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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.

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Trout production is a growing activity in recent years but requires new alternative sources of feed to be sustainable over time. The objective of this research was to determine the apparent digestibility coefficient (ADC) of dry matter (DM), organic matter (OM), crude protein (CP) and digestible energy (DE) of kañiwa (Chenopodium pallidicaule Aellen), kiwicha (Amaranthus caudatus L), quinoa (Chenopodium quinoa Willd), beans (Phaseolus vulgaris L.), sacha inchi, (Plukenetia volubilis L) and jumbo squid (Dosidicus gigas) meal in juvenile rainbow trout. The experimental diets were composed of a 70% basal diet and 30% of any raw materials. The ADC was determined by the indirect method using insoluble ash as a non-digestible marker. Jumbo squid, sacha inchi and quinoa showed the highest values of ADC (%) of DM (84.5, 73.5 and 69.7), OM (89.1, 78.4 and 72.9), CP (93.2, 98.0 and 90.3), and DE (4.57, 4.15 and 2.95 Mcal/kg DM), respectively. The ADC values for kañiwa, kiwicha and bean were significantly lower. In conclusion, quinoa meal and jumbo squid meal have an acceptable digestibility but sacha inchi meal is a potential alternative for rainbow trout feeding in the future.

Butyrivibrio fibrisolvens forms part of the gastrointestinal microbiome of ruminants and other mammals, including humans. Indeed, it is one of the most common bacteria found in the rumen and plays an important role in ruminal fermentation of polysaccharides, yet, to date, there is no closed reference genome published for this species in any ruminant animal. We successfully assembled the nearly complete genome sequence of B. fibrisolvens strain INBov1 isolated from cow rumen using Illumina paired-end reads, 454 Roche single-end and mate pair sequencing technology. Additionally, we constructed an optical restriction map of this strain to aid in scaffold ordering and positioning, and completed the first genomic structure of this species. Moreover, we identified and assembled the first chromid of this species (pINBov266). The INBov1 genome encodes a large set of genes involved in the cellulolytic process but lacks key genes. This seems to indicate that B. fibrisolvens plays an important role in ruminal cellulolytic processes, but does not have autonomous cellulolytic capacity. When searching for genes involved in the biohydrogenation of unsaturated fatty acids, no linoleate isomerase gene was found in this strain. INBov1 does encode oleate hydratase genes known to participate in the hydrogenation of oleic acids. Furthermore, INBov1 contains an enolase gene, which has been recently determined to participate in the synthesis of conjugated linoleic acids. This work confirms the presence of a novel chromid in B. fibrisolvens and provides a new potential reference genome sequence for this species, providing new insight into its role in biohydrogenation and carbohydrate degradation.