降解多菌灵和啶虫脒残留的复合菌剂研发及初步应用

针对菜田土壤中多菌灵和啶虫脒的残留问题，以多菌灵降解菌株Rhodococcusqingshengiidjl-6和啶虫脒降解菌株Pigmentiphaga D-2作为材料，进行降解菌剂的复配，研究了使用复合降解菌剂对复合农药残留土壤的修复效果及微生态效应。研究结果表明：(1)复配降解菌株djl-6与D-2的最适体积比为5∶3，复合降解菌剂在3 d内对无机盐培养基中50μg/ml的多菌灵降解率为87.14%，对50μg/ml的啶虫脒降解率为96.10%。(2)初始接种量为7%时，复合降解菌剂可在6 d内将复合农药污染土壤中5 mg/kg多菌灵降解74.40%，将5 mg/kg啶虫脒降解95.87%。土壤含水率为25%时，复合降解菌剂对复合农药污染土壤中5 mg/kg多菌灵的降解率为80.80%，对5 mg/kg啶虫脒的降解率为97.87%。(3)复合农药污染土壤中10 mg/kg多菌灵和10 mg/kg啶虫脒即可对小青菜的生长产生明显的药害，小青菜生长24d时根长仅为空白对照的66.56%、茎叶长为58.35%、鲜重为45.13%。在7%的接种量条件下，复合降解菌剂可解除药害，使小青菜的生...     

除草剂苄嘧磺隆降解菌Hansschlegelia zhihuaiae S113菌剂的研发

磺酰脲类除草剂苄嘧磺隆残留期较长，造成农田土壤的残留污染问题。以苄嘧磺隆高效降解菌株Hansschlegelia zhihuaiae S113为材料制备降解菌剂，优化液体菌剂的保护剂种类与配比，筛选固体菌剂的最佳载体，初步应用固体菌剂并评价修复效果。研究结果表明：（1）向液体菌剂中添加合适的保护剂（0.30% 柠檬酸钠、0.20% 羧甲基纤维素、0.10% CaCl2）可使活菌数提高37.25%，保存30 d的液体菌剂对50 mg/L苄嘧磺隆的降解率为94.25%。（2）筛选出猪粪有机肥为固体菌剂的最佳载体，保存60 d时固体菌剂活菌数为7.45×107 cfu/g，对土壤中10 mg/kg苄嘧磺隆的降解率为91.22%。（3）固体菌剂的添加可有效减轻土壤中苄嘧磺隆残留对玉米的药害。     

二苯醚类除草剂降解菌Bacillus sp. Za微生物制剂的研发与初步应用

以二苯醚类除草剂高效降解菌株Bacillus sp. Za为材料制备微生物制剂，优化液体制剂保护剂的物质配比，筛选固体制剂的最适材料，对固体制剂进行初步应用并评价其降解效果。研究结果表明：（1）液体制剂保护剂（0.20%柠檬酸钠、0.20%羧甲基纤维素、0.30% KCl）可使活菌数提高35.71%，保存30 d的液体制剂对50 mg/L乳氟禾草灵的降解率为83.50%。（2）筛选得到猪粪有机肥作为固体制剂的最适材料，保存60 d时固体制剂活菌数为8.26×108 cfu/g，对土壤中10 mg/kg乳氟禾草灵的降解率为85.52%。（3）添加固体制剂可有效缓解乳氟禾草灵残留对玉米所产生的药害。     

畜禽粪便堆肥作为功能微生物载体的研究

以堆肥作为3株功能芽胞细菌液体菌剂的载体,通过优化载体含水量、温度和接种浓度等关键影响因子,以不同时间载体中有效活菌数的变化为指标,探讨堆肥代替草炭作为功能微生物载体的可行性和最适条件。研究结果表明,载体C（鸡粪）、P（猪粪）、M（1∶1鸡粪猪粪）和TP（猪粪＋草炭）在72 h内的有效活菌数均显著低于草炭;混合载体TC2（50%草炭＋50%鸡粪）和TM1（25%草炭＋75%1∶1鸡粪猪粪）的有效活菌数随着时间的延长而增加,其中72 h时TC2的有效活菌数达到11.40×109 cfu.g-1,TM1的有效活菌数达到2.64×109 cfu.g-1,均与草炭无显著差异,因此适宜代替草炭作为功能微生物的载体。采用单因素实验,载体TC2和TM1的最优化影响因子为含水量30%、吸附温度30℃、菌液接种浓度为108cfu.mL-1。     

花生化感物质降解菌和抗连作拮抗菌复合菌剂应用效果的研究

为了提高花生化感物质降解菌对化感物质降解率以及连作障碍拮抗菌对植物病原菌的拮抗效果。利用优化培养后制备的复合菌剂进行大田试验(复合菌剂中降解菌有效活菌数总数达3×1010cfuml~(-1),拮抗菌有效活菌数1.12×108cfuml~(-1)),研究了复合菌剂对花生病害、降解特性及土壤微生物活性等的影响。结果表明:复合菌剂及菌剂加倍处理对化感物质的降解率分别达66.7%和89.4%;菌剂的施入对花生网斑病、焦斑病和褐斑病有明显的抑制作用,特别是菌剂加倍对网斑病抑制效果2年分别能达到56.1%和72.1%;花生生育期内,微生物生物量碳氮总体呈先增加后减少趋势,菌剂处理比空白分别能提高6.2%~34.5%和4.9%~31.1%;复合菌剂处理能明显提高土壤脲酶、酸性磷酸酶和过氧化氢酶活性;花生产量2年皆以菌剂加倍最高,分别为3734.2 kg hm~(-2)和3708.6 kg hm~(-2),复合菌剂在花生整个生育期稳定性良好。     

土霉素降解菌筛选及降解特性研究

【目的】 近年来,抗生素在畜禽及水产养殖中的使用量增加,导致固体废弃物和污水中存在大量的抗生素和耐药菌。土霉素作为用于养殖业主要的抗生素之一,在畜禽粪便和污水中的残留含量较高,因此,筛选并鉴定了能降解残留土霉素的微生物。 【方法】 采用富集驯化法,以菌肥、药渣和畜禽粪便为原料,采用摇床震荡的方法进行微生物培养,采用高效液相色谱法 (HPLC) 进行土霉素含量测定,筛选出能够高效降解土霉素的微生物。本研究还对降解菌在不同温度、pH、转速和接种量条件下的土霉素降解效果进行优化,并最终利用16S rDNA的方法鉴定菌种。 【结果】 筛选出一株能够高效降解土霉素的菌株T4菌,经16S rDNA测序鉴定该菌株为假单胞菌 (Pseudomonas sp.) ,该菌株在30℃时对土霉素的降解率最高,达到了26.75%;不同pH梯度下,T4菌在pH为7时对土霉素的降解率达到最高,为27.03%;转速为150 rpm和170 rpm时,T4菌对土霉素的降解率分别为26.18%和25.59%,考虑到摇床高转速耗能高的因素,因此选择150 rpm为优化的转速;接种量对T4菌降解土霉素的影响较小,而且二者之间呈负相关,接种量1%时降解率最高,为26.88%。优化条件下,T4菌对100 mg/L土霉素的降解率为26.29%;堆肥试验表明,添加了T4菌之后,土霉素去除率更高,为93.21%。 【结论】 本研究筛选出的菌株T4对土霉素有较好的降解能力。通过16S rDNA基因序列分析,T4菌属于假单胞菌 (Pseudomonas sp.),其降解土霉素的优化条件为温度30℃、pH 7.0、转速150 rpm、接种量1%。在堆肥中接种T4菌后,提高了对土霉素的去除作用,表明T4菌作为土霉素降解菌具有污染治理的潜力。      

一株新的多菌灵高效降解茵的筛选与降解特性分析

从长期施用多菌灵农药的土壤中,通过富集筛选,获得1株新的多菌灵高效降解菌株。通过生理生化实验和16SrDNA序列同源性分析鉴定该菌株,应用高效液相色谱法对纯培养条件下菌株的降解特性和粗酶提取液的降解性能进行了分析。结果表明,筛选所获得的菌株与Raoultella菌属的亲缘关系最近,将其命名为Raoultellasp．MBC,该菌株能在以多菌灵为唯-碳源的无机盐培养基中生长;25℃、pH7．0、200r·min。的最适生长条件下避光振荡培养72h,多菌灵的降解率达到100％;在最适培养条件下外加氮源和碳源在培养后期均可以提高多菌灵的降解率,外加氮源对多菌灵的降解效果优于外加碳源;该菌体的粗酶提取液具有降解多菌灵活性,且多菌灵降解酶为诱导酶。研究结果为多菌灵污染土壤的生物修复和酶修复提供了材料和理论依据。     

木质素降解菌的筛选及其纤维素酶基因克隆表达研究

以高效降解木质素为指标, 进行木质素降解菌的筛选和纤维素酶处理纤维材料的研究.通过测定14株白腐菌菌株在愈创木酚、苯胺兰和鞣酸培养基上生长状况和酶活分泌能力, 得到8株能产生阳性反应的菌株.以木质素和综纤维素失重的比值(SF指数)为指标, 对这几株菌进行复筛, 从中筛选出具有生长优势和强酶分泌能力的菌株平菇10969和侧耳WP1.与黄胞原毛平革菌Phanerochaete chrysosporium RP78和P.chrysosporium BKM-F-1767两株模式菌相比, 白腐菌具有良好的生长优势、强酶系分泌能力和降解的优势.从分离的白腐菌中克隆纤维素酶基因(egl2), 表达蛋白并测定酶活.用粗酶液处理不同的纤维材料, 结果表明, 其还原糖产量为综纤维素(酸解)>菌草(白腐菌处理)>未处理菌草.白腐菌的研究对草质资源的充分利用、污染物的降解、燃料乙醇的开发以及我国生态农业的持续发展等都有着重要意义.     

四环素类抗生素降解菌的分离与鉴定

为了获得四环素类抗生素的高效降解菌,从长期受四环素类抗生素污染的土壤中分离、重复驯化筛选得到6株对四环素类抗生素具有降解能力的菌株;采用高效液相色谱法研究了6株降解菌的降解效果,并通过形态、生理生化特征及16S rDNA序列分析对其进行鉴定。结果表明,菌株TJ-2#对土霉素、四环素和金霉素3种四环素类抗生素的降解效果最好,降解率分别为58.3%、63.9%和65.5%,TJ-6#其次;6株降解菌均能以四环素类抗生素作为唯一碳源生长。经形态、生理生化特征及16S rDNA鉴定,菌株TJ-2#为木糖氧化无色杆菌(Achromobacter sp.),TJ-6#为枯草芽胞杆菌(Bacillus sp.)。本研究结果对四环素类抗生素的生物降解具有一定的现实意义。     

降解玉米秸秆真菌复合菌系的构建及其降解效果评价

  【目的】  秸秆的木质纤维素含量丰富、结构复杂，在自然界中降解较慢，增加秸秆降解菌剂中菌株的多样性有利于提升还田秸秆的降解效果。探究菌株多样性水平和组成影响复合菌系秸秆降解的效果及原因，为复合菌系在秸秆降解中的应用提供理论支撑。  【方法】  通过富集驯化培养，从玉米秸秆还田土壤中筛选具有秸秆降解能力的真菌，从中挑选5株高效秸秆降解真菌进行基因间隔区序列 (ITS) 测定和物种鉴定，明确其分类地位。通过全组合构建菌株多样性为1～5的复合菌系，分别检测复合菌系的秸秆相对降解率及其滤纸酶、纤维素内切酶和木聚糖酶活性，利用方差分析和相关性分析等方法研究菌株多样性和组成对复合菌系玉米秸秆降解效果及其纤维素酶活性的影响。  【结果】  共筛选获得了15株具有秸秆降解能力的真菌，其中5株真菌的秸秆降解效果好、纤维素水解能力强。经ITS序列鉴定和系统发育分析，发现5株降解真菌的遗传差异较大，Z7-6、F7-5、F4-3、L1-1和J2-5分别与草酸青霉 (Z7-6: Penicillium oxalicum)、烟曲霉 (F7-5: Aspergillus fumigatus)、哈茨木霉 (F4-3: Trichoderma harzianum)、白囊耙齿菌 (L1-1: Irpex lacteus) 和木贼镰刀菌 (J2-5: Fusarium equiseti) 的ITS序列相似度均超过99.95%。全组合复配结果表明，复合菌系的秸秆降解能力和纤维素酶活力均高于各单一菌株，且随着菌株多样性水平的增加而提高。滤纸酶、纤维素内切酶和木聚糖酶的活力越强，复合菌系对玉米秸秆的降解效果越好，而其秸秆相对降解率主要取决于滤纸酶和纤维素内切酶的活性。抽样效应分析发现，不同菌株对复合菌系的秸秆降解效果、滤纸酶和纤维素内切酶活性的影响不同。不含菌株F7-5的复合菌系降解效果显著优于含有该菌株的组合，以Z7-6 (P. oxalicum)、F4-3 (T. harzianum)、L1-1 (I. lacteus) 和J2-5 (F. equiseti) 组合F1的玉米秸秆降解效果最佳、酶活性最高。  【结论】  秸秆降解复合菌系的构建过程需要同时考虑多样性效应和抽样效应，增加降解菌的多样性有助于增强秸秆的降解效果。本研究筛选获得的复合菌系F1在玉米秸秆降解中具有潜在的应用前景。     

Manure contaminated with the antibiotic sulfadiazine impairs the abundance of nirK- and nirS-type denitrifiers in the gut of the earthworm Eisenia fetida

The antibiotic sulfadiazine (SDZ) can affect denitrifying bacteria in soil. However, effects on denitrifiers in the gut of earthworms have not been described so far. Therefore, the influence of SDZ-contaminated manure applied to soil on denitrifiers in the gut of the earthworm Eisenia fetida was assessed by quantitative polymerase chain reaction targeting genes coding for nirK- and nirS-type nitrite reductases of denitrifiers. Gut contents of Eisenia fetida contained 2.5 × 10⁶ and 5.1 × 10⁵ gene copies of nirK and nirS, respectively, after 2 weeks in soils amended with manure only. Copy numbers of nirK and nirS in gut contents from manure treatments with SDZ were up to ten times less. Overall, the data indicate a negative impact of SDZ on denitrifiers in the gut of earthworms.     

Earthworm populations are stable in temperate agricultural soils receiving wood-based biochar

The application of decomposable organic residues such as manure and crop litter is generally beneficial to earthworms. There is an emerging interest in applying biochar, a carbonaceous product of pyrolysis, to temperate agricultural soils. The slow decomposition rate of biochar, which also contains ash and combustion byproducts, could be detrimental to earthworms. The objective of this study was to describe the earthworm populations in biochar-amended soils on a dairy farm in the St. Francis River watershed, Quebec, Canada. Earthworms were collected from replicated field plots under cereal production. Site A received three wood-based biochar types at two application rates(5 and 10 t ha~(-1) biochar) plus an unamended control, while Site B received wood-based biochar(7.5 t ha~(-1) biochar), dairy cattle slurry(10 t ha~(-1) manure), or a combination of the biochar and manure rates plus an unamended control. Earthworms were collected by hand sorting and formaldehyde expulsion from soil pits. Three species, Aporrectodea turgida, Aporrectodea tuberculata, and Lumbricus rubellus, were found at the sites, and Aporrectodea was the dominant genus. Biochar sources, rates, and application with dairy slurry did not affect the earthworm population, which had 52–218 individuals m~(-2) in 2010 and 4–96 individuals m~(-2) in 2011. The seasonal variation in earthworm population may be due to flooding in the spring of 2011, which apparently interfered with earthworm reproduction. The similarity in earthworm abundance and biomass in plots with and without wood-based biochar leads to the conclusion that earthworm populations are stable in biochar-amended soils in this cold, humid temperate region.     

菌肥对青稞根际土壤理化性质以及微生物群落的影响

应用化学分析、聚合酶链反应-变性梯度凝胶电泳(PCR-DGGE)技术和DNA测序技术,研究了西藏棕色砂壤土中微生物肥料不同施用量和施用期对青稞根际土壤理化性质和细菌群落多样性的影响。结果表明,施用谷特菌肥能显著提高土壤全氮、全磷、全钾、有机质、碱解氮、有效磷和速效钾的水平,如播前施用菌肥浓度750 ml hm-2的处理较不施用菌肥的处理上述指标分别提高13.32%、28.42%、16.20%、9.81%、21.36%、39.35%和30.48%,拔节期施用菌肥浓度2 250 ml hm-2的处理较不施用菌肥的处理分别提高7.25%、29.35%、18.04%、12.86%、15.90%、43.27%和53.99%。DGGE分析表明,相同施用方式中不同施用量土样中微生物的DGGE图谱相似。非加权组平均法(UPGMA)聚类分析将DGGE图谱分为2大类群。Shannon-Wiener指数表明,施用菌肥的土壤细菌多样性先增加后逐渐降低,播前以喷施谷特菌浓度750 ml hm-2时的细菌多样性最高;拔节期则以喷施谷特菌浓度2 250 ml hm-2处理的细菌多样性最高,且两种施用方式土壤养分的释放与Shannon指数的变化规律均为播前﹥拔节期。测序结果表明,不同施肥浓度土样微生物种群分布较为广泛,其中Actinobacteria纲细菌种类略多,少数菌种为未经培养菌种(Uncultured bacterium)。典型对应分析(CCA)表明,DGGE图谱条带分布与土壤理化性质密切相关,碱解氮、全磷和全氮是影响微生物群落的主要环境因子。研究结果表明,施用谷特菌肥可明显改善青稞根际土壤理化性状,提高土壤细菌多样性。     

Transport Of Fecal Bacteria From Poultry Litter and Cattle Manures Applied to Pastureland

Land applications of manure from confined animal systems and direct deposit by grazing animals are both major sources of bacteria in streams. An understanding of the overland transport mechanisms from land applied waste is needed to improve design of best management practices (BMPs) and modeling of nonpoint source (NPS) pollution. Plots were established on pasturelands receiving phosphorus-based livestock waste applications to measure the concentrations of Escherichia coli (E. coli), fecal coliform (FC), and Enterococcus present in overland flow at the edge of the field. The flow-weighted bacteria concentrations were highest in runoff samples from the plots treated with cowpies (1.37×10⁵ colony forming units (cfu)/100 ml of E. coli) followed by liquid dairy manure (1.84×10⁴ cfu/100 ml of E. coli) and turkey litter (1.29×10⁴ cfu/100 ml of E. coli). The temporal distribution of fecal bacterial concentrations appeared to be dependent upon both the animal waste treatment and the indicator species, with peak concentrations occurring either at the beginning of the runoff event or during peak flow rates. BMPs could be selected to reduce peak flows or first flush effects depending upon the litter or manure applied to the land. The commercial Biolog System was used to identify the dominant species of Enterococcus present in the cowpie source manure (Enterococcus mundtii 55%) and in the runoff collected from the transport plots treated with cowpies (Enterococcus faecalis 37%). The identification of predominant species of Enterococcus that are associated with specific sources of fecal pollution could greatly assist with identifying the origins of NPS pollution.     

牛粪沼渣中多重耐药细菌的环境风险

为研究牛粪厌氧发酵沼渣中多重耐药细菌的环境污染风险，选用畜禽养殖中常用的5种抗生素(红霉素、氨苄青霉素、金霉素、链霉素、环丙沙星)对牛粪沼渣中可培养抗生素抗性细菌进行筛选。结果表明：多重耐药细菌的比例高达76.5%,其中，抗5种和4种抗生素的细菌分别有11,21株。所有多重耐药细菌均对氨苄青霉素具有抗性，抗红霉素、金霉素和链霉素的细菌分别占总多重耐药细菌数的92.0%,89.3%,61.3%。通过细菌16S rRNA测序鉴定，32株具有4种以上抗性的细菌分别属于福氏志贺氏菌、摩根氏菌和假中间苍白杆菌，均为重要的临床致病菌。使用全基因组测序对7株典型多重耐药细菌携带的抗性基因进行分析，共检测出28种抗生素抗性基因，对应9种抗生素抗性类型。通过分析抗性基因及插入序列所在位点信息发现，多重耐药细菌普遍携带含有抗性基因和插入序列共存的质粒，表明抗性基因具有高度的可移动性和较强的传播风险。综上所述，牛粪沼渣中含有大量的多重耐药细菌，可视作抗生素抗性基因的储存库，应重视沼渣农业资源化利用过程中的环境风险监测与评估。     

Effect of passage through the intestinal tract of detritivore earthworms (Lumbricus spp.) on the number of selected Gram-negative and total bacteria

Laboratory experiments were carried out to investigate the fate of bacteria during and after passage through the intestinal tract of detritivore earthworms. Earthworms (Lumbricus spp.) were fed with cattle dung inoculated 7 days previously with one of five different Gram-negative bacteria. Bacterial concentrations were determined 2 days later in dung and soil, and in gut material from different parts of the earthworm intestinal tract. A high percentage (28–82%) of the total bacteria (epifluorescence direct counts) in the earthworm gut content was culturable. The concentration of total heterotrophic aerobic bacteria did not vary significantly among the five different bacterial additions and the non-inoculated control. In earthworm casts the number of total heterotrophs per gram dry matter (2.1×10⁹) was higher than in soil (1.7×10⁸), but lower than in the dung (1.5×10¹⁰). The test-bacteria, however, showed different survival patterns along the earthworm intestinal tract. The concentrations of Escherichia coli BJ 18 and Pseudomonas putida MM 1 and MM 11 in earthworm casts were lower than in the ingested dung, while concentrations of Enterobacter cloacae A 107 and Aeromonas hydrophila DMU 115 in dung and casts were similar. Ent. cloacae, and to aminor extent E. coli, were reduced in numbers by several orders of magnitude in the pharynx and/or crop. In the hind gut, however, the concentration of Ent. cloacae had increased to the same level as in the ingested dung, while the concentration of E. coli remained low. Our observations indicate that the bacterial flora of ingested food materials changes qualitatively and quantitatively during gut transit.     

不同来源有机物料对菜用蚕豆生长和品质及根际土壤性状的影响

通过盆栽试验，分析了不同来源有机物料(鸡粪、牛粪、菇渣)对菜用蚕豆生长和品质、根际土化学和生物学性状的影响。结果表明：①3种有机物料对蚕豆分枝数、荚干重、籽粒干重、大粒重、大粒蚕豆占比、籽粒淀粉含量均有积极作用，其中牛粪和菇渣能显著(P<0.05)提高大粒蚕豆占比，鸡粪能显著(P<0.05)提高二粒荚和三粒荚的占比，并且对籽粒淀粉含量的提升效果最为明显(比对照增加69.1%)；②3种有机物料增加了根际土中细菌、真菌数量，显著(P<0.05)提高了根际土中性转化酶、脲酶、酸性磷酸酶、多酚氧化酶活性(除牛粪使中性转化酶活性提高不显著外)，其中鸡粪对细菌数量、中性转化酶活性、脲酶活性的提高程度均最大；③鸡粪能显著(P<0.05)增加根际土有机碳、全氮、NH₄⁺-N含量，牛粪、鸡粪能显著(P<0.05)提高土壤pH。大粒蚕豆占比与土壤酸性磷酸酶活性、有效磷含量呈显著正相关。蚕豆淀粉含量与细菌数量、中性转化酶活性、有机碳、全氮、NH₄⁺-N含量呈显著正相关，这说明有机物料的施用可能为细菌生长提供了更多的碳源和能源，导致细菌数量增加。细菌是酶的重要产生者，细菌数量的增加可能提高了碳氮循环相关酶(中性转化酶、脲酶)的产生量和活性，从而使更多的有机氮转变为无机氮(NH₄⁺-N)，提高了土壤养分(NH₄⁺-N、全氮)水平，进而提高了蚕豆营养品质。     

Bacterial diversity as affected by application of manure in red soils of subtropical China

A plot experiment was conducted to understand the response of the soil bacterial community to manure application rates and the relationship between the composition of bacterial community and soil chemical properties. The experiment involved gradients of manure combined with chemical fertilizer in red soils from granite, red sandstone and red clay between 2013 and 2015. The soil bacterial community composition was significantly affected by different manure rates. The relative abundances of Burkholderiaceae, Micrococcaceae and Streptomycetaceae were higher at low manure rates (1.75 to 3.5 t·ha^?1·yr.^?1), whereas the relative abundance of Xanthomonadaceae was higher at high manure rates (7 to 28 t·ha^?1·yr.^?1). Manure application increased the bacterial abundance but decreased the diversity when its rates were higher than 7, 14 and 14 t·ha^?1·yr.^?1 in soils from granite, red sandstone and red clay, respectively. Redundancy analysis revealed that soils from different parent materials had different bacterial communities with soil pH and available phosphorus (AP) being determinant factors. The peanut yields exhibited significantly positive correlations with the bacterial diversity in soil, implying the importance of bacterial diversity for soil productivity. Soil AP was correlated with bacterial diversity by parabolic equations and probably AP may be an indicator of declining bacterial diversity at high manure rates. The critical value were 39.71, 65.75 and 90.16 mg·kg^?1 in soils from granite, red sandstone and red clay, respectively. This study suggests the importance of maintaining soil bacterial diversity under moderate and balanced applications of manure.