This study investigated the long-term effects (13 months) of encapsulated nitrate supplementation (ENS) on enteric methane emissions, rumen fermentation parameters, ruminal bacteria, and diversity of archaea in grazing beef cattle. We used a total of thirty-two Nellore steers (initial BW of 197 ± 15.3 kg), 12 of which were fitted with rumen cannulas. For 13 months, the animals were maintained in 12 paddocks and fed a concentrate of ground corn, soybean meals, mineral supplements, and urea (URS) or encapsulated nitrate (EN) containing 70 g of EN/100 kg of BW (corresponding to 47 g NO 3 - /100 kg BW). Encapsulated nitrate supplementation resulted in similar forage, supplement and total DMI values as URS ( P > 0.05), but ENS tended to increase (+48 g/d; P = 0.055) average daily weight gain. Daily reductions in methane emissions (-9.54 g or 18.5%) were observed with ENS when expressed as g of CH 4 /kg of forage dry matter intake (fDMI) ( P = 0.037). Lower concentrations of NH 3 -N and a higher ruminal pH were observed in ENS groups 6 h after supplementation ( P < 0.05). Total VFA rumen concentration 6 h ( P = 0.009) and 12 h after supplementation with EN resulted in lower acetate concentrations in the rumen ( P = 0.041). Steers supplemented with EN had a greater ruminal abundance of Bacteroides, Barnesiella, Lactobacillus , Selenomonas, Veillonella, Succinimonas, Succinivibrio , and Duganella sp. ( P < 0.05), but a lower abundance of Methanobrevibacter sp. ( P = 0.007). Strong negative correlations were found between daily methane emissions and Proteobacteria, Erysipelotrichaceae, Prevotellaceae, and Roseburia , Kandleria , Selenomonas , Veillonella , and Succinivibrio sp. ( P < 0.05) in the rumen of ENS steers. Encapsulated nitrate is a feed additive that persistently affects enteric methane emission in grazing steers, thereby decreasing Methanobrevibacter abundance in the rumen. In addition, ENS can promote fumarate-reducer and lactate-producer bacteria, thereby reducing acetate production during rumen fermentation.

The aim of this study was to investigate the effect of different dietary levels of concentrate on feed intake, digestibility, ruminal fermentation and microbial population in steers. Eight Nellore steers fitted with ruminal cannulas were used in a double 4 × 4 Latin square design experiment. The dietary treatments consist of four different proportions of concentrate to roughage: 30:70, 40:60, 60:40 and 80:20% in the dry matter, resulting in Diets 30, 40, 60 and 80, respectively. The roughage was corn silage, and the concentrate was composed of corn, soybean meal and urea. Apparent digestibility of organic matter and crude protein showed a linear association with concentrate proportion (p = 0.01), but the increased concentrate levels did not affect the digestibility of fibre. The lowest ruminal pH-values were observed in animals fed with Diet 80, remaining below pH 6.0 from 6 h after feeding, while in the other diets, the ruminal pH was below 6.0 not before 12 h after feeding. After feeding Diet 80, the ammonia concentration in the rumen was significantly the highest. Higher dietary concentrate levels resulted in a linear increase of propionic acid concentrations, a linear reduction of the ratio acetic acid to propionic acid (p < 0.01) and a linear increased synthesis of microbial nitrogen (p < 0.001). The predicted production of methane was lower in diets with greater amounts of concentrate (p = 0.032). The population of methanogens, R. flavefaciens and R. albus decreased with higher concentrate levels, while the population of S. ruminantium increased (p < 0.05). The results indicate that greater amounts of concentrate do not decrease ruminal pH-values as much as expected and inhibit some cellulolytic bacteria without impairing the dry matter intake and fibre digestibility in Nellore steers.

The objective of this study was to investigate three storage methods and four storage times for rumen sampling in terms of quality and yield of extracted metagenomic DNA as well as the composition of the rumen bacterial community. One Nellore steer fitted with a ruminal silicone-type cannula was used as a donor of ruminal contents. The experiment comprised 11 experimental groups: pellet control (PC), lyophilized control (LC), P-20: pellet stored frozen at -20°C for a period of 3, 6, and 12 months, P-80: pellet stored frozen at -80°C for a period of 3, 6, and 12 months, and L-20: lyophilized sample stored frozen at -20°C for a period of 3, 6, and 12 months. Metagenomic DNA concentrations were measured spectrophotometrically and fluorometrically and ion torrent sequencing was used to assess the bacterial community composition. The L-20 method could not maintain the yield of DNA during storage. In addition, the P-80 group showed a greater yield of metagenomic DNA than the other groups after 6 months of storage. Rumen samples stored as pellets (P-20 and P-80) resulted in lower richness Chao 1, ACE, and Shannon Wiener indices when compared to PC, while LC and PC were only different in richness ACE. The storage method and storage time influenced the proportions of 14 of 17 phyla identified by sequencing. In the P-20 group, the proportion of Cyanobacteria, Elusimicrobia, Fibrobacteres, Lentisphaerae, Proteobacteria, and Spirochaetes phyla identified was lower than 1%. In the P-80 group, there was an increase in the proportion of the Bacteroidetes phylum (p = 0.010); however, the proportion of Actinobacteria, Chloroflexi, SR1, Synergistetes, TM7, and WPS.2 phyla were unchanged compared to the PC group (p > 0.05). The class Clostridium was the most abundant in all stored groups and increased in its proportion, especially in the L-20 group. The rumen sample storage time significantly reduced the yield of metagenomic DNA extracted. Therefore, the storage method can influence the abundance of phyla, classes, and bacterial families studied in rumen samples and affect the richness and diversity index.

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Lipid supplementation may adversely affect rumen fermentation, microbial abundance, nutrient utilization and the duodenal flow of fatty acids (FA). In a 5 × 5 Latin square design, 10 Nellore steers with ruminal and duodenal cannulas (292 ± 28 kg BW) were fed one of the five following dietary lipid sources in the concentrate, as follow: (1) no additional fat (WF); (2) palm oil (PO) derived from the Palmaceae plant Orbignya oleifera; (3) linseed oil (LO); (4) protected fat (PF); and (5) whole soybeans (WS). The dry matter intake (DMI) was greater for the animals supplemented with WF than for those supplemented with PO, PF, and WS. The DM digestibility was 11% greater for the animals in the WF group than for those in the PO group. The animals in the PO group had greater duodenal flows of capric (C10:0), lauric (C12:0) and myristic (C14:0) saturated fatty acids (SFA) than did animals in the other groups. The amount of flows of vaccenic acid and total unsaturated acids (UFA) was greater with LO supplementation. The unsaturated fatty acid (UFA) biohydrogenation ratio was greater in the animals supplemented with WS than in those supplemented with PO, LO, and PF. The acetate: propionate ratio was lower in the animals supplemented with PO and LO, at 2.9 and 3.0, respectively. The ruminal flow of particles and rate of fiber degradation were lower with PO supplementation. The proportion of archaea was reduced with the inclusion of PO, LO and PF. PO supplementation resulted in lower total apparent NDF digestibility than did WF, PF, and WS supplementation. Therefore, LO, PF and WS can be used as dietary supplements for grazing cattle without impairing fermentation. The inclusion of PO in the diets of grazing cattle may reduce the number of methanogens and alter fiber utilization.

SUMMARYThe use of diets with increasing proportions of concentrate to fibre can ensure appropriate energy levels and result in greater efficiency in Nellore feedlot steers. It was hypothesized that higher proportions of concentrate in the diet of these Nellore steers may affect ruminal fermentation and microbiota as a consequence of ruminal pH reduction. The present study was conducted to evaluate the effect of diets with four different roughage (hay Tifton 85) : concentrate ratios on intake, digestibility, rumen fermentation and rumen microbiota of Nellore feedlot steers. Higher proportions of concentrate in the diet did not affect intake and digestibility of dry and organic matter. The concentration of N-NH3, total rumen volatile fatty acid, acetic (C2), butyric (C4), isobutyric, valeric and isovaleric acids, and microbial nitrogen did not differ among diets. However, increasing proportions of concentrate in the diet resulted in a linear reduction in average rumen pH and increased propionic acid (C3) concentration, resulting in lower relative C2 : C3. Bacterial population of Fibrobacter succinogenes, Ruminococus flavefaciens and Ruminococcus albus decreased in the rumen. However, bacteria that are consumers of lactic acid (Selenomonas ruminantium and Megasphaera elsdenii) and producers of lactic acid (Lactobacillus sp. and Streptococcus bovis) increased when animals were fed with high-concentrate diets. The total number of protozoa was similar for the different roughage : concentrate ratios. Protozoan counts were only influenced by diet for the genus Dasytricha. The findings point to diets with increasing concentrate to Tifton 85 hay ratios as inhibiting the growth of some cellulolytic bacteria and reducing fibre digestibility, and indicate Tifton 85 hay as a possible modulated rumen fermentation in the Nellore steer feedlot.

The objective of this study was to determine whether a combination of crude glycerin (CG) and soyabean oil (SO) could be used to partially replace maize in the diet of Nellore steers while maintaining optimum feed utilisation. Eight castrated Nellore steers fitted with ruminal and duodenal cannulas were used in a double 4×4 Latin square design balanced for residual effects, in a factorial arrangement (A×B), when factor A corresponded to the provision of SO, and factor B to the provision of CG. Steers feed SO and CG showed similar DM intake, DM, organic matter and neutral-detergent fibre digestibility to that of steers fed diets without oil and without glycerine (P>0·05). Both diets with CG additions reduced the acetate:propionate ratio and increased the proportion of iso-butyrate, butyrate, iso-valerate and valerate (P<0·05). Steers fed diets containing SO had less total N excretion (P<0·001) and showed greater retained N expressed as % N intake (P=0·022). SO and CG diet generated a greater ruminal abundance of Prevotella, Succinivibrio, Ruminococcus, Syntrophococcus and Succiniclasticum. Archaea abundance (P=0·002) and total ciliate protozoa were less in steers fed diets containing SO (P=0·011). CG associated with lipids could be an energy source, which is a useful strategy for the partial replacement of maize in cattle diets, could result in reduced total N excretion and ruminal methanogens without affecting intake and digestibility.