Effects of Biden Pilosa extracts on anticoccidial efficacy of Eimeria tenella of broilers

球蟲症在全球造成家禽產業巨大損失。隨著歐盟逐步禁用飼料用抗生素，使 用植化物（phytochemical）飼料添加劑 (feed additives) 的研發逐漸受到重視。 本研究探討飼糧中添加大花咸豐草萃取物 (Biden Pilosa crude extracts, BPCE) 對愛拔益加白肉雞感染球蟲後其抗球蟲能力與腸道菌相之影響。試驗分兩批次進行，四日齡愛拔益加白肉雞 84 隻，隨機分配至 7 飼糧處理組，分別為基礎飼糧 組、三種不同處理之 BPCE 添加組：BPCE control、BPCE PH 與 BPCE NaCl，各 以兩種濃度 (10ppm 與 100ppm)添加 。於 14 日齡口服感染 2.5 × 104顆 Eimeria tenella 芽孢化蟲卵，並於感染後 7 日犧牲雞隻採集樣本。結果顯示，健康雞隻於 飼糧添加 10 ppm BPCE 兩週可提升其體重與體增重。而在雞隻感染球蟲後，持 續飼以 10 ppm BPCE 則能降低球蟲對生長性狀之損害。此外，飼以 BPCE 之感 染雞隻，其感染後5-7天糞便中球蟲卵囊數 (OPG) 總排出量均較基礎飼糧組低， 其中以 BPCE control 與 BPCE PH 之效果較佳。飼糧添加 BPCE 亦可降低盲腸切 片寄生卵囊數評分。根據小腸絨毛性狀之結果顯示，100 ppm BPCE 會降低窩腺深度、減少腸道絨毛面積，可能降低養份吸收速率，並對生長性狀結果造成負面影響。腸道菌群結果顯示，BPCE 100ppm 飼糧餵飼健康雞隻兩週後增加迴腸乳桿菌數含量，然而不會提高盲腸大腸菌群與產氣莢膜梭菌數量。於感染雞隻中餵飼咸豐草萃取物則有提升盲腸乳酸桿菌數含量、降低大腸菌群與產氣莢膜梭菌數量之效果，初步推論 BPCE 具有調控腸道菌群的能力。綜上所述，球蟲感染與雞 隻週齡兩因子影響 BPCE 飼糧處理雞隻之生長性狀。雞隻餵飼 BPCE 10ppm 於兩 週可提升生長性狀，受感染後飼糧添加 BPCE 10ppm 亦能幫助雞隻對抗球蟲症， 且 BPCE 飼糧具有調控小腸及盲腸菌群的潛力。Coccidiosis causes the loss of poultry industry in the world. With the feed antibiotics being gradually banned in recent years, the development of alternative phytochemical feed additives gains more attention. The purpose of this study was to evaluate the effects of Biden Pilosa crude extracts (BPCE) to ameliorate coccidiosis and cecal microbial ecology of Arbor Acres broilers by Eimeria tenella infection. The experiment was conducted in two batches. Eighty-four day-old chicks were randomly arranged into seven floor-pens with treatment diets containing basal diet、three types of BPCE supplemented diets：BPCE control, BPCE PH and BPCE NaCl at two concentrations (10 ppm and 100 ppm). Birds were challenged orally with 2.5 x 104 sporulated oocysts of E. tenella at 14-d of age. Some birds from each group were dissected at 7-d post-infection in order to collect samples. Results showed that dietary supplementation with BPCE at 10 ppm significantly increased growth performance in the second week of age. Feeding 10 ppm BPCE reduced the damage of coccidiosis. Diet supplement with BPCE had a positive dose effect on oocysts per gram (OPG), suggesting that using of 10ppm could be effective. Diet supplemented with BPCE could reduce the histological parasite score with positive dose effect, and there was no significant difference between different types of BPCE supplement. It suggested that BPCE could effectively reduce the infection or the propagation of oocysts in the intestine. In the results of intestinal mucosal morphometry, high concentration of BPCE decreased the depth of crypt, reduced the villus area and perhaps inhibited the nutrients uptake as well, which was consistent with the retarded growth performance. Results of gut microflora showed that BPCE inclusion increased the number of Lactobacillus spp. in the ilium, but affect the number of Coliforms and Clostridium perfringens in the cecum. After infection by Et, BPCE inclusion increased the number of Lactobacillus spp., reduced the number of Coliform and Clostridium perfringens. In summary, infection by Et and chicken age were two factors affecting the growth performance on chicken fed with BPCE diets. Dietary supplementation with BPCE 10ppm had a positive effect in coccidiosis, and BPCE diet had a potential to regulate small intestine morphology and cecal microflora.中文摘要………………………………………………………………………………i 英文摘要………………………………………………………………………………ii 目次…………………………………………………………………………………. iv 表次…………………………………………………………………………………vi 圖次…………………………………………………………………………………..vii 第一章、 緒言………………………………………………………………………..1 第二章、 文獻探討………………………………………………………………….2 一、雞球蟲症………………………………………………………...…………..2 (一)、球蟲症對家禽產業之影響……………………………………………..2 (二)、球蟲種類、球蟲生活史與致病機理…………………………………..2 二、藥用植物飼料添加劑之抗球蟲效果…………………………………….....8 (一)、治療球蟲症之方法比較………………………………………………..8 (二)、具抗球蟲效用的藥用植物…………………………………………….10 (三)、咸豐草之介紹…………………………………………………………13 三、家禽腸道菌相……………………………………………………………...18 (一)、腸道微生物所扮演之角色…………………………………………..18 (二)、健康雞隻腸道微生物分布…………………………………………..18 (三)、球蟲感染對腸道菌相之影響………………………………………..22 第三章、材料與方法………………………………………………………………..24 一、咸豐草萃取物之製備與取得……………………………………………...24 二、動物試驗設計………………………………………………………………24 (一)、雞隻來源與飼養管理………………………………………………..24 (二)、試驗分組………………………………………………………..........24 三、飼糧配方與營養成分測定…………………………………………………25 (一)、飼料配方…………………………………………………..…………25 (二)、總能測定…………………………………………………..…………25 (三)、粗蛋白測定…………………………………..………………………26 (四)、乙醚抽出物測定……………………………………………………..26 (五)、鈣及磷測定…………………………………………………………..26 四、生長性狀測定………………………..……………………………………..29 五、抗球蟲能力指標測定項目………………………………………………...29 (一)、每克糞便中所含之卵囊數 (O.P.G.) 計數………………………….29 (二)、盲腸組織病變評分 (Intestinal lesion scores)……………………….29 (三)、盲腸組織切片卵囊數評分 (Histological parasite score)……….…...30 (四)、抗球蟲指數 (Anticoccidial index)…………………………….……..30 六、小腸絨毛切片分析…………………………………………………………32 七、腸道內容物樣品採集………………………………………………………..32 八、腸道微生物培養法...........………………………………..…………………32 (一)、大腸菌群 (Coliform) 培養基製作……………….………………….32 (二)、MRS 培養基製作……………………………………………………..33 (三)、TSC 培養基製作………………………………………………………33 (四)、腸道內容物樣本系列稀釋、抹菌與培養……………………………33 九、統計分析……………………………………………………………………34 第四章、 結果討論………………………………………………………...……….35 一、飼糧添加咸豐草萃取物對愛拔益加白肉雞抗球蟲能力之評估………...35 (一)、飼糧添加咸豐草萃取物對愛拔益加白肉雞生長性狀之影響…….....35 (二)、飼糧添加咸豐草萃取物對愛拔益加白肉雞小腸絨毛形態學之影響.39 (三)、飼糧添加咸豐草萃取物對愛拔益加白肉雞抗球蟲能力之影響……42 二、飼糧添加咸豐草萃取物對愛拔益加白肉雞腸道菌相之影響…………...52 (一)、飼糧添加咸豐草萃取物對不同日齡愛拔益加白肉雞腸道菌相之影 響……………………………………………………………………………..52 (二)、飼糧添加咸豐草萃取物對感染球蟲白肉雞腸道菌相之影響……….54 第五章、結論………………………………………………………………………..56 參考文獻……………………………………………………………………………..5