A functional screen of 60,672 fosmid metagenomic clones amplified from marine sediment obtained from the Dokdo islets in Korea identified the gene EstES1, whose product, EstES1, displayed lipolytic properties on tributyrin-supplemented media. EstES1 is a 576 amino acid protein with a predicted molecular weight of 59.4 kDa including 37 N-terminal leader amino acids. EstES1 exhibited the highest sequence similarity (44%) to a carboxylesterase found in Haliangium ochraceum DSM14365. Phylogenetic analysis indicated that EstES1 belongs to a currently uncharacterized family of lipases. Within the conserved domain, EstES1 retains the catalytic triad that consists of the consensus penta-peptide motif, GESAG. EstES1 demonstrated a broad substrate specificity toward the long acyl group of ethyl esters (C2-C12), and its optimal activity was recorded toward p-Nitrophenyl butyrate (C4) at pH 9.0 and $40^{\circ}C$ (specific activity of 255.4 U/mg). The enzyme remained stable in the ranges of $60-65^{\circ}C$ and pH 9.0-10.5 and in the presence of methanol, ethanol, isopropanol, and dimethyl sulfoxide. Therefore, EstES1 has potential for use in industrial applications involving high temperature, organic solvents, and/or alkaline conditions.
Journal of The Korean Society of Grassland and Forage Science
/
v.41
no.2
/
pp.102-109
/
2021
In this study, the effect of forage sources in the total mixed ration (TMR) on in vitro goat rumen fermentation was investigated. Rice straw (RS), Italian ryegrass (IRG), timothy (TIM), and alfalfa (ALF) were used as forage sources. Each forage source was mixed with a commercial goat concentrate diet in the ratio of 1:1. Total 4 TMR were prepared. Rumen simulated in vitro fermentation using goat rumen fluid collected from the slaughterhouse was conducted until 72th. For fermentation parameters, gas production (GP), volatile fatty acids (VFAs), and ammonia nitrogen (NH3-N) were examined. All assays were performed at 24th, 48th, and 72th h of incubation individually. Contents of crude protein and non-fibrous carbohydrate were greater in the order of RS < IRG < TIM < ALF. Significant treatment effects were found in valerate and NH3-N at 24th h of incubation (p<0.05). ALF showed the greatest contents of them and RS was the lowest. At 48th incubation, a significant effect was detected at GP (p<0.05) and RS was greater than others. However, GP of RS was lower than others at 72th. Significant effects on Total VFA, butyrate, and valerate productions were found at 72th h of incubation (p<0.05). ALF showed the greatest production. Methane production from all treatments was not significantly different for each incubation time (p>0.05). The present study provided primary information on how goat rumen fermentation responds to different nutrient contents and forage sources of TMR. And the information could be used for the design or optimizing economical diet formulation for goats.
Khezri, A.;Rezayazdi, K.;Mesgaran, M. Danesh;Moradi-Sharbabk, M.
Asian-Australasian Journal of Animal Sciences
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v.22
no.5
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pp.651-658
/
2009
Four multiparous lactating Holstein cows fitted with rumen cannulae were fed diets varying in the amount and source of rumen-degradable carbohydrates (starch vs. sucrose) to examine their effects on rumen fermentation, nitrogen metabolism and lactation performance. A $4{\times}4$ Latin square with four diets and four periods of 28 days each was employed. Corn starch and sucrose were added to diets and corn starch was replaced with sucrose at 0 (0 S), 2.5 (2.5 S), 5.0 (5.0 S) 7.5% (7.5 S) of diet dry matter in a total mixed ration (TMR) containing 60% concentrate and 40% forage (DM basis). Replacing corn starch with sucrose did not affect (p>0.05) ruminal pH which averaged 6.41, but the ruminal pH for 7.5 S decreased more rapidly at 2 h after morning feeding compared with other treatments. Sucrose reduced ($p{\leq}0.05$) ruminal $NH_3-N$ concentration (13.90 vs. 17.09 mg/dl) but did not affect peptide-N concentration. There was no dietary effect on total volatile fatty acids (110.53 mmol/L) or the acetate to propionate ratio (2.72). No differences (p>0.05) in molar proportion of most of the individual VFA were found among diets, except for the molar proportion of butyrate that was increased ($p{\leq}0.05$) with the inclusion of sucrose. Total branched chain volatile fatty acids tended to increase ($p{\geq}0.051$) for the control treatment (0 S) compared with the 7.5 S treatment. Dry matter intake, body weight changes and digestibility of DM, OM, CP, NDF and ADF were not affected by treatments. Sucrose inclusion in the total mixed ration did not affect milk yield, but increased milk fat and total solid percentage ($p{\leq}0.05$). Sucrose tended ($p{\geq}0.063$) to increase milk protein percentage (3.28 vs. 3.05) and reduced ($p{\leq}0.05$) milk urea nitrogen concentration (12.75 vs. 15.48 mg/dl), suggesting a more efficient utilization of the rapidly available nitrogen components in the diet and hence improving nitrogen metabolism in the rumen.
Chen, G.J.;Song, S.D.;Wang, B.X.;Zhang, Z.F.;Peng, Z.L.;Guo, C.H.;Zhong, J.C.;Wang, Y.
Asian-Australasian Journal of Animal Sciences
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v.28
no.12
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pp.1736-1741
/
2015
The objective of this study was to determine the effect of forage: concentrate ratio (F:C) on growth performance, ruminal fermentation and blood metabolites of housing-feeding yaks. Thirty-two Maiwa male yaks (initial body weight = $207.99{\pm}3.31kg$) were randomly assigned to four dietary treatments (8 yaks per treatment). Experimental diets were: A, B, C, D which contained 70:30, 60:40, 50:50 and 40:60 F:C ratios, respectively. Dry matter intake and average daily gain in yaks fed the C and D diets were greater (p<0.05) than yaks fed the A and B diets. No differences were found in ruminal $NH_3-N$, total volatile fatty acids, acetate, butyrate, valerate, and isovalerate concentrations. The propionate concentration was increased (p<0.05) in the C and D groups compared with the A and B diets. In contrast, the acetate to propionate ratio was decreased and was lowest (p<0.05) in the C group relative to the A and B diets, but was similar with the D group. For blood metabolites, no differences were found in serum concentrations of urea-N, albumin, triglyceride, cholesterol, low density lipoprotein, alanine aminotransferase, and aspartate aminotransferase (p>0.05) among treatments. Treatment C had a higher concentration of total protein and high density lipoprotein (p<0.05) than A and B groups. In addition, there was a trend that the globulin concentration of A group was lower than other treatments (p = 0.079). Results from this study suggest that increasing the level of concentrate from 30% to 50% exerted a positive effect on growth performance, rumen fermentation and blood metabolites in yaks.
This study evaluated the effect of Candida norvegensis (C. norvegensis) viable yeast culture on in vitro ruminal fermentation of oat straw. Ruminal fluid was mixed with buffer solution (1:2) and anaerobically incubated with or without yeast at $39^{\circ}C$ for 0, 4, 8, 16, and 24 h. A fully randomized design was used. There was a decrease in lactic acid (quadratic, p = 0.01), pH, (quadratic, p = 0.02), and yeasts counts (linear, p<0.01) across fermentation times. However, in vitro dry matter disappearance (IVDMD) and ammonia-N increased across fermentation times (quadratic; p<0.01 and p<0.02, respectively). Addition of yeast cells caused a decrease in pH values compared over all fermentation times (p<0.01), and lactic acid decreased at 12 h (p = 0.05). Meanwhile, yeast counts increased (p = 0.01) at 12 h. C. norvegensis increased ammonia-N at 4, 8, 12, and 24 h (p<0.01), and IVDMD of oat straw increased at 8, 12, and 24 h (p<0.01) of fermentation. Yeast cells increased acetate (p<0.01), propionate (p<0.03), and butyrate (p<0.03) at 8 h, while valeriate and isovaleriate increased at 8, 12, and 24 h (p<0.01). The yeast did not affect cellulolytic bacteria (p = 0.05), but cellulolytic fungi increased at 4 and 8 h (p<0.01), whereas production of methane decreased (p<0.01) at 8 h. It is concluded that addition of C. norvegensis to in vitro oat straw fermentation increased ruminal fermentation parameters as well as microbial growth with reduction of methane production. Additionally, yeast inoculum also improved IVDMD.
Objective: This study was to examine in vivo digestibility, nitrogen balance and ruminal fermentation of tamarind (Tamarind indica) kernel powder extract residue (TKPER) compared to soybean products and by-products in wethers. Methods: Four wethers with initial body weight (BW) of $51.6{\pm}5.5kg$ were assigned in a $4{\times}4$ Latin square design to investigate nutritional characteristics of TKPER, dry heat soybean (SB), dry soybean curd residue (SBCR) and soybean meal (SBM) feeding with ryegrass straw (R) at a ratio of 1:1 at 2% of BW in dry matter (DM) on a daily basis. Results: The digestibility of DM, crude protein, and ether extract (EE) of TKPER-R diet were 57.0%, 87.0%, and 86.0%, respectively. Higher non-fiber carbohydrates digestibility was observed in TKPER-R diet (83.2%) than in SB-R diet (73.9%, p<0.05). Wethers fed the TKPER-R diet had lower retention of nitrogen (N) and ruminal ammonia nitrogen ($NH_3-N$) contents at 4 h after feeding than those fed the SBM-R diet (p<0.05), which had values similar to the SB-R or SBCR-R diet. The TKPER feeding had higher propionate (C3) and lower butyrate content, as well as lower acetate to propionate ratio (C2:C3) in rumen fluid than SBM feeding at 4 h after feeding (p<0.05). Conclusion: TKPER did not bring any side effect to the wethers although it was lack of fiber, and could be used as a high protein and energy ingredient in concentrate with appropriate roughage to meet the fiber requirement for ruminants.
The Blue-green algae collected from whole coasts of Korea were investigated morphotaxonomically in order to list up Korean marine Cyanophyta and clarify their taxnomic position. As a result, 36 species, 20 genus, 6 families belonging to 3 orders were identified. Among these, 14 species were recorded for the first time in Korea. They are Chroococcus minutus (K$\"{u}$tzing) N$\"{a}$geli, Merismopedia punctata Meyen, Microcystis ichtyoblabe K$\"{u}$zing, Dermocarpa leibleiniae (Reinsch) Born. et Thur., Hydrocoleum confluens (Setchell et Gardner) Drouet, Lyngbya sordida (Zanard.) Gomont, Phormidium forveolarum (Mont.) Gomont, Phormidium hansgieri Schmidle, Skujaella hildebrandtii (Gomont.) de Toni, Sphaeronema lithophila (Ercegovic) Umezaki, Spirulina tenerrima K$\"{u}$tzing, Hormothamnion enteromorphoides Grunow, Michrochaete aeruginea Batters, Michrochaete grisea Thuret ex Born. et flah.. Using the phase contrast microscope and the Nomarski interference micrope, we made photomicrographs of minute blue green algae. The cellular inclusions especially PHB(poly-$\SS$-hydroxy-butyrate) granules of the blue-green algae identified were investigated. The species clearly characteriged to have PHB granule were Lyngbya confervoides, L. semiplena, Phormidium corium, Sirocoleum kurzii, Hormothamnion enteromorphoides and Calothrix crustacea. These result would be fundamental data for estabilishing phylogenetic system of blue-green algae based on physio-biochemical characteristics in future.
Kim, Dong-Hoon;Han, Sun-Kee;Kim, Sang-Hyoun;Bae, Byung-Uk;Shin, Hang-Sik
Journal of the Korea Organic Resources Recycling Association
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v.12
no.1
/
pp.49-57
/
2004
The effect of gas sparging on continuous fermentative $H_2$ production was investigated using external gases ($N_2$, $CO_2$) with various flow rates (100, 200, 300, 400 ml/min). Gas sparging showed a higher $H_2$ yield than no sparging, indicating that the decrease of $H_2$ partial pressure by gas sparging had a good effect on $H_2$ fermentation. Especially, $CO_2$ sparging was more effective in the reactor performance than $N_2$ sparging. The composition of butyrate, the main metabolic product of $H_2$ fermentation by Clostridium sp., was much higher in $CO_2$ sparging. $H_2$ production increased with increasing flow rate only in $CO_2$ sparging. The best performance was obtained by $CO_2$ sparging at 300 ml/min, resulting in the highest $H_2$ yield of 1.65 mol $H_2/mol$ hexoseconsumed and the maximum $H_2$ production of 6.77 L $H_2/g$ VSS/day. Compared to $N_2$ sparging, there could be another beneficial effect in $CO_2$ sparging apart from lowering down the $H_2$ partial pressure. High partial pressure of $CO_2$ had little effect on $H_2$ producing bacteria but inhibitory effect on other microorganisms like lactic acid bacteria and acetogens which were competitive with $H_2$ producing bacteria.
Journal of The Korean Society of Grassland and Forage Science
/
v.36
no.4
/
pp.309-317
/
2016
The study was conducted to evaluate the effects of microbial culture supplements on ruminal fermentation and fermentative quality of Italian ryegrass silage (IRGS) both in vitro and in situ. Three species of microbes (Lactobacillus casei (LC), Bacillus subtilis (BS), and Saccharomyces cerevisiae (SC)) were used in this study. They were applied to IRGS at 30 days after silage manufacture. Various items were measured using in vitro and in situ incubation technique after each microbial supplement was inoculated into IRGS at $0.5{\times}10^4CFU/g$. In the first experiment, in vitro ruminal fermentation characteristics of IRGS were evaluated at 0, 12, 24, 48, and 72 hours after microbes were inoculated into IRGS. In the second experiment, in situ fermentation characteristics were investigated at 0, 1, 3, and 5 days after the inoculation of each microbial supplement. In vitro ruminal $NH_3-N$ content was significantly (p<0.05) increased in LC-, BS-, and SC-IRGS at 12 hrs post incubation compared to that in control IRGS. In vitro ruminal total VFA concentration and dry matter digestibility (DMD) of IRGS were not significantly difference among LC-, BS-, and SC-IRGS, although they were numerically increased in LC-IRGS than those of the other IRGS. In addition, this study evaluated the fermentation characteristics and in situ DMD of IRGS with the lapse of incubation time up to 5 days. Throughout the incubation times from 1 day to 5 days, the pH value was significantly (p<0.05) lower in BS-, LC-, and SC-IRGS than that in control IRGS. Lactate was significantly (p<0.05) higher, and significantly (p<0.05) butyrate was lower in LC-IRGS than that in other treatments at 0 day. It was higher (p<0.05) in control IRGS than that of BS-, LC-, and SC-IRGS at 1-5 days. In situ DMD tended to increase in BS-, LC-, and SC-IRGS compared to that in control IRGS. Especially, DMD was higher in SC-IRGS than that in other treatments at 0 day. It tended to be higher in LC-IRGS at all incubation time. Taken together, these results suggest that it might be useful to select a microorganism by considering the feeding time of IRGS to ruminants because organic acids and DMD of IRGS were affected by the incubation time of each microorganism with IRG silage, especially for L. casei decreased the content of acetate and butyrate in IRGS.
Metabolic trial with 3 fistulated sheep was conducted in a 3 $\times$ 3 Latin square design and feeding trial with 24 Hanwoo steers in 12 month of age for 20 months was conducted to investigate the replacing effect of rice straw with fermented spent mushroom (Flammuliua velutipes) compost (FSMC) on fermentation characteristics, ruminal effective degradabilty and whole tract digestibility of nutrients in sheep, and to examine the growth performance of Hanwoo steers. Experimental diets for the metabolic trial with sheep were commercial concentrates and rice straw in the ratio of 70 : 30 (CON, DM basis). Same concentrate with 30% FSMC and 70% rice straw (FSMC-30) and 60% FSMC and 40% rice straw(FSMC-60). Diets for Hanwoo steers in three treatments were same as for metabolic trial in replacing ratio of rice straw with FSMC. pH of rumen fluid in sheep was not affected by FSMC. Ammonia-N content in the rumen fluid was highest in the sheep fed FSMC-60 at 3h (P<0.045). The CON diet increased (P<0.001) acetate proportion at 1h and 3h post feeding compared to FSMC-60 diet while propionate proportion was highest in the sheep fed FSMC-60 diet for all the sampling times (P<0.027~P<0.002). Increased proportion of butyrate was observed at 30 min prior to feeding (P<0.031), and 1h (P<0.011) and 6h(P<0.039) post feeding from sheep fed FSMC-30 diet compared to those from sheep fed other diets. Effective degradability in the rumen was not influenced by experimental diets. Whole tract digestibility of crude protein (P<0.031) and neutral detergent fiber (P<0.006) tended to be increased in the sheep fed CON diet while corresponding values were lowest in the sheep fed FSMC-60 diet. Total body weight gain of Hanwoo steers for 8 months was not different among diets, thus daily body gain was not influenced by the experimental diets.
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