Rumen bacterial diversity in relation to nitrogen retention in beef cattle.

This study aimed to characterize the rumen bacterial diversity of beef steers differing in the efficiency of nitrogen retention (ENR). Eight castrated steers and fitted with ruminal silicone - and duodenal T-type cannulas were used in a cross-over design with three consecutive periods and three diets. During each experimental period, nitrogen balance was measured, and based on the efficiency of N utilization data, steers were split into three ENR groups: high (HNR, 56.6% ± 3.3%, n = 10), medium (MNR, 45.8% ± 2.2%, n = 6), and low (LNR, 37.7% ± 1.9%, n = 8) using the NbClust package version 2.0.4 in R. Prevotellaceae, Lactobacillaceae, Leuconostocaceae, Clostridiales_Incertae_Sedis_XIII, Lachnospiraceae, and Peptostreptococcaceae were more abundant in LNR (P < 0.05) compared to HNR or MNR. Negative correlations were found between N retention and Mogibacterium, Anaerofustis, Butyrivibrio, Coprococcus, Hespellia, Lactonifactor and Lachnospiraceae (r ≤ -0.61; P ≤ 0.05). Prevotella, Hespellia, Lactonifactor, Lachnospiraceae_other, and Anaerobiospirillum were positively correlated between urinary N excretion (r > 0.55; P < 0.01), and negative correlations were found with Elusimicrobia, Victivallis and Treponema (r < -0.41; P < 0.05). The adjustment of the rumen bacterial community differed significantly between the N use retention groups. The high N retention in beef cattle was associated with less abundant bacteria in the rumen; however, N fixation capacity and uncharacterized rumen microorganisms need to be elucidated in future studies. In contrast, lower N utilization was associated with high abundance of bacteria that promote greater urinary N excretion through ruminal protein degradation.

Effect of soybean oil availabilities on rumen biohydrogenation and duodenal flow of fatty acids in beef cattle fed a diet with crude glycerine.

Soybean oil with different ruminal availability (whole soybeans (WS), soybean oil (SO) and calcium salts (CS)) was used to evaluate the fatty acid (FA) intake, rumen biohydrogenation (BH) and duodenal flow of FA in Nellore steers fed diets with crude glycerine (CG). Eight castrated Nellore steers were fitted with a ruminal and duodenal silicone cannula, and distributed in a double, simultaneous, Latin square 4 × 4 design with four diets and four experimental periods. Concentrates contained ground maize, urea, mineral salts, CG (100 g/kg DM) and soybean products with different availability of soybean oil: (1) no additional fat (CO), (2) WS, (3) SO or (4) CS. Fat supplementation was fixed to obtain 50 g ether extract/kg DM. Experimental treatments had no effect on DM intake, DM duodenal flow or ruminal turnover rate of C:16 FA. However, fat addition increased C:18 and turnover rates of total FA rumen (p < 0.05). CS resulted in lower C:18 turnover rates and lower ruminal BH of monounsaturated and unsaturated FA (UFA) than WS (p < 0.05). SO resulted in a greater duodenal flow of C18:0 (stearic acid), C18:1t-11 (vaccenic acid) and saturated FA than the WS and CS diets (p < 0.05). CS resulted in a higher duodenal flow of C18:3n-3 (linolenic acid) than WS (p < 0.05). The association of CG and calcium salts in Nellore steers was the best nutritional strategy to increase duodenal flow of healthier UFA, which may increase the deposition of these FA in meat. However, SO associated with CG association increased the duodenal flow of vaccenic acid, which is main precursor of endogenous synthesis of conjugated linoleic acids in tissues.

Efficiency of Amino Acid Utilization in Nellore Cattle Grazing Low-Quality Forage Supplemented with Different Sources of Nitrogen.

This study aimed to evaluate the effects of supplementation with non-protein nitrogen (NPN) or ruminal undegradable protein (RUP) on intake, digestibility, and amino acid (AA) use efficiency of Nellore cattle grazing during the dry season. Eight Nellore steers (12 ± 2 months old) were used in quadruplicate Latin squares (2 × 2). The animals were placed on Urochloa brizantha cv. Xaraés under continuous grazing. The treatments included the following: (1) urea supplementation (NPN) and (2) supplementation of corn gluten meal 60 (CGM, RUP). Animals supplemented with CGM showed higher intakes of dry matter (DM) supplement, total AA, essential AA, and individual AA. The supplementation did not affect the total AA digestibility, total AA flux, and the AA fluxes of microbial origin and RUP from the diet (p > 0.05). The ruminal microorganism origin flux of total AA to the duodenum was 44.5% and 52.7% for animals supplemented with NPN and CGM, respectively. Animals supplemented with CGM showed an increase in blood concentrations of isoleucine (+19.09 µmol/L), cystine (+27.29 µmol/L), and albumin (+0.11 g/dL) (p < 0.05), but this increase was not accompanied by an improvement in N use efficiency of steers (p > 0.05). RUP supplementation via CGM can be an efficient nutritional strategy to enhance the intake and absorption of AA by Nellore cattle grazing low-quality forage during the dry season.

Changes in the Lipid Metabolism of the Longissimus thoracis Muscle in Bulls When Using Different Feeding Strategies during the Growing and Finishing Phases.

The objective was to evaluate the supplementation strategy's effect on beef cattle during the growing phase and two systems during the finishing phase. One hundred and twenty young bulls were randomly divided in a 2 × 2 factorial design to receive either mineral (ad libitum) or protein + energy (3 g/kg body weight (BW)/day) during the growing phase and pasture plus concentrate supplementation (20 g/kg BW/day) or feedlot (25:75% corn silage:concentrate) during the finishing phase. Feedlot-fed bulls had meat (Longissimus thoracis-LT) with a higher content of lipids and saturated and monounsaturated fatty acids and a greater upregulation of stearoyl-CoA desaturase and sterol regulatory element-binding protein-1c than animals that fed on pasture (p < 0.05). On the other hand, pasture-fed bulls had meat with a higher content of α-linoleic acid, linolenic acid, and n6 and a greater n6:n3 ratio compared to the feedlot-fed group (p < 0.05). In addition, meat from pasture-fed bulls during the finishing phase had 17.6% more isocitrate dehydrogenase enzyme concentration than the feedlot group (p = 0.02). Mineral-fed and pasture-finished bulls showed down-regulation of peroxisome proliferator-activated receptor gamma (p < 0.05), while the bulls fed protein + energy and finished in the feedlot had higher carnitine palmitoyltransferase 2 expression (p ≤ 0.013). In conclusion, mineral or protein + energy supplementation in the growing does not affect the fatty acid composition of intramuscular fat of LT muscle. In the finishing phase, feeding bulls in the feedlot upregulates the lipogenic genes and consequently improves the intramuscular fat content in the meat.

Effects of rumen undegradable protein sources on nitrous oxide, methane and ammonia emission from the manure of feedlot-finished cattle.

The effects of sources of rumen undegradable protein (RUP) in diets on methane (CH4), nitrous oxide (N2O) and ammonia (NH3) emissions from the manure of feedlot-finished cattle were evaluated. We hypothesized that the use of different RUP sources in diets would reduce N loss via urine and contribute to reduced N2O, CH4 and NH3 emissions to the environment. Nellore cattle received different diets (18 animals/treatment), including soybean meal (SM, RDP source), by-pass soybean meal (BSM, RUP source) and corn gluten meal (CGM, RUP source). The protein source did not affect the N and C concentration in urine, C concentration in feces, and N balance (P > 0.05). The RUP sources resulted in a higher N2O emission than the RDP source (P = 0.030), while BSM resulted in a higher N2O emission than CGM (P = 0.038) (SM = 633, BSM = 2521, and CGM = 1153 g ha-2 N-N2O); however, there were no differences in CH4 and NH3 emission (P > 0.05). In conclusion, the use of RUP in diets did not affect N excretion of beef cattle or CH4 and NH3 emission from manure, but increased N2O emission from the manure.

Body weight and sex effects on digesta mean retention time in growing Saanen goats.

Despite the important role of digesta mean retention time (MRT) on digestive efficiency of ruminants, it is poorly investigated in total gastrointestinal tract (GIT) of growing ruminants, especially in goats. The objective of this study was to evaluate the effect of body weight (BW) and sex on GIT MRT of particles and solutes in growing Saanen goats. A dataset from two studies, comprising 103 individual records of castrated males (n = 36), females (n = 34), and intact males (n = 33) Saanen goats slaughtered at 15, 22, 30, 37, and 45 kg BW, was used. Goats were fed basically with total mixed ration composed by dehydrated corn plant (Zea mays) milled to pass a 10-mm screen, cracked corn grain, and soybean (Glycine max) meal. Variables evaluated were BW, feed intake, feed intake level, composition of ingested diet, wet weight of GIT tissues, wet digesta pool size, digesta composition (dry matter and neutral detergent fiber [NDF]), indigestible NDF:NDF ratio of ingested diet and GIT digesta, MRT of particles (MRTiNDF) and solutes (MRTCr), and reticulorumen selectivity factors (large particles/solutes). Reticulorumen, omasum, abomasum, small intestine, cecum, and colon-rectum segments were evaluated. The dataset was analyzed as mixed models considering sex, BW, and sex × BW interaction as fixed effects, and study and residual error as random effects. Sex did not affect MRTiNDF in any GIT segments. Females and intact males presented similar reticulorumen MRTCr (5.6 h; P = 0.92) and they presented lower reticulorumen MRTCr than castrated males (7.0; P ≤ 0.04). Total GIT MRTCr was similar between castrated males and females (15.7 h; P = 0.11) and between females and intact males (14.2 h; P = 0.76). Body weight (BW) did not affect MRTiNDF in reticulorumen and colon-rectum and total GIT MRTCr (P ≥ 0.11). Reticulorumen and omasum MRTCr increased as BW increased (P < 0.01), and abomasum MRTCr decreased as BW increased (P = 0.02). Feed intake, and wet tissues and wet pool size of all GIT segments increased as BW increased, except abomasum wet pool size (P ≤ 0.01). The mechanism related to sex effect on MRT has to be elucidated. Reticulorumen MRTiNDF and total GIT MRTCr were modulated by intake and capacity of reticulorumen and GIT, respectively. On the other hand, reticulorumen MRTCr seemed to be regulated by reticulo-omasal orifice opening and saliva secretion.

Long-Term Encapsulated Nitrate Supplementation Modulates Rumen Microbial Diversity and Rumen Fermentation to Reduce Methane Emission in Grazing Steers.

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 NO3 -/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 CH4/kg of forage dry matter intake (fDMI) (P = 0.037). Lower concentrations of NH3-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.

Corrigendum: Long-Term Encapsulated Nitrate Supplementation Modulates Rumen Microbial Diversity and Rumen Fermentation to Reduce Methane Emission in Grazing Steers.

[This corrects the article DOI: 10.3389/fmicb.2019.00614.].

Studies on bacterial community composition are affected by the time and storage method of the rumen content.

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.

Ingestive behavior of supplemented Nellore heifers grazing palisadegrass pastures managed with different sward heights.

Three sward heights (15, 25 and 35 cm) and three supplement types (energy, energy-protein, and a mineral mix supplement) were evaluated in a 3 × 3 factorial arrangement distributed in a completely randomized design to study changes in forage search patterns in Nellore heifers in a continuous grazing system. Pasture data were collected using two replicates (paddocks) per treatment over four periods during the rainy season. The behavior assessments were made in the first and fourth grazing seasons. It was hypothesized that supplements and pasture management would modify ingestive behavior, considering that animals would require less time grazing if they had energy requirements met through higher digestibility of better managed paddocks, or use of supplements high in energy. Total and green forage masses along with green : dead material ratio were greater in treatments managed with higher sward heights. Sward managed with 35 cm height resulted in lower leaf : stem ratio compared with 15 cm sward height treatments. The animals on the 15 cm pastures spent more time grazing overall and during each meal, but there were no differences observed in meal numbers in comparison to 35 cm treatments. Heifers fed protein and/or energy supplements spent less time grazing in the early afternoon, but overall grazing time was the same for all animals.

Suplementação protéica energética no desempenho de novilhas em pastejo durante a fase de terminação / Energetic protein supplementation on heifers performance in grazing during the finish phase

Para estudar o efeito da suplementação da dieta sobre o desempenho de novilhas na fase de terminação mantidas em pastagem de capim-marandu sob lotação intermitente, durante a estação chuvosa, foram utilizadas 20 novilhas cruzadas (» Nelore, » Santa Gertrudes, ½ Braunvieh) com peso corporal médio de 300 kg e 22 meses de idade. Os tratamentos constituíram-se de dois tipos de suplementação: sal mineral (SM) ad libtum e suplemento protéico energético (SPE) fornecido a 0,3 por cento do peso corporal (PC) por dia com avaliações realizadas a cada 28 dias, no período entre dezembro de 2006 e março de 2007 Foram avaliados o desempenho animal e as características de carcaça medidas por ultrassom. Observou-se o efeito (P=0,057) da suplementação sobre o ganho de peso dos animais suplementados, 0,700 kg/dia e 0,587 kg/dia dos animais que receberam sal-mineral. Houve efeito da suplementação na área de olho de lombo e profundidade do músculo Gluteus medius. As demais características analisadas pela técnica de ultrassom não diferiram entre si em função da suplementação. A suplementação protéica energética da dieta permite ganhos adicionais, o que reflete em aumento da deposição de músculo de novilhas mantidas em pastagem de capim-marandu durante o período das águas.

This study was conducted to evaluate the effect of the diet supplementation on the heifers performance in finishing phase, maintained in a marandu grass pasture managed in intermittent grazing system during the rainy season. Twenty crossbreed (» Nelore, » Santa Gertrudes, ½ Braunvieh) heifers with average initial body weight of 300 kg and 22 months old were used. The treatments consisted of two types of supplements: mineral (SM) ad libtum and energy protein supplementation (PES) provided 0.3 percent of the body weight daily. The evaluated variables were: animal performance and carcass traits measured by ultrasound each 28 days. Supplementation affected daily weight gain (P=0.057), 0.700 kg/day for supplemented animals against 0.587 kg/day for animals receiving salt-mineral. There were no supplementation effect on the loin eye area and depth of the Gluteus medius muscle. The other characteristics examined by the ultrasound technique did not differ according to the treatments. Protein and energy supplementation of the diet allows additional gains that reflected in increased deposition of muscle of heifers grazing marandu grass during the wet season.

Degradação in situ da matéria seca e da fração fibrosa do capim marandu obtido por diferentes métodos de amostragem, no período seco do ano / Dry matter and fibrous fraction in situ degradability of palissade grass sampled through different methods in the dry season

Este trabalho foi realizado na Faculdade de Ciências Agrárias Veterinárias - Unesp - Jaboticabal e foi conduzido, com o objetivo de avaliar a influência do método de amostragem do pasto sobre a degradabilidade in situ da matéria seca e da fração fibrosa de capim Marandu, colhido no período seco dos anos de 2003 e 2005. O experimento foi instalado em delineamento de blocos casualizados com parcela subdividida, com três repetições, representadas pelos piquetes amostrados. Nas parcelas, foram avaliados cinco métodos de amostragens de forragem (método do quadrado metálico; método de avaliação através de extrusa de bovino da raça Nelore; método de avaliação por meio de extrusa de bovino Cruzado (Red Angus x Nelore); método de avaliação por meio do pastejo, simulando bovino da raça Nelore; método de avaliação por meio do pastejo, simulando bovino Cruzado (Red Angus x Nelore) e as subparcelas foram constituídas pelos anos de amostragem, 2003 e 2005. Foram determinadas as frações da cinética ruminal: solúvel "A"; insolúvel potencialmente degradável "B"; taxa de degradação "Kd"; degradação potencial (DP) e fração não degradável "C" da MS e degradação potencial (DP), fração não degradável "C" e taxa de degradação "Kd" da FDN e da FDA. De acordo com os resultados obtidos, observou-se que o método do quadrado metálico subestimou as características da degradação do capim. Os métodos do pastejo simulado se assemelharam muito ao das extrusas, no entanto, a prática do simulador é que assegurou a amostragem eficiente, conforme foi constatado pelos dados obtidos no ano de 2003 e 2005.

This experiment was carried out at the Faculdade de Ciências Agrárias Veterinárias - Unesp - Jaboticabal during dry season, to evaluate the effects of sampling methods on the Marandu grass dry matter and cell wall in situ degradability. The data were analyzed according to a randomized block design with three replications (paddock), in a split plot scheme. Five sampling methods were evaluated on the parcels: metallic square method; Nellore steers extrusa, Red Angus x Nellore crossbreeding steers extrusa, simulated grazing of the Nellore, simulated grazing of Red Angus x Nellore crossbreeding, and two years (2003 and 2005) on the subparcels. The following rumen kinetic fractions were determined: soluble "A", degradable potential "B", degradability rate "Kd", the potential degradability (DP) and no degradable fraction "C" of the FDN, and FDA. According to the results obtained, the metallic square method underestimated the characteristics of the forage degradation. The results of the simulated grazing method resembled the extrusas method; however, it was the simulator practice that assured efficient sampling, as shown by evidence from data obtained between 2003 and 2005.