黑水虻幼虫处理餐厨垃圾过程中体内养分组成与消化酶活性变化规律的研究

黑水虻Hermetia illucens作为一种新型资源环境昆虫,其幼虫可以处理餐厨垃圾、畜禽粪便、蔬菜残体等各种有机废弃物。幼虫富含蛋白质和油脂,可以作为水产饲料的蛋白来源。本研究探索黑水虻幼虫处理餐厨垃圾过程中其养分组成与消化酶活性变化之间的关系。通过黑水虻幼虫自由取食餐厨垃圾,每日采集样品用于物质养分和消化酶活性的测定。结果显示：黑水虻幼虫粗蛋白含量呈现先下降后上升的变化规律,而幼虫总糖含量呈现先上升后下降的变化规律,粗脂肪含量维持上升的趋势。幼虫处理餐厨垃圾过程中,体内蛋白酶在初期迅速上升,第6天后逐渐下降,而淀粉酶呈现出先缓慢上升再迅速上升最后下降的变化规律。脂肪酶在黑水虻幼虫处理餐厨垃圾前期保持较高的活性然后缓慢下降。同时,通过相关性分析,黑水虻幼虫粗蛋白含量变化与蛋白酶活性没有相关性,而总糖与粗脂肪含量变化分别与淀粉酶、脂肪酶有相关性。因此,部分消化酶活性变化与黑水虻幼虫养分组成具有一定的联系,本研究结果为工厂化养殖黑水虻提供一定的理论基础。     

亮斑扁角水虻卵共生菌Bacillus velezensis的鉴定和产酶特征及其对幼虫分解餐厨垃圾效率的影响

[目的] 从亮斑扁角水虻（Hermetia illucens L.）卵表筛选得到一株产多种酶的卵表共生菌,对该菌进行鉴定,并探究其最适生长条件、产酶特性及其对幼虫分解餐厨垃圾效率的影响。[方法] 通过多种选择性培养基筛选得到产多种酶的菌株。通过单因素实验方法确定其最适生长条件、产酶特性及其对幼虫分解餐厨垃圾效率的影响。[结果] 经过形态学观察、生理生化鉴定和16S rDNA序列分析,将该株亮斑扁角水虻卵表共生菌命名为贝莱斯芽胞杆菌（Bacillus velezensis EEAM 10B）。最适摇瓶培养条件为：40℃,200 r/min,pH 7.0,酵母浸粉10 g/L,葡萄糖10 g/L,培养16 h活菌数达3.1×10⁹ CFU/mL。进入稳定期后开始形成单端生芽胞,24 h后芽胞形成率95.8%。使用产酶筛选培养基培养结果表明：B.velezensis EEAM 10B菌株产木聚糖酶活性最强,其次是蛋白酶、纤维素酶、果胶酶、淀粉酶和植酸酶。按照1×10⁶ CFU/g用量添加B.velezensis EEAM 10B芽胞制剂到餐厨垃圾中饲养亮斑扁角水虻,B.velezensis EEAM 10B芽胞菌剂能够显著（P<0.05）提高亮斑扁角水虻幼虫对灭菌和非灭菌餐厨垃圾的转化效率,分别为13.4%和13.54%,但物料减少率没有显著差异（P>0.05）;显著提高灭菌餐厨垃圾中幼虫存活率至95%,提高非灭菌餐厨垃圾饲养幼虫的预蛹单重0.1437 g/只,化蛹率92.57%。[结论] B.velezensis EEAM 10B菌株能够产多种酶,且在亮斑扁角水虻处理餐厨废弃物中有潜在应用价值。     

虻粪二次堆肥生化特性的动态变化趋势及其微生物演替规律

[目的] 通过培养试验动态观察了虻粪在后续二次堆肥过程中生化特性的动态变化趋势,研究了堆体微生物生态的动态演替规律,讨论了虻粪二次堆肥的稳定化机理。[方法] 采取盛有虻粪的圆桶埋入大条垛发酵堆体的动态培养试验,在为期49 d的测试期内间隔7 d动态采样,采用16S rRNA高通量测序开展虻粪堆体样品的微生物多样性与群落结构分析以及代谢功能预测。[结果] 虻粪堆体经历2周快速升温至68.7℃后,逐步降温至第49天的37.7℃,总有机碳、溶解性有机质（DOM）、pH分别达到67.3%、18.4 g/kg、8.0。在堆体熟腐过程中,Shannon指数、Simpson指数和Chao1指数等微生物多样性水平均随堆肥进程而快速增加,而后缓慢下降至结束时的7.45、0.98和985。16S rRNA测序发现,Bacteroidetes（拟杆菌门）、Firmicutes（厚壁菌门）、Proteobacteria（变形菌门）与Actinobacteria（放线菌门）等是堆体微生物群落的主要菌群。与此同时,Corynebacteriaceae（棒状杆菌科）从初始的35.7%下降至结束时的6.5%,Flavobacteriaceae（黄杆菌科）则从3.22%逐步增长至12.6%,Porphyromonadaceae（单胞菌科）快速增加至11.5%之后缓慢下降至与初始水平相当的5.1%。PICRUSt预测发现,在整个虻粪堆肥过程中,氨基糖和核苷酸糖代谢、碳固定通路、甲烷代谢等功能平均相对丰度保持较强水平,且均在堆肥中期（第15–35天）出现峰值,后续均呈现下降态势。脂肪代谢、不饱和脂肪酸生物合成代谢、多环芳烃降解以及硫代谢等平均相对丰度较弱的代谢功能呈现先降后升的特征。[结论] 虻粪二次堆肥过程中,物料含水量与DOM快速下降、有机质降解逐步完成以及物料微生物结构与功能趋于稳定等,堆制腐熟后的虻粪生化性质稳定,有利于虻粪有机肥农田再利用。     

应用HPLC一反相色谱法测定米根霉乳酸发酵液中的有机酸

提出了一种利用高效液相色谱法分析米根霉乳酸发酵液中有机酸的方法,应用反相Wakosil-II 5C18RS色谱柱,以0．01mol/L磷酸（pH2\5)作为流动相,发酵液经稀硫酸预处理后直接进样分离定量,在10min内把其中的乳酸,苹果酸,富马酸等完全分离定量,各种酸回收率大于97％。经多次实验结果证明：本方法是测定乳酸发酵液中各有机酸的快速,有效的定量测定方法。     

The structure and physical properties of glucuronic acid oligomers produced by a Gluconacetobacter hansenii strain using the waste from beer fermentation broth

The aim of this work was to study the chemical structure and physical properties of water-soluble oligosaccharides (WSOS) produced by Gluconacetobacter hansenii PJK using the waste from beer fermentation broth as a basal medium. The analysis of the hydrolyzed products and the spectroscopic studies of the native WSOS showed that it is a mixture of oligomers all having a single sugar -linked glucuronic acid as building blocks with an O-acetyl and O-methyl group, in the terminal unit of the non-reducing end. The thermal studies displayed a progressive degradation of WSOS without a major weight loss throughout a range of temperatures. The melting point and pyrolysis temperature were found to be 130.16 and 275.25 °C, respectively. The optimum concentration of WSOS for a maximum emulsifying ability was found to be 0.10% (w/v). The resulting emulsions, however, did not demonstrate a noteworthy stability.     

Improvement of phage defence in Lactococcus lactis by introduction of the plasmid encoded restriction and modification system LlaAI

AIMS: To study the ability of the plasmid-encoded restriction and modification (R/M) system LlaAI to function as a bacteriophage resistance mechanism in Lactococcus lactis during milk fermentations. METHODS AND RESULTS: Plasmid pAIcat4, carrying the R/M system LlaAI and a chloramphenicol resistance cassette, was introduced into the plasmid-free strain L. lactis MG1614 and the industrial strain L. lactis 964. By measuring changes in conductivity the influence of different phage on the growth was determined. CONCLUSIONS: The plasmid-encoded R/M system LlaAI significantly improves the bacteriophage resistance of L. lactis during milk fermentations. SIGNIFICANCE AND IMPACT OF THE STUDY: It is essential to determine the potential of a phage defence mechanism in L. lactis starter culture strains during growth in milk before steps are taken to improve starter cultures. This study shows that LlaAI is useful for improvement of starter cultures.