Feeding cashew nut shell liquid decreases methane production from feces by altering fecal bacterial and archaeal communities in Thai local ruminants

The effect of feeding cashew nut shell liquid (CNSL) on fecal fermentation products and microbiota was investigated in Thai native cattle and swamp buffaloes. Four of each animal were fed rice straw and concentrate diet with control pellets without CNSL for 4 weeks, followed by the same diet with pellets containing CNSL for another 4 weeks, so that CNSL was administered at a level of 4 g/100 kg body weight. Feces were collected the last 2 days in each feeding period. CNSL alkyl phenols were recovered from feces (16%–28%) in a similar proportion to those in the diet, indicating that most functional anacardic acid was not selectively removed throughout the digestive tract. In vitro production of gas from feces, particularly methane, decreased with CNSL feeding. The proportion of acetate in feces decreased with CNSL feeding, whereas that of propionate increased, without affecting total short-chain fatty acid concentration. CNSL feeding changed fecal microbial community, particularly in swamp buffaloes, which exhibited decreases in the frequencies of Treponema, unclassified Ruminococcaceae, and Methanomassiliicoccaceae. These results suggest that CNSL feeding alters not only rumen fermentation but also hindgut fermentation via modulation of the microbial community, thereby potentially attenuating methane emission from the feces of ruminant animals.     

Characterization of bovine ruminal epithelial bacterial communities using 16S rRNA sequencing, PCR-DGGE, and qRT-PCR analysis

Currently, knowledge regarding the ecology and function of bacteria attached to the epithelial tissue of the rumen wall is limited. In this study, the diversity of the bacterial community attached to the rumen epithelial tissue was compared to the rumen content bacterial community using 16S rRNA gene sequencing, PCR-DGGE, and qRT-PCR analysis. Sequence analysis of 2785 randomly selected clones from six 16S rDNA (～1.4kb) libraries showed that the community structures of three rumen content libraries clustered together and were separated from the rumen tissue libraries. The diversity index of each library revealed that ruminal content bacterial communities (4.12/4.42/4.88) were higher than ruminal tissue communities (2.90/2.73/3.23), based on 97% similarity. The phylum Firmicutes was predominant in the ruminal tissue communities, while the phylum Bacteroidetes was predominant in the ruminal content communities. The phyla Fibrobacteres, Planctomycetes, and Verrucomicrobia were only detected in the ruminal content communities. PCR-DGGE analysis of the bacterial profiles of the rumen content and ruminal epithelial tissue samples from 22 steers further confirmed that there is a distinct bacterial community that inhibits the rumen epithelium. The distinctive epimural bacterial communities suggest that Firmicutes, together with other epithelial-specific species, may have additional functions other than food digestion.     

The impact of rumen cannulation on the microbial community of goat rumens as measured using 16S rRNA high‐throughput sequencing

The main objective of this study was to determine the impact of rumen cannulation on the microbial community of goat rumens using 16S rRNA high‐throughput sequencing. Twelve Boer crossbred goats were used in the experiment: six goats were surgically fitted with rumen cannula, and the other six were used as controls. All goats were fed the same diet for 20 days, after which their rumen digesta were sampled once per week for three consecutive weeks. Total microbial DNA was extracted from the collected rumen fluid and was used as a template to amplify the V4 hypervariable region of the 16S rRNA gene. High‐throughput sequencing was performed using an Illumina MiSeq platform, and the sequences were analyzed primarily using the Quantitative Insights into Microbial Ecology pipeline software. The results showed that the Chao 1 index, the observed species index and the Shannon‐Wiener index were not significantly different (p > 0.05) between the two groups. Bacteroidetes, Firmicutes, Tenericutes were the predominant phylum in both groups, and their relative abundance was 60.63%, 29.48%, 2.24% (n = 6, CT group) and 61.17%, 26.92%, 1.66% (n = 6, RC group) respectively. At the phylum level, the relative abundance of Proteobacteria was significantly higher (p < 0.001) in the microbial communities of RC goats, and Planctomycetes and Chloroflexi were significantly lower (p = 0.02). The abundances of other phyla were not significantly different between treatments. A total of 19 lower‐level taxa also exhibited significant differences (p < 0.05) in relative abundance between the groups. In addition, there were 18 genera shared within the control group, 26 shared within the rumen‐cannulated group, and 16 shared by both groups. Prevotella was the most abundant shared genus, although its abundance was not significantly different (p > 0.05) between the groups. In conclusion, although the most abundant microbes kept stable, rumen cannulation had the potential to significantly change rumen microbial communities in goats.     

Effects of dietary Greek oregano (Origanum vulgare ssp. hirtum) supplementation on rumen fermentation,enzyme profile and microbial communities in goats

This study was conducted to examine in vivo long‐term effects of dietary dried oregano (Origanum vulgare ssp. hirtum) whole plant on rumen fermentation, enzyme profile and microbial communities. For this purpose, eight healthy, adult, non‐lactating Alpine goats were kept in tie stalls equipped for individual feeding and randomly divided into two homogeneous groups: one fed 0.6 kg of a concentrate mixture and 0.6 kg of wheat straw without any supplementation and served as control group (CON) while the other group (OR) fed the same diet of CON but supplemented with 20 g of dried oregano plants (OPs) to provide daily dosage of 1 ml of essential oil (EO) per animal. The experimental period lasted 69 days and individual rumen fluid samples were obtained every 2 weeks at 0 and 4 hr after feeding. The results showed that dietary supplementation with OPs increased the protease activity (p < .001) and ammonia concentration (p < .05) in the rumen. Among the studied microbial populations, Peptostreptococcus anaerobius (p = .028) and Clostridium sticklandii (p < .001) were found to be the most sensitive to oregano at the current dosage. Furthermore, the total methanogen population significantly decreased (p < .05). It is concluded that a long‐term dietary administration of OPs can suppress specific rumen micro‐organisms and modify rumen fermentation favourably at least by means of suppressing methanogens.     

柴胡中草药对奶牛瘤胃菌群多样性及纤维分解菌的影响

通过DGGE和RT-PCR技术研究日粮中添加不同剂量的柴胡中草药对奶牛瘤胃细菌多样性和主要纤维分解菌(琥珀酸丝状杆菌、黄色瘤胃球菌和白色瘤胃球菌)的影响。根据产奶量(37.5±1.8) kg/d、泌乳天数(75±15) d以及胎次(1.7±0.4)等相近的原则,将40头健康的中国荷斯坦泌乳奶牛随机分为4组(n=10),分别饲喂4种不同的处理日粮,即在基础日粮中分别添加0,0.25,0.50和1.00 g/kg的柴胡中草药(DM基础)。试验持续10周,并在第6周口腔采集瘤胃液,通过变性梯度凝胶电泳(DGGE)和实时定量PCR(RT-PCR)对瘤胃细菌进行分析。DGGE图谱显示,中草药添加组和空白对照组并没有显著的差异条带,但其指纹图谱相似性并不高,均低于0.54,且1.00 g/kg的添加量显著降低了瘤胃液细菌香农多样性指数,而0.50和1.00 g/kg的添加量则提高了菌群优势度指数(P<0.05);RT-PCR结果显示,柴胡中草药对奶牛瘤胃主要纤维分解菌并没有显著的影响。因此,柴胡中草药在一定程度上影响了瘤胃细菌的多样性,但差异并不显著,可能由于瘤胃微生物适应了柴胡中草药的添加。     

Comparison of feed intake,digestion and rumen function among domestic ruminant species grazing in upland vegetation communities

This study aimed to compare feed intake, digestion, rumen fermentation parameters and bacterial community of 5 beef cows, 12 crossed ewes and 12 goats grazing together in spring–early summer on heather–gorse vegetation communities with an adjacent area of improved pasture. Organic matter intake (OMI) and digestibility (OMD) were estimated using alkane markers. Ruminal fluid samples were collected for measuring fermentation parameters, and studying the bacterial community using terminal restriction fragment length polymorphism (T‐RFLP). Spot samples of urine were taken to determine purine derivative (PD) and creatinine concentrations to estimate microbial protein synthesis in the rumen. Herbaceous species were the main dietary component in all animal species. Cattle had higher (p < 0.05) daily OMI (g/kg LW^0.75) and OMD, whereas sheep and goats showed similar values. The highest ammonia concentration was observed in sheep. Total VFA, acetate and butyrate concentrations were not influenced by animal species, while propionate concentrations in goats were 1.8 times lower (p < 0.05) than in sheep. Acetate:propionate ratio was greater (p < 0.05) in goats, whereas cattle excreted more allantoin (p < 0.05). Estimated supply of microbial N was higher in cows (p < 0.01), whereas the efficiency of microbial protein synthesis was lower (p < 0.01) in this animal species. Hierarchical clustering analysis indicated a clear effect of animal species on rumen bacterial structure. Differences among animal species were also observed in the relative frequency of several T‐RFs. Certain T‐RFs compatible with Lachnospiraceae, Proteobacteria and Clostridiales species were not found in goats, while these animals showed high relative frequencies of some fragments compatible with the Ruminococcaceae family that were not detected in sheep and cattle. Results suggest a close relationship between animals’ grazing behaviour and rumen bacterial structure and its function. Goats seem to show a greater specialization of their microbial populations to deal with the greater fibrous and tannin content of their diet.     

Microbial population in the rumen of swamp buffalo (Bubalus bubalis) as influenced by coconut oil and mangosteen peel supplementation

Four, rumen fistulated swamp buffalo bulls were used to study microbial populations in the rumen when supplemented with coconut oil and mangosteen peel. Animals were randomly assigned to a 4 × 4 Latin square design. Four treatments were un‐supplemented (Control), supplementation with coconut oil at 50 g/kg (CO5), supplementation with mangosteen peel at 30 g/kg (MP3) and supplementation with CO5 and MP3 (COM), of total DM intake. Animals received concentrate at 10 g/kg of BW, and rice straw was given ad libitum. Abundance of total bacteria was increased by CO5 supplementation, whereas populations of protozoa and Fibrobacter succinogenes were reduced by CO5 and COM supplementation. Dietary supplementation did not affect methanogen, Ruminococcus flavefaciens or Ruminococcus albus abundances. Dietary treatments changed denaturing gradient gel electrophoresis (DGGE) band patterns of methanogens and protozoa when compared with the control group, especially when supplemented with MP3. Supplementation of COM resulted in the greatest difference in pattern of DGGE bands for total bacteria compared with the control. Coconut oil and mangosteen peel supplementation resulted in changing of rumen microbial abundances and communities; however, combination of them could be more benefit to improve rumen fermentation of swamp buffalo fed on rice straw.     

不同形式的植物油对奶牛生产性能和瘤胃微生物区系的影响

试验研究了日粮中分别补充0(对照组)、2%亚麻籽油(亚麻籽油组)、2%大豆油(大豆油组)和等同2%植物油的10%膨化大豆(膨化大豆组)对奶牛生产性能和瘤胃微生物区系的影响。结果表明:饲喂亚麻油、大豆油和膨化大豆组奶牛的奶产量显著(P<0.05)高于对照组,而乳脂含量显著低于对照组(P<0.05),校正乳(FCM)产量各组间差异不显著(P>0.05)。饲喂亚麻油、大豆油或膨化大豆对奶牛瘤胃微生物图谱条带数量和区系的相似性指数无显著影响(P>0.05)。各处理组的白色瘤胃球菌、溶纤维丁酸弧菌和产琥珀酸丝状杆菌比例无显著性差异(P>0.05)。研究结果显示,日粮补充适宜水平的不同形式植物油可改善奶牛生产性能,但对瘤胃微生物区系无显著影响。     

Analysis of methanogenic archaeal communities of rumen fluid and rumen particles from Korean black goats

Molecular diversity of methanogens in the rumen of Korean black goats was investigated with 16S rRNA gene clone libraries using methanogen‐specific primers. The libraries were composed of rumen fluid‐associated methanogens (FAM) and rumen particle‐associated methanogens (PAM) from rumen‐fistulated Korean black goats. Among the 141 clones of the FAM library, the sequences were mostly related to two phyla, the Methanobacteriaceae family (77.3%) and the Thermoplasmatales family (22.7%); and among the 68 clones of the PAM library, sequences were also mainly clustered in the two phyla, the Thermoplasmatales family (63.24%) and the Methanobacteriaceae family (35.29%). Most of the sequenced clones in the two libraries were closely related to uncultured methanogenic archaeon. Quantitative real‐time PCR revealed that PAM (8.97 log 10) had significantly higher (P < 0.01) density of methanogens by the methanogenic 16S rRNA gene copies than FAM (7.57 log 10). The two clone libraries also showed difference in Shannon index (FAM library 1.70 and PAM library 1.59) and Chao 1 estimator (FAM library 18 and PAM library 17 operational taxonomic units). Apparent differences found in the microbial community from the two 16S rRNA gene libraries could be a result of such factors as the chemical and physical nature of the target material surface, types or component of diets, the interaction between the methanogens and other microbes, and age of the experimental goats.     

ORIGINAL ARTICLE: Description of development of rumen ecosystem by PCR assay in milk‐fed,weaned and finished lambs in an intensive fattening system

This study examined the reticulo‐rumen characteristics of the microbial community and its fermentative characteristics in milk‐fed, at weaning and finished lambs in a conventional fattening system. Five lambs were assigned to each of three groups: milk‐fed lambs slaughtered at 30 days (T30), weaned lambs slaughtered at 45 days (T45) and ‘finished lambs’ slaughtered at 90 days (T90). At slaughter, rumen size, fermentation parameters (pH, volatile fatty acids and microbial enzyme activity) and protozoal counts were recorded. Quantitative PCR was used to quantify the genes encoding 16S and 18S ribosomal DNA of the rumen bacterial and protozoal populations, respectively, and the sequential colonization of the rumen by cellulolytic (Ruminococcus albus, Ruminococcus flavefaciens) and amylolytic (Prevotella ruminicola, Streptococcus bovis) bacteria, and protozoa (Entodinium sp.). Denaturing gradient gel electrophoresis was used to study the development of rumen microbiota biodiversity. Intake of solid food before weaning caused a significant increase in rumen weight (p < 0.0001) and bacterial DNA (p < 0.05) and volatile fatty acid analysis concentration (p < 0.01), whereas pH declined. In milk‐fed lambs, cellulolytic bacteria were evident after 30 days. Thereafter, in the 45‐day and 90‐day groups, the proportions of R. flavefaciens decreased and R. albus increased. Amylolytic bacteria were present in milk‐fed lambs; the proportion of P. ruminicola increased in fattening lambs and S. bovis was the least abundant species. Protozoal concentrations were irregular; milk‐fed lambs had a significant number of protozoa species from Entodinium and subfamily Isotrichiidae, but they disappeared at weaning. Lamb rumen were refaunated in some individuals at 90 days (Entodinium and subfamily Diplodiniinae spp.), although individual concentrations were variable.     

Microbial and chemical composition of liquid‐associated bacteria in goats' rumen and fermenters

This study was undertaken to investigate the relationship between chemical composition and microbial profile of rumen liquid‐associated bacteria (LAB) in vivo (Murciano‐Granadina goats) and in a rumen simulation system (single‐flow continuous‐culture fermenters). To achieve this aim, analyses of purine bases along with some molecular techniques (quantitative PCR to assess abundance and DGGE to identify biodiversity and bacterial profile) were carried out. A control diet (AHC) based on alfalfa hay (AH) and concentrate (C) in a 1:1 ratio and two experimental diets (AHCBI and AHCBII), in which concentrate was partially replaced with multinutrient blocks, were used. Diets AHCBI and AHCBII included multinutrient blocks differing in the relative amount of two‐stage olive cake and the source of protein (sunflower meal vs. fava beans). We aimed to investigate the effect of these blocks on rumen microbiota to evaluate their potential as safe substitutes of cereal‐based concentrates. Similar patterns of response to diet were found for chemical composition, microbial abundances and diversity in LAB isolated from goat's rumen and fermenters. Whereas bacterial density (log₁₀ gene copies/g FM: 11.6 and 9.4 for bacteria and methanogens, respectively, in rumen) and diversity indexes (Shannon index: 3.6) were not affected by diet, DGGE analyses showed that bacterial community profile was affected. The cluster analysis suggested differences in bacterial profile between LAB pellets isolated from the rumen of goat and fermenters. A relationship between chemical composition and bacterial community composition in LAB pellets seems to exist. Changes in the former were reflected in the bacterial community profile. Further research is needed to clarify the relationship between chemical and microbial composition of ruminal bacterial pellets with diets of different quality.     

Effect of cashew nut shell liquid on metabolic hydrogen flow on bovine rumen fermentation

Effect of cashew nut shell liquid (CNSL), a methane inhibitor, on bovine rumen fermentation was investigated through analysis of the metabolic hydrogen flow estimated from concentrations of short‐chain fatty acids (SCFA) and methane. Three cows were fed a concentrate and hay diet without or with a CNSL‐containing pellet. Two trials were conducted using CNSL pellets blended with only silica (trial 1) or with several other ingredients (trial 2). Methane production was measured in a respiration chamber system, and energy balance and nutrient digestibility were monitored. The estimated flow of metabolic hydrogen demonstrated that a part of metabolic hydrogen was used for hydrogen gas production, and a large amount of it flowed into production of methane and SCFA in both trial 1 and 2, when CNSL was administered to the bovine rumen. The results obtained by regression analyses showed that the effect of CNSL supply on methane reduction was coupled with a significant (P < 0.01) decrease of acetate and a significant (P < 0.01) increase of propionate and hydrogen gas. These findings reveal that CNSL is able to reduce methane and acetate production, and to increase hydrogen gas and propionate production in vivo.     

Effects of nitrate addition to a diet on fermentation and microbial populations in the rumen of goats,with special reference to Selenomonas ruminantium having the ability to reduce nitrate and nitrite

This study investigated the effects of dietary nitrate addition on ruminal fermentation characteristics and microbial populations in goats. The involvement of Selenomonas ruminantium in nitrate and nitrite reduction in the rumen was also examined. As the result of nitrate feeding, the total concentration of ruminal volatile fatty acids decreased, whereas the acetate : propionate ratio and the concentrations of ammonia and lactate increased. Populations of methanogens, protozoa and fungi, as estimated by real‐time PCR, were greatly decreased as a result of nitrate inclusion in the diet. There was modest or little impact of nitrate on the populations of prevailing species or genus of bacteria in the rumen, whereas Streptococcus bovis and S. ruminantium significantly increased. Both the activities of nitrate reductase (NaR) and nitrite reductase (NiR) per total mass of ruminal bacteria were increased by nitrate feeding. Quantification of the genes encoding NaR and NiR by real‐time PCR with primers specific for S. ruminantium showed that these genes were increased by feeding nitrate, suggesting that the growth of nitrate‐ and nitrite‐reducing S. ruminantium is stimulated by nitrate addition. Thus, S. ruminantium is likely to play a major role in nitrate and nitrite reduction in the rumen.     

Provision of beta‐glucan prebiotics (cellooligosaccharides and kraft pulp) to calves from pre‐ to post‐weaning period on pasture

We conducted two feeding experiments to evaluate the effects of supplementation with either cellooligosaccharide or kraft pulp on growth performance in grazing beef calves (Japanese Black) from 4 weeks pre‐weaning to 12 to 16 weeks post‐weaning. In Experiment 1 (20‐week duration), nine calves (2.9‐month‐old females) were assigned to either a control group (CON) or an experimental group (CEL) fed cellooligosaccharide at a rate of 10 g/day mixed with concentrate. Average daily weight gain tended to be greater in CEL than in CON, especially after 1 month of weaning. In Experiment 2 (16‐week duration), 10 calves (2.0‐month‐old females) were assigned to either a control group or an experimental group (KRA) fed kraft pulp at a rate of 10% replacement of total digestible nutrients with concentrate. The proportion of fibrolytic bacteria increased and that of methanogenic Archaea decreased in the rumen microbial community composition of KRA calves in Experiment 2, whereas the decrease in Fibrobacter and Archaea was observed in CEL calves at first 4 weeks in Experiment 1. We conclude that beta‐glucan prebiotic supplementation to grazing calves at pre‐weaning would affect rumen microbial composition and modified rumen fermentation characteristics, leading to a better rumen environment via different means.     

Effects of supplementing an active dry yeast product on rumen microbial community composition and on subsequent rumen fermentation of lactating cows in the mid‐to‐late lactation period

The effects of supplementing feed of cows in mid‐to‐late lactation with an active yeast product (Actisaf Sc 47) were evaluated using 15 Holstein cows in a replicated 3 × 3 Latin square design. The animals were fed a mixed ration with 33% neutral detergent fiber, consisting of timothy hay (29.8%), a commercial concentrate (70.0%) and commercial calcium triphosphate (0.2%), twice daily to meet 105% of their energy requirement. Yeast supplement was set at 0, 5 and 10 g per day over 21‐day periods, each of which consisted of 14 days for adaptation followed by 7 days of data collection. Milking performance, plasma metabolite parameters, rumen volatile fatty acids, lipopolysaccharide and microbial properties were measured. Although there were no significant differences in feeding and milking performance or blood parameters associated with supplementation, the acetate to propionate ratio in the rumen fluid tended to decrease (P = 0.08). The population of Bacteroidetes tended to be less prominent (P = 0.07) and the fibrolytic bacterium Fibrobacter significantly increased (P < 0.05) in the rumen fluid of the yeast 10 g group compared with that of the control. These data suggest that effects of supplementing live yeast to cows in mid‐to‐late lactation may be limited to microbial composition and fermentation characteristics in the rumen.     

Effect of supplemental Bacillus cultures on rumen fermentation and milk yield in Chinese Holstein cows

Two experiments were conducted to study the effect of supplemental 100 g/day of live Bacillus cultures (2 × 10¹¹ cell of Bacillus subtilis and Bacillus licheniformis) on rumen fermentation as well as milk yield and composition in Chinese Holstein cows. In experiment 1, investigating 3 × 10 cows, milk yield and milk protein were increased by using B. licheniformis (p < 0.05) in comparison with an unsupplemented group and the B. subtilis group. Body weight was not significantly affected by Bacillus culture supplementation (p > 0.05). Percentage of milk fat and lactose was not significantly different between treatments (p > 0.05). But milk protein increased with B. licheniformis supplementation (p < 0.05). In experiment 2, carried out with three non‐lactating ruminally and duodenally fistulated cows, results showed that B. licheniformis supplementation increased microbial crude protein flow into duodenum (p < 0.05) and decreased the ammonia nitrogen concentration in ruminal fluid at 0.5 h, 1 h, 3 h, 6 h after morning feeding (p < 0.05). Bacillus licheniformis supplementation increased total VFA and acetate concentration in ruminal fluid at 0.5 h, 1 h, 3 h, 6 h after morning feeding (p < 0.05). Bacillus subtilis had no significant effect on rumen fermentation characteristics, duodenal microbial N flow and ruminal apparent nutrient digestibility (p > 0.05). Bacillus licheniformis increased ruminal apparent nutrient digestibility of neutral detergent fibre, acid detergent fibre, and organic matter (p < 0.05).     

Effect of non‐starch‐polysaccharide‐degrading enzymes as feed additive on the rumen bacterial population in non‐lactating cows quantified by real‐time PCR

The effects of non‐starch‐polysaccharide‐degrading enzymes, added to a maize silage‐ and grass silage‐based total mixed ration (TMR) at least 14 h before feeding, on the rumen bacterial population were investigated. Six non‐lactating Holstein Friesian cows were allocated to three treatment groups using a duplicate 3 × 3 Latin square design with three 31‐day periods (29 days of adaptation and 2 days of sampling). Treatments were control TMR [69% forage and 31% concentrates on a dry matter (DM) basis] or TMR with 13.8 or 27.7 ml/kg of feed DM of Roxazyme G2 liquid with activities (U/ml enzyme preparation) of xylanase 260 000, β‐glucanase 180 000 and cellulase 8000 (DSM Nutritional Products, Basel, Switzerland). The concentrations of 16S rDNA of Anaerovibrio lipolytica, Fibrobacter succinogenes, Prevotella ruminicola, Ruminococcus flavefaciens, Selenomonas ruminantium and Treponema bryantii, and their relative percentage of total bacteria in rumen samples obtained before feeding and 3 and 7 h after feeding and from two rumen fractions were determined using real‐time PCR. Sampling time had only little influence, but bacterial numbers and the composition of the population differed between the transition layer between rumen fluid and the fibre mat (fraction A) and the rumen fluid (fraction B) highlighting the importance to standardize sampling. The 16S rDNA copies of total bacteria and the six bacterial species as well as the population composition were mainly unaffected by the high levels of exogenous enzymes supplemented at all sampling times and in both rumen fractions. Occasionally, the percentages of the non‐fibrolytic species P. ruminicola and A. lipolytica changed in response to enzyme supplementation. Some increases in the potential degradability of the diet and decreases in lag time which occurred collaterally indicate that other factors than changes in numbers of non‐particle‐associated bacteria are mainly responsible for the effects of exogenous enzymes.     

Metagenomic profiles of the rumen microbiota during the transition period in low‐yield and high‐yield dairy cows

We investigated potential relationships between rumen microbiota and milk production in dairy cows during the transition period. Twelve dairy cows were divided into a low‐yield (LY) or high‐yield (HY) group based on their milk yield. Rumen samples were taken from dairy cows at 3 weeks before parturition, and at 4, 8, and 12 weeks after parturition. 16S rDNA‐based metagenomic analysis showed that diversities of rumen microbiota in both groups were similar and the number of operational taxonomic units (OTUs) was lower in the postpartum than prepartum period in both groups. The abundance of Bacteroidetes and ratio of Bacteroidetes:Firmicutes was higher in the HY than the LY group. OTUs assigned to Prevotella bryantii, Fibrobacter succinogenes, Ruminococcus albus, Butyrivibrio fibrisolvens, and Succinivibrio sp. were abundant in the HY group. These OTUs were significantly related to the propionate molar proportion of rumen fluids in the HY group. OTUs assigned to Lachnospiraceae, Bifidobacterium sp. and Saccharofermentans were dominant in the LY group. Predictive functional profiling revealed that abundance of gene families involved in amino acid and vitamin metabolism was higher in the HY than the LY group. These results suggest that the community structure and fermentation products of rumen microbiota could be associated with milk production of dairy cows.     

Characteristics of various fibrolytic isozyme activities in the rumen microbial communities of Japanese Black and Holstein Friesian cattle under different conditions

Rumen microorganisms produce various fibrolytic enzymes and degrade lignocellulosic materials into nutrient sources for ruminants; therefore, the characterization of fibrolytic enzymes contributing to the polysaccharide degradation in the rumen microbiota is important for efficient animal production. This study characterized the fibrolytic isozyme activities of a rumen microbiota from four groups of housed cattle (1, breeding Japanese Black; 2, feedlot Japanese Black; 3, lactating Holstein Friesian; 4, dry Holstein Friesian). Rumen fluids in all cattle groups showed similar concentrations of total volatile fatty acids and reducing sugars, whereas acetic acid contents and pH were different among them. Predominant genera were commonly detected in all cattle, although the bacterial compositions were different among cattle groups. Zymograms of whole proteins in rumen fluids showed endoglucanase activities at 55 and 57 kDa and xylanase activity at 44 kDa in all cattle. Meanwhile, several fibrolytic isozyme activities differed among cattle groups and individuals. Treponema, Succinivibrio, Anaeroplasma, Succiniclasticum, Ruminococcus, and Butyrivibrio showed positive correlations with fibrolytic isozyme activities. Further, endoglucanase activity at 68 kDa was positively correlated with pH. This study suggests the characteristics of fibrolytic isozyme activities and their correlations with the rumen microbiota.     

Effects of yeast cultures with different fermentation times on the growth performance,caecal microbial community and metabolite profile of broilers

The objective of this study was to investigate the effects of yeast culture (YC) on the growth performance, caecal microbial community and metabolic profile of broilers. A total of 350 1-day-old healthy Arbor Acres broilers were randomly assigned to seven treatment groups. The first group received a basal diet without YC supplementation, whereas the remaining groups received a basal diet supplemented with either YC fermented for 12, 24, 36, 48 or 60 hr, or a commercial YC product (SZ2). MiSeq 16S rRNA high-throughput sequencing was used to investigate the bacterial community structure, and gas chromatography–mass spectrometry was used to identify the metabolites in the caeca of broilers. The broilers that received a diet supplemented with YC had a higher average daily gain and average daily feed intake than those received YC-free or SZ2-enriched diets. The feed conversion ratio (FCR) of YCs fermented for 24 hr resulted in the best feed efficiency, whereas the FCR of YC fermented for 60 hr resulted in poor feed efficiency (p < .05). In the caeca of broilers, the bacterial communities were well separated, as determined by principal component analysis, and the proportions of the eight genera were significantly different among the seven groups (p < .05). The genus Akkermansia was the most abundant when the diet supplemented with YC fermented for 24 hr (p < .05). Furthermore, the Firmicutes/Bacteroidetes ratio was positively correlated with the FCR in the caecum (r = .47, p < .005). Five differentially expressed metabolites (i.e., L-alanine, benzeneacetic acid, D-mannose, D-arabitol and cholesterol) were identified in the caeca of broilers that received diets supplemented with YCs fermented for 24 or 60 hr. In summary, the different fermentation times of the YCs can markedly improve the growth performance and FCR of broilers by altering the caecal microbial community, and the growth performance which is related to the changes in key metabolic pathways.