The effects of the nonionic surfactant Tween 80 and a mixture of fibrolytic enzymes on total tract digestion, in situ disappearance (ISD) and ruminal fermentation characteristics of orchardgrass hay and barley grain were investigated in a 4${\times}$4 Latin square experiment with 4 non-lactating Holstein cows and 4 diets in 4 periods. Cows were offered a total mixed ration of 50% rolled barley grain and 50% orchardgrass hay treated with either 1) water (control), 2) 0.2% (vol/wt) Tween 80, 3) 0.2% (vol/wt) hydrolytic enzyme, or 4) 0.2% hydrolytic enzyme plus 0.2% Tween 80. Total tract digestibility coefficients of DM, nitrogen, NDF and ADF were not affected (p>0.05) by dietary treatment. Compared to the control, the rate of ISD of DM from orchardgrass hay was faster (p<0.05) in cows receiving diets treated with the enzyme alone or with enzyme plus Tween 80 (0.06/h vs. 0.076 and 0.069/h). The rate of digestion was lower (p<0.05) as compared to control when barley grain was treated with these additives. Ruminal fluid pH and concentrations of total VFA, acetate, isobutyrate and butyrate were not affected (p>0.05) by treatments. Cows that consumed diets treated with enzyme plus Tween 80 had higher (p<0.05) ruminal concentrations of propionate and isovalerate, and lower (p<0.05) acetate:propionate ratios. Compared to the control, microbial protein synthesis tended (p = 0.13) to increase with the addition of enzyme to the diet while nonammonia nitrogen flow to the duodenum increased (p<0.05) with both enzyme and Tween 80 treatments. The study indicated that fibrolytic enzymes alone or in combination with Tween 80 could enhance ISD of orchardgrass hay and ruminal concentrations of propionate, valerate and iso-valerate, but do not affect total tract digestibility.
An experiment was carried out to study the effect of monensin administration on mammary functions in crossbred Holstein cattle. Fourteen non-pregnant late lactating crossbred Holstein cattle, approximately 270 days postpartum, were selected for the experiment. They were divided into two groups of 7 animals each. Seven animals in the treated group were given sodium monensin orally in a slow-release capsule. Animals in both control and treated groups were fed the similar diet to maintain milk production and body score at 2.5. Rice straw was fed as a source of dietary fiber throughout the experimental period. After monensin administration, a significant increase in the molar percent of ruminal propionate (p<0.05) and a significant decrease in the molar percent of ruminal acetate (p<0.05) were apparent in comparison to the pretreated period. The ratio of acetate to propionate concentration decreased significantly after monensin administration (p<0.05), while it was maintained at the similar level throughout the period of experiment in the control group. Monensin did not affect the molar percent of ruminal butyrate and valerate. The concentration of milk allantoin between the control group and monensin treated group was not different. An excretion rate of allantoin in milk decreased in animals treated with monensin (p<0.05). Mammary blood flow did not show significant difference between control and monensin treated groups. The plasma glucose concentration, arteriovenous concentration difference and mammary gland uptake of glucose remained constant in both groups. Milk yield of the later stage of lactation in the control group declined during lactation advance while a tendency to increase in the milk yield was apparent after 21 days monensin administration. Milk compositions for concentration of lactose, fat and protein in both control group and monensin treated group did not change throughout the experimental periods. From these results, it can be concluded that the action of monensin could affect the ruminal fermentation pattern. Monensin could not increase milk yield in the late lactating period.
Kim, Yu-Sam;An, Jae-Hyung;Yang, Bu-Hyun;Kim, Kyu-Wan
BMB Reports
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v.29
no.4
/
pp.277-285
/
1996
In Pseudomonas fluorescens grown on malonate as sole carbon source, acetyl-CoA synthetase was induced, suggesting that malonate is metabolized through acetate and then acetyl-CoA. Acetyl-CoA synthetase was purified 18.6-fold in 4 steps to apparent homogeneity. The native molecular mass of the enzyme estimated by a native acrylamide gel electrophoresis was 130 kDa. The enzyme was composed of two identical subunits with a molecular mass of 67 kDa. Optimum pH was 70. The acetyl-CoA synthetase showed typical Michaelis-Menten kinetics for the substrates, acetate, ATP and CoA, whose $K_m$ values were calculated to be 33.4, 74.8, and 40.7 mM respectively. Propionate. butyrate and pentanoate were also used as substrates by the enzyme, but the rate of the formation of the CoA derivatives was decreased in the order of the increase in carbon number. The enzyme was inhibited by the group-specific reagents diethylpyro-carbonate, 2,3-butanedione, pyridoxal-5'-phosphate and N-bromosuccinimide. In the presence of substrates the inactivation rate of the enzyme, by all of the group-specific reagents mentioned above decreased, indicating the presence of catalytically essential histidine, arginine, lysine and tryptophan residues at or near the active site. Preincubation of the enzyme with ATP, $Mg^{2+}$ resulted in the increase of its susceptibility to diethylpyrocarbonate, suggesting that ATP, $Mg^{2+}$ may induce a conformational change in the active site exposing the essential histidine residue to diethylpyrocarbonate. The enzyme was acetylated in the presence of acetyl-CoA, indicating that this is one of acyl-enzyme.
Acetyl xylan esterase was produced by E. coli HB101 harboring a recombinant plasmid pKMG6 which contained the estI gene of Bacillus stearothermophilus. The maximum production was observed when the E. coli strain was grown at 37$\circC for 12 hours in the medium containing 0.5% acetyl xylan, 1.0% tryptons, 1.0% sodium chloride, and 0.5% yeast extract. The esterase produced was purified to homogeneity using a combination of ammonium sulfate fractionation, DEAE Sepharose CL-6B ion exchange chromatography and Sephacryl S-200 gel filtration. The native enzyme had an apparent molecular mass of 60 kd and was composed of two identical subunits of 29 kd. The N-terminal amino acid sequence of the polypeptide was Ala-X-Leu-Gln- Ile-Gln-Phe-X-X-Gln. The acetyl esterase displayed a pH optimum of 6.5 and a temperature opti- mum of 45$\circC. The heavy metal ions such as Ag$^{++}$, Hg$^{++}$ and Cu$^{++}$ inhibited nearly completely the activity of the esterase, and no specific metal ion was found to be required for the enzyme activity. The enzyme readily cleaved MAS, $\beta$-D-glucose pentaacetate, $\alpha$-naphthyl acetate, $\rho$-nitrophenyl acetate as well as acetyl xylan, but had no activity on $\rho$-nitrophenyl propionate, $\beta$-nitrophenyl butyrate or $\beta$-nitrophenyl valerate. The Km and Vmax values for MAS were 2.87 mM and 11.55 $\mu$mole/min, respectively. Synergistic behavior was demonstrated with a combination of xylanase and esterase from B. stearothermophilus in hydrolyzing acetyl xylan.
Journal of The Korean Society of Grassland and Forage Science
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v.33
no.3
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pp.197-205
/
2013
An in vitro study was conducted to determine the effects of defaunation (removal of protozoa) and forage sources (rice straw, ryegrass and tall fescue) on ruminal fermentation characteristics, methane ($CH_4$) production and degradation by rumen microbes. Sodium lauryl sulfate, as a defaunation reagent, was added into the mixed culture solution to remove ruminal protozoa at a concentration of 0.375 mg/ml. Pure cellulose (0.64 g, Sigma, C8002) and three forage sources were incubated in the bottle of culture solution of mixed rumen microbes (faunation) or defaunation for up to 24 h. The concentration of ammonia-N was high under condition of defaunation compared to that from faunation in all incubations (p<0.001). Total VFA concentration was increased at 3, 6 and 12 h (p<0.05~p<0.01) but was decreased at 24 h incubation (p<0.001) under condition of defaunation. Defaunation decreased acetate (p<0.001) and butyrate (p<0.001) proportions at 6, 12 and 24 h incubation times, but increased propionate (p<0.001) proportion at all incubation times for forages. Effective degradability of dry matter was decreased by defaunation (p<0.001). Defaunation not only decreased total gas (p<0.001) and $CO_2$ (p<0.01~0.001) production at 12 and 24 h incubations, but reduced $CH_4$ production (p<0.001) at all incubation times for all forages. The $CH_4$ production, regardless of defaunation, in order of forage sources were rice straw > tall fescue > ryegrass > cellulose (p<0.001) up to 24 h incubation.
Mahesh, Malleswarapu;Bharathi, Munugala;Reddy, Mooli Raja Gopal;Kumar, Manchiryala Sravan;Putcha, Uday Kumar;Vajreswari, Ayyalasomayajula;Jeyakumar, Shanmugam M.
Preventive Nutrition and Food Science
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v.21
no.3
/
pp.171-180
/
2016
Non-alcoholic fatty liver disease (NAFLD) is one of the most prevalent liver diseases associated with an altered lifestyle, besides genetic factors. The control and management of NAFLD mostly depend on lifestyle modifications, due to the lack of a specific therapeutic approach. In this context, we assessed the effect of carrot juice on the development of high fructose-induced hepatic steatosis. For this purpose, male weanling Wistar rats were divided into 4 groups, fed either a control (Con) or high fructose (HFr) diet of AIN93G composition, with or without carrot juice (CJ) for 8 weeks. At the end of the experimental period, plasma biochemical markers, such as triglycerides, alanine aminotransferase, and ${\beta}$-hydroxy butyrate levels were comparable among the 4 groups. Although, the liver injury marker, aspartate aminotransferase, levels in plasma showed a reduction, hepatic triglycerides levels were not significantly reduced by carrot juice ingestion in the HFr diet-fed rats (HFr-CJ). On the other hand, the key triglyceride synthesis pathway enzyme, hepatic stearoyl-CoA desaturase 1 (SCD1), expression at mRNA level was augmented by carrot juice ingestion, while their protein levels showed a significant reduction, which corroborated with decreased monounsaturated fatty acids (MUFA), particularly palmitoleic (C16:1) and oleic (C18:1) acids. Notably, it also improved the long chain n-3 polyunsaturated fatty acid, docosahexaenoic acid (DHA; C22:6) content of the liver in HFr-CJ. In conclusion, carrot juice ingestion decreased the SCD1-mediated production of MUFA and improved DHA levels in liver, under high fructose diet-fed conditions. However, these changes did not significantly lower the hepatic triglyceride levels.
Objective: The objective of this study was to investigate the effect of bamboo grass (Tiliacora triandra, Diels) pellet (Bamboo-Cass) supplementation on feed intake, nutrient digestibility, rumen microbial population and methane production in Thai native beef cattle. Methods: Four Thai native beef cattle bulls ($190{\pm}2kg$) were randomly allotted to four respective dietary treatments in a $4{\times}4$ Latin square design. Treatments were the varying levels of Bamboo-Cass supplementation at 0, 50, 100, and 150 g/head/d, respectively. Rice straw was fed ad libitum and the concentrate offered at 0.5% of body weight. Results: Under this experiment, the findings revealed that acetate and butyrate production were decreased (p<0.05), propionate increased (p<0.05), whilst ruminal $NH_3-N$ concentration was decreased (p<0.05) by supplementation of Bamboo-Cass at 150 g/head/d. Moreover, rice straw intake, and microbial population were linearly increased (p<0.05), while methane production was decreased (p<0.05). Conclusion: The results from the present study indicate that supplementation of Bamboo-Cass at 150 g/head/d significantly enhanced feed intake, decreased protozoa and increased bacterial population, rumen fermentation efficiency while decreased methane production. Therefore, Bamboo-Cass as a supplement is promising as a rumen enhancer in beef cattle fed on rice straw.
Majewska, Malgorzata P.;Miltko, Renata;Belzecki, Grzegorz;Kedzierska, Aneta;Kowalik, Barbara
Animal Bioscience
/
v.34
no.7
/
pp.1146-1156
/
2021
Objective: The aim of the study was to compare the effect of two plant additives, rich in polyphenolic compounds, supplemented to sheep diets on microorganisms and carbohydrate fermentation in rumen. Methods: In the experiment, 6 ewes of the Polish Mountain breed were fitted with ruminal cannulas. Sheep were divided into three feeding groups. The study was performed in a cross-over design of two animals in each group, with three experimental periods (n = 6 per each group). The animals were fed a control diet (CON) or additionally received 3 g of dry and milled lingonberry leaves (VVI) or oak bark (QUE). Additionally, plant material was analyzed for tannins concentration. Results: Regardless of sampling time, QUE diet increased the number of total protozoa, as well as Entodinium spp., Diplodinium spp. and Isotrichidae family, while decreased bacterial mass. In turn, a reduced number of Diplodinium spp. and increased Ophryoscolex spp. population were noted in VVI fed sheep. During whole sampling time (0, 2, 4, and 8 h), the number of protozoa in ruminal fluid of QUE sheep was gradually reduced as opposed to animals receiving CON and VVI diet, where rapid shifts in the protozoa number were observed. Moreover, supplementing sheep with QUE diet increased molar proportions of butyrate and isoacids in ruminal fluid. Unfortunately, none of the tested additives affected gas production. Conclusion: The addition of VVI or QUE in a small dose to sheep diets differently affected rumen microorganisms and fermentation parameters, probably because of various contribution of catechins in tested plant materials. However, it is stated that QUE diet seems to create more favorable conditions for growth and development of ciliates. Nonetheless, the results of the present study showed that VVI and QUE additives could serve as potential natural modulators of microorganism populations and, consequently, carbohydrate digestion in ruminants.
The effects of two types of protein, soybean meal (SBM) and fish meal (FM); and two types of energy supplements, corn flour (CF) and paper pulp (PP), on intake of guinea grass (Panicum maximum), fibre digestion and microbial activities in four Merino rams with an average weight of $54.4{\pm}4.5kg$ were studied. Each animal was fitted with a ruminal cannula and a duodenal cannula at the proximal position. The animals were fed twice daily with chopped guinea grass (5 cm) ad libitum and one of the four dietary supplements: 170 g FM+268 g PP; 170 g FM+268 g CF; 200 g SBM+200 g PP or 200 g SBM+200 g CF. All the supplements were mixed with 100 g molasses. In sacco and in vivo digestibilities, digesta flow rates, fermentation and microbial population were studied in a $4{\times}4$ Latin square design with a $2{\times}2$ factorial arrangement of dietary treatments. The effects of energy or protein sources were not significant on grass intake of sheep. The potential degradabilities of NDF and ADF were not significantly affected by any of the supplements. However, the energy and protein sources had significant efects on disappearance rate of NDF and ADF. The disappearance rate of both NDF and ADF were significantly (p < 0.05) higher in animals fed PP when compared to animals fed CF. Animals fed FM also showed significantly (p < 0.03) higher disappearance rate of ADF than those fed SBM. Animals fed PP showed better digestion in the rumen and total tract. Total flow of NDF and ADF through the duodenum was not significantly affected by the various supplements. The mean rumen pH values (5.8-6.1) were not significantly different among the four different diets. The concentration of rumen ammonia was significantly (p < 0.0001) higher in animals fed SBM (235-266.4 mg N/L) supplement than in animals fed FM (174.9-179.7 mg N/L), while total VFA concentration was not significantly affected by both energy and protein supplements. Mean values of total VFA ranged from 72.5-82.3 mM. Molar proportions of acetate, propionate and butyrate were typical of a roughage type fermentation. Molar proportion of acetate was significantly (p < 0.0001) higher in sheep fed PP when compared to sheep fed CF. Animals fed FM had higher total viable bacterial counts, while animals fed CF showed higher protozoal numbers. Proportions of cellulolytic bacteria were only slightly higher in animals fed SBM or PP.
This experiment was conducted to study the effect of addition of Lucerne juice (LJ) obtained by mechanical extraction of freshly harvested crop on the nutritive value of rice straw silage. Rice straw (RS) was ensiled with intact, NaOH or $NH_3$ treated LJ at 3:7 ratio on fresh weight basis (LJ RS, LJ NaOH RS and LJ $NH_3$ RS, respectively). Each alkali was mixed with fresh juice at a level of 4% of rice straw dry matter just before ensiling. Rice straw ensiled with water was prepared as the control (W RS). In the digestion trial, goats were allocated in a $4{\times}4$ Latin-square design and fed the diet containing three parts of RS silage and one part of wheat bran (DM basis). For the goats receiving the control silage, urea was supplemented at feeding time so as to adjust the nitrogen intake except for goats on LJ $NH_3$ RS silage. Crude protein content of RS silage was increased from 5.2 to 9.1% (DM basis) by the addition of intact LJ and to about 24% by $NH_3$ treated LJ. The control W RS silage contained only trace amount of lactic acid and was dominated by acetic and butyric acid. The addition of intact LJ reduced butyric acid content and $NH_3-N/TN$ of the silage whereas the addition of alkalized LJ increased those values and shifted to a butyrate type fermentation. Nutrient digestibilities and nitrogen balance of goats were almost the same when they were fed W RS and LJ RS silage indicating the addition of intact LJ did not improve the nutritive value. The addition of alkalized LJ significantly increased the fiber digestibilities of RS silage and $NH_3$ treatment was more effective than NaOH treatment. Postprandial ruminal $NH_3-N$ and blood urea nitrogen (BUN) concentrations were decreased by feeding LJ NaOH RS silage suggesting ruminal protein synthesis was enhanced along with the increase of energy supply for supply for rumen microbes by the alkali treatment. The advantageous fiber digestibilities of LJ $NH_3$ RS silage compared with those of LJ NaOH RS silage might be attributable to a sufficient nitrogen supply for microbial fiber digestion in the rumen.
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