Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/98063
標題: 綠膿桿菌粗酵素液配合微生物處理生雞糞之開發與應用
Development and application of Pseudomonas aeruginosa crude enzyme and microorganism to treat raw chicken manure
作者: 李柏毅
Po-Yi Lee
關鍵字: 硝化菌
液化澱粉芽孢桿菌
木黴菌
氨氣
大腸桿菌
吲哚
3-甲基 吲哚
Nitrobacteria
Bacillus amyloliquefaciens
Trichoderma
NH3
E. coli
Indole
3-methylindole
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摘要: 雞糞臭味的主要來源是氨氣,是由微生物分解雞糞中尚未被利用之營養物質所產生。臭味除了造成空氣汙染之外,也容易吸引蚊蟲而影響生態環境平衡。本試驗以篩選之微生物與酵素處理生雞糞,以改善蛋雞現場之臭味與病原菌之濃度。首先,自堆肥當中篩選出具有降氨能力之硝化菌 (CB1-4-1、CA2-1、CB5H4-5、CB5H4-7),挑選具有產生蛋白酵素能力之液化澱粉芽孢桿菌 (P2、Y2、T4、T6、T7) 以及能夠產生抗菌素之木黴菌 (S5-1、S9-1、TC、Y1) 並測試其對於降低氨氣濃度效果以及抑制大腸桿菌及沙門氏菌之效果。試驗以抗菌以及降氨能力評估結果,挑選出硝化菌 CB1-4-1 (1 x 1010 CFU/ml)、液化澱粉芽孢桿菌 Y2 (1 x 1010 CFU/ml)、木黴菌 S9-1 (1 x 108 spore/ml),三種微生物以及蛋白酵素 (來自綠膿桿菌,100 U/ml) 作為試驗材料,使用酵素並配合不同微生物組合,分兩階段處理生雞糞,評估其抗菌以及除臭效果。依據試驗結果,蛋白酵素配合 CB1-4-1 以及 S9-1 組合處理 (PCBS9-1) 對於降低氨氣排放、大腸桿菌數、吲哚以及3-甲基吲哚(糞臭素)濃度之效果最為顯著。而為了評估 PCBS9-1 於養雞現場減少臭味以及降低病原菌數之效果,田間試驗分別選擇蛋雞舍以及肉雞舍作為試驗場進行試驗。試驗以兩階段方式處理雞糞,第一階段先以蛋白酵素 (100 U/ml) 均勻噴灑於雞糞表面,3 小時後(第二階段)噴灑 CB1-4-1 及 S9-1 混合微生物液,採樣時刻為噴灑前和噴灑後第 3、6、9、12、24、48、72、96 小時。分析項目包含氨態氮 (NH3-N)、氨氣、硫化氫、大腸桿菌數、吲哚、3-甲基吲哚。蛋雞試驗中,處理組相較對照組氨態氮、氨氣、大腸桿菌數、吲哚以及3-甲基吲哚結果數據均呈現下降情形,其中氨態氮及氨氣抑制效果可持續 24 小時,大腸桿菌、吲哚、3-甲基吲哚則可持續 9 小時。肉雞墊料試驗中,處理組相較對照組氨態氮、氨氣、大腸桿菌數、吲哚、3-甲基吲哚結果數據均呈現下降趨勢,其中氨態氮以及氨氣抑制效果可持續 72 小時,而大腸桿菌、吲哚以及3-甲基吲哚抑制效果依序為:12、24、96 小時。綜合以上結果,於雞舍中,使用蛋白酵素配合微生物以兩階段噴灑方式具有降低自雞糞中排放臭味分子濃度及抑制病原菌數之潛力。
The main source of chicken feces odor is ammonia (NH3), comes from nutrients decomposed by microorganisms. Odor not only produce air pollution but also attract pests that affect the ecological balance. The experiment used selected bacterial strains and enzyme sprayed on chicken feces to improved environment odor and bacteria level. First, selected Nitrobacteria which had the ability to reduce ammonia concentration from compost bacteria (CB1-4-1, CA2-1, CB5H4-5 and CB5H4-7). Chose Bacillus amyloliquefaciens which could produce protease (P2, Y2, T4, T6 and T7), Trichoderma which could secrete antibiotics (S5-1, S9-1, TC and Y1) and then evaluated their ability to reduce NH3 concentration and inhibit number of E. coli and Salmonella. With result, experiment chose Nitrobacteria CB 1-4-1 (1 x1010 CFU/ml), Bacillus amyloliquefaciens Y2 (1 x1010 CFU/ml) and Trichoderma spore solution S9-1 (1 x108 spore/ml) three kinds of microorganism and protease from Pseudomonas Aeruginosa (100 U/ml) as tested solutions for next experiment. Next experiment, was to evacuate the effect of protease with different combinations of microorganism on odor emission and number of pathogens on hen litter by two-stages-spray method. With the results, the combination of protease, CB1-4-1 and S9-1 treatment (PCBS9-1) showed the best ability to reduce NH3, E. coli, Indole and 3-methylindole concentration. In field test, in order to evacuate PCBS9-1 effect of reducing odor and pathogen on chicken farm. We chose layer farm and broiler farm for the experiment. Experiments using two stages-spraying method with PCBS9-1 for poultry excreta treatment. First stage was using protease (100 U/ml) sprayed on poultry excreta surface. The second stage (after 3 hours) using CB1-4-1 and S9-1 complex bacterials solution sprayed on poultry excreta surface. Sampling before spraying and 3, 6, 9, 12, 24, 48, 72 and 96 hours after spraying. Analyze the NH3-N, NH3, H2S, E. coli, Indole and 3-methylindole of poultry excreta sample. Results of layer showed that treatment group compare to control group had decline on NH3-N, NH3, E. coli, Indole and 3-methylindole production. The inhibition effect of NH3-N and NH3 could maintain for 24 hours, the E. coli, Indole and 3-methylindole were 9 hours. Results of broiler showed that treatment group compare to control group had decline on NH3-N, NH3, E. coli, Indole and 3-methylindole production. The inhibition effect of NH3-N and NH3 could maintain for 72 hours, and the E. coli, Indole and 3-methylindole were 12, 24 and 96 hours, repetitively. In conclusion, using protease and complex bacterial solution spray on poultry excreta with two-stages method has the potentially effect on reducing odor concentration and number of pathogen bacterium from chicken manure.
URI: http://hdl.handle.net/11455/98063
文章公開時間: 2019-08-22
Appears in Collections:動物科學系

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