Rumen-protected methionine during heat stress alters mTOR, insulin signaling, and 1-carbon metabolism protein abundance in liver, and whole-blood transsulfuration pathway genes in Holstein cows.

We investigated effects of rumen-protected Met (RPM) during a heat stress (HS) challenge on (1) hepatic abundance of mTOR, insulin, and antioxidant signaling proteins, (2) enzymes in 1-carbon metabolism, and (3) innate immunity. Holstein cows (n = 32; mean ± standard deviation, 184 ± 59 d in milk) were randomly assigned to 1 of 2 environmental groups, and 1 of 2 diets [total mixed ration (TMR) with RPM (Smartamine M; 0.105% dry matter as top-dress) or TMR without (CON); n = 16/diet] in a split-plot crossover design. There were 2 periods with 2 phases. During phase 1 (9 d), all cows were in thermoneutral conditions (TN; temperature-humidity index = 60 ± 3) and fed ad libitum. During phase 2 (9 d), half the cows (n = 8/diet) were exposed to HS using electric heat blankets. The other half (n = 8/diet) remained in TN, but was pair-fed to HS counterparts. After a 14-d washout and 7-d adaptation period, the study was repeated (period 2) and environmental treatments were inverted relative to phase 2, but dietary treatments were the same. Blood was collected on d 6 of each phase 2 to measure immune function and isolate whole-blood RNA. Liver biopsies were performed at the end of each period for cystathione ß-synthase (CBS) and methionine adenosyltransferase activity, glutathione concentration, and protein abundance. Data were analyzed using PROC MIXED in SAS. Abundance of CUL3, inhibitor of antioxidant responses, tended to be downregulated by HS suggesting increased oxidative stress. Heat-shock protein 70 abundance was upregulated by HS. Phosphorylated mTOR abundance was greater overall with RPM, suggesting an increase in pathway activity. An environment × diet (E × D) effect was observed for protein kinase B (AKT), whereas there was a tendency for an interaction for phosphorylated AKT. Abundance of AKT was upregulated in CON cows during HS versus TN, this was not observed in RPM cows. For phosphorylated AKT, tissue from HS cows fed CON had greater abundance compared with all other treatments. The same effect was observed for EIF2A (translation initiation) and SLC2A4 (insulin-induced glucose uptake). An E × D effect was observed for INSR due to upregulation in CON cows during HS versus TN cows fed CON or RPM. There was an E × D effect for CBS, with lower activity in RPM versus CON cows during HS. The CON cows tended to have greater CBS during HS versus TN. An E × D effect was observed for methionine adenosyltransferase, with lower activity in RPM versus CON during HS. Although activity increased in CON during HS versus TN, RPM cows tended to have greater activity during TN. Neutrophil and monocyte oxidative burst and monocyte phagocytosis decreased with HS. An (E × D) effect was observed for whole-blood mRNA abundance of CBS, SOD1 and CSAD; RPM led to upregulation during TN versus HS. Regardless of diet, CDO1, CTH, and SOD1 decreased with HS. Although HS increased hepatic HSP70 and seemed to alter antioxidant signaling, feeding RPM may help cows maintain homeostasis in mTOR, insulin signaling, and 1-carbon metabolism. Feeding RPM also may help maintain whole-blood antioxidant response during HS, which is an important aspect of innate immune function.

Review: Alternative and novel feeds for ruminants: nutritive value, product quality and environmental aspects.

Ruminant-based food production faces currently multiple challenges such as environmental emissions, climate change and accelerating food-feed-fuel competition for arable land. Therefore, more sustainable feed production is needed together with the exploitation of novel resources. In addition to numerous food industry (milling, sugar, starch, alcohol or plant oil) side streams already in use, new ones such as vegetable and fruit residues are explored, but their conservation is challenging and production often seasonal. In the temperate zones, lipid-rich camelina (Camelina sativa) expeller as an example of oilseed by-products has potential to enrich ruminant milk and meat fat with bioactive trans-11 18:1 and cis-9,trans-11 18:2 fatty acids and mitigate methane emissions. Regardless of the lower methionine content of alternative grain legume protein relative to soya bean meal (Glycine max), the lactation performance or the growth of ruminants fed faba beans (Vicia faba), peas (Pisum sativum) and lupins (Lupinus sp.) are comparable. Wood is the most abundant carbohydrate worldwide, but agroforestry approaches in ruminant nutrition are not common in the temperate areas. Untreated wood is poorly utilised by ruminants because of linkages between cellulose and lignin, but the utilisability can be improved by various processing methods. In the tropics, the leaves of fodder trees and shrubs (e.g. cassava (Manihot esculenta), Leucaena sp., Flemingia sp.) are good protein supplements for ruminants. A food-feed production system integrates the leaves and the by-products of on-farm food production to grass production in ruminant feeding. It can improve animal performance sustainably at smallholder farms. For larger-scale animal production, detoxified jatropha (Jatropha sp.) meal is a noteworthy alternative protein source. Globally, the advantages of single-cell protein (bacteria, yeast, fungi, microalgae) and aquatic biomass (seaweed, duckweed) over land crops are the independence of production from arable land and weather. The chemical composition of these feeds varies widely depending on the species and growth conditions. Microalgae have shown good potential both as lipid (e.g. Schizochytrium sp.) and protein supplements (e.g. Spirulina platensis) for ruminants. To conclude, various novel or underexploited feeds have potential to replace or supplement the traditional crops in ruminant rations. In the short-term, N-fixing grain legumes, oilseeds such as camelina and increased use of food and/or fuel industry by-products have the greatest potential to replace or supplement the traditional crops especially in the temperate zones. In the long-term, microalgae and duckweed of high-yield potential as well as wood industry by-products may become economically competitive feed options worldwide.

Whole Mitogenomes Reveal the History of Swamp Buffalo: Initially Shaped by Glacial Periods and Eventually Modelled by Domestication.

The newly sequenced mitochondrial genomes of 107 Asian swamp buffalo (Bubalus bubalis carabensis) allowed the reconstruction of the matrilineal divergence since ~900 Kya. Phylogenetic trees and Bayesian skyline plots suggest a role of the glacial periods in the demographic history of swamp buffalo. The ancestral swamp-buffalo mitogenome is dated ~232 ± 35 Kya. Two major macro-lineages diverged during the 2nd Pleistocene Glacial Period (~200-130 Kya), but most (~99%) of the current matrilines derive from only two ancestors (SA1'2 and SB) that lived around the Last Glacial Maximum (~26-19 Kya). During the late Holocene optimum (11-6 Kya) lineages differentiated further, and at least eight matrilines (SA1, SA2, SB1a, SB1b, SB2a, SB2b, SB3 and SB4) were domesticated around 7-3 Kya. Haplotype distributions support an initial domestication process in Southeast Asia, while subsequent captures of wild females probably introduced some additional rare lineages (SA3, SC, SD and SE). Dispersal of domestic buffaloes created local population bottlenecks and founder events that further differentiated haplogroup distributions. A lack of maternal gene flow between neighboring populations apparently maintained the strong phylogeography of the swamp buffalo matrilines, which is the more remarkable because of an almost complete absence of phenotypic differentiation.

Tropical legume supplementation influences microbial protein synthesis and rumen ecology.

Four rumen-fistulated male swamp buffaloes, 5-year-old with initiated live weight at 360 ± 12 kg, were randomly assigned according to a 4 × 4 Latin square design to investigate the effect of feeding high level of dried Leucaena leaf (DLL) on feed intake, fermentation efficiency and microbial protein synthesis. The dietary treatments were the feeding levels of DLL at 0, 2, 4 and 6 kg/head/day. All buffaloes were supplemented with concentrate mixtures at 0.1% of body weight, and rice straw was fed ad libitum with the availability of water and mineral block at all time. The results revealed that the total feed intake and nutrient digestibility were significantly improved with the increasing levels of DLL feeding, and the highest was in the buffaloes consuming DLL at 6 kg/head/day. Feeding high levels of DLL did not affect on ruminal pH and temperature, while ammonia nitrogen, blood urea nitrogen and volatile fatty acid concentration were significantly enhanced. Moreover, methane production was dramatically reduced by increasing levels of DLL feeding. Total direct counts of the micro-organism population were increased with the increasing levels of DLL feeding. According to the application of quantitative PCR to quantity cellulolytic bacteria (16S rRNA) targets, it was found that the population of total bacteria and Fibrobactor succinogenes was affected by treatments, while Ruminococcus flavefaciens and methanogen population were significantly decreased as buffaloes were fed with DLL. The nitrogen balance and microbial nitrogen supply were remarkably improved with the increasing levels of DLL feeding. Based on this study, it could be concluded that high levels of DLL feeding at 6 kg/head/day could enhance feed intake, nutrient digestibility, rumen fermentation efficiency and microbial protein synthesis in swamp buffaloes fed on rice straw without any adverse effect.

Effect of Grape Pomace Powder, Mangosteen Peel Powder and Monensin on Nutrient Digestibility, Rumen Fermentation, Nitrogen Balance and Microbial Protein Synthesis in Dairy Steers.

This study was designed to investigate the effect of grape pomace powder (GPP), mangosteen peel powder (MPP) and monensin on feed intake, nutrients digestibility, microorganisms, rumen fermentation characteristic, microbial protein synthesis and nitrogen balance in dairy steers. Four, rumen fistulated dairy steers with initial body weight (BW) of 220±15 kg were randomly assigned according to a 4×4 Latin square design to receive four treatments. The treatments were as follows: T1 = control, T2 = supplementation with monensin at 33 mg/kg diet, T3 = supplementation with GPP at 2% of dry matter intake, and T4 = supplementation with MPP at 30 g/kg diet. The steers were offered the concentrate diet at 0.2% BW and 3% urea treated rice straw (UTRS) was fed ad libitum. It was found that GPP supplemented group had higher UTRS intake and nutrient digestibility in terms of neutral detergent fiber and acid detergent fiber than those in control group (p<0.05). Ammonia nitrogen (NH3-N) and blood urea-nitrogen concentration were higher in monensin, GPP and MPP supplemented groups (p<0.05). Total volatile fatty acids and propionate in the GPP group were higher than those in the control group (p<0.05) while acetate concentration, and acetate to propionate ratio were decreased (p<0.01) when steers were supplemented with GPP, monensin, and MPP, respectively. Moreover, protozoal populations in GPP, MPP, and monensin supplementation were significantly lower than those in the control group (p<0.05), while cellulolytic bacterial population was significantly higher in the control group (p<0.05). Nitrogen retention, microbial crude protein and efficiency of microbial nitrogen synthesis were found significantly higher in steers that received GPP (p<0.05). Based on this study it could be concluded that the GPP has potential as an alternative feed supplement in concentrate diets which can result in improved rumen fermentation efficiency, digestibility and microbial protein synthesis in steers fed on treated rice straw.

Optimal Cultivation Time for Yeast and Lactic Acid Bacteria in Fermented Milk and Effects of Fermented Soybean Meal on Rumen Degradability Using Nylon Bag Technique.

The objectives of this study were to determine an optimal cultivation time for populations of yeast and lactic acid bacteria (LAB) co-cultured in fermented milk and effects of soybean meal fermented milk (SBMFM) supplementation on rumen degradability in beef cattle using nylon bag technique. The study on an optimal cultivation time for yeast and LAB growth in fermented milk was determined at 0, 4, 8, 24, 48, 72, and 96 h post-cultivation. After fermenting for 4 days, an optimal cultivation time of yeast and LAB in fermented milk was selected and used for making the SBMFM product to study nylon bag technique. Two ruminal fistulated beef cattle (410±10 kg) were used to study on the effect of SBMFM supplementation (0%, 3%, and 5% of total concentrate substrate) on rumen degradability using in situ method at incubation times of 0, 2, 4, 6, 12, 24, 48, and 72 h according to a Completely randomized design. The results revealed that the highest yeast and LAB population culture in fermented milk was found at 72 h-post cultivation. From in situ study, the soluble fractions at time zero (a), potential degradability (a+b) and effective degradability of dry matter (EDDM) linearly (p<0.01) increased with the increasing supplemental levels and the highest was in the 5% SBMFM supplemented group. However, there was no effect of SBMFM supplement on insoluble degradability fractions (b) and rate of degradation (c). In conclusion, the optimal fermented time for fermented milk with yeast and LAB was at 72 h-post cultivation and supplementation of SBMFM at 5% of total concentrate substrate could improve rumen degradability of beef cattle. However, further research on effect of SBMFM on rumen ecology and production performance in meat and milk should be conducted using in vivo both digestion and feeding trials.

Effects of Condensed Tannins in Mao (Antidesma thwaitesianum Muell. Arg.) Seed Meal on Rumen Fermentation Characteristics and Nitrogen Utilization in Goats.

Mao seed is a by-product of the wine and juice industry, which could be used in animal nutrition. The current study was designed to determine the effect of supplementation of mao (Antidesma thwaitesianum Muell. Arg.) seed meal (MOSM) containing condensed tannins (CT) on rumen fermentation, nitrogen (N) utilization and microbial protein synthesis in goats. Four crossbred (Thai Native×Anglo Nubian) goats with initial body weight (BW) 20±2 kg were randomly assigned to a 4×4 Latin square design. The four dietary treatments were MOSM supplementation at 0%, 0.8%, 1.6%, and 2.4% of total dry matter (DM) intake, respectively. During the experimental periods, all goats were fed a diet containing roughage to concentrate ratio of 60:40 at 3.0% BW/d and pangola grass hay was used as a roughage source. Results showed that supplementation with MOSM did not affect feed intake, nutrient intakes and apparent nutrient digestibility (p>0.05). In addition, ruminal pH and ammonia nitrogen (NH3-N) were not influenced by MOSM supplementation, whilst blood urea nitrogen was decreased quadraticly (p<0.05) in goats supplemented with MOSM at 2.4% of total DM intake. Propionate was increased linearly with MOSM supplementation, whereas acetate and butyrate were remained the same. Moreover, estimated ruminal methane (CH4) was decreased linearly (p<0.05) when goats were fed with MOSM at 1.6% and 2.4% of total DM intake. Numbers of bacteria and protozoa were similar among treatments (p>0.05). There were linear decreases in urinary N (p<0.01) and total N excretion (p<0.01) by MOSM supplementation. Furthermore, N retention was increased linearly (p<0.05) when goats were fed with MOSM supplementation at 1.6% and 2.4% of total DM intake. Microbial protein synthesis were not significantly different among treatments (p>0.05). From the current study, it can be concluded that supplementation of MOSM at 1.6% to 2.4% of total DM intake can be used to modify ruminal fermentation, especially propionate and N utilization in goats, without affecting the nutrient digestibility, microbial populations and microbial protein synthesis.

Improvement of Nutritive Value and In vitro Ruminal Fermentation of Leucaena Silage by Molasses and Urea Supplementation.

Leucaena silage was supplemented with different levels of molasses and urea to study its nutritive value and in vitro rumen fermentation efficiency. The ensiling study was randomly assigned according to a 3×3 factorial arrangement in which the first factor was molasses (M) supplement at 0%, 1%, and 2% of crop dry matter (DM) and the second was urea (U) supplement as 0%, 0.5%, and 1% of the crop DM, respectively. After 28 days of ensiling, the silage samples were collected and analyzed for chemical composition. All the nine Leucaena silages were kept for study of rumen fermentation efficiency using in vitro gas production techniques. The present result shows that supplementation of U or M did not affect DM, organic matter, neutral detergent fiber, and acid detergent fiber content in the silage. However, increasing level of U supplementation increased crude protein content while M level did not show any effect. Moreover, the combination of U and M supplement decreased the content of mimosine concentration especially with M2U1 (molasses 2% and urea 1%) silage. The result of the in vitro study shows that gas production kinetics, cumulation gas at 96 h and in vitro true digestibility increased with the increasing level of U and M supplementation especially in the combination treatments. Supplementation of M and U resulted in increasing propionic acid and total volatile fatty acid whereas, acetic acid, butyric acid concentrations and methane production were not changed. In addition, increasing U level supplementation increased NH3-N concentration. Result from real-time polymerase chain reaction revealed a significant effect on total bacteria, whereas F. succinogenes and R. flavefaciens population while R. albus was not affected by the M and U supplementation. Based on this study, it could be concluded that M and urea U supplementation could improve the nutritive value of Leucaena silage and enhance in vitro rumen fermentation efficiency. This study also suggested that the combination use of M and U supplementation level was at 2% and 1%, respectively.

Effects of Supplementation of Eucalyptus (E. Camaldulensis) Leaf Meal on Feed Intake and Rumen Fermentation Efficiency in Swamp Buffaloes.

Four rumen fistulated swamp buffaloes were randomly assigned according to a 4×4 Latin square design to investigate the effects of Eucalyptus (E. Camaldulensis) leaf meal (ELM) supplementation as a rumen enhancer on feed intake and rumen fermentation characteristics. The dietary treatments were as follows: T1 = 0 g ELM/hd/d; T2 = 40 g ELM/hd/d; T3 = 80 g ELM/hd/d; T4 = 120 g ELM/hd/d, respectively. Experimental animals were kept in individual pens and concentrate was offered at 0.3% BW while rice straw was fed ad libitum. The results revealed that voluntary feed intake and digestion coefficients of nutrients were similar among treatments. Ruminal pH, temperature and blood urea nitrogen concentrations were not affected by ELM supplementation; however, ELM supplementation resulted in lower concentration of ruminal ammonia nitrogen. Total volatile fatty acids, propionate concentration increased with the increasing level of EML (p<0.05) while the proportion of acetate was decreased (p<0.05). Methane production was linearly decreased (p<0.05) with the increasing level of ELM supplementation. Protozoa count and proteolytic bacteria population were reduced (p<0.05) while fungal zoospores and total viable bacteria, amylolytic, cellulolytic bacteria were unchanged. In addition, nitrogen utilization and microbial protein synthesis tended to increase by the dietary treatments. Based on the present findings, it is suggested that ELM could modify the rumen fermentation and is potentially used as a rumen enhancer in methane mitigation and rumen fermentation efficiency.

Improving the quality of rice straw by urea and calcium hydroxide on rumen ecology, microbial protein synthesis in beef cattle.

Four rumen-fistulated beef cattle were randomly assigned to four treatments according to a 4 × 4 Latin square design to study the influence of urea and calcium hydroxide [Ca(OH)2 ] treatment of rice straw to improve the nutritive value of rice straw. Four dietary treatments were as follows: untreated rice straw, 50 g/kg urea-treated rice straw, 20 g/kg urea + 20 g/kg calcium hydroxide-treated rice straw and 30 g/kg urea + 20 g/kg calcium hydroxide-treated rice straw. All animals were kept in individual pens and fed with concentrate at 0.5 g/kg of BW (DM), rice straw was fed ad libitum. The experiment was conducted for four periods, and each period lasted for 21 days. During the first 14 days, DM feed intake measurements were made while during the last 7 days, all cattle were moved to metabolism crates for total faeces and urine collections. The results revealed that 20 g/kg urea + 20 g/kg calcium hydroxide-treated rice straw improved the nutritive value of rice straw, in terms of dry matter intake, digestibility, ruminal volatile fatty acids, population of bacteria and fungi, nitrogen retention and microbial protein synthesis. Based on this study, it could be concluded that using urea plus calcium hydroxide was one alternative method to improve the nutritive value of rice straw, rumen ecology and fermentation and thus a reduction of treatment cost.

Effect of tannins and saponins in Samanea saman on rumen environment, milk yield and milk composition in lactating dairy cows.

The objective of this study was to investigate the effects of tannins and saponins in Samanea saman on rumen fermentation, milk yield and milk composition in lactating dairy cows. Four multiparous early-lactating dairy cows (Holstein-Friesian cross-bred, 75%) with an initial body weight (BW) of 405 ± 40 kg and 36 ± 8 day in milk were randomly assigned to receive dietary treatments according to a 4 × 4 Latin square design. The four dietary treatments were unsupplemented (control), supplemented with rain tree pod (S. saman) meal (RPM) at 60 g/kg, supplemented with palm oil (PO) at 20 g/kg, and supplemented with RPM at 60 g/kg and PO at 20 g/kg (RPO), of total dry matter (DM) intake. Cows were fed with concentrate diets at a ratio of concentrate to milk yield of 1:2, and chopped 30 g/kg of urea-treated rice straw was fed ad libitum. The RPM contained condensed tannins and crude saponins at 88 and 141 g/kg of DM respectively. It was found that supplementation with RPM and/or PO to dairy cows diets did not show negative effect on ruminal pH, blood urea nitrogen and milk urea nitrogen concentration (p > 0.05). However, supplementation with RPM resulted in lower ammonia nitrogen (NH3 -N) concentration (p < 0.05). In addition, propionic acid and milk production increased while acetic acid, acetic to propionic ratio, methane production, methanogens and protozoal population decreased with RPM and/or PO supplementation. Furthermore, addition of PO and RPO in the diets increased milk fat while supplementation of RPM resulted in greater milk protein and Fibrobacter succinogenes numbers (p < 0.05). The population of Ruminococcus flavefaciens and Ruminococcus albus were not affected by any treatments. The findings on the present study showed that supplementation with RPM and RPO to diets of cows improved the rumen environment and increased milk yield, content of milk protein and milk fat.

Influence of Yeast Fermented Cassava Chip Protein (YEFECAP) and Roughage to Concentrate Ratio on Ruminal Fermentation and Microorganisms Using In vitro Gas Production Technique.

The objective of this study was to determine the effects of protein sources and roughage (R) to concentrate (C) ratio on in vitro fermentation parameters using a gas production technique. The experimental design was a 2×5 factorial arrangement in a completely randomized design (CRD). Factor A was 2 levels of protein sources yeast fermented cassava chip protein (YEFECAP) and soybean meal (SBM) and factor B was 5 levels of roughage to concentrate (R:C) ratio at 80:20, 60:40, 40:60, 20:80, and 0:100, respectively. Rice straw was used as a roughage source. It was found that gas production from the insoluble fraction (b) of YEFECAP supplemented group was significantly higher (p<0.05) than those in SBM supplemented group. Moreover, the intercept value (a), gas production from the insoluble fraction (b), gas production rate constants for the insoluble fraction (c), potential extent of gas production (a+b) and cumulative gas production at 96 h were influenced (p<0.01) by R:C ratio. In addition, protein source had no effect (p>0.05) on ether in vitro digestibility of dry matter (IVDMD) and organic (IVOMD) while R:C ratio affected the IVDMD and IVOMD (p<0.01). Moreover, YEFECAP supplanted group showed a significantly increased (p<0.05) total VFA and C3 while C2, C2:C3 and CH4 production were decreased when compared with SBM supplemented group. In addition, a decreasing R:C ratio had a significant effect (p<0.05) on increasing total VFA, C3 and NH3-N, but decreasing the C2, C2:C3 and CH4 production (p<0.01). Furthermore, total bacteria, Fibrobacter succinogenes, Ruminococcus flavefaciens and Ruminococcus albus populations in YEFECAP supplemented group were significantly higher (p<0.05) than those in the SBM supplemented group while fungal zoospores, methanogens and protozoal population remained unchanged (p>0.05) as compared between the two sources of protein. Moreover, fungal zoospores and total bacteria population were significantly increased (p<0.01) while, F. succinogenes, R. flavefaciens, R. albus, methanogens and protozoal population were decreased (p<0.01) with decreasing R:C ratio. In conclusion, YEFECAP has a potential for use as a protein source for improving rumen fermentation efficiency in ruminants.

Effects of eucalyptus crude oils supplementation on rumen fermentation, microorganism and nutrient digestibility in swamp buffaloes.

This study was conducted to investigate the effects of eucalyptus (E. Camaldulensis) crude oils (EuO) supplementation on voluntary feed intake and rumen fermentation characteristics in swamp buffaloes. Four rumen fistulated swamp buffaloes, body weight (BW) of 420±15.0 kg, were randomly assigned according to a 2×2 factorial arrangement in a 4×4 Latin square design. The dietary treatments were untreated rice straw (RS) without EuO (T1) and with EuO (T2) supplementation, and 3% urea-treated rice straw (UTRS) without EuO (T3) and with EuO (T4) supplementation. The EuO was supplemented at 2 mL/h/d in respective treatment. Experimental animals were kept in individual pens and concentrate mixture was offered at 3 g/kg BW while roughage was fed ad libitum. Total dry matter and roughage intake, and apparent digestibilites of organic matter and neutral detergent fiber were improved (p<0.01) by UTRS. There was no effect of EuO supplementation on feed intake and nutrient digestibility. Ruminal pH and temperature were not (p>0.05) affected by either roughage sources or EuO supplementation. However, buffaloes fed UTRS had higher ruminal ammonia nitrogen and blood urea nitrogen as compared with RS. Total volatile fatty acid and butyrate proportion were similar among treatments, whereas acetate was decreased and propionate molar proportion was increased by EuO supplementation. Feeding UTRS resulted in lower acetate and higher propionate concentration compared to RS. Moreover, supplementation of EuO reduced methane production especially in UTRS treatment. Protozoa populations were reduced by EuO supplementation while fungi zoospores remained the same. Total, amylolytic and cellulolytic bacterial populations were increased (p<0.01) by UTRS; However, EuO supplementation did not affect viable bacteria. Nitrogen intake and in feces were found higher in buffaloes fed UTRS. A positive nitrogen balance (absorption and retention) was in buffaloes fed UTRS. Supplementation of EuO did not affect nitrogen utilization. Both allantoin excretion and absorption and microbial nitrogen supply were increased by UTRS whereas efficiency of microbial protein synthesis was similar in all treatments. Findings of present study suggested that EuO could be used as a feed additive to modify the rumen fermentation in reducing methane production both in RS and UTRS. Feeding UTRS could improve feed intake and efficiency of rumen fermentation in swamp buffaloes. However, more research is warranted to determine the effect of EuO supplementation in production animals.

Manipulation of ruminal fermentation and methane production by supplementation of rain tree pod meal containing tannins and saponins in growing dairy steers.

Four rumen-fistulated dairy steers were used in a 2 × 2 factorial arrangement in a 4 × 4 Latin square design. The main factors were two roughage-to-concentrate ratios (R:C, 60:40 and 40:60) and two supplementation levels of rain tree pod meal (RPM) [0 or unsupplemented and 60 g/kg of total dry matter (DM) intake]. Chopped 30 g/kg of urea-treated rice straw was used as a roughage source. All animals received dietary according to respective R:C ratios at 25 g/kg body weight. The RPM contained condensed tannins and crude saponins at 84 and 143 g/kg of DM respectively. It was found that total volatile fatty acids (VFAs) and propionate concentrations were increased (p < 0.01), while acetate concentration, acetate-to-propionate ratio, CH4 production and protozoal numbers were decreased (p < 0.01) when steers were supplemented with RPM and 600 g/kg of concentrate. Allantoin excretion was found different by both R:C ratio and supplementation of RPM, with the highest value at R:C of 40:60 with 60 g/kg RPM (123.6 mmol/day) (p < 0.05). Allantoin absorption and microbial crude protein were increased (p < 0.05) with an increasing concentrate ratio. Moreover, efficiency of microbial protein synthesis was increased (p < 0.05) by feeding a higher ratio of concentrate (R:C 40:60) and supplementation of RPM. Based on this study, it is suggested that supplementation of RPM was beneficial for dairy cows fed on high roughage ratio, which could improved rumen fermentation by reducing fermentation gas loss, thus improving VFA profiles and thus enhancing efficiency of microbial protein synthesis.

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.

Using Plant Source as a Buffering Agent to Manipulating Rumen Fermentation in an In vitro Gas Production System.

The objective of this study was to investigate the effect of banana flower powder (BAFLOP) supplementation on gas production kinetics and rumen fermentation efficiency in in vitro incubation with different ratios of roughage to concentrate in swamp buffalo and cattle rumen fluid. Two male, rumen fistulated dairy steers and swamp buffaloes were used as rumen fluid donors. The treatments were arranged according to a 2×2×3 factorial arrangement in a Completely randomized design by using two ratios of roughage to concentrate (R:C; 75:25 and 25:75) and 3 levels of BAFLOP supplementation (0, 2 and 4% of dietary substrate) into two different kinds of rumen fluid (beef cattle and swamp buffalo). Under this investigation, the results revealed that the rumen ecology was affected by R:C ratio. The pH declined as a result of using high concentrate ratio; however, supplementation of BAFLOP could buffer the pH which led to an improvement of ruminal efficiency. BAFLOP supplementation affected acetic acid (C2) when the proportion of concentrate was increased. However, there were no effect on total volatile fatty acid (TVFA) and butyric acid (C4) by BAFLOP supplementation. The microbial community was affected by BAFLOP supplementation, especially the bacterial population. As revealed by real-time PCR, the populations of F. succinogenes and R. albus were reduced by the high concentrate treatments while that of R. flavafaciens were increased. The populations of three dominant cellulolytic bacteria were enhanced by BAFLOP supplementation, especially on high concentrate diet. BAFLOP supplementation did not influence the ammonia nitrogen (NH3-N) concentration, while R:C did. In addition, the in vitro digestibility was improved by either R:C or BAFLOP supplementation. The BAFLOP supplementation showed an effect on gas production kinetics, except for the gas production rate constant for the insoluble fraction (c), while treatments with high concentrate ratio resulted in the highest values. In addition, BAFLOP tended to increase gas production. Based on this study, it could be concluded that R:C had an effect on rumen ecology both in buffalo and cattle rumen fluid and hence, BAFLOP could be used as a rumen buffering agent for enhancing rumen ecology fed on high concentrate diet. It is recommended that level of BAFLOP supplementation should be at 2 to 4% of total dry matter of substrate. However, in vivo trials should be subsequently conducted to investigate the effect of BAFLOP in high concentrate diets on rumen ecology as well as ruminant production.

Changes of microbial population in the rumen of dairy steers as influenced by plant containing tannins and saponins and roughage to concentrate ratio.

The objective of this study was to investigate microbial population in the rumen of dairy steers as influenced by supplementing with dietary condensed tannins and saponins and different roughage to concentrate ratios. Four, rumen fistulated dairy steers (Bos indicus) were used in a 2×2 factorial arrangement in a 4×4 Latin square design. The main factors were two roughage to concentrate ratios (R:C, 60:40 and 40:60) and two supplementations of rain tree pod meal (RPM) (0 and 60 g/kg of total DM intake). Chopped 30 g/kg urea treated rice straw was used as a roughage source. All animals received feed according to respective R:C ratios at 25 g/kg body weight. The RPM contained crude tannins and saponins at 84 and 143 g/kg of DM, respectively. It was found that ruminal pH decreased while ruminal temperature increased by a higher concentrate ratio (R:C 40:60) (p<0.05). In contrast, total bacterial, Ruminococus albus and viable proteolytic bacteria were not affected by dietary supplementation. Numbers of fungi, cellulolytic bacteria, Fibrobactor succinogenes and Ruminococus flavefaciens were higher while amylolytic bacteria was lower when steers were fed at 400 g/kg of concentrate. The population of Fibrobactor succinogenes, was found to be higher with RPM supplementation. In addition, the use of real-time PCR technique indicated that the population of protozoa and methanogens were decreased (p<0.05) with supplementation of RPM and with an increasing concentrate ratio. Supplementation of RPM and feeding different concentrate ratios resulted in changing the rumen microbes especially, when the animals were fed at 600 g/kg of concentrate and supplemented with RPM which significantly reduced the protozoa and methanogens population.

Effects of physical form and urea treatment of rice straw on rumen fermentation, microbial protein synthesis and nutrient digestibility in dairy steers.

This study was designed to determine the effect of physical form and urea treatment of rice straw on rumen fermentation, microbial protein synthesis and nutrient digestibility. Four rumen-fistulated dairy steers were randomly assigned according to a 2 (2 factorial arrangement in a 4 (4 Latin square design to receive four dietary treatments. Factor A was roughage source: untreated rice straw (RS) and urea-treated (3%) rice straw (UTRS), and factor B was type of physical form of rice straw: long form rice straw (LFR) and chopped (4 cm) rice straw (CHR). The steers were offered the concentrate at 0.5% body weight (BW) /d and rice straw was fed ad libitum. DM intake and nutrient digestibility were increased (p<0.05) by urea treatment. Ruminal pH were decreased (p<0.05) in UTRS fed group, while ruminal ammonia nitrogen (NH3-N) and blood urea nitrogen (BUN) were increased (p<0.01) by urea treatment. Total volatile fatty acid (VFA) concentrations increased (p<0.01) when steers were fed UTRS. Furthermore, VFA concentrations were not altered by treatments (p>0.05), except propionic acid (C3) was increased (p<0.05) in UTRS fed group. Nitrogen (N) balance was affected by urea treatment (p<0.05). Microbial protein synthesis (MCP) synthesis were greater by UTRS and CHR group (p<0.05). The efficiency of microbial N synthesis was greater for UTRS than for RS (p<0.05). From these results, it can be concluded that using the long form combined with urea treatment of rice straw improved feed intake, digestibility, rumen fermentation and efficiency of microbial N synthesis in crossbred dairy steers.

Effect of carbohydrate sources and levels of cotton seed meal in concentrate on feed intake, nutrient digestibility, rumen fermentation and microbial protein synthesis in young dairy bulls.

The objective of this study was to investigate the effect of levels of cottonseed meal with various carbohydrate sources in concentrate on feed intake, nutrient digestibility, rumen fermentation and microbial protein synthesis in dairy bulls. Four, 6 months old dairy bulls were randomly assigned to receive four dietary treatments according to a 2×2 factorial arrangement in a 4×4 Latin square design. Factor A was carbohydrate source; cassava chip (CC) and cassava chip+rice bran in the ratio of 3:1 (CR3:1), and factor B was cotton seed meal levels in the concentrate; 109 g CP/kg (LCM) and 328 g CP/kg (HCM) at similar overall CP levels (490 g CP/kg). Bulls received urea-lime treated rice straw ad libitum and were supplemented with 10 g of concentrate/kg BW. It was found that carbohydrate source and level of cotton seed meal did not have significant effects on ruminal pH, ammonia nitrogen concentration, microbial protein synthesis or feed intake. Animals which received CC showed significantly higher BUN concentration, ruminal propionic acid and butyric acid proportions, while dry matter, organic matter digestibility, populations of total viable bacteria and proteolytic bacteria were lower than those in the CR3:1 treatment. The concentration of total volatile fatty acids was higher in HCM than LCM treatments, while the concentration of butyric acid was higher in LCM than HCM treatments. The population of proteolytic bacteria with the LCM treatments was higher than the HCM treatments; however other bacteria groups were similar among the different levels of cotton seed meal. Bulls which received LCM had higher protein digestibility than those receiving HCM. Therefore, using high levels of cassava chip and cotton seed meal might positively impact on energy and nitrogen balance for the microbial population in the rumen of the young dairy bull.

Effect of plants containing secondary compounds with palm oil on feed intake, digestibility, microbial protein synthesis and microbial population in dairy cows.

The objective of this study was to determine the effect of rain tree pod meal with palm oil supplementation on feed intake, digestibility, microbial protein synthesis and microbial populations in dairy cows. Four, multiparous early-lactation Holstein-Friesian crossbred (75%) lactating dairy cows with an initial body weight (BW) of 405±40 kg and 36±8 DIM were randomly assigned to receive dietary treatments according to a 4×4 Latin square design. The four dietary treatments were un-supplementation (control), supplementation with rain tree pod meal (RPM) at 60 g/kg, supplementation with palm oil (PO) at 20 g/kg, and supplementation with RPM at 60 g/kg and PO at 20 g/kg (RPO), of total dry matter intake. The cows were offered concentrates, at a ratio of concentrate to milk production of 1:2, and chopped 30 g/kg of urea treated rice straw was fed ad libitum. The RPM contained condensed tannins and crude saponins at 88 and 141 g/kg of DM, respectively. It was found that supplementation with RPM and/or PO to dairy cows diets did not show negative effects on feed intake and ruminal pH and BUN at any times of sampling (p>0.05). However, RPM supplementation resulted in lower crude protein digestibility, NH3-N concentration and number of proteolytic bacteria. It resulted in greater allantoin absorption and microbial crude protein (p<0.05). In addition, dairy cows showed a higher efficiency of microbial N supply (EMNS) in both RPM and RPO treatments. Moreover, NDF digestibility and cellulolytic bacteria numbers were highest in RPO supplementation (p<0.05) while, supplementation with RPM and/or PO decreased the protozoa population in dairy cows. Based on this study, supplementation with RPM and/or PO in diets could improve fiber digestibility, microbial protein synthesis in terms of quantity and efficiency and microbial populations in dairy cows.