Ruminal Fermentation Research Articles

287 Effects of crude glycerin supplementation of beef cattle grazing wheat pasture on forage intake and characteristics of rumen fermentation

Abstract Crude glycerin (CG), mainly composed of glycerin, is a by-product of biodiesel production. Glycerin can be utilized as a substrate of gluconeogenesis by either rapid fermentation to propionate in the rumen or can be directly absorbed through the rumen epithelial wall, resulting in the conversion to glucose in the liver. Therefore, CG supplementation may increase the energy intake of cattle grazing wheat pasture. Ruminally cannulated Angus-crossbred cows [n = 9; 564.4 ±101.6 kg initial body weight (BW)] grazing wheat pasture were utilized in a split-plot design with the objective of evaluating the effects of CG supplementation and forage maturity on forage intake and rumen fermentation characteristics. Treatment was the main plot, and stage of forage maturity was the subplot. Treatments were unsupplemented control (CON) and crude glycerin supplementation (SUP; 37.2% glycerin, DM basis) dosed at 0.11% BW (DM basis). Supplementation levels correspond to 5% of the diet, assuming forage intake of 2.2% of BW on a DM basis. The stages of wheat maturity pasture were March (MAR; vegetative stage) and April (APR; early reproductive stage). Cows grazed a single wheat pasture and were supplemented directly through the rumen cannula twice daily at 0700 and 1900 h. Forage intake in kg/d and as % of BW was greater (P = 0.04) for SUP than CON. Ruminal pH, ammonia, and VFA concentrations, as well as proportions of VFAs, were not affected by CG supplementation (P ≥ 0.34; Table 1). However, ruminal pH (5.77 and 5.99 ± 0.049) and ammonia concentration (0.67 and 3.37 ± 0.307 mM) were greater, while total VFA concentration (136.3 and 110.8 ± 5.23 mM) was less (P = 0.01) for APR compared with MAR. Acetate, propionate, and butyrate molar proportions, as well as acetate: propionate ratio, were not affected (P ≥ 0.27) by the stage of wheat pasture maturity. Results indicate that crude glycerin used as an ingredient in supplementation strategies for cattle grazing wheat pasture may increase cattle energy intake without negatively affecting forage intake and ruminal fermentation.

154 Forage intake, digestibility, and ruminal fermentation characteristics of beef cows supplemented with a Bacillus-based probiotic

Abstract The aim of this study was to evaluate the influence of a Bacillus-based probiotic supplementation on intake, digestibility, and ruminal fermentation parameters of beef cows consuming a low-quality forage diet (7.0% CP, 60% NDF, 39% ADF, DM basis). Ruminally fistulated Angus × Hereford cows [n = 15; age = 6.1 ± 1 yr; body weight (BW) = 655 ± 57] were used in two identical complete randomized designs, housed in individual pens with water, and forage intake monitored daily. Treatments consisted of 1) 500 g of protein supplement (dry distillers grain + mineral mix) without probiotic (CON n = 15), and 2) 500 g of protein supplement + 3 g of a probiotic containing Bacillus licheniformis and B. subtilis (BOV; n = 15; 2.2 ×109 CFU of the mixture/g; Bovacillus, Novonesis, Hørsholm, Denmark). Each experimental period lasted 33 d, consisting of 21 d of adaptation, 12 d of collection, and 30 d of wash-out between periods. From d 22 to 26, fecal samples were collected to determine intake and nutrient digestibility using titanium dioxide as an indigestible marker. On d 27, ruminal fluid was collected at h 0 (before feeding), 4, 8, 12, 16, and 20 post-feeding to evaluate short-chain fatty acid (SCFA) and N-NH3 concentration. From d 28 to 32, hay samples (4 g) were fitted in Dracon bags and incubated in the rumen for 0, 12, 24, 48, 72, 96, and 120 h to determine nutrient digestibility. Cows receiving BOV had greater (P < 0.001) dry matter intake compared with the CON group (Table 1). Also, cows supplemented with BOV had greater (P < 0.02) dry matter and protein digestibility and a tendency (P ≤ 0.07) for greater NDF and ADF forage digestibility compared with CON. A tendency was also detected (P = 0.09) for the degradability of forage dry matter, which was improved in BOV vs. CON. No effects were observed on rumen fermentation characteristics (P ≥ 0.14; Table 2).). In conclusion, adding bacillus-based probiotics enhanced the intake, nutritional digestibility, and degradability of beef cows fed medium-quality forage diets.

401 Effect of protein and starch supplementation level on intake and digestibility by steers consuming low-quality forage: I. Bos taurus

Abstract A deficiency of ruminally available N (RAN) caused by a basal diet of low-quality forage depresses forage utilization. Supplemental protein improves forage utilization by increasing RAN, resulting in increased intake and potentially improving digestion. Addition of starch to a supplement can increase total digestible organic matter (OM) intake (TDOMI); however, too much starch decreases RAN and pH potentially decreasing forage utilization. Our objective was to determine the effect of starch supplementation on intake and digestion in Bos taurus indicus steers. Accordingly, five ruminally cannulated Brahman steers [body weight (BW) = 420 ± 55 kg) were used in a 4 × 4 Latin square to evaluate the effect of decreasing supplemental starch when consuming King Ranch Bluestem hay (3.5% CP, 73% NDF). Steers received four isonitrogenous (130 mg N/ kg BW) supplements providing increasing levels of starch (2% starch = 100% soybean meal; 20% starch = 26.3% corn, 73% soybean meal, 0.7% urea; 38% starch = 51.6% corn, 47% soybean meal, 1.4% urea; 56% starch = 78.6% corn, 19% soybean meal, 2.4% urea). Five 14-d periods were conducted, consisting of 9 d adaptation and 4 d to measure intake and digestion, and 1 d for ruminal fermentation. As designed, supplement OM intake (SOMI) increased linearly (P < 0.01) as starch level increased. Other measures of intake, including forage OM intake (FOMI), total OM intake (TOMI), and total neutral detergent fiber intake (TNDFI) decreased in a cubic manner in response to increasing starch (P ≤ 0.04). Forage OMI also responded with a cubic decrease (72.2, 68.9, and 69.5 g/kg BW0.75) when the protein supplement contained 2, 20, and 38% starch, respectively, but decreased to 61.2 g/kg BW0.75 when the 56% starch supplement was provided. Similarly, TOMI decreased 5.3% when starch inclusion increased from 38 to 56%. Total tract OM digestion and NDF digestion were not significantly affected (P ≥ 0.86) by starch level in the supplement; therefore, despite TOMI and TNDFI decreasing, there was no effect on total digestible OM intake and (TDOMI) and total digestible NDF intake (TDNDFI). In conclusion, providing supplemental starch over 38% decreased forage utilization. Decreases in FOMI with starch inclusion may be attributed to competition for RAN between fiber fermenting and starch fermenting bacteria.

Sward type alters enteric methane emissions, nitrogen output and the relative abundance of the rumen microbial ecosystem in sheep.

Observed improvements in animal and sward performance, coupled with a desire for more sustainable pasture-based feeding systems, has triggered a surge in the implementation of more botanically diverse pastures. However, thus far, there has been limited research investigating the effects of botanically diverse sward types on enteric methane (CH4) or nitrogen (N) excretion, alongside the ruminal microbiota and fermentation profile, in sheep. Hence, this study investigates the effect of sward type on CH4 production and N excretion, in addition to assessing the rumen microbiome, volatile fatty acid proportions, and ammonia nitrogen (NH3-N) concentration in sheep. A 5 × 5 Latin square design experiment was implemented to investigate 5 dietary treatments; perennial ryegrass (Lolium perenne L.; PRG) only or PRG plus white clover (Trifolium repens L.; PRG + WC), red clover (Trifolium pratense L.; PRG + RC), chicory (Chicorium intybus L.; PRG + Chic) or plantain (Plantago lanceolata L.; PRG + Plan). Diets were mixed at a ratio of 75% PRG and 25% of the respective companion forage and 100% PRG for the PRG treatment, on a dry matter basis. Twenty castrated male sheep were housed in metabolism crates across 5 feeding periods. Methane measurements were acquired utilizing portable accumulation chambers. Rumen fluid was harvested using a transoesophageal sampling device. Microbial rumen DNA was extracted and subjected to 16S rRNA amplicon sequencing and fermentation analysis. Data were analyzed using PROC MIXED in SAS. Results show that animals consuming PRG + WC ranked lower for CH4 production (g/d) than sheep offered PRG, PRG + Chic or PRG + Plan (P < 0.01) while the addition of any companion forage ranked CH4 yield (g/kg dry matter intake (DMI)) lower (P < 0.001) than PRG. There was a moderate positive correlation between DMI and CH4 (g/d; r = 0.51). Ruminal NH3-N was lowest in animals consuming the PRG diet (P < 0.01). There was a greater abundance of Methanobrevibacter and reduced abundance of Methanosphaera (P < 0.001) in sheep offered PRG, compared with any binary sward. On average, herb diets (PRG + Chic or PRG + Plan) reduced the urinary nitrogenconcentration of sheep by 34% in comparison to legume diets (PRG + WC or PRG + RC) and 13% relative to the PRG diet (P < 0.001). Sheep offered PRG + Chic had a greater dietary nitrogen use efficiency than PRG + RC (P < 0.05). This study demonstrates the potential for sward type to influence rumen function and the microbial community, along with CH4 and N output from sheep.

Multi-omics analysis reveals the effects of host-rumen microbiota interactions on growth performance in a goat model.

The growth rate of young ruminants has been associated with production performance in later life, with recent studies highlighting the importance of rumen microbes in supporting the health and growth of ruminants. However, the specific role of rumen epithelium bacteria and microbiota-host interactions in influencing the early life growth rate of ruminants remains poorly understood. In this study, we investigated the rumen fermentation pattern, microbiota characteristics, and global gene expression profiles of the rumen epithelium in 6-month-old goats with varying growth rates. Our results showed that goats with high average daily gain (HADG) exhibited higher rumen propionate concentrations. Goats with low average daily gain (LADG) had the higher relative abundances of rumen epithelium bacteria genera U29-B03 and Quinella, while exhibiting a lower relative abundance of Lachnospiraceae UCG-009. In the rumen fluid, the relative abundances of bacteria genus Alloprevotella were lower and Desulfovibrio were higher in LADG goats compared to HADG goats. Additionally, the relative abundance of fungal genus Symmetrospora was lower in LADG goats compared to HADG goats. Transcriptome analysis showed that 415 genes were differentially expressed between LADG and HADG goats, which were enriched in functions related to cell junction and cell adhesion, etc. Correlation analysis revealed that rumen epithelium bacteria genera UCG-005 and Candidatus Saccharimonas were negatively associated, while Lachnospiraceae NK3A20 group and Oscillospiraceae NK4A214 group were positively associated with average daily gain (ADG) and genes related to barrier function. The rumen fluid bacteria genus Alloprevotella was positively correlated, while Desulfovibrio was negatively correlated with rumen propionate and ammoniacal nitrogen (NH3-N) concentrations, as well as genes related to barrier function and short chain fatty acids (SCFAs) transport. In summary, our study reveals that the higher ruminal fermentation efficiency, improved rumen epithelial barrier functions, and enhanced SCFAs transport in HADG goats could be attributed to the rumen microbiota, particularly the rumen epithelium bacteria, such as Lachnospiraceae and Oscillospiraceae NK4A214 group.

Effects of protein grass hay as alternative feed resource on lamb's fattening performance and meat quality

Protein grass hay (PGH) was used as a new feed source for lambs to study its effect on fattening performance and meat quality. Fifty-six male lambs were allotted to four experimental groups and fed for eight weeks either alfalfa hay (AH)-based diet (control) or diets in which AH was replaced with 33 %, 66 %, or 99 % PGH. The inclusion of PGH did not affect final body weight, dry matter intake, average daily gain, feed conversion ratio, or carcass weight. Moreover, substituting AH with PGH at any level did not influence the ruminal fermentation or serum biochemical parameters, meat color, water holding capacity, shear force, or amino acid profile. However, relative liver weight was increased with 66 % substitutions. Furthermore, replacing 99 % AH with PGH decreased the meat's pH at 24 h. Higher levels of C18:3n-3, C20:5n-3, and total n-3 PUFA and a lower ratio of n-6: n-3 PUFA were also observed in meat from lambs fed PGH at 99 %. These findings suggest that PGH could be incorporated into the lamb's diet up to 99 % without compromising fattening performance and body health while improving their meat n-3 PUFA deposition.

The effects of dietary cation-anion difference and dietary buffer for lactating dairy cattle under mild heat stress with night cooling

The objective of this study was to investigate the interactive effect of dietary cation-anion difference (DCAD) and dietary buffer supply on DMI, ruminal fermentation, milk and milk component yields, and gastrointestinal tract (GIT) permeability in lactating dairy cattle exposed to mild heat stress. Sixteen lactating Holstein cows, including 8 ruminally cannulated primiparous (80 ± 19.2 DIM) and 8 non-cannulated multiparous (136 ± 38.8 DIM) cows, were housed in a tie-stall barn programmed to maintain a temperature-humidity index (THI) between 68 and 72 from 0600 h to 1600 h followed by natural night cooling. The experimental design was a replicated 4 × 4 Latin rectangle (21-d periods) with a 2 × 2 factorial treatment arrangement. Diets contained a low DCAD (LD; 17.5 mEq/100g of DM) or high DCAD (HD; 39.6 mEq/100g of DM) adjusted using NH4Cl and Na-acetate, with low (LB; 0% CaMg(CO3)2) or high buffer (HB; 1% CaMg(CO3)2). In addition to measurement of feed intake, ruminal fermentation, and milk and milk component yields, a ruminal dose of Cr-EDTA and an equimolar abomasal dose of Co-EDTA were used to evaluate total and post-ruminal gastrointestinal tract permeability, respectively. Treatments had no effect on DMI, ruminal short-chain fatty acid concentrations, or ruminal pH. Feeding HD improved blood acid-base balance, increased urine volume by 4 ± 1.5 kg/d, and increased milk fat by 0.14 ± 0.044 percentage units and milk fat yield by 36.5 ± 16.71 g/d. HB reduced milk fat percentage by 0.11 ± 0.044 percentage units and had no effect on milk fat yield. The HB treatments reduced urinary excretion of Co by 27% and tended to reduce urinary Cr excretion by 10%. Across all treatments, 72% of the Cr recovery was represented by Co suggesting that much of the permeability responses were post-ruminal during mild heat stress. In conclusion, increasing DCAD through greater Na supply during mild heat stress improved blood acid-base balance and may increase milk fat yield. Dietary inclusion of CaMg(CO3)2 improved post-ruminal GIT barrier function despite a lack of low ruminal pH. As there appeared to be a limited interactive effect between DCAD and buffer, increased DCAD and provision of buffer seem to independently influence physiological and performance responses in lactating dairy cows exposed to mild heat with night cooling.

Changes in ruminal fermentation and rumen bacteria population in feedlot cattle during a high lipid diet adaptation

Abstract This study aimed to investigate changes in feed intake and ruminal environmental parameters during a high-lipid diet transition in cattle. Eight Nellore steers were fed a control diet composed of 30% hay and 70% concentrate for 21 days, followed by the inclusion of 60 g/kg dry matter of soybean oil for 21 days. The DM intake expressed as a percentage of BW 21 days after lipid inclusion was lower (1.75% BW) than that observed during the control diet feeding (1.81% BW) (P&lt;0.01). Steers fed the control diet had a lower pH than the ruminal pH recorded on days 7, 14, and 21 after lipid inclusion (P=0.034). Lower total short-chain fatty acid production in the rumen and lower microbial nitrogen synthesis were observed on day 7 after lipid inclusion compared to values found when steers were fed the control diet and on days 14 and 21 after lipid inclusion (P=0.041). Lipid inclusion in the diet decreased the population of protozoa on days 7, 14, and 21 (P&lt;0.001). The abundances of R. albus and F. succinogenes were higher when steers were fed the control diet than the abundance observed on days 7, 14, and 21 after lipid inclusion in the diet (P&lt;0.05). The first seven days of lipid diet inclusion are considered the most critical for ruminal adaptation, involving reductions in fibrolytic bacteria and changes in fermentation parameters. After 14 days the rumen showed signs of recovery and adaptation.

Essential oils and capsaicin in the diet of Jersey cows at early lactation and their positive impact on anti-inflammatory, antioxidant and immunological responses.

The objective of this work was to determine whether the addition of phytogenic compounds based on essential oils (carvacrol, eugenol, cinnamaldehyde) and resinous pepper oil (capsaicin) to the diet of Jersey cows at the beginning of lactation affects anti-inflammatory, antioxidant and immunomodulatory responses, as well as whether there are effects of EO on blood metabolites, ruminal fermentation, digestibility and milk production and composition. Six primiparous cows (370.00 ± 17kg body weight (BW); 13.02kg dry matter intake (DMI); 21 days of lactation and average milk production of 20 ± 2L per day) were allocated to crossed experimental design (2 × 2) with two experimental periods of 28 days and two treatments. Blood, milk and rumen fluid were collected and, at the end of each period, feed and feces samples were collected to evaluate the apparent digestibility of nutrients. The groups were control (CLT) without supplementation and treated (BEO) with the addition of 150mg/kg of dry matter of the phytogenic to the concentrated portion of the diet. Cows in the BEO group had lower numbers of leukocytes (P ≤ 0.05) and lymphocytes (P ≤ 0.02), but total protein and globulin levels were higher on days 21 and 28 (P ≤ 0.01). In the BEO group, the levels of immunoglobulin A, immunoglobulin heavy chain and transferrin were higher (P ≤ 0.05). The levels of ceruloplasmin, haptoglobin and C-reactive protein were lower in the BEO group (P ≤ 0.05). Lipid peroxidation levels and protein carbonyl content were lower in the BEO group. The total antioxidant capacity (P ≤ 0.09) and the activity of glutathione S-transferase (P ≤ 0.03) and glutathione peroxidase (P ≤ 0.05) were higher in the BEO group. Cows in the BEO group had lower pH (P ≤ 0.05), acetic acid concentrations (P ≤ 0.01) and higher protozoa counts (P ≤ 0.01). Our results suggest that phytogenic supplementation has positive effects on the health of Jersey cows in early lactation, characterized by immunostimulant, antioxidant and anti-inflammatory effects.

Effects of High-Grain Diet on Performance, Ruminal Fermentation, and Rumen Microbial Flora of Lactating Holstein Dairy Cows.

The objectives of the current study were to evaluate the fluctuations in production performance, rumen fermentation, and microbial community in lactating dairy cows fed a high-grain diet (HG). In this study, 16 healthy Holstein lactating dairy cattle with similar milk yields of 16.80 ± 4.30 kg/d, days in milk 171.44 ± 23.25 days, and parity 2.2 ± 1.5 times were selected and randomly allocated into two groups. One group was fed a low-grain diet (LG; 40% concentrate, DM basis; n = 8), and the other group was fed a high-grain diet (HG; 60% concentrate, DM basis; n = 8). The experiment lasted 6 weeks, including 1 week for adaptation. The experimental results showed that the milk fat content in the milk of lactating cows in the HG group was significantly reduced (p < 0.05), and the milk urea nitrogen (MUN) content showed an increasing trend (0.05 < p < 0.10) compared with the LG group. Compared with the LG group, rumen fluid pH was significantly decreased after feeding a high-grain diet, and contents of total volatile fatty acids (TVFA), acetate, propionate, and butyrate were significantly increased (p < 0.05). The acetate/propionate significantly decreased (p < 0.05). HG group significantly increased the abundance of Prevotella and Bacteroides in rumen fluid while significantly reducing the abundance of Methanobrevibacter and Lachnospiraceae ND3007_group (p < 0.05). Microorganisms with LDA scores > 2 were defined as unique, with the bacterial genus Anaerorhabdus_furcosa_group identified as a biomarker for the LG group, and the unique bacterial genus in the HG group were Prevotella, Stenotrophomonas, and Xanthomonadaceae. The prediction results of microbial function showed that a total of 18 KEGG differential pathways were generated between the two treatment groups, mainly manifested in metabolic pathways, signal transduction, and the immune system. In conclusion, the HG group promoted rumen fermentation by altering the microbial composition of lactating cows. Our findings provide a theoretical basis for the rational use of high-grain diets to achieve high yields in intensive dairy farming.

Increasing concentrations of low SMCO swedes in the diet of dairy cows improves performance and ruminal metabolism without affecting dairy cow health

This study aimed to determine dry matter intake, ruminal fermentation, health-related blood metabolites, and production responses of early-lactation dairy cows fed two inclusion levels (30 and 45 %) of low SMCO swedes in the diet. Twelve pregnant multiparous lactating Holstein-Friesian dairy cows (24.1 ± 0.72 kg milk/d, 540 ± 7.3 kg BW, and 142 ± 3.9 DIM at the start of the study) were used in this study. Cows were randomly allocated to three dietary treatments in a replicated 3 ×3 Latin square design. The experiment lasted 63 d and was divided into three 21-d periods. Inclusion of swedes at 30 and 45 % in the diet had no effect on DMI, but increased milk, protein and fat yields, as well as CP concentration in milk. This was due to changes in VFA patterns with greater molar proportions of propionate and butyrate, as well as a greater utilisation of dietary N that was converted to microbial N. No negative effects on haematological and biochemical blood parameters, except for increased BHB concentrations. Fatty acids composition in milk was modified, with a greater proportion of saturated FA and lower n-3 FA. Overall, low SMCO swedes can be included up to 45 % in the diet of pasture fed dairy cows during winter.

Assessing the effects of high-carvacrol oregano oil on rumen microbial fermentation, gas production, and methane production in vitro

The objective of this in vitro study was to assess the effects of high-carvacrol oregano oil (ORE; 50 g/kg oregano oil, 800-802 g/kg carvacrol), on microbial fermentation and CH4 production. In the experiment (a complete randomized block design), treatments included a negative control (CTL, no additive), a positive control (monensin, MON, 10 mg/L), and ORE (20, 40, 80, 120, 200, 160, and 1000 mg/L). Compared with CTL, MON shifted (p &lt; 0.05) volatile fatty acid (VFA) profile from acetate and butyrate to propionate production, thereby reducing (p &lt; 0.05) CH4 production (-26%). Monensin decreased (p &lt; 0.05) NH3 and branched-chain VFA concentrations. Of the doses evaluated, only the highest dose (1000 mg/L) affected ruminal microbial fermentation and CH4 production. At this dose, ORE reduced (p &lt; 0.05) gas production, total VFA, acetate, propionate and NH3 concentrations, and CH4 production (-22%). The reduced gas production and total VFA is an indication of feed digestion inhibition. These results suggest that ORE may decrease CH4 production and improve ruminal N utilization. However, these findings need to be validated in vivo to determine the optimal dose to benefit from the positive effects while avoiding the negative impact of ORE on feed digestion.

Effect of exogenous cellulase pretreatment of straw-type fermented total mixed ration on digestibility and ruminal fermentation in growing beef cattle.

This study aimed to assess the impact of corn straw-based unfermented and fermented total mixed rations (TMR) supplemented with exogenous cellulase on the in vitro fermentation characteristics, growth performance, feeding behavior, apparent digestibility, rumen fermentation and digestive enzyme activities of Chinese Simmental bulls. Unfermented (direct spraying of exogenous cellulase onto TMR, TMR) and fermented (exogenous cellulase fermentation for more than 7 d, fermented total mixed rations [FTMR]) TMR were collected, dried, powdered and used as fermentation substrates. The fermentation liquid was ruminal fluid collected from Chinese Simmental bulls. The artificial rumen culture fluid were continuously cultured in vitro for 48 h. Based on the diets they were fed, 24 healthy Chinese Simmental bulls (average weight of 495.93 ±10.89 kg) were randomly divided into two groups, with 12 bulls in each group, which were fed TMR or FTMR. The study lasted 56 d. In in vitro experiments, the neutral detergent fiber degradability and total volatile fatty acid, propionate, iso-butyrate, iso-valerate and valerate concentrations were greater in the FTMR group (p<0.05) than in the TMR group. However, the methane production, pH and A/P of the FTMR group tended to be lower (p<0.05) than those of the TMR group. In the in vivo experiments, the average daily gain, eating rate, and feed efficiency of the FTMR groups were greater (p<0.05) than those of the TMR group. Similarly, the NDF degradability of the FTMR group was greater (p<0.05) than that of the TMR group. Compared to those in the TMR group, the concentrations of total volatile fatty acids, iso-butyrate, propionate and butyrate were greater in the FTMR group (p<0.05), and the A/P ratio was lower (p<0.05). Similarly, cellulase, xylanase, and β-glucosidase activities were greater (p<0.05) in the FTMR group than in the TMR group. Corn straw-based fermented total mixed rations supplemented with exogenous cellulase play a vital role in decreasing the structural carbohydrate content of TMR and ruminal methane production in vitro, improving nutrient digestion and absorption, optimizing rumen fermentation, and improving the growth performance of beef cattle.

The Effects of Different Doses of 3-NOP on Ruminal Fermentation Parameters, Methane Production, and the Microbiota of Lambs In Vitro

The Effects of Different Doses of 3-NOP on Ruminal Fermentation Parameters, Methane Production, and the Microbiota of Lambs In Vitro

Modulations in gastrointestinal microbiota during postpartum period fulfill energy requirements and maintain health of lactating Tibetan cattle.

Postpartum period of dairy cattle is an important phase of their life mainly associated with the changes in physiology, rumen function, and energy metabolism. Studies have shown that gut microbial composition undergoes drastic changes during the postpartum period. However, little is known about the temporal variations in digestive tract microbiota in postpartum Tibetan cattle. The aim of this study was to investigate the temporal variations in blood metabolites, ruminal fermentation, and microbial community of oral, rumen, and gut in lactating Tibetan cattle during postpartum. We collected blood, saliva, rumen fluid, and fecal samples from lactating Tibetan cattle during 1st week (1 W), the 2nd week (2 W), the 1st month (1 M), and the 2nd month (2 M) of the postpartum period. The microbiota of saliva, rumen fluid, and fecal samples were assessed using 16S rRNA sequencing. The rumen volatile fatty acid and blood parameters were also quantified. The content of volatile fatty acids (VFAs) and blood parameters showed opposite tendency to each other and reached to stability at 2 M. Rumen microbiota showed the highest alpha diversity compared to other two sites. At phylum level, the oral cavity was dominated by Proteobacteria, while most dominant phylum in rumen and feces were Firmicutes and Bacteroidetes, respectively. The dominant genera in oral cavity were Moraxella and Bibersteinia, while genera Prevotella 1 and Ruminococcaceae UCG-005 were dominant in rumen and fecal samples, respectively. Microbial network analysis revealed that most of the active genera in all networks belonged to phylum Firmicutes, indicating the importance of this phyla during postpartum period of lactating cattle. The functional analysis revealed distinct division of labor among three gastrointestinal sites associated with defense, fatty acid synthesis, and maintaining health of host. All in all, our findings provide insights into the metabolic and microbial changes of lactating Tibetan cattle and help to the improvement of the management strategies.

Dynamic Changes in the Nutrient Digestibility, Rumen Fermentation, Serum Parameters of Perinatal Ewes and Their Relationship with Rumen Microbiota.

Changes in physiological and biochemical parameters are crucial for the reproductive performance and health of perinatal ewes. This study investigated the temporal variations in feed intake, nutrient digestibility, serum parameters, and ruminal fermentation on days 21, 14, and 7 before lambing (Q21, Q14, and Q7) and days 3, 7, and 14 after lambing (H3, H7, and H14). The results showed that dry matter intake (DMI) and glucose (Glu) gradually decreased (p < 0.05) before lambing and increased (p < 0.05) after lambing. The digestibility of dry matter (DMD), crude protein (CPD), and acid detergent fiber (ADFD) increased (p < 0.05) before lambing, then decreased (p < 0.05) on day H3, and then increased (p < 0.05) on day H14. The rumen pH, NH3-N, and triglycerides (TG) gradually increased (p < 0.05) before lambing and were higher (p < 0.05) on day Q7 than after lambing. The concentrations of acetate, butyrate, and total volatile fatty acids (T-VFA) were lower (p < 0.05) on day Q7 than those on days Q21 and Q14, then increased (p < 0.05) after lambing. Total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) concentrations gradually decreased (p < 0.05) in perinatal ewes. BHBA and NEFA concentrations were lower (p < 0.05) on day Q21 than those from days Q14 to H14. The rumen microbiota compositions were different (p < 0.05) in perinatal ewes, and g_Anaerovibrio, g_Lachnobacterium, and g_Schwartzia were positively correlated (p < 0.05) with DMI, Glu, acetate, propionate, and T-VFA, and negatively correlated (p < 0.05) with LDL-C. g_Bacillus was negatively correlated (p < 0.05) with DMI, Glu, acetate, propionate, butyrate, and T-VFA, but positively correlated (p < 0.05) with rumen pH and LDL-C. In summary, the DMI, nutrient digestibility, rumen fermentation, and serum parameters changed during the perinatal period of ewes, and the changes in DMI, serum glucose, acetate, propionate, and T-VFA were related to the rumen bacteria.

Inhibiting methanogenesis by targeting thermodynamics and enzymatic reactions in mixed cultures of rumen microbes in vitro.

Mitigation of enteric methane (CH4) emissions from ruminant livestock represents an opportunity to improve the sustainability, productivity, and profitability of beef and dairy production. Ruminal methanogenesis can be mitigated via two primary strategies: (1) alternative electron acceptors and (2) enzymatic inhibition of methanogenic pathways. The former utilizes the thermodynamic favorability of certain reactions such as nitrate/nitrite reduction to ammonia (NH3) while the latter targets specific enzymes using structural analogs of CH4 and methanogenic cofactors such as bromochloromethane (BCM). In this study, we investigated the effects of four additives and their combinations on CH4 production by rumen microbes in batch culture. Sodium nitrate (NaNO3), sodium sulfate (Na2SO4), and 3-nitro-1-propionate (3NPA) were included as thermodynamic inhibitors, whereas BCM was included as a enzymatic inhibitor. Individual additives were evaluated at three levels of inclusion in experiments 1 and 2. Highest level of each additive was used to determine the combined effect of NaNO3 + Na2SO4 (NS), NS + 3NPA (NSP), and NSP + BCM (NSPB) in experiments 3 and 4. Experimental diets were high, medium, and low forage diets (HF, MF, and LF, respectively) and consisted of alfalfa hay and a concentrate mix formulated to obtain the following forage to concentrate ratios: 70:30, 50:50, and 30:70, respectively. Diets with additives were placed in fermentation culture bottles and incubated in a water bath (39°C) for 6, 12, or 24h. Microbial DNA was extracted for 16S rRNA and ITS gene amplicon sequencing. In experiments 1 and 2, CH4 concentrations in control cultures decreased in the order of LF, MF, and HF diets, whereas in experiments 3 and 4, CH4 was highest in MF diet followed by HF and LF diets. Culture pH and NH3 in the control decreased in the order of HF, MF, to LF as expected. NaNO3 decreased (p < 0.001) CH4 and butyrate and increased acetate and propionate (p < 0.03 and 0.003, respectively). Cultures receiving NaNO3 had an enrichment of microorganisms capable of nitrate and nitrite reduction. 3NPA also decreased CH4 at 6h with no further decrease at 24 h (p < 0.001). BCM significantly inhibited methanogenesis regardless of inclusion levels as well as in the presence of the thermodynamic inhibitors (p < 0.001) while enriching succinate producers and assimilators as well as propionate producers (p adj < 0.05). However, individual inclusion of BCM decreased total short chain fatty acid (SCFA) concentrations (p < 0.002). Inhibition of methanogenesis with BCM individually and in combination with the other additives increased gaseous H2 concentrations (p < 0.001 individually and 0.028 in combination) while decreasing acetate to propionate ratio (p < 0.001). Only the cultures treated with BCM in combination with other additives significantly (padj < 0.05) decreased the abundance of Methanobrevibacter expressed as log fold change. Overall, the combination of thermodynamic and enzymatic inhibitors presented a promising effect on ruminal fermentation in-vitro, inhibiting methanogenesis while optimizing the other fermentation parameters such as pH, NH3, and SCFAs. Here, we provide a proof of concept that the combination of an electron acceptor and a methane analog may be exploited to improve microbial efficiency via methanogenesis inhibition.

Mealworm larvae and black soldier fly larvae as novel protein supplements for cattle consuming low-quality forage.

The global population is projected to increase, indicating that there will be greater demand for animal protein to meet the associated food needs. This demand will place additional pressure on livestock systems to increase output while also minimizing natural resource inputs. Insect protein has emerged as a potential alternative to conventional protein feeds, such as soybean meal. Mealworm larvae (MWL; Tenebrio molitor) have been studied in poultry and swine as an alternative protein source; however, there is no research currently evaluating MWL for cattle. Black soldier fly larvae (BSFL; Hermetia illucens) have also received attention for their potential use in livestock feed due to their scalability and nutritional value, but research in cattle is limited. The objective of this study was to evaluate the effects of whole-dried MWL and defatted BSFL as protein supplements for cattle consuming forage. Five ruminally cannulated steers were utilized in a 5 × 5 Latin square experiment to determine how MWL and BSFL supplementations affect forage utilization. Steers consuming ad libitum low-quality forage (76.5% neutral detergent fiber [NDF], 4.2% crude protein) were provided one of the five treatments each period: 1) control with no supplement (CON), 2) soybean meal (CONV), 3) BSFL, 4) MWL, or 5) 50/50 by-weight blend of BSFL and MWL (MIX). All treatments were provided at 100 mg N/kg BW and periods included 8 d for treatment adaptation, 5 d for intake and digestion, and 1 d for ruminal fermentation measures. Protein supplementation stimulated forage organic matter intake (FOMI; P ≤ 0.01) relative to CON (3.28 kg/d). There was a significant difference in FOMI (P ≤ 0.01) between BSFL (4.30 kg/d) and CONV (4.71 kg/d), but not between CONV and MWL (P = 0.06, 4.43 kg/d). Total digestible OM intake (TDOMI) was also stimulated by the provision of protein (P ≤ 0.01), from 1.94 kg/d for CON to an average of 3.24 kg/d across protein supplements. Organic matter digestibility (OMD) and NDF digestibility (NDFD) were not affected by treatment (P ≥ 0.37), for an average OMD of 66.5% and NDFD of 62.7%. There was also no treatment effect on ruminal volatile fatty acid (P = 0.96) or ammonia-N (P = 0.22) concentrations. These data indicate that MWL may stimulate forage utilization by beef cattle to a greater extent than BSFL, but both are viable protein supplements.

Feeding or pen surface application of clinoptilolite with different particle sizes: impact on nitrogen utilization and manure ammonia emissions in feedlot cattle.

This study investigated the effects of feeding clinoptilolite (CLN; 2.5% of diet dry matter) with a particle size of either 30- or 400-µm on ruminal fermentation characteristics, measures of nitrogen (N) utilization, and manure ammonia-N (NH3) emissions in feedlot cattle. The impact of directly applying 30- or 400-µm CLN to the pen surface (2,250kg/ha) on manure NH3-N emissions was also evaluated. Six beef heifers were used in a replicated 3 × 3 Latin square design with 21-d periods. Dietary treatments were 1) finishing ration with no supplement (CON), 2) CON + 30-µm CLN (CLN-30), and 3) CON + 400-µm CL (CLN-400). Intake was measured daily. To evaluate fermentation characteristics, ruminal fluid was collected on day 19. Indwelling pH loggers were used to measure ruminal pH from days 15 to 21. Blood was collected 3-h post-feeding on day 21 for metabolite analysis. Fecal grab and urine spot samples were also collected from days 19 to 21 to measure nutrient digestibility, route of N excretion, and in vitro NH3 emissions. There was no diet effect (P ≥ 0.12) on nutrient intake and apparent total tract digestibility, and ruminal short-chain fatty acid profile and pH. Ruminal NH3 concentration, which was lower (P = 0.04) for CLN-30 than CON heifers, did not differ between CON and CLN-400 heifers. Although there was no diet effect (P = 0.50) on plasma urea-N (PUN) concentration, proportion of urea-N excreted in urine was lower (P = 0.01) for CLN-30 than CON and CLN-400 heifers. Urinary NH3-N excretion, which was greater (P ≤ 0.04) for CLN-400 than CON heifers, did not differ between CLN-30 and CLN-400 heifers. Feeding CLN also increased (P ≤ 0.02) fecal excretion of potassium (K) and iron (Fe) and reduced (P = 0.01) urinary excretion of calcium (Ca). There was a treatment × time interaction (P = 0.01) for NH3 emission rate, which was greatest within the first 36h of incubation and was lower for manure from CLN-400 compared to CON and CLN-30 heifers and pen surface application treatments. Cumulative NH3 emissions were lower (P < 0.01) for manure from CLN-400 compared to CON and CLN-30 heifers and the pen surface application treatments. Although surface application was ineffective, feeding 400-µm CLN to finishing cattle could result in a beneficial decrease in manure NH3 emissions. However, changes in fecal and urine excretion of minerals like K and Ca, which suggest a decrease in bioavailability, need to be considered when feeding CLN in finishing cattle diets.

Effect of Different Slow-Release Urea on the Production Performance, Rumen Fermentation, and Blood Parameter of Angus Heifer.

This study investigated the effect of replacing part of the dietary soybean meal with either polymer-coated urea or gelatinized starch urea on the production performance, blood indexes, and ruminal fermentation of Angus heifers. A total of 210 purebred Angus cattle (BW = 314.26 kg) were divided into three groups: the no urea group (CON), the polymer-coated urea group (PCU), and the gelatinized starch urea group (GSU); 20 g/kg polymer-coated urea or 25 g/kg gelatinized starch urea was used to replace part of soybean meal in the concentrate feed, according to the principle of isometabolic energy and isonitrogenous. The result showed that the PCU group had higher ADG and ADF apparent digestibility, while it had a lower feed-weight ratio. On the 86th day, the serum albumin (ALB) content in the PCU group was significantly higher than that in the CON group. In rumen, compared with the CON group, the contents of acetic acid and total volatile fatty acid were significantly higher in the PCU group, whereas butyric acid and propionic acid were significantly higher in the PCU group and GSU group. Ruminal bacterial diversity analysis found that the abundance of Firmicutes was higher in the PCU group at the phylum level, and an inverse result was observed in Bacteroidetes. The abundance of Paraprevotella was higher in the PCU group, whereas higher abundance of Prevotella was found in the GSU group at the genus level. These results indicate that slow-release urea can replace part of soybean meal in the diet, and the amount of substitution in this trial had no diverse effect on the performance of Angus heifers.