First characterization of the probiotic potential of lactic acid bacteria isolated from Costa Rican pineapple silages.

BACKGROUND: Agro-industrial waste from tropical environments could be an important source of lactic acid bacteria (LAB) with probiotic potential. METHODS: Twelve LAB isolates were isolated from pineapple silages. The species identification was carried out considering 16S rRNA and pheS genes. Experiments to evaluate the probiotic potential of the isolates included survival under simulated gastrointestinal environment, in vitro antagonistic activity (against Salmonella spp. and Listeria monocytogenes), auto-aggregation assays, antibiotic susceptibility, presence of plasmids, adhesiveness to epithelial cells, and antagonistic activity against Salmonella in HeLa cells. RESULTS: Lacticaseibacillus paracasei, Lentilactobacillus parafarraginis, Limosilactobacillus fermentum, and Weissella ghanensis were identified. Survival of one of the isolates was 90% or higher after exposure to acidic conditions (pH: 2), six isolates showed at least 61% survival after exposure to bile salts. The three most promising isolates, based on survivability tests, showed a strong antagonistic effect against Salmonella. However, only L. paracasei_6714 showed a strong Listeria inhibition pattern; this isolate showed a good auto-aggregation ability, was resistant to some of the tested antibiotics but was not found to harbor plasmids; it also showed a high capacity for adhesion to epithelial cells and prevented the invasion of Salmonella in HeLa cells. After further in vivo evaluations, L. paracasei_6714 may be considered a probiotic candidate for food industry applications and may have promising performance in acidic products due to its origin.

Silage quality and bacterial diversity of silages inoculated with Listeria monocytogenes and Lacticaseibacillus paracasei_6714

Introduction: King grass (Cenchrus purpureus (Schumach.) Morrone, syn. Pennisetum purpuphoides) and pineapple peel (Ananas comosus) silages are food alternatives for livestock in conditions of feed shortage. Objective: To describe the dynamics of the microbiota present in king grass and pineapple silage during the fermentation process using next generation sequencing (NGS) and to evaluate the protective effect of Lacticaseibacillus paracasei_6714 as a silage inoculum against Listeria monocytogenes. Methods: We used an unrestricted randomized design to characterize the microbiota present in silages made from king grass harvested 70 days after regrowth and pineapple peel. We inoculated mixtures of grass and peel with L. paracasei_6714 or L. monocytogenes, or both, with a non-inoculated treatment as control. The nutritional and fermentative profile was evaluated after 30 days. After 15 and 30 days of fermentation, we used 16S rRNA analysis to determine the dynamics and diversity of the microbiota in the inoculated and control silages. Result: Dry matter content and digestibility did not differ significantly; however, there were differences in crude protein, pH and organic acids. We obtained 4432 amplicon sequence variants of Proteobacteria, Firmicutes, Bacterioidetes, Actinobacteria, Verrucomicrobia, Planctomycetes and Patescibacteria. The relative abundance of each phylum varied depending on the material and fermentation period. Phylum similarity was over 70 % (but not greater than 50 % with Bray-Curtis at the species level). Conclusion: These bacterial communities seem to have an important role during silage fermentation. Proper management of silage processing can reduce or eliminate pathogenic bacteria.

Introducción: Los ensilajes del pasto king grass (Cenchrus purpureus (Schumach.) Morrone, syn. Pennisetum purpuphoides) y cáscaras de piña (Ananas comosus) son alternativas de alimento para ganado en condiciones de escasez alimentaria. Objetivo: Describir las dinámicas de la microbiota presente en los ensilajes de king grass y piña durante el proceso de fermentación usando secuenciación de próxima generación (NGS) y evaluar el efecto de protección de Lacticaseibacillus paracasei_6714 como inoculante de ensilaje ante Listeria monocytogenes. Métodos: Usamos un diseño aleatorio no restringido para caracterizar la microbiota presente en ensilajes de king Grass cosechados 70 días después de rebrote y de cáscaras de piña. Inoculamos mezclas de pasto y cáscara con L. paracasei_6714 o L. monocytogenes, o ambos, con un tratamiento control sin inocular. El perfil nutricional y de fermentación fue evaluado luego de 30 días. Después de 15 y 30 días de fermentación, usamos un análisis de para determinar la dinámicas y diversidad de la microbiota en los ensilajes inoculados y control. Resultados: Los contenidos de materia seca y digestibilidad, no difirieron significativamente; sin embargo, hubo diferencias en proteína cruda, pH y ácidos orgánicos. Obtuvimos 4 432 secuencias variantes de amplicon de Proteobacteria, Firmicutes, Bacterioidetes, Actinobacteria, Verrucomicrobia, Planctomycetes y de Patescibacteria. La abundancia relativa de cada filo vario dependiendo del material y periodo de fermentación. Similitudes de filo fueron mayores al 70 % (pero no mayor que 50 % con Bray-Curtis a nivel de especie). Conclusión: Estas comunidades bacterianas parecen cumplir un papel importante durante la fermentación del ensilaje. Un manejo apropiado del proceso de ensilaje puede reducir o eliminar baterías patogénicas.

Effects of coffee processing residues on anaerobic microorganisms and corresponding digestion performance.

The objective of this study was to delineate the effects of different coffee processing residues on the anaerobic microbes and corresponding digestion performance. The results elucidated that mucilage-rich feed enhanced the accumulation of methanogens, which consequently led to better digestion performance of biogas production. Fifty percent more methane and up to 3 times more net energy (heat and electricity) output were achieved by the digestion of the mucilage-rich feed (M3). The microbial community and statistical analyses further elucidated that different residues in the feed had significant impact on microbial distribution and correspondingly influenced the digestion performance.

Factores edáficos y nemátodos entomopatógenos en un agroecosistema neotropical de banano / Edaphic factors and entomopathogenic nematodes in a neotropical banana agroecosystem

Introducción: En Costa Rica el banano es uno de los cultivos más importantes en términos de producción y exportaciones, que requiere la aplicación de agroquímicos para controlar el efecto de plagas como el picudo negro, Cosmopolites sordidus. Debido a los efectos dañinos en la salud humana y ambiental, y la posibilidad de que las plagas presenten resistencia a estas sustancias, es necesario investigar alternativas amigables con el medio ambiente como potenciales controladores biológicos. Los nemátodos entomopatógenos (NEPs) de los géneros Heterorhabditis y Steinernema presentan una relación simbiótica con bacterias de los géneros Photorhabdus y Xenorhabdus respectivamente y este complejo ha sido ampliamente estudiado como controlador biológico debido a su capacidad de eliminar insectos hospederos en horas. Objetivo: Determinar la presencia de nemátodos de los géneros Heterorhabditis y Steinernema en suelos de plantaciones de banano del Caribe costarricense y zonas aledañas y caracterizar su hábitat. Metodología: Se tomaron muestras de suelo en plantaciones y áreas circundantes, a partir de las cuales, se llevó cabo aislamiento de NEPs y análisis químicos, físicos y microbiológicos (pH, acidez, materia orgánica, Ca, K, Mg, P, Fe, Cu, Zn, Mn, textura del suelo, biomasa y respiración microbiana). Se encontraron diferencias entre sitios de muestreo, y muestras con presencia y ausencia de NEPs, y se realizó un análisis de componentes principales utilizando las variables con mayor influencia en ambas categorías para encontrar combinaciones de las mismas que explicaran la presencia según condiciones del hábitat. Se efectuó un análisis discriminante para determinar la eficiencia de las variables seleccionadas en el diagnóstico de presencia. Resultados: Se mostró que las variables relacionadas con la presencia de NEPs corresponden a materia orgánica, biomasa microbiana de carbono y respiración microbiana. Se encontró que el 38.4 % de las muestras fueron positivas con respecto a la presencia de los mismos, de las cuales el 95 % corresponde al género Steinernema y el 5 % restante a Heterorhabditis. Según la ecuación discriminante cuadrática, de un 81.2 % de las muestras sin nemátodos, seis deberían pertenecer al grupo con presencia de los mismos; mientras que para aquellas que contenían NEPs, un 55.0 % (nueve de 20) al grupo de ausencia. Conclusiones: No se observaron diferencias estadísticas entre los hábitats de muestreo seleccionados (P = 0.4296), por lo que se determinó que las tres variables mencionadas anteriormente influyen en la presencia de NEPs y valores altos indican condiciones favorables para su persistencia.

Introduction: In Costa Rica the banana is one of the most important crops in terms of production and exports; the application of pesticides is hence necessary to control the effect of pests on the plantation, such as the Cosmopolites sordidus black weevil. Because of the harmful effects of pesticides on human and environmental health, and the possibility that pests become resistant to these substances, alternatives compatible with the environment, as potential biological controllers, have been investigated to complement the effect of agrochemicals. The entomopathogenic nematodes (EPN) of genera Heterorhabditis and Steinernema have become a focus of study in the field of biological control because of their symbiotic relation with bacteria of genera Photorhabdus and Xenorhabdus, which are a complex capable of eliminating host insects in hours. Objective: The objective was to determine the presence of EPN of genera Heterorhabditis and Steinernema in soil samples taken from banana plantations in the Caribbean region of Costa Rica, and to characterize their habitat. } Methods: Sampling points were selected within the plantations and surrounding areas; physicochemical and microbiological analyses were undertaken (pH, acidity, organic matter, Ca, K, Mg, P, Fe, Cu, Zn, Mn, texture, biomass and microbial respiration of the soil, EPN isolation). Variables were analyzed statistically to determine differences between them, in sampling sites, and samples with presence and absence of EPN. A principal-component analysis (PCA) was undertaken using the variables of greater influence in both categories to find their combinations that would explain their presence according to habitat conditions. A discriminant analysis was undertaken to determine the efficiency of the variables selected in the diagnosis of nematode occurrence. Results: The variables related to the presence of EPN were shown to correspond to organic matter, carbon biomass and microbial respiration. Of the samples 38.4 % were found to be positive with respect to the presence of EPN, of which 95 % correspond to genus Steinernema and the remaining 5 % to Heterorhabditis. According to the quadratic discriminant equation, from 81.2 % of the samples without nematodes, six belong to the group with their presence, whereas for those with EPN, 55.0 % (nine of 20) belong to the absence group. Conclusions: No statistical difference was found between the selected sampling habitats (P= 0.4296); the three variables mentioned above were thus determined have an influence on the presence of EPN; their large values indicate favorable conditions for their persistence.