耐高温亚硝酸盐降解菌的分离、鉴定及降低雪茄烟叶亚硝胺的应用

【背景】烟草特有亚硝胺(tobacco-specific nitrosamines, TSNAs)是烟草于调制和发酵阶段产生的一类致癌物质,由烟草生物碱与氮氧化物发生亚硝化反应生成,生物碱和亚硝酸盐是TSNAs的直接前体物质。【目的】发掘适用雪茄高温发酵且显著降低TSNAs形成与积累的微生物。【方法】以TSNAs前体物质亚硝酸盐的高效降解为目标,对从雪茄烟叶分离得到的烟草源微生物菌株进行高温培养、亚硝酸盐降解及亚硝酸盐耐受能力研究,得到可于50℃高效降解亚硝酸盐及耐受高浓度亚硝酸盐的微生物菌株,将菌株应用于雪茄烟叶高温发酵35 d,对发酵前后亚硝酸盐、TSNAs、常规化学成分和中性香味成分含量进行测定,分析菌株在雪茄烟叶发酵中对TSNAs含量及烟叶品质的影响。【结果】获得了3株于50℃高效降解亚硝酸盐的菌株NY7、NY8和NY9,分别鉴定为莫海威芽孢杆菌(Bacillus mojavensis) NY7、耐盐芽孢杆菌(Bacillus halotolerans) NY8和枯草芽孢杆菌(Bacillus subtilis) NY9,其中B. halotolerans NY8亚硝酸盐降解能...     

朱鹮肠道微生物多样性与产酶活性

【背景】朱鹮是我国国家一级保护动物,属于世界上最濒危的鸟类之一。对朱鹮肠道微生物的多样性和产胞外酶活性进行分析,可为朱鹮种群数量恢复提供思路。【目的】了解朱鹮肠道微生物的多样性,测定其产胞外酶活性。【方法】采用纯培养的方法获得朱鹮肠道微生物,通过革兰氏染色和生理生化鉴定,结合16S rRNA基因扩增和序列分析对细菌进行鉴定。使用水解圈法筛选产淀粉酶、蛋白酶、纤维素酶、脂肪酶的菌株。【结果】从人工喂养的朱鹮新鲜粪便中共分离到296株细菌,共计2个门11个属。变形菌门(Proteobacteria) 236株,占分离总数的79.73%,分别为：埃希氏菌属(Escherichia) 137株,占分离总数的46.28%;哈夫尼亚菌属(Hafnia) 39株,占分离总数的13.18%;变形菌属(Proteus) 28株,占分离总数的9.46%;柠檬酸杆菌属(Citrobacter) 23株,占分离总数的7.77%;气单胞菌属(Aeromonas) 6株,占分离总数的2.03%;肠杆菌属(Enterobacter) 1株,占分离总数的0.34%;志贺菌属(Shigella) 1株,占分离总数的0.34%;克雷伯菌属(Klebsiella) 1株,占分离总数的0.34%。厚壁菌门(Firmicutes) 60株,占分离总数的20.27%,分别为：肠球菌属(Enterococcus) 33株,占分离总数的11.15%;库特氏菌属(Kurthia) 14株,占分离总数的4.73%;芽孢杆菌属(Bacillus) 13株,占分离总数的4.39%。优势菌群为变形菌门(Proteobacteria)中的埃希氏菌属(Escherichia),占细菌总数的46.28%。经过生理生化鉴定,每个菌株生理生化鉴定出的种属与各自的16S rRNA基因鉴定出的种属相一致。产酶活力分析结果显示有238株产蛋白酶、25株产脂肪酶、24株产淀粉酶、15株产纤维素酶,分别占分离总数的80.41%、8.45%、8.11%和5.07%。【结论】朱鹮肠道微生物分离出的细菌可分为2门11属,优势菌群为变形菌门(Proteobacteria)中的埃希氏菌属(Escherichia),占细菌总数的46.28%;产酶活性分析显示,80.41%的菌株具有产蛋白酶能力。     

根际微生态调控白菜根肿病发生的机制研究进展

白菜根肿病是由芸薹根肿菌(Plasmodiophora brassicae Woron)引起的一种常见土传病害,主要危害白菜的根部。根际是土壤-植物-微生物相互作用最活跃的关键微域,根际微生态系统中的微生物失衡是导致土传病害的重要因素,深入探究根际微生态与土传病害互作机制,有利于从根际微生物、抑病物质和功能代谢等方面挖掘防控土传病害安全高效的方法。本文综述了根际微生态与白菜根肿病的发生机制关系,从该病害的危害、发生的根际微生态机制及生防菌防治研究等方面综合分析了根际微生物调控白菜根肿病发生的机制,以期为白菜根肿病防控、促进土壤健康和维持根际微生态系统稳定提供理论依据。     

松萝内生酿酒酵母菌株耐酸生理特性与分子机制探究

【目的】对我国西藏地区来源的不同酵母菌株进行有机酸发酵性能测试,此外,对具有良好产酸性能的分离自松萝内部的酿酒酵母菌株Saccharomy cescerevisiae 2-2进行耐酸性能分析,并探究其耐酸较强的分子机制。【方法】比较不同糖浓度培养基液体发酵培养过程中pH的变化,并比较低pH胁迫条件下菌株的生长,检测酿酒酵母菌株的产酸潜力和耐酸特性;对菌株2-2和模式酵母菌株S288C进行比较基因组分析,并利用实时荧光定量聚合酶链式反应(real-time fluorescence quantitative polymerase chain reaction,RT-qPCR)分析关键基因的转录,探究菌株2-2耐酸分子机制。【结果】松萝内生酿酒酵母2-2在所有检测的菌株中产酸潜力较大,耐酸性能较好。在菌株2-2中与胁迫耐受性相关的基因PDR15、PDR12和SUR1在低pH胁迫条件下存在显著的上调或下调,但这些基因转录变化趋势与菌株S288C相反。【结论】松萝内生酿酒酵母2-2是一株产酸耐酸性能较好的菌株,对其独特的调节机制进行深入分析,有希望选育性能更好的产酸酵母菌株。     

Light and electron microscopy studies of the midgut and salivary glands of second and third instars of the horse stomach bot,Gasterophilus intestinalis

A morphological study of the midgut and salivary glands of second and third instars of Gasterophilus intestinalis (De Geer) (Diptera: Oestridae) was conducted by light, scanning and transmission electron microscopy. The midgut is anteriorly delimited by a proventriculus, without caeca, and is composed of posterior foregut and anterior midgut tissue from which a double‐layered peritrophic matrix is produced. The midgut can be divided into anterior, median and posterior regions on the basis of the structural and physiological variations of the columnar cells which occur along its length. Two other types of cell were identified: regenerative cells scattered throughout the columnar cells, and, more rarely, endocrine cells of two structural types (closed and open). Different secretion mechanisms (merocrine, apocrine and microapocrine) occur along the midgut epithelium. Abundant microorganisms are observed in the endoperitrophic space of the anterior midgut. The origin and nature of these microorganisms remain unknown. No structural differences are observed between the second and third instar midguts. The salivary glands of G. intestinalis second and third instars consist of a pair of elongated tubular structures connected to efferent ducts which unite to form a single deferent duct linked dorsally to the pharynx. Several intermediate cells, without cuticle, make the junction with the salivary gland epithelium layer. Cytological characteristics of the gland epithelial cells demonstrate high cellular activity and some structural variations are noticed between the two larval stages.     

Microbial cycling of iron and sulfur in sediments of acidic and pH-neutral mining lakes in Lusatia (Brandenburg, Germany)

A vast number of lakes developed in the abandoned opencast lignite mines of Lusatia (East Germany) contain acidic waters (mmolSm^–2a^–). Potential Fe(III) reduction measured by the accumulation of Fe(II) during anoxic incubation yielded similar rates in both types of sediments, however, the responses towards the supplementation of Fe(III) and organic carbon were different. Sulfate reduction rates estimated with ³⁵S-radiotracer were much lower in the slightly acidic sediment than in the pH-neutral sediment (156 v.s. 738mmolSO₄ ^2–m^–2a^–1). However, sulfate reduction rates were increased by the addition of organic carbon. Severe limitation of sulfate-reducing bacteria under acidic conditions was also reflected by low most probable numbers (MPN). High MPN of acidophilic iron- and sulfur-oxidizing bacteria in acidic sediments indicated a high reoxidation potential. The results show that potentials for reductive processes are present in acidic sediments and that these are determined mainly by the availability of oxidants and organic matter.     

Effectiveness of phytoremediation as a secondary treatment for polycyclic aromatic hydrocarbons (PAHs) in composted soil

A greenhouse study was conducted over a 12-month period to investigate the fate of polycyclic aromatic hydrocarbons (PAHs) in soil using phytoremediation as a secondary treatment. The soil was pretreated by composting for 12 weeks, then planted with tall fescue (Festuca arundinacea), annual ryegrass (Lolium multiflorum), and yellow sweet clover (Melilotus officinalis). Two sets of unvegetated controls also were evaluated, one fertilized and one unfertilized. Total PAH concentrations decreased in the tall fescue, annual ryegrass, and yellow sweet clover treatments by 23.9%, 15.3%, and 9.1%, respectively, whereas the control was reduced by less than 5%. The smaller two- and most of the three-ringed compounds--naphthalene, acenaphthylene, acenaphthene, fluorene, and anthracene--were not found in detectable concentrations in any of the treatments. The most probable number analysis for microbial PAH degraders did not show any statistically significant differences among treatments. There were significant differences among treatments (p < 0.05) for the residual concentrations of five of the target PAHs. Root surface area measurements indicated that tall fescue and annual ryegrass both had significantly higher root surface area than yellow sweet clover, although the two species were not significantly different from each other. The tall fescue treatment resulted in the highest root and shoot biomass, followed by annual ryegrass and yellow sweet clover, and also had the highest percent of contaminant removal after 12 months. These results imply a positive relationship between plant biomass development and PAH biodegradation.     

Functional diversity and community structure of microorganisms in rhizosphere and non-rhizosphere Canadian arctic soils

Functional diversities of microorganisms in arctic soil samples at three incubation temperatures were assessed using sole-carbon-source-utilization (SCSU). Soil samples from four sites were collected from the rhizosphere and non-rhizosphere soils. Microorganisms were extracted from samples and inoculated into ECO-Biolog plates and incubated at 4, 10 and 28 °C. Calculations of Shannon–Weaver diversity and Shannon–Weaver evenness were based on the substrate utilization in the Biolog plates. Shannon–Weaver diversities (H) in rhizosphere samples were significantly greater ( _H = 3.023 ± 0.197; P < 0.005) than in non-rhizosphere samples ( _H = 2.770 ± 0.154). Similarly, the evenness (E) of the inoculated microbial cells exhibited significant differences (P < 0.005) between the rhizosphere and non-rhizosphere soil samples ( _E = 0.880 ± 0.057 for soils with rhizosphere; _E = 0.807 ± 0.044 for non-rhizosphere samples). Higher microbial diversity and evenness were observed in samples incubated at 4 °C than at 28 °C [least significant difference (lsd) = 0.29], and evenness indices were higher in rhizosphere samples than in non-rhizosphere soils incubated at all three temperatures (lsd = 0.02). Principal component analysis (PCA) of the multivariate data set differentiated the soil samples on the relatively gross scale of microbial communities isolated from rhizosphere and non-rhizosphere soils at all three temperatures.     

Lactones. 6. Microbial lactonization of gamma,delta-epoxy esters

The ability of 19 microorganisms to perform the enantioselective lactonization of racemic gamma,delta-epoxy ester 3a and its 7-methyl homolog 3b was checked. It was found that Rhodotorula rubra preferentialy transformed both substrates to (-)-enantiomers of trans delta-hydroxy-gamma-lactones with ee 76% for 3a and 24% for 3b. The best efficiency (20-30%) and enantioselectivity (ee 60-100%) of formation of (-)-gamma-hydroxy-delta-lactones 6a and 6b was observed for lactonization by Botrytis cinerea and Fusarium semitectum, respectively.     

Effect of electrical charges and fields on injury and viability of airborne bacteria

In this study, the effects of the electric charges and fields on the viability of airborne microorganisms were investigated. The electric charges of different magnitude and polarity were imparted on airborne microbial cells by a means of induction charging. The airborne microorganisms carrying different electric charge levels were then extracted by an electric mobility analyzer and collected using a microbial sampler. It was found that the viability of Pseudomonas fluorescens bacteria, used as a model for sensitive bacteria, carrying a net charge from 4100 negative to 30 positive elementary charges ranged between 40% and 60%; the viability of the cells carrying >2700 positive charges was below 1.5%. In contrast, the viability of the stress-resistant spores of Bacillus subtilis var. niger (used as simulant of anthrax-causing Bacillus anthracis spores when testing bioaerosol sensors in various studies), was not affected by the amount of electric charges on the spores. Because bacterial cells depend on their membrane potential for basic metabolic activities, drastic changes occurring in the membrane potential during aerosolization and the local electric fields induced by the imposed charges appeared to affect the sensitive cells' viability. These findings facilitate applications of electric charging for environmental control purposes involving sterilization of bacterial cells by imposing high electric charges on them. The findings from this study can also be used in the development of new bioaerosol sampling methods based on electrostatic principles.