Analysis of the Intestinal and Faecal Bacterial Microbiota of the Cervidae Family Using 16S Next-Generation Sequencing: A Review.

The Cervidae family has a wide distribution due to its adaptation to numerous ecological environments, which allows it to develop a diverse microbial community in its digestive tract. Recently, research has focused on the taxonomic composition and functionality of the intestinal and faecal microbiota of different cervid species worldwide, as well as their microbial diversity and variation under different associated factors such as age, sex, diet, distribution, and seasonal variation. In addition, there is special interest in knowing how cervids act as reservoirs of zoonotic pathogenic microorganisms, which represent a threat to public health. This review provides a synthesis of the growing field of microbiota determination in cervids worldwide, focusing on intestinal and faecal samples using 16S next-generation sequencing. It also documents factors influencing microbial diversity and composition, the microorganisms reported as pathogenic/zoonotic, and the perspectives regarding the conservation of these species. Knowing the interactions between bacteria and cervid health can drive management and conservation strategies for these species and help develop an understanding of their evolutionary history and the interaction with emerging disease-causing microorganisms.

Effect of waste mango silage on the in vitro gas production, in situ digestibility, intake, apparent digestibility, and ruminal characteristics in calf diets.

Background and Aim: Mexico is the fifth largest producer of mangoes in the world. For the conservation of agro-industrial waste and crop residues, the ensiling technique has shown good results. This study aimed to evaluate the effect of increasing the level of mango silage (86% waste mango and 14% pangola grass hay) in calf diets on in vitro gas production, in situ digestibility, intake, apparent digestibility, and ruminal characteristics. Materials and Methods: The diets contained 0 (T0), 30 (T1), 45 (T2), and 60% (T3) mango silage. The partial (24, 48, and 72 h) and cumulative (72 h) biogas, CH4 production, and degradation were determined in the in vitro evaluation. In situ digestibility and estimators of fermentation kinetics of dry matter (DM) and organic matter (OM) were determined. Intake, apparent nutrient digestibility, and rumen parameters of calves (200 kg) were evaluated in a 4 × 4 Latin square design. Response to increased mango silage was calculated by linear and quadratic orthogonal contrasts. Results: In vitro partial and cumulative biogas production decreased linearly (p < 0.05), and the partial and cumulative CH4 production did not show linear or quadratic contrast (p > 0.05); in vitro DM degradation, in vitro neutral detergent fiber degradation, and in vitro acid detergent fiber degradation showed a linear increase (p < 0.05). In situ dry matter digestibility (DMDis), in situ organic matter digestibility (OMDis), b, a + b, c, and effective digestibility (ED) of DMDis, a, a + b, c, and ED of OMDis increased linearly (p < 0.05). Dry matter intake, OM intake, and crude protein intake showed a linear increase (p < 0.05); NDF intake and ADF intake presented a quadratic behavior (p < 0.05). Apparent digestibility of DM, OM, CP, and hemicellulose, pH, N-NH3, total bacterial count, acetate, propionate, butyrate, volatile fatty acids, acetate: propionate ratio, cellulolytic bacteria, and protozoa did not present a linear or quadratic orthogonal effect (p > 0.05). Conclusion: The in vitro, in situ, and in vivo variables demonstrated that up to 60% mango silage can be used for the intensive fattening of calves in confinement.

Effect of parota (Enterolobium cyclocarpum) pod protein supplement on feed intake and digestibility and calf ruminal characteristics.

The objective of this study was to evaluate feed intake and digestibility and ruminal characteristics of development-stage calves fed mulatto II (Brachiaria sp.) grass hay (MGH) and a protein supplement (PS) consisting of increasing levels of the parota (Enterolobium cyclocarpum) pod (PP). We used eight Swiss-zebu calves in growth stage with an average age of 11 months and initial average weight of 157.6 ± 8.5 kg. They were distributed in a repeated 4 × 4 Latin square design with 4 treatments (period 30 days): 0% (PP0), 25% (PP25), 50% (PP50), and 75% (PP75) of the PP. Calves in the PP0 and PP25 treatments had higher intake of PS and MGH as dry matter (DM) than those in the PP50 and PP75 treatments (p < 0.05). Organic matter intake (OMI) of the PP75 calves was lower than that of PP0 and PP25 calves. Crude protein intakes (CPI) of PP0 and PP25 calves were higher than those of PP50 and PP75 calves (p < 0.05). Apparent digestibility of crude protein was higher in the PP0, PP25, and PP50 treatments compared with that in treatment PP75 (p < 0.05). The treatments did not affect total bacterial count, cellulolytic bacterial count, cellulase enzymatic activity, volatile fatty acids, or the acetate/propionate ratio (p > 0.05). Rumen pH in the PP0 calves was higher than that of the PP25 calves, whereas the protozoa count and ammonia content were higher in PP0 calves than in PP75 (p < 0.05). In conclusion, the inclusion of 25% PP in the PS for forage-fed calves is a feeding alternative.

Inclusion of ripe mango as a source of energy in diets for Creole lambs in the dry tropics.

The aim of this investigation was to evaluate the use of ripe mango as a source of energy in an integral diet consisting of regional products for fattening lambs. Twenty-seven Creole lambs were used and given a 10-day period to adapt to the diet. Three treatments were evaluated: T1 = 40% mango; T2 = 50% mango; and T3 = control diet. In the in vitro analysis, partial and accumulated production of biogas and methane, degradation of dry matter, and neutral detergent fiber were determined. In the in vivo analysis, dry matter intake (DMI), daily weight gain (DWG), apparent digestibility of nutrients, and ruminal characteristics were estimated. The experimental design was completely random. The in vitro results helped hypothesize that the diets with low fiber content would not affect potential dry matter intake. With the exception of the protozoa count, ruminal characteristics were not different among diets (p > 0.05). In productive response, T3 was 13.73% higher DMI than T1, which was 32.24% higher than T2. T3 promoted 61.22% more DWG than T1 and T2. T2 displayed better digestibility of dry matter, organic matter, neutral detergent fiber, and crude protein than T3 (p < 0.05). Fattening lambs fed rations that contain 40% and 50% mango reduces the dry matter intake and daily weight gain relative to the control, although apparent digestibility is higher than for the control diet.

Uso de disacáridos y carbón activado para preservar consorcios de bacterias ruminales celulolíticas liofilizadas / Use of disaccharides and activated carbon to preserve cellulolytic ruminal bacterial consortiums lyophilized

RESUMEN Objetivo. Determinar la fermentación in vitro de consorcios bacterianos ruminales celulolíticos (CBC) conservados por liofilización usando carbón activado, maltosa y lactosa como preservadores. Materiales y métodos. Un CBC se aisló de fluido ruminal de una búfala de agua en medios selectivos celulolíticos. Los CBC se liofilizaron con carbón activado (CA), lactosa (LA) o maltosa (MA) como preservadores y sin preservador (SP). El diseño experimental fue completamente al azar para medir biogás a diferentes intervalos de tiempo; así como, un diseño completamente al azar con arreglo factorial 4x3, los factores fueron preservadores (SP, CA, LA y MA) y tiempo de fermentación (24, 48 y 72 h) para pH, nitrógeno amoniacal (N-NH3), degradación de materia seca (DMS) y de fibra detergente neutro (DFDN), actividad enzimática celulasas y la población de bacterias totales. Resultados. LA produjo mayor biogás acumulado a las 72 h y parcial a partir de las 12 h (p≤0.05). SP no mostró diferencias (p>0.05) en celulasas, conteo de bacterias total, DMS y DFDN en los tiempos de fermentación evaluados con el resto de los preservadores. Conclusiones. La producción de biogás parcial y acumulada, el aumento en la tasa de degradación de 8.3 y 91.1 % en la DMS y DFDN de las 24 a 72 h (p≤0.05) con el preservador LA, muestran que la lactosa puede usarse como preservador de bacterias celulolíticas ruminales.

ABSTRACT Objective. To determine in vitro fermentation of cellulolytic ruminal bacterial consortia (CBC) preserved by lyophilization using activated carbon, maltose and lactose as preservatives. Materials and methods. A CBC was isolated from the ruminal fluid of a female water buffalo in selective cellulolytic media. The CBC were lyophilized without preservative (SP), activated carbon (CA), lactose (LA) o maltose (MA) as preservatives. The experimental design was completely random to measure biogas at different time intervals; as well as completely random with 4x3 factorial arrangement, factors were preservative [SP, CA, LA and MA] and fermentation time (24, 48 and 72 h) for pH, ammoniacal nitrogen (NH3-N), dry matter degradation (DMD), neutral detergent fiber degradation (NDFD), enzymatic activity cellulases and total bacteria population. Results. LA produced higher accumulated biogas at 72 h and partial biogas after 12 h (p≤0.05). SP did not show differences (p>0.05) in cellulases, total bacteria population, DMD and NDFD in the fermentation times evaluated with the rest of the preservative. Conclusions. The production of partial and accumulated biogas, the increase in the degradation rate of 8.3 and 91.1% in the DMD and NDFD from 24 to 72 h (p≤0.05) in the LA preservative, show that lactose can be used as a preservative of ruminal cellulolytic bacteria.

In vitro fermentation of fibrous substrates by water buffalo ruminal cellulolytic bacteria consortia / Fermentación in vitro de consorcios bacterianos celulolíticos ruminales de búfalos de agua en sustratos fibrosos

ABSTRACT Objective. To measure the in vitro fermentation variables of a cellulolytic bacteria consortium (CBC) isolated from a water buffalo rumen in coculture with total ruminal bacteria (TRB) on two fibrous substrates. Materials and methods. A CBC was isolated from the ruminal fluid of a female water buffalo in selective cellulolytic media. The experimental design was completely random with a 3x2 factorial arrangement; factors were treatments [TRB, CBC, and coculture (TRB + CBC)] and substrates (cobra grass and corn stover). Total gas and methane (CH4) production were measured at different time intervals. At 72 h, measurements were taken of pH, ammoniacal nitrogen (NH3-N), dry matter degradation (DMD), neutral detergent fiber degradation (NDFD) and total bacteria population. Results. Gas production with both substrates was highest (p≤0.05) in the coculture at 3, 6 and 24 h. At 48 and 72 h, gas production in the cobra grass was highest (p≤0.05) in the coculture. The coculture and TRB did not differ (p>0.05) in terms of CH4, DMD and NDFD values at 48 and 72 h. With the cobra grass, NH3-N concentration was higher (p≤0.05) in the coculture than in the TRB. Conclusions. The gas production and dry matter degradation values of the water buffalo rumen cellulolytic bacteria consortia indicate them to be a promising alternative for improving cobra grass structural carbohydrates degradation when in coculture with bovine ruminal bacteria.

RESUMEN Objetivo. Determinar las variables fermentativas in vitro de un consorcio bacteriano celulolítico (CBC) aislado de una búfala de agua en cocultivo con bacterias ruminales totales (BRT) en sustratos fibrosos. Materiales y métodos. Un CBC se aisló de fluido ruminal de una búfala de agua en medios selectivos celulolíticos. El diseño experimental fue un diseño completamente al azar con arreglo factorial 3x2, los factores fueron tratamientos [BRT, CBC y un cocultivo (BRT + CBC)] y sustratos (pasto pangola y rastrojo de maíz). La producción de gas total y metano (CH4) se midieron a diferentes intervalos de tiempo. Además, se estimó pH, nitrógeno amoniacal (N-NH3), degradación de materia seca (DMS) y de fibra detergente neutro (DFDN), y la población de bacterias totales a 72 h de incubación. Resultados. El cocultivo produjo mayor (p≤0.05) cantidad de gas a las 3, 6 y 24 h en ambos sustratos. A las 48 y 72 h, el cocultivo produjo mayor (p≤0.05) gas en pasto cobra. El cocultivo y las BRT no presentaron diferencias (p>0.05) en la producción de CH4 a 48 y 72 h, y en DMS y DFDN (p>0.05). En el pasto cobra, la concentración de N-NH3 con el cocultivo fue mayor (p≤0.05) que con BRT. Conclusiones. La producción de gas y degradación de materia seca de los consorcios bacterianos celulolíticos procedentes del rumen de una búfala de agua muestran que son una alternativa para mejorar la fermentación de carbohidratos estructurales del pasto cobra cuando se cocultivan con bacterias ruminales bovinas.

Fribrinolytic activity and gas production by Pleurotus ostreatus-IE8 and Fomes fomentarius - EUM1 in bagasse cane / Actividad fibrolítica y producción de gas por Pleurotus ostreatus-IE8 y Fomes fomentarius-EUM1 en bagazo de caña

Objective. To characterize the fibrolytic enzymatic activity of Pleurotus ostreatus-IE8 and Fomes fomentarius-EUM1 in sugarcane bagasse (BCA); to evaluation of the kinetics of in vitro production of BCA treated by solid fermentation (FS), crude enzyme extract (ECE) of P. ostreatus-IE8 and Fibrozyme®. Materials and methods. In fungi measured radial growth rate ( Vcr ) and biomass production in two culture media (with or without nitrogen source); activity of xylanases, cellulases and FS on BCA at 0, 7 and 15 d. The chemical analysis and kinetic analysis of in vitro gas production in 4 treatments (ECE adding enzymes obtained from the direct addition FS or FS ), witness (Fibrozyme®) and a control without addition and analyzed by a was completely randomized design. Results. Xylanases (7 d ) showed 6.32 and 5.50 UI g-1 initial substrate dry weight (SSi) for fungi P. ostreatus-IE8 and F. fomentarius-EUM1 , respectively ; P. ostreatus-IE8 scored higher activity of laccases (10.65 g-1 UI SSi) and F. fomentarius-EUM1 (1.90 UI g-1 SSi) cellulases. The ECE of P. ostreatus-IE8 and commercial enzyme did not differences (p>0.05). In the chemical composition or the gas production kinetics. The 4 treatments evaluated decreased values of the variables measured in the kinetics of gas production compared to the control (p≤0.05). Conclusions. The ECE of P. ostreatus-IE8 was similar to commercial enzyme degradation in vitro, so it is feasible to use pre-digest high fiber products.

Objetivos. Caracterizar la actividad enzimática fibrolítica de Pleurotus ostreatus-IE8 y Fomes fomentarius-EUM1 en bagazo de caña de azúcar (BCA) y evaluar la cinética de producción de gas in vitro del BCA por fermentación sólida (FS) o con extractos crudos enzimáticos (ECE) de P. ostreatus-IE8 o Fibrozyme®. Materiales y métodos. En los hongos de estudio se evaluó la velocidad de crecimiento radial (Vcr) y producción de biomasa en dos medios de cultivo (con o sin fuente de nitrógeno); actividad de xilanasas, celulasas y lacasas de la FS sobre BCA a 0, 7 y 15 d. El análisis químico y cinética de producción de gas in vitro en 4 tratamientos (proceso de FS o adición de enzimas obtenidas de ECE de la FS), un testigo (Fibrozyme®) y un control sin adición de enzimas, todo ello se analizó en un diseño completamente al azar. Resultados. Las xilanasas (7 d) mostraron 6.32 y 5.50 UI g-1 sustrato seco inicial (SSi) en P. ostreatus-IE8 y F. fomentarius-EUM1, respectivamente. P. ostreatus-IE8 mostró mayor actividad lacasa (10.65 UI g-1 SSi) y F. fomentarius-EUM1 (1.90 UI g-1 SSi) de celulasas. El ECE de P. ostreatus-IE8 y Fibrozyme® no presentaron diferencias (p>0.05) en la composición química ni en la cinética de producción de gas. Los 4 tratamientos evaluados disminuyeron los valores de las variables medidas en la cinética de producción de gas in vitro respecto al testigo (p≤0.05). Conclusiones. El ECE de P. ostreatus-IE8 fue similar a Fibrozyme® en la degradación in vitro, indicando su viabilidad y uso para pre-digerir subproductos altos en fibra.