Efecto de la formulación de poso de café en pienso sobre el rendimiento productivo, calidad de leche y emisiones de metano de origen entérico en vacas lecheras

El objetivo del estudio fue determinar el efecto de formular poso de café (PC) en el pienso sobre el rendimiento productivo, calidad de la leche, fermentación ruminal, emisiones de metano de origen entérico y la capacidad antioxidante de la leche. Se utilizó un diseño experimental en cross over 2x2. Se formuló un 10% de PC en pienso y éste se diseñó para que aportara cantidades similares de energía, proteína y grasa a las aportadas por un pienso control (CTR). Un total de 4 vacas lecheras Holstein Friesian y 12 vacas lecheras Parda Suizas fueron emparejadas según la raza, el número de partos, los días en leche y la producción diaria. Todas las vacas fueron alimentadas con pienso CTR durante 15 días (covariable), y una vez finalizado, cada vaca fue asignada aleatoriamente dentro de la pareja al pienso CTR o PC. Las dos vacas que conformaron una pareja recibieron la misma cantidad de concentrado y además dispusieron de mezcla forrajera a libre voluntad. Cada período duró 44 d; de los cuales los primeros 14 d se destinaron a la adaptación al pienso, tomándose medidas durante los siguientes 30 d. Para evitar un efecto de arrastre de los efectos del primer periodo en el segundo los animales recibieron durante un periodo de tiempo de 30 d mezcla forrajera a libre voluntad y concentrado CTR. No se encontró un efecto significativo de formular PC en el pienso sobre la producción de ácidos grasos volátiles totales o sobre sus proporciones individuales. Tampoco se encontró un efecto significativo sobre la producción de leche (19,7 vs.19, 3 kg/d, P=0,338), el contenido en grasa (4,58 vs. 4,71 %, P=0,419), proteína (3,50 vs.3, 47 %, P=0,632) o lactosa (4,92 vs. 4, 86 %, P=0,218). En consecuencia, no se encontró un efecto significativo sobre la leche corregida por grasa y proteína (21,0 vs. 19,8 kg/d, P=0,348). Tampoco se encontraron diferencias significativas sobre el contenido total de polifenoles (0,365 vs 0,378 mg/g; P=0,844) o la capacidad antioxidante de la leche (0,060 vs. 0,057 μmol trolox/g; P=0,941). Formular PC en el pienso no redujo ni las emisiones totales de CH4 (440 vs. 461 g/d, P=0,222) ni su intensidad de emisión (19,7 vs. 21,3 g/kg, P=0,321). En conclusión, se puede revalorizar el PC mediante su inclusión en piensos para vacuno lechero con una inclusión de hasta un 10% sin que ello conlleve una pérdida de rendimiento productivo. Sin embargo, dicho nivel de inclusión no ha permitido cambiar el patrón de fermentación ruminal hacia rutas energéticamente más eficientes y en consecuencia reducir las emisiones de metano de origen entérico.Palabras clave: posos de café-desechos-emisiones de metano-contaminación-subproductos-alimentación animal.<br /

Evaluación de parámetros de fermentación ruminal y emisiones de metano en raciones representativas de diferentes sistemas de producción de ovino lechero en la Comunidad Autónoma del País Vasco

El objetivo de este estudio fue: i) Evaluar in vitro los parámetros de fermentación ruminal, la digestibilidad y emisiones de metano de las dietas empleadas en explotaciones de ovino lechero con diferente nivel de intensificación y representativas de los principales sistemas de producción existentes en la Comunidad Autónoma del País Vasco (CAPV); ii) así como proponer estrategias de mitigación de emisiones de metano para este sistema ganadero. Se monitorizaron y caracterizaron 16 rebaños en fase de preparto y 14 rebaños al inicio de lactación. Para cada caso, mediante encuesta a los pastores se identificaron características de raza empleada, manejo nutricional y tipo de forraje suministrado. Se tomaron muestras individuales de los ingredientes que conformaron las raciones suministradas en cada uno de los rebaños para realizar el análisis químico laboratorial, calcular el aporte nutricional de la ración total y realizar incubaciones in vitro para determinar parámetros de la cinética de producción de gas, digestibilidad, producción de ácidos grasos volátiles y de metano. La proporción media forraje/concentrado empleado en las raciones de preparto fue de 70:30, siendo heno de pradera de diferente calidad el forraje suministrado en la mayoría de los rebaños. Las raciones de inicio de lactación emplearon una proporción media forraje/concentrado de 60:40, ofreciendo heno de pradera en la mitad de los rebaños en conjunto con otro tipo de forrajes. Las raciones de inicio de lactación presentaron con respecto a las de preparto presentaron una menor producción potencial de gas (268,11 vs. 312.98 ml/g MOI, P<0,001), menor producción de gas/día por gramo de materia orgánica incubada (159,87 vs. 169,23 ml/g, MOI, P=0,043) y una mayor digestibilidad (64 vs. 59 g/100 g, P=0,005); la eficiencia de los procesos fermentativos también fue mayor, siendo superiores el factor de partición a 24 horas (4,04 vs. 3,66 mg MO/ml, P=0,009), la síntesis de biomasa microbiana a 24 horas (126,08 vs. 104,37 mg, P=0,010) y la eficiencia de dicha síntesis microbiana a 24 horas (45 vs. 39 g/100g, P=0,002), mientras que la producción de gas/día por gramo de materia orgánica fermentable (248,18 vs. 278,77 ml/g, P<0,001) y el ratio ácido graso volátil : materia orgánica fermentable (17,33 vs. 19,84 mmol/g MOF, P=0,005) fueron menores. Las diferencias dentro de las raciones de preparto sólo se presentaron para el parámetro que describe el tiempo en el que se alcanza la mitad del valor potencial de gas, siendo menor para la ración que empleó heno de alfalfa con un valor relativo forrajero de segunda como único aporte forrajero en una proporción forraje/concentrado 69/31 y un contenido en ración total de FND y FAD de 353 y 276 g/kg respectivamente, con respecto a 9 raciones con proporciones similares donde en uno de los casos el único componente forrajero fue paja de cereal y para el resto fue heno de pradera de diferente calidad. El promedio de producción de metano/día por gramo de materia orgánica fermentable e incubada en las raciones de preparto fue de 5,97 mmol y 3,56 mmol y en inicio de lactación de 5,65 mmol y 3,57 mmol respectivamente. De las raciones de lactación temprana, aquella que menos producción de metano presentó, empleó heno de alfalfa y de pradera con un valor relativo forrajero excelente en una proporción forraje/concentrado 72/28 y un contenido en ración total de FND y FAD de 267 y 198 g/kg respectivamente, en sentido opuesto con una ración 50/50 que empleó heno de alfalfa y ensilaje de maíz con un valor relativo forrajero de primera y un contenido de FND y FAD de 404 y 253 g/kg respectivamente. De esta parte se concluye que existe una amplia variabilidad en el manejo alimenticio en los dos estados fisiológicos monitorizados, que repercutió de igual forma en la variabilidad observada dentro de las raciones de cada estado fisiológico, así como en la observada en la comparación entre estados fisiológicos. Las estrategias de mitigación evaluadas en laboratorio consideraron la inclusión de distintas fuentes de grasa en la dieta, diferentes niveles de inclusión en la ración y henos de diferente calidad. Se realizó un diseño multifactorial con 2 henos de diferente calidad (alta y baja) x 3 tipos de grasa (torta de girasol, de colza y grasa de palma) x 2 niveles de grasa en ración total (3% y 5%). Se realizaron incubaciones in vitro para determinar los parámetros de la cinética de producción de gas, digestibilidad de la materia orgánica, producción de AGV’S y producción de metano. La interacción entre la calidad del heno y el nivel de grasa redujo la emisión de gas por gramo de materia orgánica fermentable (275,87 vs. 236,55 ml/g MOF, P=0,001), la proporción de ácido acético (0,71 vs. 0,69 g/g del total de AGV, P=0,030), el ratio ácido acético/propiónico (4,06 vs. 3,94, P=0,040) y la producción de metano por gramo de materia orgánica fermentable (5,98 vs. 4,44 mmol día, P=0,006) en la incubación del heno de alta calidad y un nivel de grasa del 5%. De otro modo, la interacción entre el tipo de grasa y la calidad del heno afectó la digestibilidad in vitro de la materia orgánica, siendo mayor para el heno de alta calidad cuando se incubó con torta de colza (64 g/100g vs. 51 g/100g, P=0,001), torta de girasol (64 g/100g vs. 50 g/100g, P=0,001) y grasa de palma (65 g/100g vs. 47 g/100g, P=0,001). El efecto simple de incubar con torta de girasol redujo la producción de gas por g/materia orgánica incubada con respecto a la torta de colza (142,30 vs. 149,90, p=0,003). Del mismo modo, la torta de girasol incrementó el factor de partición a 24 horas (4,03 vs. 3,78 mg MO/ml, P=0,035) y la síntesis de biomasa microbiana a 24 horas (105,56 mg vs. 93,08, P=0,049) con respecto a las raciones con grasa de palma. El heno de alta calidad en relación con el de baja redujo la producción potencial de gas (258,17 vs. 308,83 ml/g MO, P<0,001), incrementó el factor de partición a 24 horas (4,06 vs. 3,71 mg MO/ml, P=0,003), la síntesis de biomasa microbiana a 24 horas (119,74 vs. 80,20 mg, P<0,001) y la eficiencia de síntesis microbiana a 24 horas (46 vs. 41 g/100g, P<0,001 respectivamente), y redujo el ratio ácido graso volátil : materia orgánica fermentable (20,94 vs. 25,25 mmol/g MOF, P=0,002). Desde el punto de vista de la mitigación de emisión de metano se concluye que la combinación de un heno de alta calidad de características composicionales como las del incubado en este ensayo, con un nivel de grasa del 5% en la ración, reduce la emisión de gas diaria y de metano por gramo de materia. Por su parte, la producción potencial de gas se redujo al incubar con un heno de alta calidad con respecto al de baja

Vegetable by-products as alternative and sustainable raw materials for ruminant feeding: nutritive evaluation and their inclusion in a novel ration for calf fattening

This research aimed to evaluate the nutritional composition, in vitro digestibility, and gas production kinetics of 15 vegetable by-products generated by the agri-food industry compared with corn silage as a reference raw material. Nutritional characterization and in vitro ruminal fermentation tests were performed to determine in vitro organic matter digestibility and digestible energy values, short-chain fatty acids, and the gas production profile. Results indicate that vegetable by-products were more degradable, more extensively fermented, and fermented at a faster rate than corn silage. Going one step further in the valorization of these by-products in animal feed, the second part of the research aimed to compare the novel ration designed for calf fattening with a conventional one. An artificial rumen unit was used to obtain nutrient disappearance, rumen fermentation parameters, and gas production of rumen digesta. Very slight differences were observed between both experimental rations, with their composition being the main difference. Most of the unitary vegetable by-products and all mixes, as real examples of by-product generation in the agri-food industry, have higher digestibility and a greater nutritional value than corn silage. These by-products showed the potential to be used in ruminant-ensiled rations and could replace part of the ingredients in conventional diets.This research was funded by the Government of Navarra and the European Regional Development Fund (ERDF) in the framework of the call for R & D projects with BEEF + project “Carne saludable a través de la economía circular” (0011-1365-2020-000288), I.G. Industrial PhDs 2020 (0011-1408-2020-000009) and S.L.-E. Universidad Pública de Navarra post-graduate scholarships (UPNA-2022)

Evaluation of Valorisation Strategies to Improve Grape Stems’ Nutritional Value as an Ingredient for Ruminants’ Diets

Grape stems are a by-product of wine production which is managed as a waste. Animal feeding arises as a potential alternative. However, its practical use may be compromised by its high lignin content. In this sense, hydrolysis emerges as a strategy to increase fibre digestibility. In addition, due to its high and variable moisture content, it should be dried to prevent microbial deterioration and a washing pre-treatment to reduce sugar content becomes necessary to minimize drying problems due to sugar melting at high temperatures. Within this framework, this study assessed the effect of washing and three different hydrolysis on the nutritive value of grape stems. A factorial design was carried out, with washing (with or without) and hydrolysis (without, enzymatic, alkali, and alkali-enzymatic) as factors. When the washing pre-treatment was not applied, only the alkali hydrolysis process maintained in vitro digestibility, but at the expense of a lower fermentation efficiency. When the washing pre-treatment was applied, fibre contents were similar among hydrolysis processes, but the alkali hydrolysis improved in vitro digestibility with similar fermentation efficiency. In conclusion, the alkali hydrolysis maintained or improved the grape stem nutritive value depending on whether the washing pre-treatment was applied or not.This paper is supported by the PRIMA program under grant agreement No 2013, project NEWFEED. The PRIMA program is supported by the European Union

Fungal and ciliate protozoa are the main rumen microbes associated with methane emissions in dairy cattle

14 Pág. Departamento de Mejora Genetica AnimalMitigating the effects of global warming has become the main challenge for humanity in recent decades. Livestock farming contributes to greenhouse gas emissions, with an important output of methane from enteric fermentation processes, mostly in ruminants. Because ruminal microbiota is directly involved in digestive fermentation processes and methane biosynthesis, understanding the ecological relationships between rumen microorganisms and their active metabolic pathways is essential for reducing emissions. This study analysed whole rumen metagenome using long reads and considering its compositional nature in order to disentangle the role of rumen microbes in methane emissions.This research was financed by RTA2015-00022-C03-02 (METALGEN) project from the National Plan of Research, Development and Innovation 2013–2020 and the Department of Economic Development and Competitiveness (Madrid, Spain). A.L.G. was funded by FPI-INIA grant with reference FPI-SGIT2016-06.Peer reviewe

Comparison of Mothur and QIIME for the Analysis of Rumen Microbiota Composition Based on 16S rRNA Amplicon Sequences

Background: Microbiome studies need to analyze massive sequencing data, which requires the use of sophisticated bioinformatics pipelines. Up to date, several tools are available, although the literature is scarce on studies that compare the performance of different bioinformatics pipelines on rumen microbiota when 16S rRNA amplicons are analyzed. The impact of the pipeline on the outcome of the results is also unknown, mainly in terms of the output from studies using these tools as an intermediate phenotype (pseudophenotypes). This study compares two commonly used software (Quantitative Insights Into Microbial Ecology) (QIIME) and mothur, and two microbial gene data bases (GreenGenes and SILVA) for 16S rRNA gene analysis, using metagenome read data collected from rumen content of a cohort of dairy cows.Results: We compared the relative abundance (RA) of the identified OTUs at the genus level. Both tools presented a high degree of agreement at identifying the most abundant genera: Bifidobacterium, Butyrivibrio, Methanobrevibacter, Prevotella, and Succiniclasticum (RA &gt; 1%), regardless the database. There were no statistical differences between mothur and QIIME (P &gt; 0.05) at estimating the overall RA of the most abundant (RA &gt; 10%) genera, either using SILVA or GreenGenes. However, differences were found at RA &lt; 10% (P &lt; 0.05) when using GreenGenes as database, with mothur assigning OTUs to a larger number of genera and in larger RA for these less frequent microorganisms. With this database mothur resulted in larger richness (P &lt; 0.05), more favorable rarefaction curves and a larger analytic sensitivity. These differences caused significant and relevant differences between tools at identifying the dissimilarity of microbiotas between pairs of animals. However, these differences were attenuated, but not erased, when SILVA was used as the reference database.Conclusion: The findings showed that the SILVA database seemed a preferred reference dataset for classifying OTUs from rumen microbiota. If this database was used, both QIIME and mothur produced comparable richness and diversity, and also in the RA of most common rumen microbes. However, important differences were found for less common microorganisms which impacted on the beta diversity calculated between pipelines. This may have relevant implications at studying global rumen microbiota

Solid State Fermentation as a Tool to Stabilize and Improve Nutritive Value of Fruit and Vegetable Discards: Effect on Nutritional Composition, In Vitro Ruminal Fermentation and Organic Matter Digestibility

This research aimed to evaluate in vitro organic matter digestibility, fermentation characteristics and methane production of fruit and vegetable discards processed by solid state fermentation (SSF) by Rhizopus sp. Mixtures were composed of approximately 28% citric fruits, 35% other fruits and 37% vegetables. Fruit and vegetables were processed and fermented to obtain a stabilized product. Nutritional characterization and in vitro ruminal fermentation tests were performed to determine the effect of fungal bioconversion on digestibility, end products and gas production kinetics. Results indicate that SSF reduced organic matter and reducing sugars, while it increased crude protein and neutral detergent fiber, acid detergent fiber and neutral detergent insoluble protein. The in vitro gas production showed that SSF led to a reduction of the organic matter digestibility (p &lt; 0.001), short chain fatty acids (SCFA; p = 0.003) and CH4 (p = 0.002). SSF reduced the gas production from the insoluble fraction (p = 0.001), without modifying the production rate (p = 0.676) or the lag time (p = 0.574). Regarding SCFA profile, SSF increased acetic (p = 0.020) and decreased propionic (p = 0.004) and butyric (p = 0.006) acids proportions, increasing acetic to propionic (p = 0.008) and acetic plus butyric to propionic (p = 0.011) ratios. SSF succeeded in obtaining a stabilized material enriched in protein, but at the expense of a reduction of protein availability and organic matter digestibility. These changes should be considered before including them in a ruminant’s rations

Comparison of Mothur and QIIME for the Analysis of Rumen Microbiota Composition Based on 16S rRNA Amplicon Sequences

Background: Microbiome studies need to analyze massive sequencing data, which requires the use of sophisticated bioinformatics pipelines. Up to date, several tools are available, although the literature is scarce on studies that compare the performance of different bioinformatics pipelines on rumen microbiota when 16S rRNA amplicons are analyzed. The impact of the pipeline on the outcome of the results is also unknown, mainly in terms of the output from studies using these tools as an intermediate phenotype (pseudophenotypes). This study compares two commonly used software (Quantitative Insights Into Microbial Ecology) (QIIME) and mothur, and two microbial gene data bases (GreenGenes and SILVA) for 16S rRNA gene analysis, using metagenome read data collected from rumen content of a cohort of dairy cows. Results: We compared the relative abundance (RA) of the identified OTUs at the genus level. Both tools presented a high degree of agreement at identifying the most abundant genera: Bifidobacterium, Butyrivibrio, Methanobrevibacter, Prevotella, and Succiniclasticum (RA > 1%), regardless the database. There were no statistical differences between mothur and QIIME (P > 0.05) at estimating the overall RA of the most abundant (RA > 10%) genera, either using SILVA or GreenGenes. However, differences were found at RA < 10% (P < 0.05) when using GreenGenes as database, with mothur assigning OTUs to a larger number of genera and in larger RA for these less frequent microorganisms. With this database mothur resulted in larger richness (P < 0.05), more favorable rarefaction curves and a larger analytic sensitivity. These differences caused significant and relevant differences between tools at identifying the dissimilarity of microbiotas between pairs of animals. However, these differences were attenuated, but not erased, when SILVA was used as the reference database. Conclusion: The findings showed that the SILVA database seemed a preferred reference dataset for classifying OTUs from rumen microbiota. If this database was used, both QIIME and mothur produced comparable richness and diversity, and also in the RA of most common rumen microbes. However, important differences were found for less common microorganisms which impacted on the beta diversity calculated between pipelines. This may have relevant implications at studying global rumen microbiota

Changes in microbiological, chemical, rheological and sensory characteristics during ripening of vacuum packaged Manchego cheese

The effects of cheese age when packaged and of packaging material permeability on Manchego cheese characteristics were studied throughout a 6‐month ripening period. Cheese age when packaged had no significant effect on microbial levels, proteolysis, lipolysis or flavor. However, moisture content and rheological characteristics were significantly different for cheeses packaged 1, 2 or 3 wk after salting. Packaging material had no influence on microbiological, chemical, rheological or sensory characteristics of Manchego cheese. Packaged cheese had a lower flavor intensity than nonpackaged cheese, but no significant differences in flavor quality between packaged and non‐packaged cheese were recorded. Weight losses after 6 months were undetectable in packaged cheese, but they averaged 15.16% in non‐packaged cheese. Copyright © 1986, Wiley Blackwell. All rights reserve

The effect of ripening and cooking temperatures on proteolysis and lipolysis in Manchego cheese

The influence of ripening temperature on proteolysis and lipolysis was studied on four lots of raw ewe's milk Manchego cheese held for 60 days at 5, 10, 15 or 20°C. Mean levels of pH 4·6, trichloroacetic acid and phosphotungstic acid soluble N in 20°C cheeses were 52%, 78% and 95% higher than the respective levels in 5°C cheeses at the end of the ripening period. Free fatty acids content after 60 days was 90% higher in 20°C cheeses than in 5°C cheeses. Significant effects of the cooking temperature of the curd (30, 36, 38 or 40°C) on pH, moisture and NaCl content were recorded, but levels of nitrogenous fractions or free fatty acids in 60-day cheeses were not affected. © 1986