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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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Foot and mouth disease is an economically devastating disease of livestock that mainly effect cloven-hoofed animals i.e. sheep, goat, cattle, pig, buffalo, deer etc. The aim of this study was to determine the serotypes circulating in the region during 2016. Sampling was done from different outbreaks initially on the basis of clinical signs and later reverse transcriptase-polymerase chain reaction (RT-PCR) was employed for the confirmation of FMDV genome. Out of total 72 samples, 65 were found positive which were then serotyped into type O (n=30), Asia1 (n=19) and A (n=5). Some samples (n=5) were found positive for more than one serotype that were subjected to reverse transcriptase loop-mediated isothermal amplification assay (RT-LAMP) for serotype determination.

Antibiotics are routinely used in commercial poultry farms for the treatment of economically important bacterial diseases. Repeated use of antibiotics, usually administered in the feed or drinking water, may also result in the selection of resistant bacteria in animal feces, able to transfer their antimicrobial-resistance genes ( ARG ), residing on mobile elements, to other microorganisms, including human pathogens. In this study, single and multiplex PCR protocols were performed to detect tetracycline-, lincomycin-, chloramphenicol-, aminoglycoside-, colistin-, vancomycin-, and carbapenem-resistance genes, starting from 38 litter samples collected from 6 poultry and 2 turkey Italian flocks. The ARG were confirmed for all investigated classes of antimicrobials, except for colistin ( mcr-1, mcr-2, mcr-3,mcr-4 mcr-5 ) and carbapenem ( IMP, OXA-48, NDM , KPC ), while the vanB gene was only detected for vancomycin. The highest positivity was obtained for tetracycline ( tet[L], tet[M] , tet[K], tetA[P] ] and aminoglycoside ( aadA2 ) ARG, confirming the predominant use of these antimicrobials in the veterinary practice and their potential to enhance the resistance patterns also in humans as a consequence of environmental contamination. On the contrary, the dissemination by poultry of ARG for critically important antimicrobials seems to be of minor concern, suggesting a negligible environmental dissemination by these genes in the Italian poultry industry. Finally, the molecular screening performed in this study using a noninvasive sampling method represents a simple and rapid tool for monitoring the ARG patterns at the farm level.

Antimicrobial resistance is a complex and widespread problem threatening human and animal health. In poultry farms, a wide distribution of resistant bacteria and their relative genes is described worldwide, including in Italy. In this paper, a comparison of resistance gene distribution in litter samples, recovered from four conventional and four antibiotic-free broiler flocks, was performed to highlight any influence of farming systems on the spreading and maintenance of resistance determinants. Conventional PCR tests, targeting the resistance genes related to the most used antibiotics in poultry farming, along with some critically important antibiotics for human medicine, were applied. In conventional farms, n. 10 out of n. 30 investigated genes were present in at least one sample, the most abundant fragments being the tet genes specific for tetracyclines, followed by those for aminoglycosides and chloramphenicol. All conventional samples resulted negative for colistin, carbapenems, and vancomycin resistance genes. A similar trend was observed for antibiotic-free herds, with n. 13 out of n. 30 amplified genes, while a positivity for the mcr-1 gene, specific for colistin, was observed in one antibiotic-free flock. The statistical analysis revealed a significant difference for the tetM gene, which was found more frequently in the antibiotic-free category. The analysis carried out in this study allowed us to obtain new data about the distribution of resistance patterns in the poultry industry in relation to farming types. The PCR test is a quick and non-expensive laboratory tool for the environmental monitoring of resistance determinants identifying potential indicators of AMR dissemination.