中藥草為具有特定生物活性之天然植物，常以複方的形式使用達到全身性之調節。本試驗以常用於治療人類腹瀉、下痢、厭食等症狀之中藥草做為飼料添加物，進行離乳仔豬飼養試驗，以期改善仔豬離乳期間的下痢及生長緩慢等問題。先以體外試驗（in vitro）評估中藥草複方對豬隻血液PMN（polymorphonuclear neutrophils, PMN）免疫能力之影響，再進行動物試驗（in vivo），以中藥草複方粉末做為飼料添加物，評估其對離乳仔豬生長表現及下痢發生之改善，並且分析仔豬糞便、腸道及血液樣品，以探討中藥草複方對離乳仔豬腸道健康及免疫生理之影響。In vitro 試驗結果顯示，本試驗所探討之中藥草複方水萃液，具有抑制Escherichia coli O157:H7及Salmonella typhimurium E29菌株生長之效果，並可顯著提升豬隻血液PMN呼吸爆發強度、NO產量及殺滅沙門氏菌（P < 0.05）。動物試驗以 240 頭 28 日齡之三品種雜交離乳仔豬（平均體重為7公斤），逢機分配至四飼糧處理組，分別為控制組（CT）、0.1%中藥草（LC）、0.3%中藥草（HC）及抗生素（AB）組。結果顯示，LC 及 HC 組仔豬離乳後2週之生長表現均顯著較 CT 組為佳（P < 0.05），且HC組仔豬離乳後10日內之下痢指數顯著低於其他各組。試驗第7日測定各組仔豬血液PMN呼吸爆發強度及殺滅沙門氏菌之能力，LC及HC組皆顯著較 CT 組為高（P < 0.05）。LC及HC 組仔豬離乳後4週糞便之 coliform 菌數與AB組無顯著差異，而顯著低於CT組（P < 0.05）。試驗第14日進行腸道組織採樣，結果顯示添加中藥草使仔豬腸道 pH 值降低（P=0.099），並顯著增加結腸 lactobacilli 菌數（P < 0.05）；HC 組仔豬小腸澱粉酶及蛋白酶活性顯著高於 CT 組（P < 0.05），與AB組無顯著差異；LC、HC 組仔豬小腸絨毛高度與腺窩深度比皆顯著高於 CT 組（P < 0.05），而HC組與AB組則無顯著差異，且中藥草處理組仔豬之小腸壁肌肉層較薄。試驗第25日餵飼含 Cr2O3 飼糧，結果顯示 HC 組仔豬對乾物質、粗蛋白質及總能之消化率皆顯著高於 CT 組（P < 0.05），達到與AB組無顯著差異之水準。综上所述，於飼糧中添加0.1%中藥草複方可提升離乳仔豬免疫能力並改善腸道健康，添加0.3%則可進一步減緩下痢之發生，改善體增重，有助於仔豬度過離乳障礙。Diarrhea and growth lag has been a handicap in pig industry. Chinese medicine herbs were natural plates with particular bio-activities. They have been used as a prescription to an all over cure. The aim of this study is to evaluate a Chinese medicine prescription (CMH), combined with six species, supplementation replace antibiotic on weanling piglets. In vitro experiments were done to evaluate the bio-effects in this prescription on pig polymorphonuclear leukocytes (PMN). Then, weanling piglets were raised and sacrificed to evaluate their gastrointestinal status in feeding experiment with Chinese medicine herb supplement diet compare with antibiotic. In in vitro trail, the water extract of CMH inhibited Escherichia coli O157:H7 and Salmonella typhimurium E29. It significantly promoted PMN' respiratory burst, nitric oxide production, and Salmonella-killing ability. Two hundred fifty six 28-day old Landrace×Yorkshire×Duroc weanling piglets (average body weight 7 kg) were allotted to 4 diets supplemented with 0.1% CMH (low concentrate, LC), 0.3% CMH (high concentrate, HC), antibiotic (AB) , and basal diet control (CT). The results showed that, on day 14, average daily gain of HC and AB group were higher than LC and CT group, and feed conversion in LC, HC, and AB group were better than CT group. HC group had the least diarrhea score in ten days after weanling. Furthermore, we investigated the PMN' respiratory burst and Salmonella-killing ability in HC and LC group were significantly better than CT and AB group (P < 0.05). Fecal coliform in LC and HC group were not different from AB group, but less than CT group (P < 0.05), at 4th week of experiment. We also found that CMH-supplemented groups had lower gastrointestinal pH value, and more lactobacilli in colon (P < 0.05). The intestinal amylase and protease activities of HC group were not different from AB group, but higher than CT (P < 0.05). In intestinal histological morphology, CMH-supplemented groups were higher in villi/crypt ratio than CT group (P < 0.05), and HC group was not different from AB group. HC group had higher dry matter, crude protein and gross energy digestibilities than CT group (P < 0.05), and which were not different from AB group. In conclusion, CMH supplement promoted immunity in weanling piglets and improved gastrointestinal tract to overcome weanling check.第壹章 前言.............................................1
第貳章 文獻檢討.........................................2
第参章 In vitro 試驗評估中藥草複方水萃液之抑菌作用及其對豬隻PMN（polymorphonuclear neutrophils, PMN）免疫活性之影響.................................................... 14
一、前言...............................................14
二、材料與方法.........................................14
(一)中藥草水萃液之製備.................................14
(二)對病原菌之抑菌作用.................................14
(三)對豬隻Polymorphoneuclear neutrophil（PMN）免疫活性之影響.....................................................14
1.分離豬隻血液PMN......................................14
2.NO產量之測定.........................................15
3.呼吸爆發之測定.......................................15
4.PMN之殺菌能力測定....................................15
(四)統計分析...........................................15
三、結果與討論.........................................16
(一)對病原菌之抑菌作用.................................16
(二)呼吸爆發...........................................18
(三)NO產量.............................................19
(四)殺菌能力...........................................21
四、結論...............................................22
第肆章 以動物試驗探討中藥草複方做為飼料添加物對早期離乳仔豬生長、免疫及腸道性狀之影響...........................23
一、前言...............................................23
二、材料與方法.........................................23
(一)試驗設計...........................................23
(二)生長與下痢指數之觀察...............................25
(三)糞便及血液樣品採集.................................25
(四)豬隻犧牲與腸道取樣.................................25
(五)分析項目與方法.....................................25
1.血液PMN免疫活性測定..................................27
(1)分離血液PMN.........................................27
(2)呼吸爆發之測定......................................27
(3)PMN之殺菌能力.......................................27
2.脾臟單核球增生能力測定...............................27
3.血清免疫球蛋白immunoglobulin G（IgG）及immunoglobulin A（IgA）濃度測定........................................27
4.糞便及腸道內容物菌相.................................28
(1)Coliform 菌數.......................................28
(2)Lactobacilli 菌數...................................28
5.製作小腸組織切片.....................................28
6.小腸消化酵素活性.....................................28
(1)粗酵素液之製備......................................28
(2)澱粉酶之測定........................................28
(3)蛋白酶之測定........................................29
(4)蛋白質濃度測定......................................29
(六)消化率之測定.......................................29
1.乾物質之分析.........................................30
2.粗蛋白質之分析.......................................30
3.總能之分析...........................................30
4.三氧化二鉻之分析.....................................30
(七)統計分析...........................................30
三、結果與討論.........................................31
(一)中藥草作為飼料添加物對早期離乳仔豬生長性狀之影響...31
(二)中藥草作為飼料添加物對早期離乳仔豬下痢指數之影響...33
(三)中藥草作為飼料添加物對早期離乳仔豬免疫性狀之影響...34
1.血液PMN免疫活性......................................35
(1)呼吸爆發............................................35
(2)PMN對Salmonella typhimurium殺菌能力.................36
2.脾臟單核免疫細胞增生能力.............................38
3.血清抗體IgA及IgG濃度.................................39
(四)中藥草飼糧對仔豬腸道性狀之影響.....................41
1.腸道重量.............................................41
2.腸道pH值.............................................42
3.腸道菌相.............................................43
4.糞便菌相.............................................44
5.小腸組織學型態之觀察.................................46
6.小腸消化酵素活性.....................................48
7.中藥草作為飼料添加物對仔豬飼糧消化率之影響...........50
四、結論...............................................51
第伍章 總結論........................................52
第陸章 參考文獻......................................5
