The fate of recombinant plasmids during composting of organic wastes

Composting was investigated as a means for safe disposal of organic waste containing bacteria that carry transgenes in recombinant plasmids. To generate model recombinant plasmids, a mobile IncQ plasmid, RSF1010, and a non-mobile plasmid, pGFP, were genetically modified to carry a DNA segment encoding both green fluorescent protein and kanamycin resistance and were designated as RSF1010-GFPK and pGFPK. Escherichia coli (E. coli) C600 harboring these plasmids were inoculated into chicken manure specimens that were placed in compost at 20 and 60 cm from the bottom of a 1.0-m high compost bin. Control specimens were held at ambient temperature. By day 10, compost temperatures at the lower and upper levels of the bin had reached 45.3 and 61.5°C, respectively, and at both levels the target E. coli had been inactivated and the plasmids had lost their capacity to be transformed or mobilized. Furthermore, based on real time Polymerase chain reaction (PCR), the transgene fragments along with the host chromosomal DNA fragment from specimens at the upper level had been degraded beyond the detection limit. However, at the lower level where temperatures remained below 48°C these fragment persisted to day 21. At ambient temperatures (0–8°C), the E. coli, plasmids and the transgene fragments persisted in manure specimens throughout the 21 day test period. The study showed the potential for composting as a safe procedure for disposal of bacteria carrying transgenes in recombinant plasmids.     

Influences of adding easily degradable organic waste on the minimization and humification of organic matter during straw composting

To find a better composting process with low greenhouse gas emission and high humus production, the effect of adding kitchen waste on reduction and humification of organic matter during straw composting was studied. Three processes were compared, consisting of different ratios of straw and kitchen waste (1:2, 1:1, and 2:1). At four time points over a 62-d incubation, the reduction and humification of compost was evaluated by measuring the total mass, carbon content, and humic material content of the compost. Treatment 1 (straw/kitchen waste ratio of 1:2) reduced the total mass of compost the most. Treatment 2 (straw/kitchen waste ratio of 1:1) reduced the total carbon content the most, reflecting the highest emission of greenhouse gas. Treatment 3 produced the most humic acid material and released the lowest amount of carbon. Hence, from the point of view of reducing greenhouse gas emissions and increasing stable organic matter such as humus and humic acid during composting, treatment #3 was optimal. The three treatments resulted in significant differences in microbial biomass and enzyme activity during composting. The highest amount of active microbial biomass was associated with the largest reduction in compost mass (treatment 1). Higher proportions of straw (treatments 2 and 3), which contains more lignin, were associated with greater β-glycosidase activity, which may generate more humus that can improve soil quality. Dehydrogenase activity seemed to be the most important microbial factor in organic carbon catabolism or humification.     

微生物生理群在猪粪秸秆高温堆肥碳氮转化中的作用

在自制的强制通风静态堆肥反应箱中,猪粪与秸秆以鲜重7∶1的比例进行了堆肥化实验,在堆制的23 d里根据堆温变化分阶段采集堆肥样品,利用MPN法测定了堆料中纤维素分解菌和氮素微生物生理群的数量变化,同时测定了相应的碳、氮含量。结果表明,纤维素分解菌在稳定腐熟阶段较多,对于后期有机碳的降解和腐殖质含量的增大起了很大的作用,在堆制的23 d里,腐殖质增加了2.4%。整个堆制过程中,氨化细菌的数量最大且与氨气释放浓度和铵态氮含量呈显著正相关,都在高温期增加,降温期后减少,氨化细菌的数量在高温期的增加率远高于降温期后的减少率,而铵态氮在高温期的增加率远低于在降温后期的减少率,铵态氮总体上减少了74.1%;亚硝化细菌数量与硝态氮呈正相关;反硝化细菌数量在降温期上升幅度较大,堆制结束时为堆制初期的13倍,且与堆肥中硝态氮含量呈正相关;硝态氮含量增加了87.5%;堆肥后期硝态氮的增加可能与堆肥中存在能进行硝化作用的反硝化细菌有关。固氮菌数量在堆制结束时达堆制初期的2.61倍,主要在降温期增加较多,对堆肥中有机氮的形成起很大作用。     

利用城市污泥堆肥及建筑弃土种植麦冬研究

将污泥堆肥产品分别与建筑弃土按0%、10%、20%、30%、40%和50%(湿重)的比例配制营养土,以麦冬为例进行180 d的栽培实验,分析了营养土对麦冬生长特性、植物体内及栽培土壤中营养学指标和重金属含量的变化规律,并通过模糊综合评价方法选择营养土较佳配制方案。实验结果表明:堆肥比例在50%时麦冬生物量增加率、新生芽数及地表部分比重为最大值,在30%时纺锤体肉质数减少率为最大值;随着营养土中堆肥比例增加,麦冬体内养分含量、重金属积累量随之增加,均在50%时达到最大值;栽培土壤养分越多损失量越多,各指残留率越低,同时全磷、全氮及K含量降低率较有效磷、碱解氮小得多,Cr降低率在5.20%～8.34%之间,Zn降低率在3.27%～17.35%之间;堆肥比例在40%以上的营养土对麦冬生长效果好,但较佳配制方案为堆肥产品占20%。     

牛粪堆肥升温菌剂的筛选及其作用机制

以糖、淀粉、蛋白质、纤维素培养基及菌株增殖倍数、菌株性能等为筛选方法,从牛粪低温堆肥中筛选不同原料重要功能菌并组成功能菌剂;以不同类别原料菌及单菌株发酵实验对升温机制进行研究。结果表明,芽孢杆菌、假单胞菌、纤维单胞菌及小孢霉菌等为重要功能菌;菌剂起温快,升温效果好,48 h堆体即由低温进入中温,5 d达到高温,腐熟时间可缩短至约17~18 d;糖功能菌升温效果最为显著,淀粉及蛋白质功能菌次之,纤维素功能菌升温能力较弱;菌株利用各种原料能力越强,增殖速度越快,对升温作用越大。各菌株充分利用基质各成分,紧密协同和促进,是低温堆体发酵升温的主要作用机制。     

利用污泥熟肥作为高含水率污泥堆肥调理剂

采用静态强制通风好氧堆肥的方法,以木屑作为对比,考察了利用污泥熟肥作为调理剂对污泥堆肥过程的影响。结果表明,与以木屑作为调理剂的污泥堆体(对照组)相比,以污泥熟肥作为调理剂的污泥堆体(实验组)升温快,高温阶段(>50℃)持续时间长达10 d,满足粪便无害化卫生标准的要求,而对照组仅持续了2 d;实验组腐熟的堆肥含水率从60%降到39%,下降了21%,pH维持在7.5～8.5范围内,微生物活性较强,而对照组含水率仅下降15%,pH始终低于7.5;实验组种子发芽指数(GI)在第14天就回升到80%以上,基本上去除了植物毒性,而对照组GI在第22天才回升到50%。总体而言,污泥熟肥能显著改善堆肥中微生物的微环境,促进有机物的降解,缩短堆肥腐熟时间,是一种优质的调理剂。     

添加氮损失抑制剂对蓝藻泥堆肥质量的影响

对脱水蓝藻进行大型生产性堆肥实验。研究堆肥过程中,氮损失抑制剂过磷酸钙的添加对堆肥物料各类理化性质的影响。研究结果表明,过磷酸钙加入的处理组与未加过磷酸钙的对照组堆制37 d后堆肥物料均可腐熟,且堆肥肥效均符合NY525-2002的相关要求。添加过磷酸钙处理组有机质含量为490 g/kg,全N、全P和全K含量分别为20.75、10.02和11.32 g/kg,总养分含量达到9.77%,堆肥品质明显优于对照处理。同时,在微生物的作用下,对照组中微囊藻毒素MC-RR与微囊藻毒素MC-LR去除率分别达到89.8%与78.3%。值得一提的是,添加过磷酸钙后,MC-RR和MC-LR的去除率得到进一步提高,分别达到了92.96%和100%,较好地保证了蓝藻堆肥农用的安全性,为脱水蓝藻好氧堆肥化提供了可行性依据。     

添加不同量的腐殖酸对猪粪堆肥中主要养分变化的影响

腐殖酸是一种可以控制堆肥氮素损失的天然固定剂,通过研究腐殖酸对猪粪堆肥过程中主要养分含量变化的影响,确定了腐殖酸的最佳加入量.结果表明,加入5%含量(占鲜猪粪重)的腐殖酸最有利于发酵反应的进行,能够促进有机质的分解,有效控制氮素损失,可使速效钾含量增加,但对有效磷的转化能力影响不大;与其他处理相比,加入5%含量的腐殖酸...     

人粪便中温好氧堆肥过程的有机物降解特性研究

利用密闭式小型堆肥反应器,以锯末作为堆肥载体,采用批量反应的操作方式,在35℃的温控条件下进行了人粪便的中温好氧堆肥实验。结果表明,在14 d的堆肥反应周期内,粪便中以COD为参数的有机物去除率可达到70%左右,且在10～12 d内基本完成有机物降解。在中温条件下,堆体容易保持适宜的含水率和pH值,维持适宜微生物繁殖生长的条件。运用磷脂法进行堆肥过程中微生物总量的分析,表明在一个堆肥周期内,微生物经历了环境适应期、稳定增殖期、生物量下降期等3个阶段,这一微生物繁衍规律与有机物的去除密切相关。与高温堆肥相比,中温堆肥能够减少设备加热的能量消耗,是缺水地区人粪便处置的可选方法之一。     

Biomethanation of plant materials and agricultural residues using dung samples as wild population of microbes and also with isolated methanogens

Recycling of plant materials and agricultural residues for biomethanation was attempted in vials. The methanogenic activities of certain sewage samples have also been tested. Both sterilized and non-sterilized biomasses were used. Biomethanation was carried out with dung samples (cow, goat, buffalo, piggery wastes and poultry wash) as wild populations of microbes and in combination with other microbial isolates (isolated in the laboratory).Biomethanation had been observed to be good in most cases and particularly with the sterilized biomass. Mixed inoculum (dung samples and poultry wash) was found to be best for biomethanation. Of the microbe isolates, isolates from buffalo, pig and paper mill wastes appear to be most effective. Pretreated sawdust and rice straw were found to be good subtrates for biomethanation. Of the different plant biomass used Spirogyra (algae), Ipomea and water hyacinth were most effective whereas Jatropa gossypifolia and Parthenium sp. were the least effective. Biomethanation of Spirogyra was carried out both in anoxic and oxic conditions. Though methane production decreased enormously under oxic conditions, definite methane production continued indicating that the biomethanation process is not exclusively anoxic. Similarly, biomethanation of sewage samples from different sewage treatment plants were carried out with and without isolated methanogens and methane production was found to be moderate.