倭蜂猴粪便微生物苯酚羟化酶和邻苯二酚1,2-双加氧酶基因多样性研究

【目的】分析倭蜂猴粪便微生物中苯酚羟化酶(Phenol hydroxylase,PH)和邻苯二酚1,2-双加氧酶(Catechol 1,2-dioxygenase,C12O)的基因多样性。【方法】利用简并引物,以倭蜂猴粪便微生物宏基因组DNA为模板,通过PCR扩增,分别构建PH和C12O基因克隆文库,并对克隆进行测序分析。【结果】倭蜂猴粪便微生物来源的PH和C12O基因序列经BLAST比对分析,与GenBank中相应酶的序列一致性分别介于92%?100%和87%?100%。系统进化树分析表明PH基因序列与Neisseria、Burkholderia、Alcaligenes、Acinetobacter 4个属来源的PH序列相关;C12O基因序列全部与Acinetobacter来源的C12O序列相关。序列比对结果表明PH序列具有LmPH (Largest subunit of multicomponent PH)中高保守的两个DEXRH结构域;C12O序列具有能被Ag+和Hg2+抑制的位点(半胱氨酸)。【结论】倭蜂猴粪便微生物来源的PH为多组分PH,其降解苯酚的中间产物邻苯二酚可以被C12O通过邻位开环途径裂解。     

应用定量微生物风险评估海水浴场人体健康风险

【背景】定量微生物风险评估作为定量评估游泳人群暴露于病原微生物后健康风险的方法,在国外已得到广泛应用,但目前国内的应用处于起步阶段且缺乏所需的游泳人群暴露数据。【目的】收集游泳人群暴露数据,并在海水浴场中进行应用,评估粪大肠菌群作为风险评估指标的可行性。【方法】通过对6个典型海水浴场的水质状况、粪大肠菌群浓度与环境因子的相关性进行分析,并发放调查问卷收集国内游泳人群的暴露数据,进而应用定量微生物风险评估方法,得出各个海水浴场的胃肠道疾病患病风险。【结果】6个海水浴场中粪大肠菌群浓度均与水温、气温及总云量具有显著相关性(P<0.01)。位于南方的海水浴场粪便污染情况较北方严重,粪大肠菌群浓度第95百分位数远高于国内“差”类水质标准的阈值。儿童、成年男性、成年女性单次沐浴事件吞下海水的体积分别为35.1 mL (95%置信区间为32.4-37.8,α=0.578,β=0.016),45.0 mL (95%置信区间为31.1-59.3,α=0.532,β=0.012),35.7 mL (95%置信区间为29.7-41.8,α=0.753,β=0.032)。6个海水浴场患胃肠道疾病的风险...     

Recent advances in polysaccharide bio-based flocculants

Natural polysaccharides, derived from biomass feedstocks, marine resources, and microorganisms, have been attracting considerable attention as benign and environmentally friendly substitutes for synthetic polymeric products. Besides many other applications, these biopolymers are rapidly emerging as viable alternatives to harmful synthetic flocculating agents for the removal of contaminants from water and wastewater. In recent years, a great deal of effort has been devoted to improve the production and performance of polysaccharide bio-based flocculants. In this review, current trends in preparation and chemical modification of polysaccharide bio-based flocculants and their flocculation performance are discussed. Aspects including mechanisms of flocculation, biosynthesis, classification, purification and characterization, chemical modification, the effect of physicochemical factors on flocculating activity, and recent applications of polysaccharide bio-based flocculants are summarized and presented.     

Fecal antibody levels as a noninvasive method for measuring immunity to gastrointestinal nematodes in ecological studies

Among‐individual variation in antibody‐associated immunity to gastrointestinal nematode parasites (GIN) is known be associated with life‐history traits and vital rates in wild vertebrate systems. To date, measurement of levels of antibodies against GIN antigens in natural populations has exclusively been based on invasive blood sampling techniques. Previous work in laboratory rodents and ruminant livestock suggests that antibody measures from feces may provide a viable noninvasive approach. We measured total and anti‐GIN antibodies of different isotypes (immunoglobulin (Ig) G, IgA and IgE) from paired samples of plasma and feces from free‐living Soay sheep of different ages and sexes. We tested the correlations among these measures as well as their associations with body mass and Strongyle nematode fecal egg counts (FEC). Significant positive correlations were present among plasma and fecal anti‐GIN antibody levels for IgG and IgA. Generally, correlations between total antibody levels in plasma and feces were weaker and not significant. No significant relationships were found between any antibody measures and body mass; however, fecal anti‐GIN antibody levels were significantly negatively correlated with FEC. Our data clearly demonstrate the feasibility of measuring anti‐GIN antibodies from fecal samples collected in natural populations. Although associations of fecal antibody levels with their plasma counterparts and FEC were relatively weak, the presence of significant correlations in the predicted direction in a relatively small and heterogeneous sample suggests fecal antibody measures could be a useful, noninvasive addition to current eco‐immunological studies.     

Multiple antibiotic resistance indexing of coliforms to identify high risk contamination sites in aquatic environment

Bacteriological analysis of the water samples collected from upstream, midstream and downstream points along the bank of the river revealed high populations of Escherichia coli, Citrobacter freundii, Citrobacter diversus, Enterobacter aerogens and Klebsiella species. All these isolates were screened against eight antibiotics to determine the prevalence of multiple antibiotic resistance among isolates at different sites of the river. The study revealed that multiple antibiotic resistance was prominently seen in coliforms at downstream sites (Average multiple antibiotic resistance index, MAR Index = 0.43) while it was low in coliforms at upstream sites (MAR Index = 0.15). These differences in MAR indices provide a method for distinguishing high risk contamination sites in aquatic environment.     

Rapid multiplex PCR based species identification of wild tigers using non-invasive samples

Conservation and management of rare and elusive species requires accurate data on presence or absence. In such cases, molecular genetics based species identification approaches can prove invaluable, especially in conjuncture with non-invasive DNA sampling. However, non-invasive sources yield DNA in low concentration that is degraded, which could result in false negatives for species identification. In this paper, we developed a set of primers for PCR-based species identification of tigers. Our results reveal high rates (upto 90%) of species identification for both fresh (less than 48 h) and old (between 7 days and 3 months) fecal samples from the field. Experiments reveal that multiplex PCR (amplifying more than one genomic region) results in an increase in conclusive species identification (and a consequent decrease in the number of false negatives) from 55% to 89% for old fecal samples. We demonstrate that this increased success is because we experimentally overcome the problems of low DNA template quantity (using the multiplex PCR kit, increases species identification from 55% to 72%) and low template DNA quality (two sets of primers increase the species identification success from 72% to 89%). We recommend that multiplex PCR based methods be used (in conjuncture with species specific primers) for other rare and elusive species since such methods will potentially significantly decrease error in species identification.     

Characterization of bacterial coliform occurrences in different zones of a drinking water distribution system

AIMS: To compare the bacterial coliforms detected from occurrences in three zones of a water distribution system supplied by two separate water sources. METHODS AND RESULTS: Conventional and standardized protocols for identifying enterobacterial populations were applied. Additional tests to confirm isolates were included. Analyses of diversity and population similarity were performed using the Phene Plate System, a miniaturized biochemical phenotyping method. Isolates were identified by the API 20E system in tandem with biochemical phenotyping. A total of 16 576 samples were taken from the water distribution system, with 1416 isolates analysed. A low number of coliform occurrences were observed (2%). Escherichia coli was not detected in either water origin or in Zone 2 samples; however, in Zones 1 and 3 a low number of cases of E. coli were recorded. The percentages of E. coli depended on the identification criteria. Eight biochemical profiles for coliform populations were defined according to the results of the confirmative tests. There was a high diversity among these populations in the three zones studied, although no significant variations in their composition (associated with occurrences in the different zones) were observed. Klebsiella oxytoca was the most commonly detected species irrespective of zone, although seven other enterobacterial genera were also found. CONCLUSIONS: Analysis of the enzymatic activity of beta-glucuronidase or application of the criteria established in the norm ISO 9308-1, in tandem with thermotolerance was needed to evaluate the occurrence of E. coli in the distribution systems. Detected occurrences of bacterial coliforms could be associated with re-growth patterns for specific sampling points in the distribution system. Seasonal differences, independent of the studied zones, were observed. SIGNIFICANCE AND IMPACT OF THE STUDY: Biochemical phenotyping of bacterial coliforms was shown to be a useful method on the characterization of occurrences in water distribution systems.     

粪便提取液毒性检测方法的建立及应用

目的建立检测人体粪便提取液对肠上皮细胞毒性的方法。方法以Caco-2细胞为体外模型,应用建立好的毒性检测方法：用MTT法（四甲基偶氮唑盐微量酶反应比色法）检测细胞毒性,用单细胞凝胶电泳技术检测遗传毒性,对糖尿患者和健康人的粪便提取液进行分析。结果糖尿病患者粪便提取液的细胞毒性显著高于健康人（n=30,P〈0.05）;其遗传毒性显著高于阴性对照PBS（P〈0.05）,与健康人相比有升高趋势,但差异无统计学意义。结论粪便提取液的毒性检测方法可以简便、有效地检测人体粪便中毒性物质对肠上皮细胞的影响,因此可作为评价肠道菌群状态的方法。     

高密度种群的社会条件对大沙鼠幼年雄鼠形态生理特征的影响:一个实例研究

在非繁殖期高密度大沙鼠(RhombomysopimusLicht)种群中,我们研究了胁迫以及幼年雄鼠的性激素浓度、腹中腺大小和体重对社群中成体(≥1年龄)存在的依赖性。用无损伤放射免疫方法,测定了于1999年秋天在野外采集的幼年雄鼠粪便样品的皮质酮和睾酮。有成体社群中的个体大于无成体社群的个体。因此,成体对幼体的影响不能独立于社群大小本身。成体的存在促进粪便睾酮的浓度和抑制了幼年雄鼠的性成熟(由雄激素依赖的较小的腹中腺来估计),但是同时促进了其生长。因此,自然存在的社群环境能影响幼年雄鼠的形态生理特征发育。     

Evaluation of the methods for enumerating coliform bacteria from water samples using precise reference standards

AIMS: To use BioBall cultures as a precise reference standard to evaluate methods for enumeration of Escherichia coli and other coliform bacteria in water samples. METHODS AND RESULTS: Eight methods were evaluated including membrane filtration, standard plate count (pour and spread plate methods), defined substrate technology methods (Colilert and Colisure), the most probable number method and the Petrifilm disposable plate method. Escherichia coli and Enterobacter aerogenes BioBall cultures containing 30 organisms each were used. All tests were performed using 10 replicates. The mean recovery of both bacteria varied with the different methods employed. CONCLUSIONS: The best and most consistent results were obtained with Petrifilm and the pour plate method. Other methods either yielded a low recovery or showed significantly high variability between replicates. SIGNIFICANCE AND IMPACT OF THE STUDY: The BioBall is a very suitable quality control tool for evaluating the efficiency of methods for bacterial enumeration in water samples.