In this study, we developed new sets of primers to detect Brucella spp. and M. avium subsp. paratuberculosis (MAP) through isothermal amplification. We selected a previously well-characterized target gene, bscp31, specific for Brucella spp. and IS900 for MAP. The limits of detection using the loop-mediated isothermal amplification (LAMP) protocols described herein were similar to those of conventional PCR targeting the same sequences. Hydroxynaphtol blue and SYBR GreenTM allowed direct naked-eye detection with identical sensitivity as agarose gel electrophoresis. We included the LAMP-based protocol in a rapid identification scheme of the respective pathogens, and all tested isolates were correctly identified within 2 to 3 h. In addition, both protocols were suitable for specifically identifying the respective pathogens; in the case of Brucella, it also allowed the identification of all the biovars tested. We conclude that LAMP is a suitable rapid molecular typing tool that could help to shorten the time required to identify insidious bacteria in low-complexity laboratories, mainly in developing countries.

The high fibrolytic activity and large biomass of strictly-anaerobic bacteria that inhabit the rumen makes them primarily responsible for the degradation of the forage consumed by ruminants. Llamas feed mainly on low quality fibrous roughages that are digested by an active and diverse microflora. The products of this fermentation are volatile fatty acids and microbial biomass, which will be used by the animals. The aim of this study was to detect the three major fiber-digesting anaerobic bacteria in the forestomach contents of llamas by PCR. In this study, we detected Ruminococcus albus, Ruminococcus flavefaciens and Fibrobacter succinogenes in the forestomach contents of eight native llamas from Argentina.

La alta actividad fibrolítica y la gran biomasa de las bacterias anaerobias estrictas que habitan el rumen las hacen las principales responsables de la degradación del forraje consumido por los rumiantes. Las llamas se alimentan sobre todo de forrajes fibrosos de baja calidad, que son digeridos por una activa y diversa microflora. Los productos de esta fermentación son ácidos grasos volátiles y biomasa bacteriana, los cuales serán utilizados por el animal. El objetivo de este estudio fue detectar las tres principales bacterias anaerobias que digieren fibra en el contenido del preestómago de llamas por PCR. En este estudio, detectamos Ruminococcus albus, Ruminococcus flavefaciens y Fibrobacter succinogenes en el contenido del preestómago de ocho llamas nativas de Argentina.