Effects of microbial inoculants on humification and enzyme activity during composting of planting and breeding wastes
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摘要:目的
研究接种自制复合菌剂和商用菌剂对牛粪、菌糠与尾菜混合堆肥中腐殖化程度和酶活性的影响，为种养废弃物的资源化利用提供高效复合菌剂。
方法以牛粪、菌糠与尾菜(湿重比5∶4∶1)为原料进行混合堆肥，初始混合堆料碳氮比约为30∶1，堆料量为1.5 t，高度为100～120 cm，条剁式堆肥。CDS菌剂主要由皮氏不动杆菌(Acinetobacter pittii)、枯草芽孢杆菌粪便亚种(Bacillus subtilis subsp. Stercoris)和高地芽孢杆菌(Bacillus altitudinis)组成，XY4菌剂主要由短小芽孢杆菌(Bacillus pumilus)、解淀粉芽孢杆菌(Bacillus amyloliquefaciens)和嗜热链霉菌(Streptomyces thermovulgaris)组成。堆肥设接种自制菌剂CDS、XY4、商用菌剂EM和无菌剂培养基对照(CK)共4个处理，以质量比为1.5%的接种量接种。每日监测堆肥温度，分别在堆肥的第1、12、28、36天进行样品采集，测定腐殖酸和黄腐酸含量、酶活性(脲酶、碱性磷酸酶、蔗糖酶和过氧化氢酶)和腐熟指标(种子发芽指数、碳氮比和pH)。
结果所有处理(CK、CDS、XY4和EM)的高温期(>50.0℃)均持续超过20天。在堆肥28天后，CDS和XY4处理的堆肥浸提液处理的种子发芽指数分别达到85.1%、89.6%，显著高于CK处理的73.8%，EM处理与CK处理无显著差异。XY4处理堆肥结束时(36天)的C/N值为14.6，显著低于CK处理的19.1。第1～36天CDS和XY4处理的黄腐酸含量始终高于CK (P<0.05)，第36天的腐殖酸含量也显著高于CK (P<0.05)。此外，CDS、XY4处理较其他处理显示了较高的酶活性：XY4处理第28天的脲酶活性高达28.0 mg/(g·d)，第12天时的蔗糖酶活性达到97.6 mg/(g·d)，显著高于其他处理组(P<0.05)；CDS处理第12天和第28天的碱性磷酸酶活性最高，分别为40.0和32.1 mg/(g·d)。相关性和冗余分析显示，相较于CK，CDS和XY4处理中过氧化氢酶对腐殖酸形成具有重要作用(解释率分别为17.3%和26.7%)。同时，CDS、XY4和EM处理腐殖酸与碱性磷酸酶活性呈显著负相关(P<0.05)。
结论由短小芽孢杆菌(Bacillus pumilus)、解淀粉芽孢杆菌(Bacillus amyloliquefaciens)和嗜热链霉菌(Streptomyces thermovulgaris)组成的微生物菌剂(XY4)，提升堆肥腐熟过程碱性磷酸酶和蔗糖酶活性的功能高于其他菌剂组合，因而加速了堆肥中有机物的快速分解和腐殖化，有效缩短了堆肥的腐熟周期，加快了堆肥的腐熟进程，显示了较好的市场前景。
Abstract:ObjectivesThe effects of inoculation of self-made and commercial compound microbial inoculants on the humification degree and enzyme activities during composting procession of planting and breeding waste were studied, so as to provide efficient compound microbial inoculants for their utilization as resources.
MethodsCow dung, mushroom chaff and tail vegetables (wet weight ratio of 5∶4∶1) were used as raw materials for mixed composting. The initial compost had a C/N ratio of about 30∶1, and was 1.5 t in weight, 100−200 cm in height. Two microbial inoculum CDS and XY4 were prepared in our lab for the experiment. CDS was mainly composed of Acinetobacter pittii, Bacillus subtilis subsp. Stercoris and Bacillus altitudinis. XY4 was mainly composed of Bacillus pumilus, Bacillus amyloliquefaciens and Streptomyces thermovulgaris. The two homemade microbial inoculum CDS, XY4, a commercial microbial inoculum (EM), and inactivated medium (CK) were used as treatments, with an inoculum ratio of 1.5% by mass. During the composting procession, compost temperature was daily monitored. On the 1st, 12th, 28th and 36th day of composting, samples were collected for the determination of seed germination index (GI), C/N ratio, pH, humic and fulvic acid content, and enzyme activities.
ResultsA high-temperature phase (>50.0℃) lasted more than 20 days in all the four treatment composts. After 28 days of composting, the compost extract from the XY4 treatment was recorded a germination index exceeding 80.0%. With the elongation of composting process, the C/N ratio decreased from 30 at initial to about 14.6. Compared with CK, CDS and XY4 treatments were tested higher fulvic acid content from 1 to 36 days, and higher humic acid content on day 36 (P<0.05). CDS and XY4 treatments were also tested higher enzyme activities than the other two treatments: the urease activity was as high as 28.0 mg/(g·d) on day 28 and the sucrase activity was as high as 97.6 mg/(g·d) on day 12 under XY4 treatment; CDS treatment exhibited the highest alkaline phosphatase activity on day 12 and 28, with values of 40.0 and 32.1 mg/(g·d), respectively. Correlation and redundancy analyses showed that catalase had an important role in humic acid formation in CDS and XY4 compared to CK, explained 17.3% and 26.7% of the difference. For all the CDS, XY4 to EM treatments, humic acid was significantly and negatively correlated with alkaline phosphatase (P<0.05).
ConclusionsCompared to commercial microbial inoculants, XY4 homemade microbial preparations could significantly enhance the activities of alkaline phosphatase and sucrase during the composting maturation process. These inoculants not only accelerate the rapid decomposition and humification of organic matter in the compost, but also increase the compost temperature and the duration of the high-temperature phase, effectively shortening the compost maturation period. Consequently, the homemade inoculants XY4 expedites the compost maturation process.
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Keywords:
- cow dung /
- composting /
- compound microbial agent /
- maturity index /
- humus /
- enzyme activity
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现代农业活动(大规模畜牧业和集约化作物生产)产生了大量未被利用的固体和液体废物[1−2]。据统计，农业废弃物的上升速度为每年5%至10%，年产量高达50亿t[3]。其中，每年产生约15亿t畜禽粪便，但其综合利用率不到60%；尾菜(蔬菜废弃物)的产量约为5亿t，未利用比例超过90%[4]；此外，种植100 kg食用菌所需的原料，收获鲜菇后产生了60 kg菌糠[5]。这些有机废弃物处理不当，不仅浪费资源，而且由于其富含有机物和营养物质，会对空气、土壤和水体等造成严重污染[6]。目前，堆肥技术被广泛认为是处理农业废弃物的一种实用且经济的方法，提高了资源化利用效率，减少了环境污染[7]。然而，单一物料堆肥存在含水率高、降解率低、碳氮比低等缺点。目前，对多组分物料混合堆肥进行了大量研究。多组分物料混合堆肥是一种利用多种有机废弃物混合堆肥的方法，堆肥产品具有生物利用度高和丰富的营养元素，其发酵分解能力优于单一堆肥[8]。然而，混合堆肥的过程较为复杂，涉及了多种物理、化学和微生物等因素的相互作用。微生物与其分泌的酶在堆肥中发挥着重要作用，将原始有机物质降解为简单物质，并将其合成为腐殖化合物[9]。虽然堆肥中原有的微生物对堆肥有降解作用，但畜禽粪便、菌糠和尾菜这类纤维素含量较高的有机废弃物，其堆肥进程均存在发酵温度上升缓慢、腐熟周期长等问题[10]，这不仅降低了堆肥的农业价值，而且成为制约好氧堆肥处理农业废弃物商业化和产业化的关键因素[11]。因此，研究畜禽粪便、菌糠和尾菜等有机质降解的措施，对实现废弃物的资源化利用具有重要意义。
目前，接种外源微生物菌剂是优化堆肥技术的有效手段，具有代谢强度高、繁殖速度快等特点[12]。接种微生物菌剂能加快堆肥温度，并延长高温持续时间，提高pH和种子发芽指数，促进有机质的碳化和腐殖化过程[13−14]。张祥永等[15]研究发现，接种菌剂增加了堆肥发酵的温度，延长了高温期，并且显著提高了酶(纤维素酶和蛋白酶)活性。Pandey等[16]研究表明，外源复合微生物接种有助于纤维素、半纤维素和木质素的分解，加速堆肥的腐殖化。李恕艳等[17]发现，在鸡粪中接种菌剂能增加堆肥腐殖质的含量，对其腐熟度和酶活性的提高有促进作用。上述研究结果表明，接种外源微生物菌剂能够提高堆肥效率并改善堆肥质量。由于堆肥原料的不同，接种的微生物菌剂也存在差异。研究表明，接种复合菌剂的效果通常优于单一菌剂[18]。目前，市售的微生物复合菌剂中很少有适应多种物料混合堆肥的菌剂，针对牛粪、菌糠渣和尾菜等原料的堆肥特性的复合菌剂研究和开发也相对较少。因此，本研究利用前期试验研究积累的降解菌，制备出两种自制的复合菌剂。以牛粪、菌糠渣和尾菜为堆肥材料，进行不同菌剂接种处理的对比研究，探讨自制菌剂对混合堆肥腐熟度、腐殖质含量和酶活性的影响，以及不同菌剂对堆肥发酵阶段动态变化的影响，旨在为接种外源复合菌剂促进混合堆肥高效腐熟提供理论支持。
1. 材料与方法
1.1 试验材料
堆肥原料包括牛粪、菌糠和尾菜。其中，牛粪和菌糠渣来自甘肃省大行农业废弃物处理有限公司，尾菜则来自周边农村菜地。尾菜经过粉碎，碎片大小约为1～2 cm。所有堆肥原材料的物理化学特性参见相关文献[19]。
1.2 微生物菌剂
试验所用菌剂CDS和XY4由本课题组研制，CDS菌剂包括皮氏不动杆菌(Acinetobacter pittii)、枯草芽孢杆菌粪便亚种(Bacillus subtilis subsp. Stercoris)和高地芽孢杆菌(Bacillus altitudinis)，XY4菌剂包括短小芽孢杆菌(Bacillus pumilus)、解淀粉芽孢杆菌(Bacillus amyloliquefaciens)和嗜热链霉菌(Streptomyces thermovulgaris)[19−20]。市售菌剂(EM)由山东绿陇生物科技有限公司生产，主要菌种包括酵母菌、乳酸菌、放线菌等。
将菌剂接种于50.0 mL蛋白胨纤维素液体培养基(peptone cellulose solution)，该培养基配方为每升蒸馏水中含5.0 g 蛋白胨、5.0 g NaCl、2.0 g CaCO3、1 g酵母粉，2.0×6.0 cm滤纸条，于28.0℃下180 r/min摇床上培养5天，制成有效活菌数均达1.0×108 CFU/mL数量级的菌液。
1.3 试验方法
堆肥试验于2021年8—9月在甘肃大行农业废弃物处理有限公司进行。牛粪、菌糠与尾菜按湿重比为5∶4∶1混合混匀，初始C/N约为30∶1，含水率约为60%。试验设计4个堆肥接种处理，3个菌剂CDS、XY4、EM以质量比为1.5%的接种量接种，以接种等量无菌剂为对照(CK)。将堆料充分混匀后堆制成长×宽×高=3.0 m×1.2 m×1.35 m的条垛式堆，采用人工翻堆的形式，每4天翻堆1次，以保证供氧和堆料腐熟一致。
在堆肥的第1、12、28、36天进行样品采集。将每个堆体分成4段，在每段上层0～30 cm、中间层30～60 cm、下层60～90 cm随机采集等量样品，充分混合后，取500.0 g鲜样[21]，分为两份：一份自然风干后用于测定腐熟指标和腐殖质含量；另一份在4.0℃下保存，用于测定堆肥中酶活性。
1.4 测定方法
1.4.1 堆体温度
利用针式数字温度仪(dot type digital thermometer)分别在堆肥顶部30、60和90 cm处进行测温，取平均值作为堆体温度，同时测定当日的环境气温，具体参考李昌宁等[21]的方法测定。
1.4.2 腐熟度、腐殖质含量和酶活性测定
种子发芽指数(germination index，GI)的测定：堆肥样品与蒸馏水按1∶10比例充分混合，取10.0 mL滤液以3000 r/min离心10 min，吸取5.0 mL上清液于铺有滤纸的培养皿中，用蒸馏水作为对照，放置30粒白菜种子，3个重复，30.0℃培养4天，计算种子的发芽率，并用游标卡尺测量种子的根长。根据下列公式，GI计算方法[21]如下：
GI=(堆肥浸提液处理种子发芽率×种子根长)/(蒸馏水处理种子发芽率×种子根长)×100%
酸碱度(pH)采用固液比1∶5浸提，用酸度计测定[22]；总有机碳(TOC)采用重铬酸钾氧化法[23]测定，全氮(TN)采用凯氏定氮法[23]测定。
腐殖酸和黄腐酸采用1000.0 mL碱性磷酸钠和氢氧化钠溶液从10.0 g样品提取，提取液在25.0℃下振荡24 h，然后12000 r/min离心15 min，以0.45 µm滤膜过滤。重复上述步骤，直到提取液几乎无色，合并所有提取液。用盐酸调节腐殖质溶液pH至1，在4.0℃下静置12 h，随后以12000 r/min离心15 min，上清液为黄腐酸，沉淀为腐殖酸。沉淀用NaHCO3溶解得到腐殖酸，最后，采用重铬酸钾氧化法，分析腐殖酸和黄腐酸的总有机碳，以得到腐殖酸和黄腐酸含量[24]。计算公式为：
腐殖酸或黄腐酸=(V0−V)×c×3×10−3×分取倍数×100/(m×f)
式中：V0、V分别为空白测定和样品测定所消耗的硫酸亚铁的体积(mL)；c为硫酸亚铁的浓度(mol/L)；m为称取干物质量(g)；f为含碳量系数(系数为1.742)。
堆肥中脲酶、碱性磷酸酶、蔗糖酶以及过氧化氢酶的活性参照关松荫[25]方法测定，脲酶活性测定以尿素为基质，根据酶促产物氨与苯酚钠−次氯酸钠作用生成蓝色的靛酚，采用比色法来测定脲酶活性；碱性磷酸酶活性的测定以磷酸苯二钠为基质，采用分光光度计比色法测定；根据堆肥因生化反应生成3-氨基-5-硝基水杨酸而呈橙黄色，采用分光光度计比色法测定蔗糖酶活性；过氧化氢酶活性采用高锰酸钾溶液滴定法测定。
1.5 数据处理与分析
采用Microsoft Excel 2010进行数据整理；用SPSS 26.0软件进行单因素方差分析和LSD法进行多重比较；并运用Canoco 5.0进行冗余分析(redundancy analysis，RDA)，用Origin 2021绘图。
2. 结果与分析
2.1 菌剂对堆肥过程中温度的影响
由图1可知，4组处理的温度变化均经历了升温期、高温期、降温期和腐熟期4个阶段，各处理高温期温度均达到50.0℃以上。堆肥第1天，XY4处理首先进入高温期，其他3个处理于堆肥第2天进入高温期。CK、CDS、XY4和EM处理的高温期分别维持了22、23、25和26天。由于堆肥过程中的翻堆，各处理高温期的温度变化幅度较大，主要集中在堆肥的4～22天。上述结果表明，XY4和EM处理有利于加速堆肥升温和延长高温持续时间。
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图 1 堆肥过程中温度的变化注：CK为未接种菌剂的处理；CDS为接种CDS菌剂的处理；XY4为接种XY4菌剂的处理；EM为接种市售菌剂EM的处理。Figure 1. Changes of temperature during composting processNote: CK is the treatment without inoculum; CDS is the treatment inoculated with CDS inoculum; XY4 is the treatment inoculated with XY4 inoculum; EM is the treatment inoculated with the commercially available inoculum EM.2.2 菌剂对堆肥腐熟指标的影响
2.2.1 菌剂对堆肥种子发芽指数(GI)的影响
如表1所示，各组的GI值随堆肥时间延长均呈上升趋势，堆肥样品对植物的毒性逐渐降低。堆肥前12天，GI值均小于80.0%；在28天时，与CK相比，XY4处理显著提高了GI值(P<0.05)，且大于80.0%。这表明XY4处理接种微生物菌剂可增强堆肥浸提液的促生能力。堆肥进行至第36天，各处理GI值均大于80.0%，表明已完全腐熟。
表 1 堆肥过程中种子发芽指数的变化(%)Table 1. Changes of germination index during composting process堆肥时间 (d)
Composting timeCK CDS XY4 EM 1 32.9±5.0 cA 34.3±6.2 dA 31.2±5.3 dA 31.2±6.9 dA 12 60.4±8.8 bA 60.1±8.6 cA 62.4±7.1 cA 60.3±6.4 cA 28 73.8±7.4 bB 85.1±5.6 bAB 89.6±5.0 bA 78.7±7.0 bAB 36 103.2±7.2 aA 115.0±13.2 aA 120.5±5.7 aA 114.7±10.2 aA 注：CK为未接种菌剂的处理；CDS为接种CDS菌剂的处理； XY4为接种XY4菌剂的处理；EM为接种市售菌剂EM的处理。同列数据后不同小写表示相同处理不同天数间差异显著；同行数据后不同大写字母相同天数不同处理间差异显著 (P<0.05)。
Note: CK is the treatment without inoculum; CDS is the treatment inoculated with CDS inoculum; XY4 is the treatment inoculated with XY4 inoculum; EM is the treatment inoculated with the commercially available inoculum EM. Different lowercase letters after data in a column indicate significant difference among days for a treatment; different capital letters in a row indicate significant difference among treatments on the same day (P<0.05).2.2.2 菌剂对堆肥pH的影响
如图2所示，前28天堆肥的pH呈现急剧上升至9.7～10.2，随后逐渐降低，在36天时降至8.3～8.7，XY4和EM处理的pH显著低于CK，符合国家有机肥标准要求。
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图 2 堆肥过程中pH的变化注：CK为未接种菌剂的处理；CDS为接种CDS菌剂的处理；XY4为接种XY4菌剂的处理；EM为接种市售菌剂EM的处理。Figure 2. Changes of pH during composting processNote: CK is the treatment without inoculum; CDS is the treatment inoculated with CDS inoculum; XY4 is the treatment inoculated with XY4 inoculum; EM is the treatment inoculated with the commercially available inoculum EM.2.2.3 菌剂对堆肥C/N的影响
如表2所示，各组堆肥C/N随堆肥时间延长均呈下降趋势。在前28天，C/N下降迅速，之后下降速度减缓并趋于稳定。CK、CDS、XY4和EM处理的C/N分别从初始的约30∶1降至36天时的19.1、16.3、14.6和15.4。其中，XY4处理的C/N较低(P<0.05)，表明菌剂接种加速了堆肥中总有机碳的降解，促进了总氮含量相对增加，最终导致堆肥的C/N降低。
表 2 堆肥过程中C/N的变化Table 2. Changes of C/N during composting process堆肥时间 (d)
Composting timeCK CDS XY4 EM 1 28.2±1.4 aA 26.6±2.1 aA 28.5±1.9 aA 27.8±0.8 aA 12 25.1±1.0 abA 24.2±1.5 abA 24.1±0.3 abA 25.7±0.3 aA 28 19.1±2.1 bA 18.3±1.0 bcA 19.2±1.7 bA 20.1±1.4 bA 36 19.1±0.4 bA 16.3±1.7 cAB 14.6±0.4 cB 15.4±0.2 cAB 注：CK为未接种菌剂的处理；CDS为接种CDS菌剂的处理； XY4为接种XY4菌剂的处理；EM为接种市售菌剂EM的处理。同列数据后不同小写字母表示相同处理不同天数间差异显著；同行数据后不同大写字母表示相同天数不同处理间差异显著 (P<0.05)。
Note: CK is the treatment without inoculum; CDS is the treatment inoculated with CDS inoculum; XY4 is the treatment inoculated with XY4 inoculum; EM is the treatment inoculated with the commercially available inoculum EM. Different lowercase letters after data in a column indicate significant difference among composting days under the same treatment; different capital letters in a row indicate significant difference among treatments in the same day (P<0.05).2.2.4 菌剂对堆肥有机碳的影响
CDS和EM处理在堆肥第12天后总有机碳含量开始明显下降，第28～36天期间保持相对稳定(图3)。相比之下，XY4和CK处理在前12天下降明显，之后下降速度逐渐减缓并趋于稳定。第36天CK总有机碳含量略微回升至276.4 g/kg，XY4和EM处理总有机碳含量分别为242.2和255.5 g/kg。最终，XY4处理的总有机碳含量最低，表明接种自制菌剂促进了总有机碳的转化。
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图 3 堆肥过程中总有机碳的变化注：CK为未接种菌剂的处理；CDS为接种CDS菌剂的处理；XY4为接种XY4菌剂的处理；EM为接种市售菌剂EM的处理。Figure 3. Changes of total organic carbon during composting processNote: CK is the treatment without inoculum; CDS is the treatment inoculated with CDS inoculum; XY4 is the treatment inoculated with XY4 inoculum; EM is the treatment inoculated with the commercially available inoculum EM.2.3 接种菌剂对堆肥过程中腐殖质的影响
2.3.1 菌剂对堆肥后期腐殖化的影响
堆肥过程中各处理的腐殖酸含量均呈增加趋势(表3)。CDS和XY4处理在堆肥28～36天，腐殖酸含量分别从25.5和25.7 g/kg上升至28.0和30.1 g/kg，表明在接种菌剂后28～36天，CDS和XY4处理使腐殖酸含量增加。此外，第36天，接种菌剂CDS、XY4处理的腐殖酸含量均显著高于CK (P<0.05)，这说明接种自制菌剂可以促进降温期和腐熟期腐殖酸的形成，加快堆肥有机质的腐殖化。
表 3 堆肥过程中腐殖酸和黄腐酸含量 (mg/g)的变化Table 3. Changes of humic acid and fulvic acid contents during composting process堆肥时间 (d)
Composting time腐殖酸 Humic acid 黄腐酸 Fulvic acid CK CDS XY4 EM CK CDS XY4 EM 1 20.6±1.4 bB 23.6±1.2 bA 24.2±1.5 bA 21.4±1.3 cB 15.7±1.4 aAB 17.5±1.0 aA 15.2±1.7 bB 16.0±1.3 bcA 12 22.9±2.0 abA 24.3±2.1 bA 24.7±2.3 bA 23.5±1.2 bcA 16.9±0.7 abC 19.3±0.9 abB 21.1±0.7 aA 20.1±1.6 aA 28 24.6±2.0 aA 25.5±1.2 abA 25.7±2.3 bA 25.2±1.8 bA 13.9±0.6 bcB 16.3±1.4 bcA 16.3±1.2 bA 17.5±1.7 bA 36 24.4±0.7 aB 28.0±2.0 aA 30.1±2.1 aA 31.1±3.4 aA 12.7±2.0 cB 15.4±1.4 cA 14.9±1.7 bA 14.0±0.9 cAB 注：CK为未接种菌剂的处理；CDS为接种CDS菌剂的处理； XY4为接种XY4菌剂的处理；EM为接种市售菌剂EM的处理。同列数据后不同小写字母表示相同处理不同天数间腐殖酸或黄腐酸含量差异显著；同行数据后不同大写字母表示相同天数不同处理间腐殖酸或黄腐酸含量差异显著 (P<0.05)。
Note: CK is the treatment without inoculum; CDS is the treatment inoculated with CDS inoculum; XY4 is the treatment inoculated with XY4 inoculum; EM is the treatment inoculated with the commercially available inoculum EM. Different lowercase letters after data in a column indicate significant difference in humic acid or fulvic acid content among composting days under the same treatment; different capital letters in a row indicate significant difference in humic acid or fulvic acid content among treatments in the same day (P<0.05).2.3.2 菌剂对堆肥过程中黄腐酸的影响
由(表3)可知，堆肥过程中各处理黄腐酸含量随堆肥时间延长呈现先上升后下降逐渐趋于稳定的趋势。堆肥第1～12天，各处理的黄腐酸含量逐渐增加至最大值，其中XY4处理含量最高，CK处理最低，两者相差4.2 g/kg (P<0.05)。在12～36天，CK、CDS、XY4和EM处理黄腐酸含量分别从16.9、19.3、21.1和20.1 g/kg降至12.7、15.4、14.9和14.0 g/kg，与CK相比，CDS和XY4处理的黄腐酸含量较高(P<0.05)。这表明CDS和XY4处理接种菌剂后促进了整个堆肥过程中黄腐酸的形成。
2.4 接种菌剂对堆肥过程中酶活性的影响
2.4.1 菌剂对堆肥中脲酶活性的影响
在堆肥过程中，随堆肥时间延长脲酶活性逐渐降低(表4)。CK、CDS、XY4和EM处理的脲酶活性分别为14.8～53.9、10.7～59.9、11.3～65.8和10.1～58.2 mg/(g·d)。在堆肥第1、12和36天，CK、CDS、XY4和EM处理中脲酶活性没有显著差异(P>0.05)。然而，在第28天，XY4处理的脲酶活性显著高于CK (P<0.05)，为28.0 mg/(g·d)，表明XY4处理在降温期有助于提高脲酶活性。
表 4 堆肥过程中脲酶和碱性磷酸酶活性[mg/(g·d)]的变化Table 4. Changes in urease and alkaline phosphatase activities during composting process堆肥时间 (d)
Composting time脲酶 Urease 碱性磷酸酶 Alkaline phosphatase CK CDS XY4 EM CK CDS XY4 EM 1 53.9±5.9 aA 59.9±3.8 aA 65.8±12.5 aA 58.2±15.1 aA 21.5±0.2 aD 42.2±0.6 aB 52.5±0.1 aA 38.6±0.04 aC 12 20.0±1.0 bA 22.8±4.7 bA 25.5±5.0 bA 22.4±1.5 bA 21.3±0.1 aD 40.0±0.1 bA 35.3±0.2 bC 38.5±0.2 aB 28 16.9±7.3 bB 19.0±1.0 bAB 28.0±5.6 bA 20.7±4.1 bAB 19.2±0.03 bC 32.1±0.2 cA 23.6±0.1 cB 18.2±0.03 bD 36 14.8±5.7 bA 10.7±2.8 cA 11.3±3.3 cA 10.1±4.2 bA 15.0±0.1 cC 19.1±0.2 dB 20.2±0.1 dA 14.8±0.1 cC 注：CK为未接种菌剂的处理；CDS为接种CDS菌剂的处理； XY4为接种XY4菌剂的处理；EM为接种市售菌剂EM的处理。同列数据后不同小写字母表示相同处理不同天数间脲酶或碱性磷酸酶活性差异显著；同行数据后不同大写字母表示相同天数不同处理间脲酶或碱性磷酸酶活性差异显著 (P<0.05)。
Note: CK is the treatment without inoculum; CDS is the treatment inoculated with CDS inoculum; XY4 is the treatment inoculated with XY4 inoculum; EM is the treatment inoculated with the commercially available inoculum EM. Different lowercase letters after data in a column indicate significant difference in urease or alkaline phosphatase activity among composting days under the same treatment; different capital letters in a row indicate significant difference in urease or alkaline phosphatase activity among treatments in the same day (P<0.05).2.4.2 菌剂对堆肥过程碱性磷酸酶活性的影响
各处理的碱性磷酸酶活性随堆肥进程呈不断下降趋势，CK、CDS、XY4和EM处理的碱性磷酸酶活性分别为15.0～21.5、19.1～42.2、20.2～52.5和14.8～38.6 mg/(g·d) (表4)。堆肥第12和第28天，CDS处理的碱性磷酸酶活性分别达到最高值(P<0.05)，分别为40.0和32.1 mg/(g·d)。在第1天(升温期)和第36天，XY4处理的碱性磷酸酶活性分别为52.5和20.2 mg/(g·d) (P<0.05)，说明在升温期和腐熟期XY4处理有效提高了碱性磷酸酶活性。
2.4.3 菌剂对堆肥过程中蔗糖酶活性的影响
各处理蔗糖酶活性随堆肥时间延长呈先下降后趋于平缓，CK、CDS、XY4和EM处理的蔗糖酶活性分别为44.6～74.3、44.1～118.2、41.7～98.6和44.6～97.1 mg/(g·d) (表5)。在堆肥第1和28天，CDS处理的蔗糖酶活性最高(P<0.05)，分别为118.2和64.5 mg/(g·d)。第12天，XY4处理的蔗糖酶活性显著高于其他处理(P<0.05)，为97.6 mg/(g·d)，表明CDS处理在升温期和降温期提升蔗糖酶活性的效果显著，XY4处理在高温期提升蔗糖酶活性的效果显著。
表 5 堆肥过程中蔗糖酶和过氧化氢酶活性[mg/(g·d)]的变化Table 5. Changes of sucrase and catalase activity during composting process堆肥时间 (d)
Composting time蔗糖酶 Sucrase 过氧化氢酶 Catalase CK CDS XY4 EM CK CDS XY4 EM 1 74.3±0.4 aD 118.2±0.6 aA 98.6±0.6 aB 97.1±0.3 aC 5.3±0.4 bB 6.9±0.3 aA 7.4±0.2 aA 5.7±0.2 aB 12 54.4±0.5 bD 93.3±0.5 bB 97.6±0.1 bA 84.4±0.3 bC 2.3±0.1 cB 1.9±0.2 dC 3.2±0.2 dA 2.1±0.1 bC 28 47.3±0.1 cD 64.5±0.2 cA 56.2±0.1 cB 53.0±0.3 cC 5.7±0.4 bAB 6.1±0.2 bA 5.3±0.2 cB 5.3±0.5 aB 36 44.6±0.4 dAB 44.1±0.3 dB 41.7±0.2 dC 44.6±0.2 dA 6.4±0.3 aA 5.1±0.1 cB 5.0±0.1 bB 5.3±0.3 aB 注：CK为未接种菌剂的处理；CDS为接种CDS菌剂的处理； XY4为接种XY4菌剂的处理；EM为接种市售菌剂EM的处理。同列数据后不同小写字母表示相同处理不同天数间蔗糖酶或过氧化氢酶活性差异显著；同行数据后不同大写字母表示相同天数不同处理间蔗糖酶或过氧化氢酶活性差异显著 (P<0.05)。
Note: CK is the treatment without inoculum; CDS is the treatment inoculated with CDS inoculum; XY4 is the treatment inoculated with XY4 inoculum; EM is the treatment inoculated with the commercially available inoculum EM. Different lowercase letters after data in a column indicate significant difference in sucrase or catalase activity among composting days under the same treatment; different capital letters in a row indicate significant difference in sucrase or catalase activity among treatments in the same day (P<0.05).2.4.4 菌剂对堆肥过程中过氧化氢酶的影响
堆肥各处理的过氧化氢酶活性随堆肥时间延长呈先下降后上升逐步趋于稳定的趋势(表5)。在堆肥第1天，CK、CDS、XY4和EM处理过氧化氢酶活性分别为5.3、6.9、7.4和5.7 mg/(g·d) (P<0.05)，表明接种菌剂有助于增加堆肥的过氧化氢酶活性，其中XY4效果较好。第12天，CK、CDS、XY4和EM处理的过氧化氢酶活性均急剧下降。第36天，CDS、XY4、EM处理的过氧化氢酶活性较CK显著降低(P<0.05)，其中XY4处理的过氧化氢酶活性最低，为5.0 mg/(g·d)。
2.5 腐熟度指标、酶活性与腐殖质之间的相互关系
通过相关性与冗余分析，分别对4个处理的腐熟指标、腐殖质和酶活性进行分析(图4 a～h)。不同菌剂致使腐殖酸和黄腐酸形成的主要驱动因素存在差异，各个处理RDA分析前两主轴的总解释率在91.1%～97.6%。相比CK，CDS和XY4处理中过氧化氢酶对腐殖酸的形成有重要作用(解释率分别为17.3%和26.7%)；相比CK和CDS处理，XY4和EM处理温度与脲酶活性是腐殖酸形成的主要解释变量。除CK外，CDS、XY4和EM处理蔗糖酶与温度显著正相关(P<0.05) (图4b、c、d)。同时，CDS和EM处理腐殖酸也与碱性磷酸酶活性均呈显著负相关(P<0.05) (图4b、d)。
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图 4 堆肥中腐熟度指标、酶活性与腐殖酸含量的相关性分析和冗余分析注：CK为未接种菌剂的处理；CDS为接种CDS菌剂的处理； XY4为接种XY4菌剂的处理；EM为接种市售菌剂EM的处理。图4a～d为相关性热图，*、**分别表示相关性达到0.05、0.01显著水平；图4e～h为冗余分析，连线长度代表环境因子与研究对象的相关性，线越长，影响越大，线条之间的夹角越小，相关性越大。TEM—温度；GI—种子发芽指数；TOC—总有机碳；HA—腐殖酸；FA—黄腐酸；URE—脲酶；ALP—碱性磷酸酶；SUR—蔗糖酶；CAT—过氧化氢酶。Figure 4. Correlation and redundancy analysis of maturity indicators, enzyme activities and humic acid contents in compostNote: CK is the treatment without inoculum; CDS is the treatment inoculated with CDS inoculum; XY4 is the treatment inoculated with XY4 inoculum; EM is the treatment inoculated with the commercially available inoculum EM. Fig. 4a–d are correlation analysis, * and ** indicate significant correlation at 0.05 and 0.01 levels. Fig. 4e–h are redundancy analysis, the longer the connecting line, the greater of the factor effect; the smaller the angle between the lines, the greater the correlation of factors. TEM—Temperature; GI—Germination index; TOC—Total organic carbon; HA—Humic acid; FA—Fulvic acid; URE—Urease; ALP—Alkaline phosphatase; SUR—Sucrase; CAT—Catalase.3. 讨论
3.1 接种微生物菌剂对堆肥温度和腐熟指标的影响
温度是堆肥中各生理生化过程顺利进行的基础，也是决定堆肥成功的关键因素[26]。本研究表明，将XY4菌剂(含有芽孢杆菌)接种到XY4处理可导致温度峰值最高，并延长高温期。Zhao等[27]研究发现，芽孢杆菌菌剂接种能促进堆肥中的纤维素和果胶等有机物的降解，形成的营养物质也使微生物大量繁殖，产生大量热量。此外，高温期堆肥翻堆，各处理的温度变化幅度较大，但堆肥回温较快，究其原因可能是翻堆改变了堆肥的孔隙度，并增加其中的氧气含量，从而提升微生物的代谢效率，加速了堆肥热量的产生[21]。
种子发芽指数(GI)可以通过评估植物毒性来估计堆肥腐熟，GI>80.0%表明堆肥对植物生长毒害较低，这是堆肥腐熟并实现无害化的基本要求之一[28]。本研究中XY4、EM处理发酵腐熟时显著提高了GI，可能是由于接种菌剂后加速堆肥中有机物的降解和无害化，使其具有良好的稳定性和较低的植物毒性，有助于植物的生长发育[22]。pH值是影响堆肥腐熟过程的关键因素之一，可以调节微生物的活性以及碳、氮等营养物质的转化，通常堆肥腐熟后pH值在8.0～9.0[29−30]。堆肥结束时，较堆肥初期pH均有所增加，可能的原因是接种菌剂后堆肥腐熟期的有机物分解加快，同时产生丰富的可挥发性有机酸，如乙酸、丁酸等[21]。C/N是评价堆肥成熟度和稳定性的重要参数，堆肥腐熟最终的C/N约为15～20[31]。李季等[32]对微生物群落组成与理化指标间的相关性进行研究后，发现厚壁菌门在低C/N堆肥中占主导地位，其中以芽孢杆菌最为活跃。本研究中XY4处理的C/N降幅最大，可能其加速了氨态氮被转化为亚硝酸盐和硝酸盐，氮素水平升高，最终使堆肥C/N下降。堆肥是微生物降解有机废弃物的生理生化过程。在这一过程中，碳具有重要作用，既是微生物的能量来源，也是有机物分解的基本元素[33]。本研究发现，接种菌剂后堆肥的有机碳含量减少，分解速度加快，其中XY4处理的效果最为显著，分解较为彻底。王义祥等[34]研究发现，在堆肥中有机碳会被降解，而碳的转化受到堆肥原料、温度、含水率和微生物等多种因素的共同影响，其中微生物对这一过程的影响最为显著，通过接种纤维素降解菌株，可以促进有机碳更有效地腐殖化，这与本研究所得结果基本一致。
3.2 接种微生物菌剂对堆肥腐殖质的影响
腐殖酸和黄腐酸是腐殖质的主要组成部分。He等[35]研究发现，接种菌剂的堆肥处理提高了微生物木质纤维素的分解能力，使合成腐殖酸的化合物增多，致使腐殖酸含量升高。李昌宁等[21]研究表明，在猪粪堆肥中接种菌剂的腐熟期能促进腐殖酸的芳香结构增加，并被微生物利用再生，合成大量稳定的腐殖酸。在接种菌剂的堆肥升温阶段，与黄腐酸合成和分解相关的微生物和土著微生物差异不显著，但在高温期和降温期增加了黄腐酸含量[23]。本研究表明，CDS和XY4处理在堆肥第1天腐殖酸含量显著高于CK和EM处理；在28～36天，CDS、XY4处理在堆肥初期促进腐殖化作用，提高了腐殖酸含量。1～12天，4组处理黄腐酸含量均达到最大值，其中XY4处理最高，提高了高温期堆肥中黄腐酸含量，这与上述研究结果基本一致。此外，在第36天，接种菌剂处理腐熟期黄腐酸含量较高，这主要是因为黄腐酸含量的变化受到矿化和腐殖化平衡效应的影响，其分子量小、结构简单、容易被微生物降解，但接种菌剂可以使黄腐酸合成大于分解，致使黄腐酸含量增加[23, 36]。
3.3 接种微生物菌剂对堆肥酶活性的影响及与堆肥腐熟的相互关系
在堆肥中脲酶是氨化过程中的关键酶，且氨化是硝化作用的关键和限速步骤[37]。4个堆肥处理脲酶活性随堆肥时间延长均呈逐渐下降趋势。其中，在堆肥1～12天，各处理脲酶活性较高，但相较于CK，接种菌剂不能有效提高堆肥升温期和高温期脲酶活性，这可能是堆肥在升温期和高温期含有大量的含氮有机物，致使脲酶活性较高，但随着温度的升高，含氮有机物分解生成氨，抑制了脲酶活性，此外高温也会使脲酶菌群死亡，从而降低脲酶活性[37]。在第28天，降温期XY4处理提高了脲酶活性，这可能是接种菌剂提高了XY4处理降温期脲酶相关菌群的代谢，加快含氮有机物的矿化和分解速度，从而提高了脲酶活性[12]。碱性磷酸酶对堆肥中含磷有机物具有分解和矿化作用[38]。本研究中，碱性磷酸酶活性随堆肥时间延长呈逐渐降低的趋势，这与堆肥在升温期存在大量有机磷化合物有关，提高了堆肥中碱性磷酸酶的活性，并增加了有机磷的转化[39]。随着堆肥的不断发酵腐熟，有机磷被分解完全，导致碱性磷酸酶活性不断降低。在第12和28天，CDS处理提高了高温期和降温期碱性磷酸酶活性，究其原因可能是接种菌剂引起的耐高温菌种丰度发生变化，导致碱性磷酸酶活性升高，从而影响了有机磷的转化途径[40]。
蔗糖酶能够分解蔗糖，生成葡萄糖、果糖等小分子物质，这些小分子物质恰好是微生物繁殖所需的碳源[37]。在堆肥1～28天，CDS、XY4与EM处理的蔗糖酶活性呈下降趋势，但均高于CK，可能是接种菌剂促进了堆肥中蔗糖酶的协同代谢，进而提高了堆肥过程中的代谢活性[41]。此外，在堆肥第12天，XY4处理的蔗糖酶活性为97.6 mg/(g·d)，有效提高了高温期蔗糖酶活性。Li等[11]研究发现，在猪粪堆肥中接种芽孢杆菌等微生物能更快到达高温期，并提高蔗糖酶活性。XY4处理的蔗糖酶贡献度高达59.4%，说明在堆肥中蔗糖酶起着至关重要的作用，同时，CDS、XY4和EM处理蔗糖酶与温度、C/N均呈极显著正相关，较高的温度有助于激发蔗糖酶活性，提高堆肥腐熟度，促进有机质的降解[37]。过氧化氢酶主要反映堆肥有机物的降解强度和对微生物的毒性影响[42−43]。本研究表明，堆肥第1天，CK、CDS、XY4和EM处理过氧化氢酶活性分别为5.3、6.9、7.4和5.7 mg/(g·d)，表明接种菌剂有助于增加堆肥的过氧化氢酶活性，其中XY4处理效果较好。研究表明，接种菌剂能加速堆肥微生物新陈代谢，分解有机物(主要是木质素)，使堆肥有机物的转化率升高[12, 44]。此外，XY4处理中较高的过氧化氢酶活性也降低了过氧化氢对微生物的毒害，促进作物生长繁殖。第36天，XY4处理的过氧化氢酶活性最低，为5.0 mg/(g·d)，说明能被微生物利用的可降解有机化合物减少，过氧化氢酶的活性也随之降低[45]，从而反映出接种菌剂加速了堆肥的快速腐熟。
微生物分泌的各种胞外酶在堆肥中发挥着重要作用，几乎参与了养分循环中所有生化反应[46]。在堆肥过程中监测酶活性可以获取与腐殖质形成相关的信息。有机物在水解酶(蔗糖酶、淀粉酶等)的作用下进行矿化反应，而在氧化酶的作用下进行腐殖化反应[47]。齐熙平[48]研究表明，接种外源菌剂时，碱性磷酸酶、过氧化物酶是影响腐殖酸组分变化的主要因素，且腐殖酸主要通过类蛋白和类木质纤维素缩合来形成，同时碱性磷酸酶对各处理腐殖酸的形成都有影响，这与本研究的结果相似。
4. 结论
包含短小芽孢杆菌(Bacillus pumilus)、解淀粉芽孢杆菌(Bacillus amyloliquefaciens)和嗜热链霉菌(Streptomyces thermovulgaris)的菌剂(XY4)较其他菌剂更有效地提高了堆肥中的碱性磷酸酶和蔗糖酶活性，促进有机质更快分解，腐殖化程度更高，缩短堆肥的腐熟时间，因此，具有很好的市场应用前景。
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图 1 堆肥过程中温度的变化
注：CK为未接种菌剂的处理；CDS为接种CDS菌剂的处理；XY4为接种XY4菌剂的处理；EM为接种市售菌剂EM的处理。
Figure 1. Changes of temperature during composting process
Note: CK is the treatment without inoculum; CDS is the treatment inoculated with CDS inoculum; XY4 is the treatment inoculated with XY4 inoculum; EM is the treatment inoculated with the commercially available inoculum EM.
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图 2 堆肥过程中pH的变化
注：CK为未接种菌剂的处理；CDS为接种CDS菌剂的处理；XY4为接种XY4菌剂的处理；EM为接种市售菌剂EM的处理。
Figure 2. Changes of pH during composting process
Note: CK is the treatment without inoculum; CDS is the treatment inoculated with CDS inoculum; XY4 is the treatment inoculated with XY4 inoculum; EM is the treatment inoculated with the commercially available inoculum EM.
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图 3 堆肥过程中总有机碳的变化
注：CK为未接种菌剂的处理；CDS为接种CDS菌剂的处理；XY4为接种XY4菌剂的处理；EM为接种市售菌剂EM的处理。
Figure 3. Changes of total organic carbon during composting process
Note: CK is the treatment without inoculum; CDS is the treatment inoculated with CDS inoculum; XY4 is the treatment inoculated with XY4 inoculum; EM is the treatment inoculated with the commercially available inoculum EM.
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图 4 堆肥中腐熟度指标、酶活性与腐殖酸含量的相关性分析和冗余分析
注：CK为未接种菌剂的处理；CDS为接种CDS菌剂的处理； XY4为接种XY4菌剂的处理；EM为接种市售菌剂EM的处理。图4a～d为相关性热图，*、**分别表示相关性达到0.05、0.01显著水平；图4e～h为冗余分析，连线长度代表环境因子与研究对象的相关性，线越长，影响越大，线条之间的夹角越小，相关性越大。TEM—温度；GI—种子发芽指数；TOC—总有机碳；HA—腐殖酸；FA—黄腐酸；URE—脲酶；ALP—碱性磷酸酶；SUR—蔗糖酶；CAT—过氧化氢酶。
Figure 4. Correlation and redundancy analysis of maturity indicators, enzyme activities and humic acid contents in compost
Note: CK is the treatment without inoculum; CDS is the treatment inoculated with CDS inoculum; XY4 is the treatment inoculated with XY4 inoculum; EM is the treatment inoculated with the commercially available inoculum EM. Fig. 4a–d are correlation analysis, * and ** indicate significant correlation at 0.05 and 0.01 levels. Fig. 4e–h are redundancy analysis, the longer the connecting line, the greater of the factor effect; the smaller the angle between the lines, the greater the correlation of factors. TEM—Temperature; GI—Germination index; TOC—Total organic carbon; HA—Humic acid; FA—Fulvic acid; URE—Urease; ALP—Alkaline phosphatase; SUR—Sucrase; CAT—Catalase.
表 1 堆肥过程中种子发芽指数的变化(%)
Table 1 Changes of germination index during composting process
堆肥时间 (d)
Composting timeCK CDS XY4 EM 1 32.9±5.0 cA 34.3±6.2 dA 31.2±5.3 dA 31.2±6.9 dA 12 60.4±8.8 bA 60.1±8.6 cA 62.4±7.1 cA 60.3±6.4 cA 28 73.8±7.4 bB 85.1±5.6 bAB 89.6±5.0 bA 78.7±7.0 bAB 36 103.2±7.2 aA 115.0±13.2 aA 120.5±5.7 aA 114.7±10.2 aA 注：CK为未接种菌剂的处理；CDS为接种CDS菌剂的处理； XY4为接种XY4菌剂的处理；EM为接种市售菌剂EM的处理。同列数据后不同小写表示相同处理不同天数间差异显著；同行数据后不同大写字母相同天数不同处理间差异显著 (P<0.05)。
Note: CK is the treatment without inoculum; CDS is the treatment inoculated with CDS inoculum; XY4 is the treatment inoculated with XY4 inoculum; EM is the treatment inoculated with the commercially available inoculum EM. Different lowercase letters after data in a column indicate significant difference among days for a treatment; different capital letters in a row indicate significant difference among treatments on the same day (P<0.05).
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表 2 堆肥过程中C/N的变化
Table 2 Changes of C/N during composting process
堆肥时间 (d)
Composting timeCK CDS XY4 EM 1 28.2±1.4 aA 26.6±2.1 aA 28.5±1.9 aA 27.8±0.8 aA 12 25.1±1.0 abA 24.2±1.5 abA 24.1±0.3 abA 25.7±0.3 aA 28 19.1±2.1 bA 18.3±1.0 bcA 19.2±1.7 bA 20.1±1.4 bA 36 19.1±0.4 bA 16.3±1.7 cAB 14.6±0.4 cB 15.4±0.2 cAB 注：CK为未接种菌剂的处理；CDS为接种CDS菌剂的处理； XY4为接种XY4菌剂的处理；EM为接种市售菌剂EM的处理。同列数据后不同小写字母表示相同处理不同天数间差异显著；同行数据后不同大写字母表示相同天数不同处理间差异显著 (P<0.05)。
Note: CK is the treatment without inoculum; CDS is the treatment inoculated with CDS inoculum; XY4 is the treatment inoculated with XY4 inoculum; EM is the treatment inoculated with the commercially available inoculum EM. Different lowercase letters after data in a column indicate significant difference among composting days under the same treatment; different capital letters in a row indicate significant difference among treatments in the same day (P<0.05).
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表 3 堆肥过程中腐殖酸和黄腐酸含量 (mg/g)的变化
Table 3 Changes of humic acid and fulvic acid contents during composting process
堆肥时间 (d)
Composting time腐殖酸 Humic acid 黄腐酸 Fulvic acid CK CDS XY4 EM CK CDS XY4 EM 1 20.6±1.4 bB 23.6±1.2 bA 24.2±1.5 bA 21.4±1.3 cB 15.7±1.4 aAB 17.5±1.0 aA 15.2±1.7 bB 16.0±1.3 bcA 12 22.9±2.0 abA 24.3±2.1 bA 24.7±2.3 bA 23.5±1.2 bcA 16.9±0.7 abC 19.3±0.9 abB 21.1±0.7 aA 20.1±1.6 aA 28 24.6±2.0 aA 25.5±1.2 abA 25.7±2.3 bA 25.2±1.8 bA 13.9±0.6 bcB 16.3±1.4 bcA 16.3±1.2 bA 17.5±1.7 bA 36 24.4±0.7 aB 28.0±2.0 aA 30.1±2.1 aA 31.1±3.4 aA 12.7±2.0 cB 15.4±1.4 cA 14.9±1.7 bA 14.0±0.9 cAB 注：CK为未接种菌剂的处理；CDS为接种CDS菌剂的处理； XY4为接种XY4菌剂的处理；EM为接种市售菌剂EM的处理。同列数据后不同小写字母表示相同处理不同天数间腐殖酸或黄腐酸含量差异显著；同行数据后不同大写字母表示相同天数不同处理间腐殖酸或黄腐酸含量差异显著 (P<0.05)。
Note: CK is the treatment without inoculum; CDS is the treatment inoculated with CDS inoculum; XY4 is the treatment inoculated with XY4 inoculum; EM is the treatment inoculated with the commercially available inoculum EM. Different lowercase letters after data in a column indicate significant difference in humic acid or fulvic acid content among composting days under the same treatment; different capital letters in a row indicate significant difference in humic acid or fulvic acid content among treatments in the same day (P<0.05).
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表 4 堆肥过程中脲酶和碱性磷酸酶活性[mg/(g·d)]的变化
Table 4 Changes in urease and alkaline phosphatase activities during composting process
堆肥时间 (d)
Composting time脲酶 Urease 碱性磷酸酶 Alkaline phosphatase CK CDS XY4 EM CK CDS XY4 EM 1 53.9±5.9 aA 59.9±3.8 aA 65.8±12.5 aA 58.2±15.1 aA 21.5±0.2 aD 42.2±0.6 aB 52.5±0.1 aA 38.6±0.04 aC 12 20.0±1.0 bA 22.8±4.7 bA 25.5±5.0 bA 22.4±1.5 bA 21.3±0.1 aD 40.0±0.1 bA 35.3±0.2 bC 38.5±0.2 aB 28 16.9±7.3 bB 19.0±1.0 bAB 28.0±5.6 bA 20.7±4.1 bAB 19.2±0.03 bC 32.1±0.2 cA 23.6±0.1 cB 18.2±0.03 bD 36 14.8±5.7 bA 10.7±2.8 cA 11.3±3.3 cA 10.1±4.2 bA 15.0±0.1 cC 19.1±0.2 dB 20.2±0.1 dA 14.8±0.1 cC 注：CK为未接种菌剂的处理；CDS为接种CDS菌剂的处理； XY4为接种XY4菌剂的处理；EM为接种市售菌剂EM的处理。同列数据后不同小写字母表示相同处理不同天数间脲酶或碱性磷酸酶活性差异显著；同行数据后不同大写字母表示相同天数不同处理间脲酶或碱性磷酸酶活性差异显著 (P<0.05)。
Note: CK is the treatment without inoculum; CDS is the treatment inoculated with CDS inoculum; XY4 is the treatment inoculated with XY4 inoculum; EM is the treatment inoculated with the commercially available inoculum EM. Different lowercase letters after data in a column indicate significant difference in urease or alkaline phosphatase activity among composting days under the same treatment; different capital letters in a row indicate significant difference in urease or alkaline phosphatase activity among treatments in the same day (P<0.05).
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表 5 堆肥过程中蔗糖酶和过氧化氢酶活性[mg/(g·d)]的变化
Table 5 Changes of sucrase and catalase activity during composting process
堆肥时间 (d)
Composting time蔗糖酶 Sucrase 过氧化氢酶 Catalase CK CDS XY4 EM CK CDS XY4 EM 1 74.3±0.4 aD 118.2±0.6 aA 98.6±0.6 aB 97.1±0.3 aC 5.3±0.4 bB 6.9±0.3 aA 7.4±0.2 aA 5.7±0.2 aB 12 54.4±0.5 bD 93.3±0.5 bB 97.6±0.1 bA 84.4±0.3 bC 2.3±0.1 cB 1.9±0.2 dC 3.2±0.2 dA 2.1±0.1 bC 28 47.3±0.1 cD 64.5±0.2 cA 56.2±0.1 cB 53.0±0.3 cC 5.7±0.4 bAB 6.1±0.2 bA 5.3±0.2 cB 5.3±0.5 aB 36 44.6±0.4 dAB 44.1±0.3 dB 41.7±0.2 dC 44.6±0.2 dA 6.4±0.3 aA 5.1±0.1 cB 5.0±0.1 bB 5.3±0.3 aB 注：CK为未接种菌剂的处理；CDS为接种CDS菌剂的处理； XY4为接种XY4菌剂的处理；EM为接种市售菌剂EM的处理。同列数据后不同小写字母表示相同处理不同天数间蔗糖酶或过氧化氢酶活性差异显著；同行数据后不同大写字母表示相同天数不同处理间蔗糖酶或过氧化氢酶活性差异显著 (P<0.05)。
Note: CK is the treatment without inoculum; CDS is the treatment inoculated with CDS inoculum; XY4 is the treatment inoculated with XY4 inoculum; EM is the treatment inoculated with the commercially available inoculum EM. Different lowercase letters after data in a column indicate significant difference in sucrase or catalase activity among composting days under the same treatment; different capital letters in a row indicate significant difference in sucrase or catalase activity among treatments in the same day (P<0.05).
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