1、中图分类号： 密级： 硕士学位论文论文题目： 高土霉素残留猪粪的高效好氧堆肥技术研究 作者姓名： 学科专业： 环境工程 研究方向：废物处置与资源化技术指导教师： 提交日期：I高土霉素残留猪粪的高效好氧堆肥技术研究摘 要本论文调查研究了浙江省规模化养猪场饲料及猪粪中土霉素的残留情况，探讨了土霉素的饲料添加量与粪便残留量的相关性，明确了土霉素残留对猪粪好氧堆肥过程的影响，在此基础上，对高土霉素残留猪粪的高效好氧堆肥技术进行了探究，得到以下主要结论：(1)对浙江省 11 个市的规模化养猪场进行小猪、母猪、肉猪的饲料样品及粪便样品的采样分析，结果表明，浙江省规模化养殖场猪饲料与粪便样品中土霉素的检出率
2、均为 100%，饲料中土霉素的平均含量为 8.76 mg/kg，粪便中土霉素的平均残留量为 2.98 mg/kg，浙江省规模化养殖场猪粪中土霉素的残留变幅明显高于德国和土耳其等欧洲国家。小猪饲料中土霉素的添加量与其粪便中土霉素的残留量呈极显著正相关，相关系数为 0.604；母猪和肉猪的土霉素饲料添加量与粪便残留量之间则没有显著相关性，根据相关性结果，提出小猪饲料中土霉素的添加量应小于 38.04 mg/kg 的参考指标。(2)不同土霉素初始残留浓度下的猪粪好氧堆肥试验结果表明，土霉素残留抑制了堆肥过程中微生物的活性，土霉素残留不利于堆体温度的上升，加速了堆温的下降，50 mg/kg、100 m
3、g/kg、150 IImg/kg 土霉素处理升温过程仅持续 4 d；100 mg/kg、150 mg/kg 土霉素处理不利于堆肥过程中水分的散失，并且会造成堆肥结束后堆体的高含水率；土霉素残留对堆肥过程中微生物利用有机碳、有机氮及硝化细菌的硝化作用存在一定的抑制作用，不同土霉素残留量的影响不同，其中 150 mg/kg 土霉素处理抑制作用最显著。高温是土霉素降解的关键，土霉素的降解主要发生在堆肥前期的升温及高温阶段，其残留量随时间的变化规律符合一级动力学方程，不同的土霉素初始残留量对应不同的降解速率常数，10 mg/kg、50 mg/kg、100 mg/kg、150 mg/kg 土霉素残留处理
4、的土霉素降解半衰期分别为 8.97 d、5.12 d、3.09 d、6.18 d。10 mg/kg、50 mg/kg、100 mg/kg、150 mg/kg 土霉素残留处理在 30 d 堆肥后土霉素的降解率分别为 27.22%、81.06%、88.3% 、82.64%。(3)从前期土霉素强化刺激后的好氧堆肥堆体物质中分离出一株能以土霉素为唯一碳源生长的菌株 TJ-1，综合其生理生化特性、形态特征及 16S rDNA 序列分析，鉴定该菌株为葡萄球菌，命名为 TJ-1 菌株( Staphylococcus sp. TJ-1)。该菌株在接种量 3%，培养温度 30 ，摇床 130 r/min 连续培
5、养 5.5 d 后，培养基中土霉素的降解率为80.7%，显著高于已经报道的菌株 Oxy2；对菌株的降解特性进一步研究后发现该菌株降解土霉素的最优条件为：pH 值 7.0，温度 30 ，接种量 5%；该菌株的最高耐受浓度为 300 mg/l。(4)对实际猪粪好氧堆肥过程中切实可行的参数指标(堆肥初始含水率、C/N、土霉素高效降解菌剂接种量、翻堆频率)的优化结果表III明，随着含水率、C/N、菌剂接种量的增大，土霉素降解率呈上升的趋势，且初始含水率、C/N 对土霉素的降解率具有极显著的影响(p0.01)，菌剂接种量对土霉素的降解率具有显著的影响(p0.05)。随着翻堆频率的增大，土霉素降解呈不断下
6、降的趋势，翻堆频率对土霉素的降解率具有显著的影响(p0.05)。猪粪好氧堆肥过程中土霉素降解的最优环境因子条件为堆肥初始含水率 70%、初始 C/N 30、菌剂 TJ-1 接种量 1%、堆肥翻堆频率 1.5 d/次。(5)土霉素高效降解堆肥工艺能显著提高土霉素的降解率(p0.05)，21 d 堆肥结束后的降解率为 82.0%，而普通堆肥工艺的降解率仅为62.7%。土霉素残留量随时间的变化规律符合一级动力学方程，普通堆肥工艺及土霉素高效降解工艺中 150 mg/kg 土霉素的降解半衰期分别为 2.75 d、0.78 d。土霉素残留不利于微生物对有机氮的分解及硝化细菌的硝化作用，但土霉素高效降解工
7、艺与普通堆肥工艺相比能显著提高堆肥高温期微生物对有机氮的利用。对堆体微生物的群落多样性分析表明，土霉素高效降解工艺对比普通堆肥工艺的堆体微生物活性高，有利于增加微生物群落的多样性，两者差异显著(p0.05)。关键词：土霉素；猪粪；土霉素降解菌；好氧堆肥IVTHE HIGHLY EFFECTIVE TECHNOLOGY OF AEROBIC COMPOST OF PIG MANURE WITH HIGH OXYTETRACYCLINE ABSTRACTIn this paper, the oxytetracyline residues in the feeds and manures of pi
8、g in scale farms in Zhejiang province were investigated to find the correlationship between the content of oxytetracyline of pig feeds and manures. The effect of oxytetracyline on pig manure composting was found , and then the highly effective technology of aerobic compost of pig manure with high ox
9、ytetracycline was discussed in this paper. Main conclutions were draw as following:(1)The oxytetracycline residues of the feeds and manures of pig in scale farms of 11 citys in Zhejiang province were analyzed, and the results showed the detection rates of oxytetracycline in pig feeds and manures sam
10、pled from scale farms in Zhejiang province were all 100%.The average content of feeds was 8.76 mg/kg, while the content of pig manures was 2.98 mg/kg averaged, the scale of oxytetracycline Vcontent of Zhejiang province was significantly higher than that of Germany, Turkey and other European countrie
11、s. The content of oxytetracycline in feeds of piglet was significantly positive correlated with the residues in its manure, the correlation coefficient was 0.604, but the similar correlationship was not founded in sow and fattening pig. According to the correlationship of piglet, the addition of oxy
12、tetracycline in the feeds of piglet must be less than 38.04 mg/kg was suggested.(2)The composting results of five different initial oxytetracycline residues treatments which was 0(CK), 10 mg/kg, 50 mg/kg, 100 mg/kg, 150 mg/kg showed that oxytetracycline residues restricted the activity of microorgan
13、isms during the composting process, oxytetracycline residues accelerated the decline of temperature and gone against the rise of temperature, 50 mg/kg, 100 mg/kg, 150 mg/kg oxytetracycline treatment just had 4 days temperature risen duration. 100 mg/kg and 150 mg/kg oxytetracycline treatment had neg
14、ative influence on the loss of water during composting and would result in the high water content after composting.Oxytetracycline restricted the activity of microbial in manure, including the utilization of organic carbon and nitrogen and the nitrification of nitrifying bacteria. Different oxytetra
15、cycline residues had different effects, the 150 mg/kg oxytetracycline treatment had the most significantly inhibition.The high temperature was the key factor of the degradation of oxytetracycline, the degradation of oxytetracycline VIoccurred mainly in the temperature-rise and high temperature perio
16、d. The changes of the concentration of OTC in pig manure with time had a trend of kinetic equation, and different concentration of oxytetracycline residues had different dynamics equation, the half-life of the 10 mg/kg, 50 mg/kg, 100 mg/kg, 150 mg/kg oxytetracycline treatment were 8.97 d, 5.12 d, 3.
17、09 d and 6.18 d. The 10 mg/kg, 50 mg/kg, 100 mg/kg, 150 mg/kg oxytetracycline treatment had the degradation rate of 27.22%, 81.06%, 88.3%, 82.64% after 30 d composting.(3)A strain TJ-1, that could utilize oxytetracycline as the sole carbon source, was isolated from pig manure compost and identified
18、as Staphylococcus sp. The oxytetracycline degrading rate was 80.7% within 5.5 d with the inoculum concentration 3%, temperature 30 , pH 7.0 and the shake bed 130 r/min, and was significantly higher than that of Oxy2 which was reported ever. The prome condition for OTC-degradation of TJ-1 was the ino
19、culum concentration 5%, pH 7.0, temperature 30 . The highest concentration of oxytetracycline that the strain could tolerate was 300 mg/l. (4)The results of optimising the pratical parameters in aerobic composting such as moisture content, C/N, bacterium inoculation and turning frequency showed that
20、 with the increase of moisture content, C/N, bacterium inoculation increased, the oxytetracycline degrading rate also increased, the oxytetracycline degrading rate was significantly effected VIIby the moisture content and C/N(p0.01) and was effected by the bacterium inoculation(p0.05).While the turn
21、ing frequency increased, the oxytetracycline degrading rate decreased adversely, the turning frequency could effect the oxytetracycline degrading rate(p0.05). The optimal environment factor conditions for oxytetracycline degradation of pig manure was initial moisture content 70%, C/N 30, 1% of bacte
22、rium agent TJ-1 inoculation amount, turning frequency 1.5 d/time.(5)The highly effective technology of aerobic compost of pig manure could significantly improve the degradation rate of oxytetracycline (p0.05), the degradation rate was 82.0% after 21 d composting, while the degradation rate of ordina
23、ry composting process was only 62.7%. The change of the concentration of OTC in pig manure with time had a trend of kinetic equation, the half-life of 150 mg/kg oxytetracycline in ordinary compost and the highly effective technology of aerobic compost were 2.75 d and 0.78 d seperately. Oxytetracycli
24、ne residues restricted the activity of microorganisms including the utilization of nitrogen and the nitrification of nitrifying bacteria during the composting process, but the highly effective technology of aerobic compost of pig manure could reduse the negative influence of oxytetracycline, and cou
25、ld significantly increase the utilization of nitrogen of microorganisms in high temperature period. The effects of oxytetracycline on the diversity of microbial community in composting VIIImaterials were explored using Biolog method. The results showed that microbial activities of the highly effecti
26、ve technology of aerobic compost was higher than ordinary compost, conductive to communities richness and functional diversity. Correlation analysis showed that the difference was significant between the highly effective technology of aerobic compost and ordinary compost (p0.05). KEYWORDS: oxytetrac
27、ycline; pig manure; oxytetracycline degrading bacterium; aerobic compistingIX目 录1 绪论 .11.1 土霉素在畜禽养殖业的应用及其残留现状 .11.1.1 土霉素的理化性质 .11.1.2 土霉素在畜禽养殖业中的应用 .11.1.3 畜禽粪便中土霉素的残留现状 .21.2 土霉素在环境中的生态毒性 .31.2.1 土霉素对微生物的生态毒性 .31.2.2 土霉素对植物的生态毒性 .41.2.3 土霉素对动物的生态毒性 .41.3 畜禽粪便的处理技术现状与分析 .41.3.1 饲料化处理技术 .51.3.2 肥料化处理技术 .61.3.3 能源化处理技术 .61.3.4 各处理技术优缺点对比 .71.4 好氧堆肥技术研究进展 .71.4.1 好氧堆肥过程的影响研究 .71.4.2 好氧堆肥腐熟度指标的研究 .91.4.3 好氧堆肥工艺的研究 .91.5 论文研究内容及意义 .101.5.1 研究目的及意义 .101.5.2 研究内容 .112 养猪场饲料及猪粪中土霉素残留情况调查 .132.1 前言 .13
