Inclusion of Camelina sativa Seeds in Ewes’ Diet Modifies Rumen Microbiota

Simple Summary Modern livestock research has focused on the evaluation of feeding strategies that led to modify the rumen microbiome to achieve optimum productivity without compromising ruminants’ physiology and health. For this reason, the supplementation of unconventional feedstuffs is extensively studied. In our study, we investigated the effect of Camelina sativa seeds supplementation on ewe’s rumen microbiota. Our results suggested that supplementing Camelina sativa seeds, especially in the highest studied level (160 g·kg−1 of concentrate), resulted in significant alterations in the relative abundance of the rumen microorganisms, with those reported in methanogens being considered the most promising. Abstract Supplementing ruminant diets with unconventional feedstuffs (Camelina sativa seeds; CS) rich in bioactive molecules such as polyunsaturated fatty acids, may prove a potential eco-efficient strategy to manipulate rumen microbiome towards efficiency. Forty-eight ewes were divided into four homogenous groups (n = 12) according to their fat-corrected milk yield (6%), body weight, and age, and were fed individually with concentrate, alfalfa hay, and wheat straw. The concentrate of the control group (CON) had no CS inclusion, whereas the treated groups were supplemented with CS at 60 (CS6), 110 (CS11), and 160 (CS16) g·kg−1 of concentrate, respectively. Rumen digesta was collected using an esophageal tube and then liquid and solid particles were separated using cheesecloth layers. An initial bacteriome screening using next-generation sequencing of 16S was followed by specific microbes targeting with a RT-qPCR platform, which unveiled the basic changes of the rumen microbiota under CS supplementation levels. The relative abundances of Archaea and methanogens were significantly reduced in the solid particles of CS11 and CS16. Furthermore, the relative abundance of Protozoa was significantly increased in both rumen fluid and solid particles of the CS6, whereas that of Fungi was significantly reduced in the rumen particle of the CS16. In rumen fluid, the relative abundance of Fibrobacter succinogens and Ruminobacter amylophilus were significantly increased in the CS6 and CS11, respectively. In the solid particles of the CS11, the relative abundance of Ruminococcus flavefaciens was significantly reduced, whereas those of Butyrivibrio proteoclasticus and Ruminobacter amylophilus were significantly increased. Additionally, the relative abundance of Selenomonas ruminantium was significantly increased in both CS11 and CS16. Consequently, the highest CS content in the concentrate reduced the relative abundance of methanogens without inducing radical changes in rumen microorganisms that could impair ruminal fermentation and ewes’ performance.

[1]  B. Prandi,et al.  The amino acid profile of Camelina sativa seeds correlates with the strongest immune response in dairy ewes. , 2022, Animal : an international journal of animal bioscience.

[2]  R. Kirkegaard,et al.  Oxford Nanopore R10.4 long-read sequencing enables the generation of near-finished bacterial genomes from pure cultures and metagenomes without short-read or reference polishing , 2022, Nature Methods.

[3]  K. Sotirakoglou,et al.  Assessing the Optimum Level of Supplementation with Camelina Seeds in Ewes’ Diets to Improve Milk Quality , 2021, Foods.

[4]  E. Flemetakis,et al.  The Effect of Forage-to-Concentrate Ratio on Schizochytrium spp.-Supplemented Goats: Modifying Rumen Microbiota , 2021, Animals : an open access journal from MDPI.

[5]  M. Hayes,et al.  Seaweed and Seaweed Bioactives for Mitigation of Enteric Methane: Challenges and Opportunities , 2020, Animals.

[6]  M. Rohde,et al.  Unsaturated Fatty Acids Control Biofilm Formation of Staphylococcus aureus and Other Gram-Positive Bacteria , 2020, Antibiotics.

[7]  Mengwei Li,et al.  Camelina sativa L. Oil Mitigates Enteric in vitro Methane Production, Modulates Ruminal Fermentation, and Ruminal Bacterial Diversity in Buffaloes , 2020, Frontiers in Veterinary Science.

[8]  Z. Klimont,et al.  Technical potentials and costs for reducing global anthropogenic methane emissions in the 2050 timeframe –results from the GAINS model , 2020, Environmental Research Communications.

[9]  P. Nichols,et al.  Enhancing Omega-3 Long-Chain Polyunsaturated Fatty Acid Content of Dairy-Derived Foods for Human Consumption , 2019, Nutrients.

[10]  P. Cotter,et al.  The rumen microbiome: a crucial consideration when optimising milk and meat production and nitrogen utilisation efficiency , 2018, Gut microbes.

[11]  R. Dewhurst,et al.  Addressing Global Ruminant Agricultural Challenges Through Understanding the Rumen Microbiome: Past, Present, and Future , 2018, Front. Microbiol..

[12]  G. Suen,et al.  Camelina Seed Supplementation at Two Dietary Fat Levels Change Ruminal Bacterial Community Composition in a Dual-Flow Continuous Culture System , 2017, Front. Microbiol..

[13]  Zhongtang Yu,et al.  Rumen methanogens and mitigation of methane emission by anti-methanogenic compounds and substances , 2017, Journal of Animal Science and Biotechnology.

[14]  M. F. Jahromi,et al.  Dietary n-6:n-3 Fatty Acid Ratios Alter Rumen Fermentation Parameters and Microbial Populations in Goats. , 2017, Journal of agricultural and food chemistry.

[15]  A. Sazili,et al.  Effects of feeding whole linseed on ruminal fatty acid composition and microbial population in goats , 2016, Animal nutrition.

[16]  A. R. Cabrita,et al.  The Potential Role of Seaweeds in the Natural Manipulation of Rumen Fermentation and Methane Production , 2016, Scientific Reports.

[17]  T. Nagaraja Microbiology of the Rumen , 2016 .

[18]  N. McEwan,et al.  The Role of Ciliate Protozoa in the Rumen , 2015, Front. Microbiol..

[19]  E. Forano,et al.  Effect of camelina oil or live yeasts (Saccharomyces cerevisiae) on ruminal methane production, rumen fermentation, and milk fatty acid composition in lactating cows fed grass silage diets. , 2015, Journal of dairy science.

[20]  D. Yáñez-Ruiz,et al.  Use of stomach tubing as an alternative to rumen cannulation to study ruminal fermentation and microbiota in sheep and goats , 2014 .

[21]  C. Martin,et al.  Influence of rumen protozoa on methane emission in ruminants: a meta-analysis approach. , 2014, Animal.

[22]  Itzhak Mizrahi,et al.  Potential Role of the Bovine Rumen Microbiome in Modulating Milk Composition and Feed Efficiency , 2014, PloS one.

[23]  S. Vaughn,et al.  Evaluating the Phytochemical Potential of Camelina: An Emerging New Crop of Old World Origin , 2014 .

[24]  R. Jetter Phytochemicals – Biosynthesis, Function and Application , 2014, Recent Advances in Phytochemistry.

[25]  Yanfen Cheng,et al.  Discovery of a novel rumen methanogen in the anaerobic fungal culture and its distribution in the rumen as revealed by real-time PCR , 2014, BMC Microbiology.

[26]  P. Moate,et al.  Effects of feeding algal meal high in docosahexaenoic acid on feed intake, milk production, and methane emissions in dairy cows. , 2013, Journal of dairy science.

[27]  M. Kreuzer,et al.  The Effect of Saturated Fatty Acids on Methanogenesis and Cell Viability of Methanobrevibacter ruminantium , 2013, Archaea.

[28]  Z. Yu,et al.  Effects of coconut and fish oils on ruminal methanogenesis, fermentation, and abundance and diversity of microbial populations in vitro. , 2013, Journal of dairy science.

[29]  S. López,et al.  Manipulation of rumen fermentation and methane production with plant secondary metabolites , 2012 .

[30]  A. Wright,et al.  Rumen fermentation and microbial population in lactating dairy cows receiving diets containing oilseeds rich in C-18 fatty acids , 2012, British Journal of Nutrition.

[31]  K. Beauchemin,et al.  Structures of free-living and protozoa-associated methanogen communities in the bovine rumen differ according to comparative analysis of 16S rRNA and mcrA genes. , 2012, Microbiology.

[32]  D. Kenny,et al.  Effect of Phenotypic Residual Feed Intake and Dietary Forage Content on the Rumen Microbial Community of Beef Cattle , 2012, Applied and Environmental Microbiology.

[33]  I. Mizrahi,et al.  Composition and Similarity of Bovine Rumen Microbiota across Individual Animals , 2012, PloS one.

[34]  J. Loor,et al.  Effect of incremental levels of fish oil supplementation on specific bacterial populations in bovine ruminal fluid. , 2012, Journal of animal physiology and animal nutrition.

[35]  S. Koike,et al.  Evidence for the possible involvement of Selenomonas ruminantium in rumen fiber digestion. , 2011, FEMS microbiology letters.

[36]  D. Kenny,et al.  The effect of dietary concentrate and soya oil inclusion on microbial diversity in the rumen of cattle , 2011, Journal of applied microbiology.

[37]  L. Barton,et al.  Microbe–Microbe Interactions , 2011 .

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

[39]  J. Edwards,et al.  Diversity and activity of enriched ruminal cultures of anaerobic fungi and methanogens grown together on lignocellulose in consecutive batch culture. , 2009, Bioresource technology.

[40]  K. Beauchemin,et al.  Crushed sunflower, flax, or canola seeds in lactating dairy cow diets: effects on methane production, rumen fermentation, and milk production. , 2009, Journal of dairy science.

[41]  Yong Guo,et al.  Effect of octadeca carbon fatty acids on microbial fermentation, methanogenesis and microbial flora in vitro , 2008 .

[42]  S. Koike,et al.  Ecological and physiological characterization shows that Fibrobacter succinogenes is important in rumen fiber digestion — Review , 2008, Folia Microbiologica.

[43]  C Hurtaud,et al.  Effects of feeding camelina (seeds or meal) on milk fatty acid composition and butter spreadability. , 2007, Journal of dairy science.

[44]  C. Gaskins,et al.  A direct method for fatty acid methyl ester synthesis: application to wet meat tissues, oils, and feedstuffs. , 2007, Journal of animal science.

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

[46]  S. Denman,et al.  Development of a real-time PCR assay for monitoring anaerobic fungal and cellulolytic bacterial populations within the rumen. , 2006, FEMS microbiology ecology.

[47]  S. Sawanon,et al.  Synergistic fibrolysis in the rumen by cellulolytic Ruminococcus flavefaciens and non-cellulolytic Selenomonas ruminantium: Evidence in defined cultures , 2006 .

[48]  R. M. Maurício,et al.  In vitro microbial inoculum: A review of its function and properties , 2005 .

[49]  M. Kreuzer,et al.  Myristic acid supports the immediate inhibitory effect of lauric acid on ruminal methanogens and methane release. , 2004, Anaerobe.

[50]  M. Kreuzer,et al.  Effect of coconut oil and defaunation treatment on methanogenesis in sheep. , 2003, Reproduction, nutrition, development.

[51]  J. Loor,et al.  Characterization of 18:1 and 18:2 isomers produced during microbial biohydrogenation of unsaturated fatty acids from canola and soya bean oil in the rumen of lactating cows. , 2002, Journal of animal physiology and animal nutrition.

[52]  P. Luton,et al.  The mcrA gene as an alternative to 16S rRNA in the phylogenetic analysis of methanogen populations in landfill. , 2002, Microbiology.

[53]  J. Zubr,et al.  Effects of growth conditions on fatty acids and tocopherols in Camelina sativa oil , 2002 .

[54]  Christine M. Williams,et al.  Dietary fatty acids and human health , 2000 .

[55]  B. White,et al.  Polysaccharide Degradation in the Rumen and Large Intestine , 1997 .

[56]  A. Demain,et al.  Gastrointestinal Microbiology , 1997, Chapman & Hall Microbiology Series.

[57]  C. Stewart,et al.  The Rumen Microbial Ecosystem , 1997, Springer Netherlands.

[58]  M. Fondevila,et al.  Interactions between Fibrobacter succinogenes, Prevotella ruminicola, and Ruminococcus flavefaciens in the digestion of cellulose from forages. , 1996, Journal of animal science.

[59]  J. Jouany,et al.  The importance of methanogens associated with ciliate protozoa in ruminal methane production in vitro , 1995, Letters in applied microbiology.

[60]  D. Johnson,et al.  Methane emissions from cattle. , 1995, Journal of animal science.

[61]  J. Costerton,et al.  Interactions between Treponema bryantii and cellulolytic bacteria in the in vitro degradation of straw cellulose. , 1987, Canadian Journal of Microbiology (print).

[62]  M. Wolin,et al.  Propionate Formation from Cellulose and Soluble Sugars by Combined Cultures of Bacteroides succinogenes and Selenomonas ruminantium , 1973 .