Differential Diet and pH Effects on Ruminal Microbiota, Fermentation Pattern and Fatty Acid Hydrogenation in RUSITEC Continuous Cultures

The aim of this study was to distinguish effects due to diet composition from those triggered by ruminal pH on fermentation patterns and microbial profiles in a continuous culture system (RUSITEC). The study followed a 2 × 2 factorial design, with two diets varying in the proportions of forage and concentrate and two pH levels in the culture medium. RUSITEC fermenters were used to simulate rumen fermentation and feed digestibility, fermentation end-products, microbial protein synthesis, microbial community, and long-chain fatty acid profiles in the digesta were determined. Multivariate analyses were applied to summarize the overall results. High concentrate (34% cereal grain, 32% hay) diets were more digestible (p < 0.05) than high forage (10% cereal grain, 78% hay) diets, resulting in a greater (p < 0.05) formation of most fermentation end-products and microbial protein in the rumen. However, there were no significant (p > 0.05) differences between diets in methane production. Ciliate protozoa, anaerobic fungi, some fibrolytic bacteria, hydrogenation of oleic acid, and relative proportion of conjugated linoleic acid were increased (p < 0.05) with high forage diets. A decline in rumen pH from 6.8 to 6.4 decreased (p < 0.05) feed digestibility, protein degradability, and the daily outputs of some fermentation end-products (gas, VFA, acetate, ammonia) but had no effect (p > 0.05) on the synthesis of microbial protein, and on the output of methane, propionate, butyrate or lactate. Minor changes in microbial community profile or the fatty acid relative proportions were observed within this pH range. The overall multivariate analysis revealed a clear discrimination between high-concentrate and high-forage diets, with subtler and less-defined pH effects on ruminal fermentation and microbial communities.

[1]  M. Zamiri,et al.  In vitro rumen fermentation pattern: insights from concentrate level and plant oil supplement. , 2023, Archives Animal Breeding.

[2]  R. Cooke,et al.  Shifts in bacterial communities in the rumen, vagina and uterus of beef heifers receiving different levels of concentrate. , 2022, Journal of Animal Science.

[3]  M. Hanigan,et al.  Effects of acetate, propionate, and pH on volatile fatty acid thermodynamics in continuous cultures of ruminal contents. , 2022, Journal of dairy science.

[4]  Tao Guo,et al.  Rumen Bacteria Abundance and Fermentation Profile during Subacute Ruminal Acidosis and Its Modulation by Aspergillus oryzae Culture in RUSITEC System , 2022, Fermentation.

[5]  Hongrong Wang,et al.  Subacute ruminal acidosis in dairy herds: Microbiological and nutritional causes, consequences, and prevention strategies , 2022, Animal nutrition.

[6]  Fei Li,et al.  Changes in the Fermentation and Bacterial Community by Artificial Saliva pH in RUSITEC System , 2021, Frontiers in Nutrition.

[7]  Simujide Huasai,et al.  Effects of dietary forage to concentrate ratio on nutrient digestibility, ruminal fermentation and rumen bacterial composition in Angus cows , 2021, Scientific Reports.

[8]  T. Park,et al.  Conditions stimulating neutral detergent fiber degradation by dosing branched-chain volatile fatty acids. III: Relation with solid passage rate and pH on prokaryotic fatty acid profile and community in continuous culture. , 2021, Journal of dairy science.

[9]  B. Wenner,et al.  Conditions stimulating neutral detergent fiber degradation by dosing branched-chain volatile fatty acids. II: Relation with solid passage rate and pH on neutral detergent fiber degradation and microbial function in continuous culture. , 2021, Journal of dairy science.

[10]  S. S. Lee,et al.  Diet Transition from High-Forage to High-Concentrate Alters Rumen Bacterial Community Composition, Epithelial Transcriptomes and Ruminal Fermentation Parameters in Dairy Cows , 2021, Animals : an open access journal from MDPI.

[11]  A. Faciola,et al.  Ruminal acidosis, bacterial changes, and lipopolysaccharides. , 2020, Journal of animal science.

[12]  Fadi Li,et al.  Ruminal cellulolytic bacteria abundance leads to the variation in fatty acids in the rumen digesta and meat of fattening lambs. , 2020, Journal of animal science.

[13]  Lijun Wang,et al.  The Effects of Different Concentrate-to-Forage Ratio Diets on Rumen Bacterial Microbiota and the Structures of Holstein Cows during the Feeding Cycle , 2020, Animals : an open access journal from MDPI.

[14]  C. García-Estrada,et al.  Dietary supplemental plant oils reduce methanogenesis from anaerobic microbial fermentation in the rumen , 2020, Scientific Reports.

[15]  Lijun Wang,et al.  Effects of High Forage/Concentrate Diet on Volatile Fatty Acid Production and the Microorganisms Involved in VFA Production in Cow Rumen , 2020, Animals : an open access journal from MDPI.

[16]  S. López,et al.  Effects of supplemental plant oils on rumen bacterial community profile and digesta fatty acid composition in a continuous culture system (RUSITEC). , 2019, Anaerobe.

[17]  O. Alzahal,et al.  Screening of live yeast and yeast derivatives for their impact of strain and dose on in vitro ruminal fermentation and microbial profiles with varying media pH levels in high-forage beef cattle diet. , 2019, Journal of the science of food and agriculture.

[18]  Yulin Chen,et al.  Effect of dietary concentrate to forage ratios on ruminal bacterial and anaerobic fungal populations of cashmere goats. , 2019, Anaerobe.

[19]  D. Pitta,et al.  Symposium review: Understanding diet-microbe interactions to enhance productivity of dairy cows. , 2018, Journal of dairy science.

[20]  Shengli Li,et al.  Effect of Dietary Forage to Concentrate Ratios on Dynamic Profile Changes and Interactions of Ruminal Microbiota and Metabolites in Holstein Heifers , 2017, Front. Microbiol..

[21]  C. García-Estrada,et al.  Effect of Sunflower and Marine Oils on Ruminal Microbiota, In vitro Fermentation and Digesta Fatty Acid Profile , 2017, Front. Microbiol..

[22]  I. Mateos,et al.  Shifts in microbial populations in Rusitec fermenters as affected by the type of diet and impact of the method for estimating microbial growth (15N v. microbial DNA). , 2017, Animal : an international journal of animal bioscience.

[23]  K. Shingfield,et al.  Manipulation of milk fatty acid composition in lactating cows: Opportunities and challenges , 2016 .

[24]  O. Choi,et al.  Production of medium-chain carboxylic acids by Megasphaera sp. MH with supplemental electron acceptors , 2016, Biotechnology for Biofuels.

[25]  S. López,et al.  Effects of the inclusion of flaxseed and quercetin in the diet of fattening lambs on ruminal microbiota, in vitro fermentation and biohydrogenation of fatty acids , 2015, The Journal of Agricultural Science.

[26]  P. B. Pope,et al.  Rumen microbial community composition varies with diet and host, but a core microbiome is found across a wide geographical range , 2015, Scientific Reports.

[27]  E. Okine,et al.  Impact of ruminal pH on enteric methane emissions. , 2015, Journal of animal science.

[28]  R. Kallenbach,et al.  Comparison of acetyl bromide lignin with acid detergent lignin and Klason lignin and correlation with in vitro forage degradability , 2015 .

[29]  A. Offner,et al.  Effects of inoculum source, pH, redox potential and headspace di-hydrogen on rumen in vitro fermentation yields. , 2014, Animal : an international journal of animal bioscience.

[30]  M. Long,et al.  Effects of the acid-tolerant engineered bacterial strain Megasphaera elsdenii H6F32 on ruminal pH and the lactic acid concentration of simulated rumen acidosis in vitro. , 2014, Research in Veterinary Science.

[31]  M. Wanapat,et al.  Changes of rumen pH, fermentation and microbial population as influenced by different ratios of roughage (rice straw) to concentrate in dairy steers , 2013, The Journal of Agricultural Science.

[32]  A. C. Fluck,et al.  Forage degradability, rumen bacterial adherence and fibrolytic enzyme activity in vitro: effect of pH or glucose concentration , 2013, The Journal of Agricultural Science.

[33]  M. Jordán,et al.  Effect of dietary carnosic acid on the fatty acid profile and flavour stability of meat from fattening lambs. , 2013, Food chemistry.

[34]  A. Strathe,et al.  Ruminal pH regulation and nutritional consequences of low pH , 2012 .

[35]  K. Schwartzkopf-Genswein,et al.  Ruminal acidosis in feedlot cattle: Interplay between feed ingredients, rumen function and feeding behavior (a review) , 2012 .

[36]  A. AbuGhazaleh,et al.  The Effect of Forage Level and Oil Supplement on Butyrivibrio fibrisolvens and Anaerovibrio lipolytica in Continuous Culture Fermenters , 2011, Asian-Australasian journal of animal sciences.

[37]  S. Calsamiglia,et al.  Effect of pH on ruminal fermentation and biohydrogenation of diets rich in omega-3 or omega-6 fatty , 2011 .

[38]  D. Yáñez-Ruiz,et al.  The effect of the feed-to-buffer ratio on bacterial diversity and ruminal fermentation in single-flow continuous-culture fermenters. , 2011, Journal of dairy science.

[39]  S. López,et al.  Decrease of ruminal methane production in Rusitec fermenters through the addition of plant material from rhubarb (Rheum spp.) and alder buckthorn (Frangula alnus). , 2010, Journal of dairy science.

[40]  M. Ranilla,et al.  Comparison of fermentation of diets of variable composition and microbial populations in the rumen of sheep and Rusitec fermenters. I. Digestibility, fermentation parameters, and microbial growth. , 2010, Journal of dairy science.

[41]  A. Belenguer,et al.  Changes in the rumen bacterial community in response to sunflower oil and fish oil supplements in the diet of dairy sheep. , 2010, Journal of dairy science.

[42]  A. Richardson,et al.  Toxicity of unsaturated fatty acids to the biohydrogenating ruminal bacterium, Butyrivibrio fibrisolvens , 2010, BMC Microbiology.

[43]  S. Calsamiglia,et al.  Effect of pH and level of concentrate in the diet on the production of biohydrogenation intermediates in a dual-flow continuous culture. , 2009, Journal of dairy science.

[44]  V. Fievez,et al.  Effects of capric acid on rumen methanogenesis and biohydrogenation of linoleic and α-linolenic acid. , 2009, Animal : an international journal of animal bioscience.

[45]  J. Edwards,et al.  Advances in microbial ecosystem concepts and their consequences for ruminant agriculture. , 2008, Animal : an international journal of animal bioscience.

[46]  C. J. Härter,et al.  Microbial colonization and degradation of forage samples incubated in vitro at different initial pH , 2008 .

[47]  S. Calsamiglia,et al.  Changes in rumen microbial fermentation are due to a combined effect of type of diet and pH. , 2008, Journal of animal science.

[48]  T. Jenkins,et al.  Board-invited review: Recent advances in biohydrogenation of unsaturated fatty acids within the rumen microbial ecosystem. , 2008, Journal of animal science.

[49]  S. Calsamiglia,et al.  Effect of the magnitude of the decrease of rumen pH on rumen fermentation in a dual-flow continuous culture system. , 2008, Journal of animal science.

[50]  T. Nagaraja,et al.  Ruminal acidosis in beef cattle: the current microbiological and nutritional outlook. , 2007, Journal of dairy science.

[51]  L. Chaudhary,et al.  Metabolism of polyunsaturated fatty acids and their toxicity to the microflora of the rumen , 2007, Antonie van Leeuwenhoek.

[52]  N. St-Pierre,et al.  Kinetics of fatty acid biohydrogenation in vitro. , 2007, Journal of dairy science.

[53]  D. Palmquist,et al.  Milk fatty acid composition in response to reciprocal combinations of sunflower and fish oils in the diet , 2006 .

[54]  A. Troegeler-Meynadier,et al.  Rates and efficiencies of reactions of ruminal biohydrogenation of linoleic acid according to pH and polyunsaturated fatty acids concentrations. , 2006, Reproduction, nutrition, development.

[55]  R. Bhatta,et al.  Influence of Temperature and pH on Fermentation Pattern and Methane Production in the Rumen Simulating Fermenter (RUSITEC) , 2006 .

[56]  M. Eastridge,et al.  Biohydrogenation of fatty acids and digestibility of fresh alfalfa or alfalfa hay plus sucrose in continuous culture. , 2005, Journal of dairy science.

[57]  A. Grandison,et al.  Effect of forage type and proportion of concentrate in the diet on milk fatty acid composition in cows given sunflower oil and fish oil , 2005 .

[58]  Hanqing Yu,et al.  Anaerobic degradation of cellulose by rumen microorganisms at various pH values , 2004 .

[59]  A. Ferlay,et al.  Dietary lipids and forages interactions on cow and goat milk fatty acid composition and sensory properties. , 2004, Reproduction, nutrition, development.

[60]  J. Loor,et al.  Biohydrogenation, duodenal flow, and intestinal digestibility of trans fatty acids and conjugated linoleic acids in response to dietary forage:concentrate ratio and linseed oil in dairy cows. , 2004, Journal of dairy science.

[61]  E. Okine,et al.  Dietary manipulation to increase conjugated linoleic acids and other desirable fatty acids in beef: A review , 2003 .

[62]  C. Bayourthe,et al.  Effects of pH and concentrations of linoleic and linolenic acids on extent and intermediates of ruminal biohydrogenation in vitro. , 2003, Journal of dairy science.

[63]  L. Chaudhary,et al.  Eubacterium pyruvativorans sp. nov., a novel non-saccharolytic anaerobe from the rumen that ferments pyruvate and amino acids, forms caproate and utilizes acetate and propionate. , 2003, International journal of systematic and evolutionary microbiology.

[64]  S. Calsamiglia,et al.  Effects of pH and pH fluctuations on microbial fermentation and nutrient flow from a dual-flow continuous culture system. , 2002, Journal of dairy science.

[65]  D. Bauman,et al.  Regulation and nutritional manipulation of milk fat. Low-fat milk syndrome. , 2001, Advances in experimental medicine and biology.

[66]  J. Russell,et al.  The importance of pH in the regulation of ruminal acetate to propionate ratio and methane production in vitro. , 1998, Journal of dairy science.

[67]  W. Bryden,et al.  Perspectives on ruminant nutrition and metabolism I. Metabolism in the Rumen , 1998, Nutrition Research Reviews.

[68]  E. Depeters,et al.  Digestion kinetics of neutral detergent fiber and chemical composition within some selected by-product feedstuffs , 1997 .

[69]  J. Russell,et al.  The effect of pH on ruminal methanogenesis , 1996 .

[70]  H. Abel,et al.  Untersuchungen zur Laktateliminierung aus dem Pansen von Schafen und durch Pansenmikroben in vitro (RUSITEC) , 1996 .

[71]  P. V. Soest,et al.  Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. , 1991, Journal of dairy science.

[72]  R. Mackie,et al.  An in vivo study of ruminal micro-organisms influencing lactate turnover and its contribution to volatile fatty acid production , 1984, The Journal of Agricultural Science.

[73]  J. Czerkawski,et al.  Design and development of a long-term rumen simulation technique (Rusitec) , 1977, British Journal of Nutrition.

[74]  R. Gruninger,et al.  Invited review: Application of meta-omics to understand the dynamic nature of the rumen microbiome and how it responds to diet in ruminants. , 2019, Animal : an international journal of animal bioscience.

[75]  A. Troegeler-Meynadier,et al.  Starch plus sunflower oil addition to the diet of dry dairy cows results in a trans-11 to trans-10 shift of biohydrogenation. , 2013, Journal of dairy science.

[76]  J. Mrázek,et al.  Diet-dependent shifts in ruminal butyrate-producing bacteria , 2008, Folia Microbiologica.

[77]  M. Song,et al.  pH affects the in vitro formation of cis-9, trans-11 CLA and trans-11 octadecenoic acid by ruminal bacteria when incubated with oilseeds , 2003 .

[78]  M. Song,et al.  Effect of Sources and Levels of Carbohydrates on Fermentation Characteristics and Hydrogenation of Linoleic Acid by Rumen Bacteria In Vitro , 2001 .

[79]  O Hammer-Muntz,et al.  PAST: paleontological statistics software package for education and data analysis version 2.09 , 2001 .

[80]  C. V. Van Nevel,et al.  Influence of pH on lipolysis and biohydrogenation of soybean oil by rumen contents in vitro. , 1996, Reproduction, nutrition, development.

[81]  E. Mcdougall Studies on ruminant saliva. 1. The composition and output of sheep's saliva. , 1948, The Biochemical journal.