纯种强化发酵细菌型水豆豉细菌菌群动态研究

采用聚合酶链式反应-变性梯度凝胶电泳（PCR-DGGE）技术对利用枯草芽孢杆菌（Bacillus subtilis）BJ3-2强化发酵的细菌型水豆豉前发酵及后发酵过程中的细菌菌群种类及数量进行动态研究。结果表明,前发酵42 h后,细菌种类及数量丰富并趋于稳定,后发酵第5天,细菌种类达到最大。整个发酵期间共鉴定出6个属15个种,包括芽孢杆菌属（Bacillus）、沙雷菌属（Serratia）、产碱菌属（Alcaligenes）、肠球菌属（Enterococcus）、不动杆菌属（Acinetobacter）及变形杆菌属（Proteus）。共检测到6种条件致病菌,分别为粘质沙雷菌（Serratia marcescens）、粪产碱菌（Alcaligenes faecalis）、蜡样芽孢杆菌（Bacillus cereus）、奇异变形杆菌（Proteus mirabilis）、鲍氏不动杆菌（Acinetobacter baumannii）、屎肠球菌（Enterococcus faecium）,其中,奇异变形杆菌首次报道存在于贵州纯种发酵细菌型水豆豉中。     

有机碳源对SNAD工艺脱氮性能及微生物种群结构的影响

为考察不同有机碳源质量浓度对亚硝化的全程自养脱氮工艺(SNAD)脱氮性能的影响,将该工艺应用到生活污水的处理中,采用MBR反应器,以葡萄糖作为有机物来源,通过逐步增大COD来实现,并运用PCR-DGGE技术研究了微生物种群结构的变化.反应器运行结果和DGGE图谱分析表明:碳氮比为0~2时,COD的增加不会抑制AOB和Anammox菌,AOB和Anammox菌的菌属种类不受影响,反而通过反硝化作用提高氮去除负荷.总氮去除率和氮去除负荷分别为67%和0.34 kg/(m3·d)左右.碳氮比为3~4及生活污水运行条件下,Anammox菌不受影响,AOB的活性受到抑制,菌属种类减少,脱氮效率下降.生活污水运行阶段,总氮去除率和氮去除负荷平均分别为73%和0.17 kg/(m3·d).Nitrosomonas和Candidatus Kuenenia stuttgartiensis一直是反应器内的优势菌属,共同完成脱氮过程.     

苯胺废水SBR工艺生物强化处理效能

应用生物强化技术,将实验室筛选出的高效苯胺降解菌投加到SBR反应器中,考察其经生物强化后对苯胺的去除效果,COD、氨氮、亚硝氮、硝氮的变化及微生物群落的演替规律.结果表明,生物强化系统可有效实现对苯胺的降解,系统达到稳定运行后,苯胺降解率为100%,COD的去除率为93%左右,氨氮的浓度随苯胺降解而升高.因水力滞留时间较短,并未驯化出亚硝化、硝化细菌.扫描电镜照片可以看出,系统稳定运行后,苯胺降解菌遍布于活性污泥上.此外,由PCR-DGGE图谱及相似性矩阵图可见,经16周期的运行,微生物群落种类、数量产生了明显的差异性,系统内微生物优势种群得到强化.     

城市污水SBR法短程生物脱氮系统硝化菌群的定量分析

为了研究实时控制对短程生物脱氮中试系统内硝化菌群结构组成的影响,在考察系统常温条件下处理城市污水时短程硝化效果的同时,采用FISH、PCR-DGGE和PCR-克隆序列(Cloning Sequencing)分子生物学方法对SBR中试系统(有效容积为54 m³)硝化菌群中的氨氧化菌(AOB)和亚硝酸氧化菌(NOB)进行定性与定量化分析.FISH结果表明,在短程脱氮系统中,AOB相比于NOB已成为明显的优势菌群,占总菌群的3%~12%;且没有检测出NOB.PCR-DGGE结果表明,SBR短程脱氮中试系统中的AOB均以Nitrosomonas-like为主.污泥样品的PCR-Cloning-Sequencing结果表明,所有的克隆相似于Nitrosomonas,其中60%以上的克隆相似于Nitro- somonas europaea.     

运用PCR-DGGE和克隆技术对串联附积床系统生物膜菌群的分析

采用PCR-DGGE﹑克隆等分子生物学手段研究了多级串联附积床同时硝化反硝化脱氮系统生物膜菌群的时间演变,并对生物膜菌群进行同源性分析和系统发育树构建,同时讨论了生物膜菌群对系统中有机物的去除及对同时硝化反硝化脱氮的贡献.结果表明,随着时间的推移,生物膜菌群发生了较大演变而且具有高度多样性.对DGGE图谱优势条带进行分析表明,优势菌群分为5个不同的细菌类群:β-变形菌类群(β-proteobacteria)﹑γ-变形菌类群(γ-proteobacteria)、未分类菌类群(unclassified bacteria)、α-变形菌类群(α-proteobacteria)、放线菌类群(Actinobacteria).β-变形菌类群不仅在数量上占有优势,而且在有机物的降解、营养物质的去除中起着重要作用.生物膜细菌中起硝化作用的主要是亚硝化单胞菌和硝化螺旋菌;起反硝化作用的主要是施氏假单胞菌.     

郫县豆瓣自然发酵过程中真菌群落的演替及黄曲霉毒素B1的消长

以四川省郫县豆瓣为研究对象,运用PCR-DGGE指纹图谱技术研究豆瓣自然发酵过程中真菌群落的演替,采用酶联免疫吸附法(ELISA)检测豆瓣样品中黄曲霉毒素B1(AFB1)含量。结果表明:郫县豆瓣自然发酵过程中的优势真菌有米曲霉(Aspergillus oryzae)、淀粉丝菌(Amylomyces rouxii)、米根霉(Rhizopus oryzae)、异常毕赤酵母(Pichiaanomala)、汉逊德巴利酵母(Debaryomyces hansenii)的近缘种,其中米曲霉是制曲阶段和发酵初期的优势真菌,淀粉丝菌和米根霉是制曲阶段的优势真菌,异常毕赤酵母和汉逊德巴利酵母是发酵初期的优势真菌。各发酵时间点样品的AFB1含量均低于国标(5μg/kg);在制曲和发酵初期阶段,AFB1的含量均不断增加,但在第14天后趋于稳定。因此,对AFB1的含量生物防治可重点集中在制曲阶段。     

Effect of grape indigenous Saccharomyces cerevisiae strains on Montepulciano d'Abruzzo red wine quality

The growth of selected, indigenous Saccharomyces cerevisiae added as starters (SRS1, MS72 and RT73) was monitored during Montepulciano d'Abruzzo wine production. In all the fermentations the addition of the starter, caused a decrease of the non-Saccharomyces yeasts. When strains MS72 and RT73 were used as starters they were detected in the first phases of fermentations, while strain SRS1 competed successfully with native yeasts during all the process. Wines obtained by fermentation with the indigenous starters showed some different characteristics, according to the chemical and sensory analyses. This study highlighted that among selected starters with high fermentative capacity, some are able to dominate better than other natural wine yeast biota, whereas some strains can interact and survive besides native yeast populations during the fermentation. As a consequence, the dominance character can have a positive or negative effect on wine quality and has to be considered in the frame of yeast selection in order to improve or characterize traditional wines. Winemakers could choose among different degrees of yeast dominance to modulate the interaction among starter and native wine yeast population.     

肉鸡屠宰过程中腐败微生物的污染情况分析

采用PCR-DGGE指纹图谱技术研究肉鸡屠宰加工过程中微生物的污染情况,以确定关键控制点。分别采取肉鸡屠宰加工过程中胴体表面、工人手表面、预冷池中的水、空气中的样品和真空包装贮藏过程中样品,然后进行PCR-DGGE指纹图谱分析。结果表明:肉鸡的屠宰加工过程包括浸烫脱毛、倒挂、去内脏、冷却和分割,不同处理会对胴体表面的污染微生物产生不同的影响,去内脏后肉鸡胴体表面微生物的种类和数量都有所增加,但预冷能有效减少肉鸡胴体表面微生物的种类和数量,空气中微生物数量随着生产时间的延长不断增加,这些操作点也会成为交叉污染的地方,尤其工人的手表面的污染是引起交叉污染最主要的原因。     

Comparative assessment of decoupling of biomass and hydraulic retention times in hydrogen production bioreactors

This study compared biological hydrogen production from glucose in two continuously stirred tank reactors (CSTRs) and two integrated biohydrogen reactor clarifier systems (IBRCSs) comprising CSTRs with gravity settlers to decouple the hydraulic retention time (HRT) from solids retention time (SRT). The four systems were operated at organic loading rates of 6.5–42.8 gCOD/L-d, and HRTs of 8–12 h. The SRT was maintained at 2 days in the two IBRCSs. The decoupling of SRT from HRT not only increased glucose conversion from 29–50% in the CSTR to 99.9% in the IBRCSs, but also the volumetric hydrogen production from 0.55–1.8 in the CSTRs to 2.4–9.6 L/L-d. Biomass yields in the two IBRCSs were 0.09 and 0.13 g VSS/g glucose converted, about 50% lower than the CSTR yields of 0.19 and 0.29 g VSS/g glucose converted. Hydrogen yield increased from 0.5–1.0 mol H₂/mol glucose converted in the CSTR to 2.8 mol H₂/mol glucose converted in the IBRCSs. The inverse relationship between hydrogen yield and biomass yield observed in this study implies that the hydrogen yield is maximized with the minimization of biomass yield, thus necessitating decoupling of SRT from HRT to ensure sufficient reactor biomass. DGGE analysis confirmed the specificity of the microbial culture in the IBRCSs with the high-hydrogen producing Clostridium species, as compared to the more diverse cultures in the CSTR.