiler's drinking water.
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