酒、盐度及酸碱度对醉泥螺中副溶血性弧菌生长的影响

醉泥螺中的副溶血性弧菌对人有较高的致病风险,为了提高醉泥螺的食用安全性,本文对其腌制工艺进行了优化,探讨了酒、盐度和酸碱度三个因素对醉泥螺中副溶血性弧菌生长状况的影响,并用描述性检验对盐度不同的醉泥螺进行了感官评价。研究结果表明,52%vol白酒,6. 6%盐度,酸碱度p H 5. 5的工艺条件能够最高效地降低醉泥螺腌制过程中副溶血性弧菌引起的食用安全风险,副溶血性弧菌的生长最受抑制。但兼顾不同盐度醉泥螺的感官评价,本研究认为52%vol白酒,4. 4%盐度,酸碱度pH 5. 5是最适合腌制醉泥螺的加工条件。     

生食贝类水产品中副溶血性弧菌的半定量风险评估

为研究上海市生食贝类水产品的食用安全性,进行了半定量风险评估。牡蛎、醉泥螺、象拔蚌、北极贝为上海市主要的生食贝类水产品,调查显示贝类水产品中副溶血性弧菌平均检出率为43.7%,其中副溶血性弧菌致病性基因Tdh+/Trh+在贝类水产品中的平均检出率为3.75%。对上海地区对生食贝类的消费情况进行了膳食调查,并利用"Risk Ranger"软件对牡蛎、醉泥螺、象拔蚌、北极贝中副溶血性弧菌进行了半定量风险评估。上海市生食贝类消费人群比例为13.86%,生食贝类的食用频率平均2.4月/餐次,平均每餐消费生食贝类水产品为49.06 g/餐。风险评估结果显示:牡蛎/副溶血性弧菌(50)属于高度风险,醉泥螺/副溶血性弧菌(46)、象拔蚌/副溶血性弧菌(44)、北极贝/副溶血性弧菌(39)均属于中度风险,根据风险排序风险大小依次为牡蛎醉泥螺象拔蚌北极贝。     

我国东部海域近岸水体中副溶血性弧菌的多元相关性分析

副溶血性弧菌（Vibrio parahemolyticus）是一种重要的水产品食源性致病菌,在我国沿海地区已成为引发细菌性食物中毒的首要风险因子。采集了我国东部海域(18.1°～38.5°N,109.3°～122.5°E)近岸水体样品,测定了水样的pH、盐度、温度、电导率、溶解性固体总量、溶解氧,以及副溶血性弧菌疑似菌含量,建立了水体环境因素与副溶血性弧菌相关性的模型。初步分析结果显示,水体的盐度是影响我国东部海域近岸水体中副溶血性弧菌丰度的主要环境因素。研究结果可为沿海地区副溶血性弧菌流行预警的研究提供数据参考。     

高通量快速测定致病性及非致病性副溶血性弧菌最大比生长速率

【目的】比较15 °C、20 °C和25 °C时, 9株不同致病性与非致病性副溶血性弧菌菌株在TSB (3% NaCl, pH 8.0)中的最大比生长速率之间的差异。【方法】应用Bioscreen C全自动微生物生长曲线分析仪测定副溶血性弧菌的最大比生长速率。【结果】15 °C、 20 °C和25 °C时, 9株副溶血性弧菌菌株间最大比生长速率的变异系数分别为20.72%、17.5%和15.98%。不同副溶血性弧菌最大比生长速率之间的差异随着温度的降低而增加。【结论】在进行定量微生物风险评估时, 应用仅基于一株菌建立的生长预测模型会给预测结果带来较大的不确定性。需建立可描述不同副溶血性弧菌菌株的最大比生长速率的随机模型来为定量微生物风险评估提供更准确有效的预测模型。     

致病性副溶血性弧菌菌落杂交检测体系的优化

对常见食源性致病菌副溶血性弧菌的杂交条件进行了优化。基于副溶血性弧菌的tdh,trh,toxR基因选用了3种探针序列,从菌落杂交样品的预处理方法、杂交时间和显色检测等方面对基于副溶血性弧菌毒力基因的原位杂交实验进行了条件优化。结果表明,采用80℃热固定2 h,37℃预杂交10 min后杂交8 h以及封闭1 h的改良方法可获得高特异性、低背景且着色清晰的实验结果。优化的菌落杂交技术检测副溶血性弧菌与传统检测方法相比,具有高效、准确、可区分致病性菌株的优点,为水产品中副溶血性弧菌致病菌株和非致病菌株的同步检测和筛选提供参考。     

基于组合胶体金纳米颗粒的免疫层析试纸条快速可视化检测海产品中副溶血性弧菌

本研究以野生型副溶血性弧菌(Vibrio parahaemolyticus)为免疫原制备单克隆抗体,并以组合胶体金纳米粒子(CP-AuNPs)为抗体标记探针,开发了一种基于双抗体夹心原理的可视化快速检测副溶血性弧菌的免疫层析试纸条方法。该方法被应用于梅子鱼、鲜虾、白蛤等海产品中副溶血性弧菌的检测,具有特异性强、检测时间短、灵敏度高的特点,其检测限可达4.77×10³CFU/mL。结果表明该方法可作为海产品中副溶血性弧菌检测的一种有效、快速的诊断工具。     

响应面分析法优化副溶血性弧菌生长条件

通过单因素分析,确定最适于副溶血性弧菌VPJ33生长的pH、温度和盐度.在此基础上,综合考虑3个因素对VPJ33生长的影响,用Design-Expert软件进行响应面分析,优化VPJ33的培养条件得到了菌体生长模型,以及取得模型最优值时各因素的水平.结果表明,VPJ33的最优培养条件为:pH 8.41、温度34.1℃和盐度2.47%;菌体生长过程中,pH和盐度以及pH和温度对VPJ33生长的交互作用显著,盐度和温度对VPJ33生长的交互作用不显著.菌体生长模型达到显著水平,可以对VPJ33在不同条件下的生长情况进行分析和预测.     

副溶血性弧菌毒力基因表达时内参基因的选择

[目的]筛选出合适的内参基因用于分析不同环境条件下副溶血性弧菌毒力基因的表达情况.[方法]本研究以虾样品中、海水样品中、过滤海水样品中以及TSB培养条件下的副溶血性弧菌为材料,利用qRT-PCR技术评价了GAPDH、pvuA、pvsA和rpoS4种常用管家基因在不同条件下的表达稳定性.[结果]4种管家基因均能特异扩增,表达稳定性排列顺序为pvuA(2.906)＞pvsA(3.197)＞GAPDH(3.746)＞rpoS(6.512),进一步通过geNorm软件分析,最终选择两个表达最为稳定的内参基因即pvuA和pvsA,以二者的几何平均值作为参照可更为准确地校正目的基因的表达.[结论]pvuA和pvsA可作为环境样品中副溶血性弧菌毒力基因表达变化研究的内参基因.     

响应面分析法优化副溶血性弧菌生长条件

通过单因素分析, 确定最适于副溶血性弧菌VPJ33生长的pH、温度和盐度。在此基础上, 综合考虑3个因素对VPJ33生长的影响, 用Design-Expert软件进行响应面分析, 优化VPJ33的培养条件得到了菌体生长模型, 以及取得模型最优值时各因素的水平。结果表明, VPJ33的最优培养条件为：pH 8.41、温度34.1℃和盐度2.47％; 菌体生长过程中, pH和盐度以及pH和温度对VPJ33生长的交互作用显著, 盐度和温度对VPJ33生长的交互作用不显著。菌体生长模型达到显著水平, 可以对VPJ33在不同条件下的生长情况进行分析和预测。     

信号分子AI-2的体外合成及其对副溶血性弧菌四环素耐药性的调控作用

【背景】抗菌药的过度使用引起细菌耐药性日益严重,作为重要的食源性致病菌,副溶血性弧菌也表现出一定程度的耐药性。群体感应系统可以调控细菌的耐药性,为研究副溶血性弧菌的耐药机制和控制技术提供新的途径。【目的】探讨群体感应信号分子AI-2 (autoinducer-2)对海产品中分离的副溶血性弧菌四环素耐药性的调控作用。【方法】通过原核表达制备AI-2合成关键酶——S-核糖同型半胱氨酸酶(S-ribosylhomocysteinase, LuxS)和S-腺苷同型半胱氨酸核苷酶(S-adenosylhomocysteinenucleosidase,Pfs),体外合成AI-2,通过菌落计数法分析AI-2对副溶血性弧菌在四环素亚抑菌浓度下耐受性的影响,采用逆转录实时荧光定量PCR法测定不同浓度AI-2对副溶血性弧菌四环素耐药基因转录水平的影响。【结果】通过原核表达获得LuxS和Pfs,作用于底物S-腺苷同型半胱氨酸能合成具有生物活性的AI-2,其荧光强度约为阳性对照的6倍。在四环素亚抑菌浓度下,AI-2能显著促进副溶血性弧菌的生长,6、15、30μmol/L浓度AI-2能不同程度地提高副溶血性弧菌四环素耐药基因的转录水平。【结论】AI-2能增强副溶血性弧菌对四环素的耐受作用,为解析副溶血性弧菌的耐药机制、研制以AI-2为靶点的副溶血性弧菌耐药性控制技术提供基础。     

超高压处理对副溶血性弧菌的影响研究

摘要：【目的】探讨超高压致死微生物的机理。【方法】本文以副溶血性弧菌为对象,研究了超高压处理对副溶血性弧菌的灭菌效果、对副溶血性弧菌细胞超微结构、细胞无机盐离子含量以及细胞膜蛋白的影响。【结果】结果表明,在20℃下分别经100、200 MPa高压处理10min后,副溶血性弧菌致死率为40%、84.7%,经300 MPa及以上的压力处理,副溶血性弧菌的致死率为100%。超高压处理对细菌细胞形态结构造成明显的损伤：局部细胞壁遭到破坏,出现缺口;胞质内含物结构紊乱,出现泄漏,细胞中部出现透电子区;细胞结构不完整     

Salinity,water use and yield of maize: Testing of the mathematical model ecosys

There is a need to establish how root water uptake should be calculated under saline conditions, and to test calculated uptake against experimental data recorded under documented site conditions. In this study, the ecosystem simulation model ecosys was expanded to include an ion transfer-equilibrium-exchange model used to calculated electrical conductivity and osmotic potential. This expanded model was tested against experimental data for maize growth and water use reported under different irrigation and salinity levels at four different sites in the western U.S. to determine if salinity effects on crop growth and water use could be modelled from the effects of salinity on soil osmotic potential. The model was able to reproduce reductions in water use and phytomass yields on salinized (10 g total salts kg^–1 water) soils that ranged from 10 to 50% of those on non-salinized controls. In general, these reductions increased with increasing irrigation deficits. These reductions arose in the model from reduced canopy water potentials and conductances caused by reduced osmotic potentials in the saline soils. The hypothesis that salinity effects on crop growth and water use are caused by salinity effects on soil osmotic potential appear to be supported under the range of conditions included in this study. Models such as ecosys that are based on general hypotheses for the effects of salinity upon biological activity may be well adapted for general use in assessing the effects of salinity on crop growth and water use with different soils, managements and climates.     

Water column dynamics of Vibrio in relation to phytoplankton community composition and environmental conditions in a tropical coastal area

Vibrio abundance generally displays seasonal patterns. In temperate coastal areas, temperature and salinity influence Vibrio growth, whereas in tropical areas this pattern is not obvious. The present study assessed the dynamics of Vibrio in the Arabian Sea, 1-2 km off Mangalore on the south-west coast of India, during temporally separated periods. The two sampling periods were signified by oligotrophic conditions, and stable temperatures and salinity. Vibrio abundance was estimated by culture-independent techniques in relation to phytoplankton community composition and environmental variables. The results showed that the Vibrio density during December 2007 was 10- to 100-fold higher compared with the February-March 2008 period. High Vibrio abundance in December coincided with a diatom-dominated phytoplankton assemblage. A partial least squares (PLS) regression model indicated that diatom biomass was the primary predictor variable. Low nutrient levels suggested high water column turnover rate, which bacteria compensated for by using organic molecules leaking from phytoplankton. The abundance of potential Vibrio predators was low during both sampling periods; therefore it is suggested that resource supply from primary producers is more important than top-down control by predators.     

Nondestructive leaf-area estimation and validation for green pepper (<Emphasis Type="Italic">Capsicum annuum</Emphasis> L.) grown under different stress conditions

Leaf area of a plant is essential to understand the interaction between plant growth and environment. This useful variable can be determined by using direct (some expensive instruments) and indirect (prediction models) methods. Leaf area of a plant can be predicted by accurate and simple leaf area models without damaging the plant, thus, provide researchers with many advantages in horticultural experiments. Several leaf-area prediction models have been produced for some plant species in optimum conditions, but not for a plant grown under stress conditions. This study was conducted to develop leaf area estimation models by using linear measurements such as lamina length and width by multiple regression analysis for green pepper grown under different stress conditions. For this purpose, two experiments were conducted in a greenhouse. The first experiment focused to determine leaf area of green pepper grown under six different levels of irrigation water salinity (0.65, 2.0, 3.0, 4.0, 5.0, and 7.0 dS m⁻¹) and the other under four different irrigation regime (amount of applied water was 1.43, 1.0, 0.75, and 0.50 times of required water). In addition to general models for each experiment, prediction models of green pepper for each treatment of irrigation water salinity and of irrigation regime experiments were obtained. Validations of the models for both experiments were realized by using the measurements belong to leaf samples allocated for validation purposes. As a result, the determined equations can simply and readily be used in prediction of leaf area of green pepper grown under salinity and water stress conditions. The use of such models enable researchers to measure leaf area on the same plants during the plant growth period and, at the same time, may reduce variability in experiments.     

Influence of salinity and organic nutrient concentration on survival and growth of Vibrio cholerae in aquatic microcosms.

Laboratory microcosms were employed to evaluate the influence of selected environmental parameters, organic nutrient concentration, and salinity on the growth and survival of a toxigenic strain of Vibrio cholerae LA4808. Over the range conditions tested, this strain of V. cholerae showed maximum response as determined by increased plate counts and direct microscopic counts in microcosms prepared with a chemically defined sea salts solution at a salinity of 25%, but with lower or higher salinity levels, the maximum population size declined. When added organic concentrations of less than 1,000 micrograms/liter were present, a marked salinity effect on the growth of V. cholerae was detected. However, at or above an organic nutrient concentration of 1,000 micrograms/liter, the need for an optimum salinity level was spared. From the results of this study, it is concluded that V. cholerae can grow under conditions of organic nutrient concentration and salinity typical of estuaries. Results obtained support the hypothesis that V. cholerae is an autochthonous member of the estuarine microbial community.     

Influence of salinity and organic nutrient concentration on survival and growth of Vibrio cholerae in aquatic microcosms

Laboratory microcosms were employed to evaluate the influence of selected environmental parameters, organic nutrient concentration, and salinity on the growth and survival of a toxigenic strain of Vibrio cholerae LA4808. Over the range conditions tested, this strain of V. cholerae showed maximum response as determined by increased plate counts and direct microscopic counts in microcosms prepared with a chemically defined sea salts solution at a salinity of 25%, but with lower or higher salinity levels, the maximum population size declined. When added organic concentrations of less than 1,000 micrograms/liter were present, a marked salinity effect on the growth of V. cholerae was detected. However, at or above an organic nutrient concentration of 1,000 micrograms/liter, the need for an optimum salinity level was spared. From the results of this study, it is concluded that V. cholerae can grow under conditions of organic nutrient concentration and salinity typical of estuaries. Results obtained support the hypothesis that V. cholerae is an autochthonous member of the estuarine microbial community.     

A mechanistic model for understanding invasions: using the environment as a predictor of population success

Aim We set out to develop a temperature‐ and salinity‐dependent mechanistic population model for copepods that can be used to understand the role of environmental parameters in population growth or decline. Models are an important tool for understanding the dynamics of invasive species; our model can be used to determine an organism’s niche and explore the potential for invasion of a new habitat. Location Strait of Georgia, British Columbia, Canada. Methods We developed a birth rate model to determine the environmental niche for an estuarine copepod. We conducted laboratory experiments to estimate demographic parameters over a range of temperatures and salinities for Eurytemora affinis collected from the Nanaimo Estuary, British Columbia (BC). The parameterized model was then used to explore what environmental conditions resulted in population growth vs. decline. We then re‐parameterized our model using previously published data for E. affinis collected in the Seine Estuary, France (SE), and compared the dynamics of the two populations. Results We established regions in temperature–salinity space where E. affinis populations from BC would likely grow vs. decline. In general, the population from BC exhibited positive and higher intrinsic growth rates at higher temperatures and salinities. The population from SE exhibited positive and higher growth rates with increasing temperature and decreasing salinity. These different relationships with environmental parameters resulted in predictions of complex interactions among temperature, salinity and growth rates if the two subspecies inhabited the same estuary. Main conclusions We developed a new mechanistic model that describes population dynamics in terms of temperature and salinity. This model may prove especially useful in predicting the potential for invasion by copepods transported to Pacific north‐west estuaries via ballast water, or in any system where an ecosystem is subject to invasion by a species that shares demographic characteristics with an established (sub)species.     

Effect of some abiotic factors on the biological activity of Gluconacetobacter diazotrophicus

AIMS: The effect of some abiotic factors, dryness, heat and salinity on the growth and biological activity of Gluconacetobacter diazotrophicus, and the influence of a salt stress on some enzymes involved in carbon metabolism of these bacteria is studied under laboratory conditions. METHODS AND RESULTS: Strain PAL-5 of G. diazotrophicus was incubated under different conditions of drying, heat and salinity. Cells showed tolerance to heat treatments and salt concentrations, and sensitivity to drying conditions. Higher NaCl dosage of 150 and 200 mmol l -1 limited its growth and drastically affected the nitrogenase activity and the enzymes glucose dehydrogenase, alcohol dehydrogenase, fumarase, isocitrate dehydrogenase and malate dehydrogenase. CONCLUSIONS: Gluconacetobacter diazotrophicus, despite its endophytic nature, tolerated heat treatments and salinity stress, but its nitrogenase activity and carbon metabolism enzymes were affected by high NaCl dosage. SIGNIFICANCE AND IMPACT OF THE STUDY: The investigation of the biological activity of G. diazotrophicus in response to different abiotic factors led to more knowledge of this endophyte and may help to clarify pathways involved in its transmission into the host plant.