• Asian-Australasian Journal of Animal Sciences
• /
• 제25권1호
• /
• pp.66-74
• /
• 2012

The effects of exogenous fibrolytic enzymes (EFE; a mixture of two preparations from Trichoderma spp., with predominant xylanase and ${\beta}$-glucanase activities, respectively) on colonization and digestion of ground barley straw and alfalfa hay by Fibrobacter succinogenes S85 and Ruminococcus flavefaciens FD1 were studied in vitro. The two levels (28 and 280 ${\mu}g$/ml) of EFE tested and both bacteria were effective at digesting NDF of hay and straw. With both substrates, more NDF hydrolysis (p<0.01) was achieved with EFE alone at 280 than at 28 ${\mu}g$/ml. A synergistic effect (p<0.01) of F. succinogenes S85 and EFE on straw digestion was observed at 28 but not 280 ${\mu}g$/ml of EFE. Strain R. flavefaciens FD1 digested more (p<0.01) hay and straw with higher EFE than with lower or no EFE, but the effect was additive rather than synergistic. Included in the incubation medium, EFE showed potential to improve fibre digestion by cellulolytic ruminal bacteria. In a second batch culture experiment using mixed rumen microbes, DM disappearance (DMD), gas production and incorporation of $^{15}N$ into particle-associated microbial N ($^{15}N$-PAMN) were higher (p<0.001) with ammoniated (5% w/w; AS) than with native (S) ground barley straw. Application of EFE to the straws increased (p<0.001) DMD and gas production at 4 and 12 h, but not at 48 h of the incubation. EFE applied onto S increased (p<0.01) $^{15}N$-PAMN at 4 h only, but EFE on AS increased (p<0.001) $^{15}N$-PAMN at all time points. Prehydrolysis increased (p<0.01) DMD from both S and AS at 4 and 12 h, but reduced (p<0.01) $^{15}N$-PAMN in the early stage (4 h) of the incubation, as compared to non-prehydrolyzed samples. Application of EFE to barley straw increased rumen bacterial colonization of the substrate, but excessive hydrolytic action of EFE prior to incubation decreased it.

• Animal Bioscience
• /
• 제36권8호
• /
• pp.1190-1198
• /
• 2023

Objective: To our knowledge, there are few studies on the correlation between internal structure of fermented products and nutrient delivery from by-products from coffee processing in the ruminant system. The objective of this project was to use advanced mid-infrared vibrational spectroscopic technique (ATR-FT/IR) to reveal interactive correlation between protein internal structure and ruminant-relevant protein and energy metabolic profiles of by-products from coffee processing affected by added-microorganism fermentation duration. Methods: The by-products from coffee processing were fermented using commercial fermentation product, called Saus Burger Pakan, consisting of various microorganisms: cellulolytic, lactic acid, amylolytic, proteolytic, and xylanolytic microbes, for 0, 7, 14, 21, and 28 days. Protein chemical profiles, Cornell Net Carbohydrate and Protein System crude protein and CHO subfractions, and ruminal degradation and intestinal digestion of protein were evaluated. The attenuated total reflectance-Ft/IR (ATR-FTIR) spectroscopy was used to study protein structural features of spectra that were affected by added microorganism fermentation duration. The molecular spectral analyses were carried using OMNIC software. Molecular spectral analysis parameters in fermented and non-fermented by-products from coffee processing included: Amide I area (AIA), Amide II (AIIA) area, Amide I heigh (AIH), Amide II height (AIIH), α-helix height (αH), β-sheet height (βH), AIA to AIIA ratio, AIH to AIIH ratio, and αH to βH ratio. The relationship between protein structure spectral profiles of by-products from coffee processing and protein related metabolic features in ruminant were also investigated. Results: Fermentation decreased rumen degradable protein and increased rumen undegradable protein of by-products from coffee processing (p<0.05), indicating more protein entering from rumen to the small intestine for animal use. The fermentation duration significantly impacted (p<0.05) protein structure spectral features. Fermentation tended to increase (p<0.10) AIA and AIH as well as β-sheet height which all are significantly related to the protein level. Conclusion: Protein structure spectral profiles of by-product form coffee processing could be utilized as potential evaluators to estimate protein related chemical profile and protein metabolic characteristics in ruminant system.

• 생명과학회지
• /
• 제18권4호
• /
• pp.515-521
• /
• 2008

본 연구는 반추위 미생물 발효에 있어서 계면활성제의 이온성 여부가 발효시간별 in vitro 건물소화율, 미생물 성장량, pH 변화, Cas 발생량 및 SEM에 의한 미생물 부착 양상을 조사하기 위해 수행되었다. 1 mm 입자도의 볏짚을 기질로 하여 Holstein 젖소 위액을 이용한 Dehority's artificial medium에 대조구를 비롯하여 비이온성 계면활성제(NIS)로서 시판되고 있는 Tween 80과 SOLFA-850 2종류, 그리고 양쪽(+/-) 이온성 계면활성제(ZIS)로서 3-(Dodecyldimethylammonio) propanesulfanate (DDAP) 1 종류를 이용하여 각각 0.05% 및 0.1% 수준으로 첨가함으로써 총 7처리를 두었다. 발효시간은 6, 12, 24, 48 및 72시간으로 설정하여 각 처리 당 3반복으로 시험을 수행하였다. In vitro 건물 소화율은 NIS인 Tween 80 첨가구에서 48시간 및 72시간 발효 시, 타 처리구에 비해 유의적(P<0.05)으로 높았으나, ZIS인 DDAP 첨가구는 발효 24시간이후 부터 대조구보다도 건물소화율이 낮게 나타났다(P< 0.05). 가스 발생량은 NIS 두 처리구 모두, 대조구나 ZIS 처리구보다 유의적(P<0.05)으로 많았으며, 발효시간의 경과함에 따라 증가하였다. 미생물 성장량은 NIS인 Tween 80 첨가구에서 가장 많았고, 다음으로 SOLFA 850 첨가구 순이었으며, ZIS인 DDAP 첨가구는 대조구보다도 적었다(P<0.05). 전자현미경으로 관찰한 미생물 부착 양상에서 NIS 첨가구는 무처리구에 비해 미생물 군집이 현저히 많았으나 ZIS첨가구는 오히려 적은 것으로 나타났다. 따라서 양쪽(+/-) 이온성 계면활성제는 반추위 발효 작용과 미생물 성장에 긍정적인 효과가 없는 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제10권4호
• /
• pp.357-363
• /
• 1997

The effect of replacement of cotton seed meal (CSM), with various levels and sources of non-protein nitrogenous (NPN), substances on in vitro ruminal fermentation were studied. Cotton seed meal, in control ration provided nitrogen equivalent to 12.5 percent crude protein while in experimental ration was replaced at 30, 50 & 70 percent levels with urea, diammonium phosphate (DAP) and biuret, respectively. The results of incubation upto 48 hours indicated an improvement in digestibility by replacement of CSM with urea and biuret upto 50 percent protein equivalent, but not with DAP. Bacterial count from cultures containing 50% nitrogen from biuret was significantly higher than DAP, urea and CSM. Various sources of nitrogen produced $NH_3-N$ in increasing order of CSM, biuret, DAP and urea. Increasing levels of NPN resulted in progressive increase in the levels of $NH_3-N$. The levels of various NPN sources had no effect on pH. However, the pH values determined for urea and CSM were higher than biuret and DAP.

• Asian-Australasian Journal of Animal Sciences
• /
• 제21권1호
• /
• pp.59-65
• /
• 2008

Six total mixed rations (TMR) containing 0, 4, 6, 8, 10, 12% tannin (TMR I-VI), using Accacia nilotica pods as a source of tannin, were used to study the effect of Acacia tannin on in vitro nutrient digestibility and gas production in goats. This study also investigated the degraded products of Acacia nilotica tannin in goat rumen liquor. Degraded products of tannins were identified using high performance liquid chromatography (HPLC) at different hours of incubation. In vitro digestibility of dry matter (IVDMD) and organic matter (IVOMD) were similar in TMR II, and I, but declined (p<0.05) thereafter to a stable pattern until the concentration of tannin was raised to 10%. In vitro crude protein digestibility (IVCPD) decreased (p<0.05) with increased levels of tannins in the total mixed rations. Crude protein digestibility was much more affected than digestibility of dry matter and organic matter. In vitro gas production (IVGP) was also reduced (p<0.05) with increased levels of tannins in the TMR during the first 24 h of incubation and tended to increase (p>0.05) during 24-48 h of incubation. Gallic acid, phloroglucinol, resorcinol and catechin were identified at different hours of incubation. Phloroglucinol and catechin were the major end products of tannin degradation while gallate and resorcinol were produced in traces. It is inferred that in vitro nutrient digestibility was reduced by metabolites of Acacia nilotica tannins and ruminal microbes of goat were capable of withstanding up to 4% tannin of Acacia nilotica pods in the TMR without affecting in vitro nutrient digestibility.

• Asian-Australasian Journal of Animal Sciences
• /
• 제22권1호
• /
• pp.82-89
• /
• 2009

Two in vitro experiments were conducted to examine the effects of propionate precursors in the dicarboxylic acid pathway on ruminal fermentatation characteristics, $CH_4$ production and degradation of feed by rumen microbes. Fumarate or malate as sodium salts (Exp. 1) or acid type (Exp. 2) were added to the culture solution (150 ml, 50% strained rumen fluid and 50% artificial saliva) to achieve final concentrations of 0, 8, 16 and 24 mM, and incubated anaerobically for 0, 1, 3, 6, 9 and 12 h at $39^{\circ}C$. For both experiments, two grams of feed consisting of 70% concentrate and 30% ground alfalfa (DM basis) were prepared in a nylon bag, and were placed in a bottle containing the culture solution. Addition of fumarate or malate in both sodium salt and acid form increased (p<0.0001) pH of culture solution at 3, 6, 9 and 12 h incubations. The pH (p<0.0001) and total volatile fatty acids (VFA, p<0.05) were enhanced by these precursors as sodium salt at 3, 6 and 9 h incubations, and pH (p<0.001) and total VFA (p<0.01) from fumarate or malate in acid form were enhanced at a late stage of fermentation (9 h and 12 h) as the addition level increased. pH was higher (p<0.001) for fumarate than for malate as sodium salt at 3 h and 6 h incubations. Propionate ($C_3$) proportion was increased (p<0.0001) but those of $C_2$ (p<0.05) and $C_4$ (p<0.01 - p<0.001) were reduced by the addition of sodium salt precursors from 3 h to 12 incubation times while both precursors in acid form enhanced (p<0.011 - p<0.0001) proportion of $C_3$ from 6h but reduced (p<0.018 - p<0.0005) $C_4$ proportion at incubation times of 1, 3, 9 and 12 h. Proportion of $C_3$ was increased (p<0.05 - p<0.0001) at all incubation times by both precursors as sodium salt while that of $C_3$ was increased (p<0.001) from 6h but $C_4$ proportion was decreased by both precursors in acid form as the addition level increased. Proportion of $C_3$ was higher (p<0.01 - p<0.001) for fumarate than malate as sodium salt from 6 h incubation but was higher for malate than fumarate in acid form at 9 h (p<0.05) and 12 h (p<0.01) incubation times. Increased levels (16 and 24 mM) of fumarate or malate as sodium salt (p<0.017) and both precursors in acid form (p<0.028) increased the total gas production, but no differences were found between precursors in both chemical types. Propionate precursors in both chemical types clearly reduced (p<0.0001 - p<0.0002) $CH_4$ production, and the reduction (p<0.001 - p<0.0001) was dose dependent as the addition level of precursors increased. The $CH_4$ generated was smaller (p<0.01 - p<0.0001) for fumarate than for malate in both chemical types. Addition of fumarate or malate as sodium type reduced (p<0.004) dry matter degradation while both precursors in both chemical types slightly increased neutral detergent fiber degradability of feed in the nylon bag.

• Asian-Australasian Journal of Animal Sciences
• /
• 제22권6호
• /
• pp.819-826
• /
• 2009

An in vitro study was conducted to investigate the effect of malate or fumarate on fermentation characteristics, and production of conjugated linoleic acid (CLA) and methane ($CH_4$) by rumen microbes when incubated with linolenic acid (${\alpha}-C_{18:3}$). Sixty milligrams of ${\alpha}-C_{18:3}$ alone (LNA), or ${\alpha}-C_{18:3}$ with 24 mM malic acid (M-LNA) or ${\alpha}-C_{18:3}$ with 24 mM fumaric acid (F-LNA) were added to the 150 ml culture solution consisting of 75 ml strained rumen fluid and 75ml McDougall's artificial saliva. Culture solution for incubation was also made without malate, fumarate and ${\alpha}-C_{18:3}$ (Control). Two grams of feed consisting of 70% concentrate and 30% ground alfalfa (DM basis) were also added to the culture solution of each treatment. In vitro incubation was made anaerobically in a shaking incubator up to 12 h at $39^{\circ}C$. Supplementation of malate (M-LNA) or fumarate (F-LNA) increased pH at 6 h (p<0.01) and 12 h (p<0.001) incubation times compared to control and linolenic acid (LNA) treatments. Both malate and fumarate did not influence the ammonia-N concentration. Concentration of total VFA in culture solution was higher for M-LNA and F-LNA supplementation than for control and LNA treatments from 6 h (p<0.040) to 12 h (p<0.027) incubation times, but was not different between malate and fumarate for all incubation times. Molar proportion of $C_3$ was increased by F-LNA and M-LNA supplementation from 6 h (p<0.0001) to 12 h (p<0.004) incubation times compared to control and LNA treatments. No differences in $C_{3}$ proportion, however, were observed between M-LNA and F-LNA treatments. Accumulated total gas production for 12h incubation was increased (p<0.0002) by M-LNA or F-LNA compared to control or LNA treatment. Accumulated $CH_4$ production for 12 h incubation, however, was greatly reduced (p<0.0002) by supplementing malate or fumarate compared to the control, and its production from M-LNA or F-LNA treatment was smaller than that from LNA treatment. Methane production from LNA, M-LNA or F-LNA treatment was steadily lower (p<0.01 - p<0.001) from 3 h incubation time than that from the control, and was also lower for M-LNA or F-LNA treatment at incubation times of 6 h (p<0.01) and 9 h (p<0.001) than for LNA treatment. Methane production from LNA, however, was reduced (p<0.01 - p<0.001) from 3 h to 9 h incubation times compared to the control. Both malate and fumarate increased concentration of trans11-$C_{18:1}$ from 3 h to 12 h incubation (p<0.01), cis9,trans11-CLA up to 6 h incubation (p<0.01 - p<0.01), trans10,cis12-CLA at 3 h (p<0.05) and 12 h (p<0.01), and total CLA for all incubation times (p<0.05) compared to corresponding values for the ${\alpha}-C_{18:3}$ supplemented treatment (LNA). In conclusion, malate and fumarate rechanneled the metabolic $H_2 pathway to production of propionate and CLA, and depressed the process of biohydrogenation and methane generation. Linolenic acid alone would also be one of the optimistic alternatives to suppress the $CH_4$ generation.

• 농업생명과학연구
• /
• 제50권5호
• /
• pp.139-152
• /
• 2016

본 연구는 국내 주요 조사료(rice straw, timothy and alfalfa) 및 쑥과 녹차의 반추위 소화율과 물리적 구조를 비교하기 위해 in vitro 와 in situ 실험을 실시하였다. 각 사료별 3, 6, 9, 12, 18, 24, 36, 48 및 72 시간 동안 배양하였고, in vitro 실험에서는 가스 발생량, 미생물 성장량, pH를 측정하였다. 가스발생량은 배양시간이 경과할 수록 모든 시간대에서 증가하였고(p<0.05), 녹차에서의 가스발생량이 가장 낮았다. 미생물 성장량은 배양시간이 경과할수록 모든 처리구에서 증가하는 경향이었으나, 유의차는 나타나지 않았고(p<0.05), 쑥과 녹차의 미생물 성장량이 다른 처리구에 비해 낮게 나타났다(p<0.05). 반추위내 pH는 배양시간 경과할 수록 감소하였고, 티모시의 pH가 다른 사료에 비해 가장 낮았으며, 볏짚의 pH가 다른 사료에 비해 높았다(p<0.05). In situ 실험에서 모든 시간대 녹차의 건물소화율(DM; Dry Matter) 및 조단백 소화율(CP; Crude Protein)은 가장 높았다(p<0.05). 또한 반추위내 통과 속도 4% 쑥에서 건물소화율이 가장 높았고, 알팔파는 조단백 소화율이 가장 높았다(p<0.05). 녹차와 쑥의 표면과 기공을 주사전자현미경(SEM; scanning electron microscope)으로 관찰한 결과, 쑥 표면에는 미생물이 존재하지 않아 반추위 내 영양소 소화율이 낮은 것으로 사료되고, 녹차는 기공에 미생물이 관찰 되었다. In vitro 및 in situ 실험의 결과를 통하여 반추동물의 사료원료로 잠재적인 가치가 있을 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제5권3호
• /
• pp.487-494
• /
• 1992

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.

• Asian-Australasian Journal of Animal Sciences
• /
• 제30권10호
• /
• pp.1416-1424
• /
• 2017

Objective: The objective of this study was to examine the effects of alfalfa flavonoids on the production performance, immunity, and ruminal fermentation of dairy cows. Methods: The experiments employed four primiparous Holstein cows fitted with ruminal cannulas, and used a $4{\times}4$ Latin square design. Cattle were fed total mixed ration supplemented with 0 (control group, Con), 20, 60, or 100 mg of alfalfa flavonoids extract (AFE) per kg of dairy cow body weight (BW). Results: The feed intake of the group receiving 60 mg/kg BW of AFE were significantly higher (p<0.05) than that of the group receiving 100 mg/kg BW. Milk yields and the fat, protein and lactose of milk were unaffected by AFE, while the total solids content of milk reduced (p = 0.05) linearly as AFE supplementation was increased. The somatic cell count of milk in group receiving 60 mg/kg BW of AFE was significantly lower (p<0.05) than that of the control group. Apparent total-tract digestibility of neutral detergent fiber and crude protein showed a tendency to increase (0.05<$p{\leq}0.10$) with ingestion of AFE. Methane dicarboxylic aldehyde concentration decreased (p = 0.03) linearly, whereas superoxide dismutase activity showed a tendency to increase (p = 0.10) quadratically, with increasing levels of AFE supplementation. The lymphocyte count and the proportion of lymphocytes decreased (p = 0.03) linearly, whereas the proportion of neutrophil granulocytes increased (p = 0.01) linearly with increasing levels of dietary AFE supplementation. The valeric acid/total volatile fatty acid (TVFA) ratio was increased (p = 0.01) linearly with increasing of the level of AFE supplementation, the other ruminal fermentation parameters were not affected by AFE supplementation. Relative levels of the rumen microbe Ruminococcus flavefaciens tended to decrease (p = 0.09) quadratically, whereas those of Butyrivibrio fibrisolvens showed a tendency to increase (p = 0.07) quadratically in response to AFE supplementation. Conclusion: The results of this study demonstrate that AFE supplementation can alter composition of milk, and may also have an increase tendency of nutrient digestion by regulating populations of microbes in the rumen, improve antioxidant properties by increasing antioxidant enzyme activities, and affect immunity by altering the proportions of lymphocyte and neutrophil granulocytes in dairy cows. The addition of 60 mg/kg BW of AFE to the diet of dairy cows was shown to be beneficial in this study.