摘要
摘要:发酵床养猪是一种新型的养殖技术，可有效缓解养猪的环境污染问题，微生物在其中起关键作用。为明确养猪发酵床发酵过程微生物群落的变化规律，为发酵床的科学管理提供依据，本研究采用磷脂脂肪酸生物标记（phospholipid fatty acids，PLFA）法分析养猪发酵床不同发酵等级垫料的微生物群落结构特征。采用色差法将垫料分为3个发酵程度等级：1级、2级和3级，采集不同发酵等级表层（0~15 cm）和里层（30~45 cm）垫料样本，测定各样本的PLFA。结果表明，共检测到61种PLFA，发酵2级垫料的PLFA种类最多，发酵3级垫料的PLFA种类最少。在各垫料中，PLFA分布量均表现为细菌>真菌>放线菌。指示细菌、真菌、放线菌、革兰氏阳性细菌（G⁺）、革兰氏阴性细菌（G^-）的PLFA及总PLFA在各发酵等级表层垫料的分布量均显著大于其在里层垫料的分布量，最大值出现在发酵1级表层垫料中。与对照（未发酵垫料）相比，发酵垫料总PLFA含量均显著增加（P < 0.05）。发酵3级表层垫料的真菌/细菌值最大，发酵2级表层垫料的G⁺/G^-值最大。多样性分析表明，Shannon指数和Pielou指数最大值出现在发酵2级垫料中，而Simpson指数最大值出现在发酵3级表层垫料中。聚类分析表明，当欧氏距离为233.15时，可将不同发酵等级垫料聚为3个类群，同一发酵级别的垫料聚在相同类群中；主成分分析表明，发酵1级表层和里层垫料单独归一类群，其他发酵等级垫料和对照垫料归另一类群中。综上，不同发酵等级垫料的微生物种群结构不同，发酵1级表层垫料微生物分布量最大，发酵2级垫料的微生物种类最多，相同发酵级别表层和里层垫料微生物群落结构相似。
Abstract:Pig-on-litter system is a new pig-raising technology that can reduce environmental pollution. In this system, micro-organisms are generally considered as the key factor. In order to determine the change in microbial community during fermentation and to set up basic data for scientific management of pig-on-litter system, microbial community of litter with different fermentation levels was analyzed using phospholipid fatty acid (PLFA) biomarkers. Fermentation levels of litters were divided into three grades (1^st, 2^nd and 3^rd) using the chromatic aberration (△E) method. Both surface (0-15 cm) and inner layer (30-45 cm) litters of each fermentation level were sampled. PLFA composition of each sample was determined by the Sherlock MIS 4.5 system. The results showed that the method used was sufficient to detect total 61 kinds of PLFA biomarkers. The most and least kinds of PLFA biomarkers occurred in litters with the 2^nd and 3^rd fermentation levels, respectively. PLFA biomarkers displayed the same order of distribution abundance in all samples-bacteria > fungi > actinomycetes. The contents of PLFAs that were referable to bacteria, fungi, actinomycetes, G⁺, G^- and total PLFA in the surface layer samples were all higher than those in the inner layer samples; being highest in surface layer litters with the 1^st fermentation level. Fermented litter had significantly higher content of total PLFA than unfermented litter (CK) (P < 0.05). The highest fungi/bacteria and G⁺/G^- ratios were for surface layer litter with the 3^rd and 2^nd fermentation levels, respectively. Diversity analyses indicated that the maximum values of the Shannon index and Pielou index were for litter of the 2^nd fermentation level and the maximum values of the Simpson index were for surface layer litter of the 3^rd fermentation level. Based on cluster analysis, the samples were clustered into three groups for Euclidean-distance of 223.15. Samples with the same fermentation level were clustered together. Also based on principal component analysis, surface layer and inner layer samples of the 1^st fermentation level were clustered into one lone group, while the other samples were clustered into other several groups. Put together, litters with different fermentation levels had different microbial community structures. The maximum values of microbial content and species were in surface layer litter with the 1^st and 2^nd fermentation levels, respectively. Moreover, surface layer and inner layer litters of the same fermentation level had similar microbial communities.



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