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Study of the kinetic characteristics of enteric methane production in dairy cattle and pigs by in vitro gas production technique
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|摘要:||甲烷為主要溫室氣體之一。過去研究顯示，畜牧業甲烷排放量佔全球排放量約20%。除此之外，反芻動物（牛和羊）及非反芻動物（豬和馬）消化道內微生物發酵作用所產生之甲烷亦可視為動物之能量損失。動物消化道內甲烷生成量受多種因素影響，為確認甲烷減量策略之有效性，需要一可快速、準確且再現性高之體外技術測定甲烷生成量。因此，本論文之主要目的為建立一套可快速評估動物消化道內甲烷生成量之體外技術，有助於深入了解消化道內甲烷動態生成情況，作為預測原料甲烷生成量之依據並提高其預測之準確性。為達成此目的，本研究使用體外氣體生產技術 (In vitro gas production technique, IVGPT) 配合氣體成分測定，評估臺灣乳牛及豬隻常用飼料原料之甲烷生成量及其動態生成特性。結果顯示，大麥、玉米、小麥、甜菜渣及大豆殼粒於IVGPT培養期間所測得之體外甲烷產量高於麩皮、玉米酒糟、大豆粕、菜籽粕、苜蓿乾草、百慕達乾草及玉米青眝料。將原料依據組成分特性分為穀物、富含纖維副產物、富含蛋白質副產物及芻料等四組後，各組原料之中洗纖維 (Neutral detergent fibre, NDF)、酸洗纖維 (Acid detergent fibre, ADF) 及酸洗木質素 (Acid detergent lignin, ADL) 含量和體外真消化率 (In vitro true digestibility, IVTD) 為影響瘤胃甲烷產量之主要因素。藉由為每組飼料選擇適合之組成分作為預測因子，可以準確預測原料之甲烷產量。由於穀物、甜菜渣及大豆殼粒可被微生物大量且快速發酵，造成大量氫氣累積於發酵瓶之頂部空間，因此這些原料發酵後可於較短時間 (tHP) 達到較其它原料高之氫氣累積量 (VFH及VHP)。藉由添加sodium-2-bromoethanesulphonate (BES) 或monensin (MON) 抑制甲烷生成作用時，BES處理組具有顯著較未添加及MON處理組高之氫氣累積量 (VFH及VHP)，其中以不可溶氫氣 (fraction y, Vhfi) 比例較高。相反地，MON處理組具有較BES處理組高之可溶性氫氣累積量 (fraction x, VHpx) 及累積之比例 (c)。於豬隻方面，飼糧纖維為不可被動物消化酵素所水解之碳水化合物，可作為後腸微生物發酵作用之主要碳水化合物來源。因此飼糧中可溶性 (SDF) 和不可溶性 (IDF) 飼糧纖維之含量為影響豬隻飼料原料於消化道內甲烷生成量 (CH4 orig）及生成速率 (Rmax) 之主要因素。富含纖維副產物由於含有大量飼糧纖維，因此具有最高之CH4 orig。豬隻腸道中CH4 orig生成量隨飼料中飼糧纖維含量提高呈線性增加。於線性方程式中使用飼糧纖維作為預測因子可顯著提升預測CH4 orig生成量之準確性 (R2)。飼料原料可藉由其SDF與IDF含量預估於豬隻消化道內之CH4 orig生成量 (CH4 orig = 0.279 + 0.054 SDF + 0.011 IDF, R2 = 0.837)。由於IDF被微生物發酵速率較SDF低，豬隻消化道內甲烷生成速率受飼料中SDF與IDF比例所影響。麩皮及富含蛋白質副產物中IDF佔總飼糧纖維 (TDF) 之比例達90％以上，因此具有較大麥、玉米、小麥、甜菜渣及大豆殼低之甲烷生成速率 (Rmax)。綜合上述結果顯示，可由飼料原料之化學組成分預估其甲烷生成量，各組成分對乳牛與豬隻消化道內甲烷生成量之影響可藉由IVGPT提供之動態生成特性解釋。此外，甲烷抑制劑作用方式之差異亦可藉由IVGPT所測得之氫氣累積模式區分。因此，IVGPT配合氣體組成分測定同時評估消化道內微生物發酵作用、甲烷及氫氣之生成量與動態生成特性，可用於快速篩選低甲烷生成量之飼料原料並評估飼糧處理降低乳牛及豬隻消化道內甲烷生成量之效果，可供未來配製低甲烷生成飼糧作為參考。|
Methane is one of the main greenhouse gases. The livestock sector contributes approximately 20% of global methane emissions. Both ruminant animals (cattle and sheep) and non-ruminants (swine and horses) produce methane as a byproduct of enteric fermentation, which is considered as energy loss to the host animals. The enteric methane production varies with several factors. To identify effective mitigation strategies, a rapid, accurate and repeatable in vitro technique is required. Therefore, the main objective of this thesis was to establish an in vitro technique for rapid evaluation of enteric methane production, providing further insight into the kinetic methane production, and to improve the accuracy for methane prediction from chemical composition of feedstuffs. To achieve this objective, in vitro gas production technique (IVGPT) was applied to determine the kinetic methane production from the feedstuffs and forages that commonly used for dairy cattle and pigs in Taiwan. The results showed that higher in vitro methane productions were determined in cereals (barley, maize and wheat), sugar beet pulp and soybean hull than those determined in wheat bran, protein-rich byproducts (dried distillers grains with solubles, soybean meal and rapeseed meal) and forages for dairy cows. After the feedstuffs were classified into four groups, including cereals, fibrous byproducts, protein-rich byproducts and forages, the content of detergent fiber fractions (NDF, ADF and ADL) and in vitro true digestibility (IVTD) of feedstuffs were the major factors affecting ruminal methane production in all groups of feedstuff. The methane production of feedstuffs can be accurately predicted from their chemical compositions and IVTD by selecting the suitable predictors for each group of feedstuffs. A large amount of hydrogen accumulated in the headspace due to the rapid and extensive fermentation of cereals, sugar beet pulp and soybean hull. The fermentation of these feedstuffs took a shorter time (tHP) to reach a higher accumulation of hydrogen (VFH and VHP) than that of forages. When methanogenesis was inhibited by the addition of sodium-2-bromoethanesulphonate (BES) and monensin (MON), a much higher level of hydrogen accumulation (VFH and VHp) in high proportion of non-dissolvable hydrogen (fraction y, Vhfi) was observed in the BES addition groups. In contrast, the addition of MON resulted in a higher amount (VHpx) and proportion (c) of dissolvable hydrogen (fraction x) accumulation than BES addition groups. For pigs, the contents of dietary fiber fractions, including soluble (SDF) and insoluble (IDF) dietary fiber, are the major factors that affecting the amount (CH4 orig) and the rate (Rmax) of methane production from the feedstuffs. The highest CH4 orig was observed in the fibrous byproducts with abundant dietary fiber content. A linear response of CH4 orig to the dietary fiber inclusion level was also observed for pig diets. The R2 for CH4 orig prediction was dramatically improved by including dietary fiber fractions in the equations. The CH4 orig of feedstuffs can be accurately predicted from the content of dietary fiber fractions in the feedstuffs and diets (CH4 orig = 0.279 + 0.054 SDF + 0.011 IDF, R2 = 0.837). The rate of methane production is decided by the proportions of dietary fiber fractions in the feedstuffs due to the different microbial accessibility between SDF and IDF. The high proportion of IDF accounted over 90% of TDF in the wheat bran and protein-rich byproducts, resulting in a lower Rmax of methane production on incubated DM basis as compared to the cereals, sugar beet pulp and soybean hull. According to these results, the methane production of feedstuffs can be predicted from their chemical composition, and the effect of chemical compositions on methane production can be interpreted by the kinetic characteristics determined by the IVGPT for dairy cattle and pigs. The difference in the mode of action between methane inhibitors can be observed in the patterns of hydrogen accumulation. Therefore, the IVGPT combined with measurement of gas composition can simultaneously determine the kinetic characteristics of microbial fermentation, methane production and hydrogen accumulation. It can be applied for rapidly screening the low methane-producing feedstuffs and evaluating the effect of dietary treatments on methane mitigation for dairy cattle and pigs.
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