Food habits of two-year-old Chinese mitten crab (Eriocheir sinensis) stocked in Lake Bao'an, China

Gastric mills of 362 specimens of two-year-old Chinese mitten crab (Eriocheir sinensis), which contained recognizable food items, from Lake Bao'an, China were examined. The food items were macrophytes, algae, arthropods, oligochaetes, fish, protozoa, rotifers, gastropods, and detritus, and the percent frequencies of occurrence (FO) for these items were 87.3%, 82.0%, 48.2%, 28.2%, 28.7%, 0.3%, 0.6%, 0.3% and 88.7%, respectively. Unidentified animal tissue was often observed and had a FO of 46.1%. In total, FO of plants (macrophytes + algae) was 87.7% and of animals was 89.8%. However, 5.8% of the gastric mills contained only animals, 5.3% had only macrophytes, and 0.3% contained only algae. There was no significant difference (p>0.05) in food habits between male and female crabs. The ratio of cell number of macrophytes to algae was about 156:1.

黄海纤毛虫的丰度和生物量

海洋浮游纤毛虫是在海洋中浮游生活的一类单细胞原生动物，主要是指寡毛类纤毛虫(Oligotrich ciliates)，隶属原生动物界(Protozoa)、纤毛门(Ciliophora)、寡毛纲(Oligotrichea)，分属于Oligotrichina 和Tintinnina两个亚纲。它们个体微小，粒径在5-200 µm之间，是微型浮游动物和海洋微食物环(Marine Microbial Food Web)的重要组成部分。 2006年4月至2007年12月，在黄海(包括胶州湾)采样分析海洋浮游纤毛虫的种类组成(砂壳纤毛虫)、丰度和生物量，分析纤毛虫在这一海区的季节变化和空间变化。 纤毛虫丰度和生物量的研究方法为：Rosette采水器(胶州湾用Niskin采水器)采集水样，取1 L水样，加Lugol’s试剂固定(终浓度1%)，Utermöhl方法100倍镜检。测量虫体的体长、体宽，按最接近的几何形状(圆柱体、球体和圆锥体)计算体积。生物量由体积乘转换系数(0.19 pgC/µm3)得到，砂壳纤毛虫的肉体体积按照壳体积的1/3近似。 本文的结果表明，胶州湾各站纤毛虫平均丰度于6月达到全年最高值6065 ind./L，12月为全年丰度最低值843 ind./L。平均生物量8月达全年最高值(18.5 µg C/L)，6月为全年最低值(0.6 µg C/L)。砂壳纤毛虫种丰富度于8月达到最高值，共发现25种砂壳纤毛虫，1月种类最少(6种)。湾内站位的纤毛虫平均丰度比湾外的高(6月和8月除外)。砂壳纤毛虫在纤毛虫总丰度中的比例较小，平均为25%，范围为8-57%，分别于1月和8月达到最低和最高值。 两次冷水团大面调查结果表明，4月表层纤毛虫平均丰度(1490 ind./L)要高于10月(972 ind./L)。10月表层纤毛虫生物量0.14-5.33 µg C/L，14194站、15694站和15894站生物量较高，为4.08-5.33 µg C/L。无壳纤毛虫优势种Laboea strobila在两个航次中均呈现斑块分布，4月航次丰度0-10000 ind./L，10月航次丰度11-350 ind./L；砂壳纤毛虫优势种Ptychocylis obtusa仅在4月航次发现，最大丰度2895 ind./L，10月航次未发现。4月航次砂壳纤毛虫有百乐拟铃虫(Tintinnopsis beroidea)，丰度为0-1920 ind./L；卡拉直克拟铃虫(Tintinnopsis karajacensis)，丰度很小(10-93 ind./L)。10月航次砂壳纤毛虫优势种Tintinnidium primitivum，丰度为35-700 ind./L；也出现了尖底类瓮虫(Amphorellopsis acuta)和网纹虫(Favella spp.)，但丰度不大(0-210 ind./L)；运动类铃虫(Codonellopsis mobilis)、筒状拟铃虫(Tintinnopsis tubulosoides)和Eutintinnus sp.丰度也较低(35-105 ind./L)；Craterella torulata丰度为0-120 ind./L，主要分布于15694站。10月航次已经出现了温跃层，位于30 m左右水层，纤毛虫主要分布于温跃层之上。 六次黄海断面航次表明：温跃层在5月已经出现，到12月消失。在有温跃层的5月、6月、8月、9月，纤毛虫主要分布于温跃层(30 m左右)之上。其中8月份航次纤毛虫丰度最高，表层平均丰度3103 ind./L。12月份纤毛虫丰度最低，表层平均丰度406 ind./L。纤毛虫生物量春夏季为0.02-5.5 µg C/L，冬季为0.04-1.99 µg C/L。小型无壳纤毛虫占优势，砂壳纤毛虫东方拟铃虫(Tintinnopsis orientalis)、筒状拟铃虫、运动类铃虫、Craterella torulata和Tintinnidium primitivum几乎在各个航次均有分布。

潮间带纤毛虫原生动物和小型底栖动物生态学研究

微型和小型底栖动物是底栖微/小食物网的重要构成。相对浮游生态系统, 迄今国际间对底栖食物网的认知极为欠缺。这一方面是由于微型生物形态和功能上的复杂性和多样性, 另一方面原因在于研究方法的障碍—主要是微型和小型底栖动物的定量提取和定性分析。本研究首先进行了方法学改良, 并应用新方法对底栖微食物网的重要功能类群—纤毛虫原生动物和小型底栖动物进行了不同生境的周年按月采样, 定性及定量研究的同时, 联系环境因子对微型和小型底栖生物的环境监测进行了探讨。 微型和小型底栖生物的定量研究首先涉及到目标生物在沉积物中的有效提取, 目前硅胶液提取是普遍使用的方法, 其中Ludox液主要应用于小型生物, 它不但价格便宜而且比重合适, 因此在常规生态研究中被广为接受。不过, Ludox易与于海水中的阳离子产生凝结而无法直接用于微型生物; 目前唯一直接应用于微型生物提取的是Percoll硅胶液, 但其昂贵的价格使其在常规生态研究中受到极大限制。本研究以价格低廉的Ludox 硅胶液结合定量蛋白银染色 (QPS) 技术开发了一种新的方法, 即Ludox-QPS法。主要流程为: 样品采集与固定、淘洗/稀释降盐、Ludox密度梯度离心、过滤浓缩和琼脂包埋, QPS染色、永久封片及鉴定计数。添加已知数量的纤毛虫至无生物底泥的重获实验表明, 该密度梯度离心的提取率大于94%; 该方法对自然沉积物中纤毛虫的提取率达97.6%, 对沙质、泥沙质和泥质中海洋线虫的提取率分别达97%、96.9% 和97.8%。对比实验表明, 经QPS制片获得的小型动物的丰度和类群数量与传统方法相当或更高, 尤其当小个体虫体占优势时, 该法显示出较传统方法 (导致数量低估) 更为明显的定量优越性。该方法除用于纤毛虫和小型动物的定量分析外, 还具有较高的分类分辨率, 染色后的纤毛虫原生动物大多类群可鉴定到属, 部分可鉴定到种, 以此可在群落水平上研究其生态作用。 根据新开发的Ludox-QPS技术, 在大沽河潮间带依据盐度梯度选定2个站位 (IIQ和营海) 进行了周年按月采样, 对底栖纤毛虫和小型底栖动物进行了定量研究。纤毛虫原生动物在IIQ和营海的年平均丰度分别为2236 inds./10 cm2 和935 inds./10 cm2 (28 inds./ml 和12 inds./ml), 平均生物量分别为119.1 gC/10 cm2和54.2 gC/10 cm2 (1.5 gC /ml 0.7 gC/ml)。丰度的季节变化趋势为: 春天 > 秋天 > 夏天 > 冬天。垂直分布上, 在营海分布于表层0-0.5 cm 的比例为57.1%, 分布于0.5-2 cm、2-4 cm和4-8 cm比例分别为23.1%、11.4% 和8.5%; 13个月中除12月份外, 4-8 cm均有一定数量的纤毛虫分布; 而在IIQ, 97% 的纤毛虫分布在0-0.5 cm, 分布在0.5-2 cm、2-4 cm和4-8 cm比例分别为2.4%、0.4%和0.2%, 4-8 cm的分布只发生在春季和秋季。纤毛虫的多样性季节变化明显, 春秋季物种丰富, 两个站点每毫升沉积物的平均物种数分别为18和6。Two-Way Crossed ANOSIM 分析表明纤毛虫群落在月份间和站点间的差异极其显著。Pseudochilodonopsis sp., Chilodontopsis sp., Euplotes sp.及Prorodon sp.是表征两个生镜中纤毛虫群落的主要类群。 同时, 发现了14个小型生物类群, 其中线虫在IIQ和营海的丰度优势度分别为97.4% 和78.6%。小型动物在IIQ和营海的年平均丰度分别为4793 inds./10 cm2和8915 inds./10 cm2 (60 inds./ml和111 inds./ml), 其生物量分别为1068.8 gC /10 cm2和1790 gC /10 cm2 (13.4 gC/ml和22.4 gC/ml)。小型底栖动物的丰度在IIQ的季节变化为: 夏季 (7888 inds./10cm2) > 秋季 (5447 inds./10cm2) > 春季 (3731 inds./10cm2) > 冬季 (2780 inds./10cm2); 在营海则完全相反: 冬季 (15579 inds./10cm2) > 春季 (10691 inds./10cm2) > 秋季 (6611 inds./10cm2) > 夏季 (4667 inds./10cm2)。小型底栖动物和纤毛虫的相对重要性存在明显的区域和季节差异。 纤毛虫原生动物、小型动物及环境因子的相关分析表明, 纤毛虫的丰度和多样性与温度和盐度及有机质含量显著相关, 与小型动物没有显著相关性; 群落结构分析表明, 温度、有机质和小型动物的丰度的组合与纤毛虫群落丰度的相关系数为0.345; 盐度、脱镁叶绿素、有机质和小型动物生物量的组合与纤毛虫群落多样性的相关系数为0.403。依据海洋线虫和桡足类的比值 (N/C) 推测, IIQ 可能存在严重的有机污染, 营海则存在明显的季节波动, 8月和9月及2月可能是污染最严重的季节, 这种状况在纤毛虫群落结构的CLUSTER聚类中得到验证。虽然目前尚没有形成有关微型底栖生物-纤毛虫原生动物的污染检测的直接依据, 但本研究说明纤毛虫群落的确对环境污染具有一定的感应度, 而且这种感应和利用小型生物的主要类群估算的污染检测 (N/C) 存在一定程度的关联。 90年代早期有关青岛湾有机污染带的研究表明, 经彻底截污后, 其环境状况向良性发展。进一步了解该湾的健康状况, 2006.5-2007.5月对该湾沙质和泥沙质的小型动物进行周年按月采样。小型动物在泥沙质和砂质沉积物中的年平均丰度分别为4853 ± 1292 inds./10 cm2和1528 ± 569 inds./10 cm2; 年平均总生物量分别为1434.1 ± 897.0 gC /10cm2和720.7 ± 353.8 gC/10cm2。沙质底小型生物的丰度季节波动明显, 6月份和12月份最高, 3月份和9月份最低; 泥沙质季节波动不明显, 6月份最高。两个站点均有48%的小型动物分布在0-0.5 cm 表层, 海洋线虫在表层的分布比例分别为48% (泥沙质) 和34% (砂质)。共检获14个小型动物类群, 其中线虫在泥沙质和砂质沉积物中的年平均丰度分别4619 ± 1255 inds./10cm2和1014 ± 376 inds./10cm2, 其丰度优势度分别为95.2%和66.4%。其它在丰度上占优势的类群, 泥沙质依次为多毛类 (1.5%)、甲壳幼体 (1.5%) 和桡足类 (0.7%); 沙质依次为: 甲壳类幼体 (12.6%)、腹毛类 (8.3%) 和 桡足类 (6.2%)。CLUSTER聚类分析表明, 泥沙质和和砂质中小型生物的丰度组成具有64%的相似性。BIOENV分析表明, 温度、盐度、中值粒径和粘土粉砂含量的组合最能解释不同月份之间和不同站位间的差异, 其相关系数为0.614。依据小型生物的丰度和类群组成, 表明泥沙质底尚存一定的有机污染。

A bioaugmentation failure caused by phage infection and weak biofilm formation ability

Two biological aerated filters (BAF) were setup for ammonia removal treatment of the circulation water in a marine aquaculture. One of the BAFs was bioaugmented with a heterotrophic nitrifying bacterium, Lutimonas sp. H10, where the ammonia removal was not improved and the massive inoculation was even followed by a nitrification breakdown from day 9 to 18. The nitrification was remained stable in control BAF operated under the same conditions. Fluorescent in situ hybridization (FISH) with rRNA-targeted probes and cultivable method revealed that Lutimonas sp. H10 almost disappeared from the bioaugomented BAF within 3 d, and this was mainly due to the infection of a specific phage as revealed by flask experiment, plaque assay and transmission electron observation. Analyses of 16S rRNA gene libraries showed that bacterial groups from two reactors evolved differently and an overgrowth of protozoa was observed in the bioaugmented BAR Therefore, phage infection and poor biofilm forming ability of the inoculated strain are the main reasons for bioaugmentation failure. In addition, gazing by protozoa of the bacteria might be the reason for the nitrification breakdown in bioaugmented BAF during day 9-18.

Summertime ciliate and copepod nauplii distributions and micro-zooplankton herbivorous activity in the Laizhou Bay, Bohai Sea, China

The abundance and biomass of ciliated protozoa and copepod nauplii were investigated at 21 grid stations and two anchored stations in the Laizhou Bay, Bohai Sea, China in June 1998. Dilution incubations were carried out to investigate micro-zooplankton grazing pressure at the anchored stations during spring tide and neap tide. The dominant species were Tintinnopsis amoyensis, T. chinglanensis, T. pallida and aloricate ciliates. A total of 13 species of tintinnids were found. The total abundance of ciliates and nauplii ranged from 30 to 2390 ind l(-1) at grid stations. Tintinnopsis amoyensis was the only ciliate found at the anchored stations and in concentrations which varied from 0 to 6700 ind l(-1). The spatial distribution of ciliates was patchy. Tintinnopsis amoyensis and T. pallida were distributed in the Weihe River mouth and Xiaoqinghe River mouth respectively. The aloricate ciliates, T. chinglanensis and Codonellopsis ostenfeldi dominated offshore in sequence. The water mixing process may affect the spatial pattern of the dominant ciliate species. The abundance and biomass of copepod nauplii were in the range of 0-140 ind l(-1) and 0-7 mu g C l(-1) respectively, with the peak appearing at grid station 15. The total biomass of ciliates and copepod nauplii was in the range of 1(.)5-25 mu g C l(-1). Water column biomass of ciliates and nauplii varied from 2(.)37 to 52(.)3 mg C m(-2). At the anchored stations, the phytoplankton growth rates ranged from undetectable to 0 21 d(-1) and micro-zooplankton grazing rates from 0 13 to 0(.)57 d(-1). The grazing pressure of micro-zooplankton were 12 to 43% of the chlorophyll standing stock and 84 to 267% of the chlorophyll (C) 2000 Academic Press.

Community structure and seasonal dynamics of planktonic ciliates along salinity gradients

The ciliate community structure and seasonal dynamics in a solar saltern of the Yellow Sea were studied based oil 4 sampling dates and 8 stations with salinities from 27.7 parts per thousand to 311.0 parts per thousand. The effects of the type and concentration of the fixative used (Lugol's and Bouin's) were tested at the first sampling date. Fixative type and fixative concentration had significant effects on ciliate abundance and blovolume, with 1% Lugol's giving the best results. A detailed investigation using live observations and protargol staining techniques revealed a total of 98 morphospecies from 8 sampling stations. There was obvious seasonal variation in species composition at most of the stations, but this tended to be less distinct with increasing salinity, as the dominant ciliate group shifted from oligotrichs to heterotrichs. Ciliate abundance varied from 4.40 x 10(1) to 2.11 x 10(5) cells l(-1) and biomass ranged between 2.39 and 9.87 x 10(3) mu g Cl-1 (at a salinity of 147.6 parts per thousand). Both abundance and biomass decreased abruptly when salinity exceeded 100-150 parts per thousand. Statistical analyses Suggested that the dynamics of ciliate abundance and biomass were regulated by both salinity and by season, but those of diversity and species richness were mainly controlled by salinity and both significantly decreased with increasing salinity. (C) 2009 Elsevier GmbH. All rights reserved.

Detection of Emerging and Re-Emerging Pathogens in Surface Waters Close to an Urban Area

Current knowledge about the spread of pathogens in aquatic environments is scarce probably because bacteria, viruses, algae and their toxins tend to occur at low concentrations in water, making them very difficult to measure directly. The purpose of this study was the development and validation of tools to detect pathogens in freshwater systems close to an urban area. In order to evaluate anthropogenic impacts on water microbiological quality, a phylogenetic microarray was developed in the context of the EU project µAQUA to detect simultaneously numerous pathogens and applied to samples from two different locations close to an urban area located upstream and downstream of Rome in the Tiber River. Furthermore, human enteric viruses were also detected. Fifty liters of water were collected and concentrated using a hollow-fiber ultrafiltration approach. The resultant concentrate was further size-fractionated through a series of decreasing pore size filters. RNA was extracted from pooled filters and hybridized to the newly designed microarray to detect pathogenic bacteria, protozoa and toxic cyanobacteria. Diatoms as indicators of the water quality status, were also included in the microarray to evaluate water quality. The microarray results gave positive signals for bacteria, diatoms, cyanobacteria and protozoa. Cross validation of the microarray was performed using standard microbiological methods for the bacteria. The presence of oral-fecal transmitted human enteric-viruses were detected using q-PCR. Significant concentrations of Salmonella, Clostridium, Campylobacter and Staphylococcus as well as Hepatitis E Virus (HEV), noroviruses GI (NoGGI) and GII (NoGII) and human adenovirus 41 (ADV 41) were found in the Mezzocammino site, whereas lower concentrations of other bacteria and only the ADV41 virus was recovered at the Castel Giubileo site. This study revealed that the pollution level in the Tiber River was considerably higher downstream rather than upstream of Rome and the downstream location was contaminated by emerging and re-emerging pathogens.

Drug delivery strategies for photodynamic antimicrobial chemotherapy: From benchtop to clinical practice

Light and photosensitizer-mediated killing of many pathogens, termed photodynamic antimicrobial chemotherapy (PACT), has been extensively investigated in vitro. A wide range of organisms from the Gram-positive Staphylococcus aureus to the Gram-negative Pseudomonas aeruginosa have been proven to be susceptible to PACT. Multidrug-resistant strains are just as susceptible to this treatment as their naive counterparts. Both enveloped and non-enveloped viruses have demonstrated susceptibility in vitro, in addition to fungi and protozoa. Significantly, however, no clinical treatments based on PACT are currently licensed. This paper provides a comprehensive review of work carried out to date on delivery of photosensitizers for use in PACT, including topical, intranasal and oral/buccal delivery, as well as targeted delivery. We have also reviewed photo-antimicrobial surfaces. It is hoped that, through a rational approach to formulation design and subsequent success in small-scale clinical trials, more widespread use will be made of PACT in the clinic, to the benefit of patients worldwide. (C) 2009 Elsevier B.V. All rights reserved.

Antifungal photodynamic therapy

In photodynamic antimicrobial chemotherapy (PACT), a combination of a sensitising drug and visible light causes selective destruction of microbial cells. The ability of light-drug combinations to kilt microorganisms has been known for over 100 years. However, it is only recently with the beginning of the search for alternative treatments for antibiotic-resistant pathogens that the phenomenon has been investigated in detail. Numerous studies have shown PACT to be highly effective in the in vitro destruction of viruses and protozoa, as well as Gram-positive and Gram-negative bacteria and fungi. Results of experimental investigations have demonstrated conclusively that both dermatomycetes and yeasts can be effectively killed by photodynamic action employing phenothiazinium, porphyrin and phthatocyanine photosensitisers. Importantly, considerable setectivity for fungi over human cells has been demonstrated, no reports of fungal resistance exist and the treatment is not associated with genotoxic or mutagenic effects to fungi or human cells. In spite of the success of cell culture investigations, only a very small number of in vivo animal. and human trials have been published. The present paper reviews the studies published to date on antifungal applications of PACT and aims to raise awareness of this area of research, which has the potential to make a significant impact in future treatment of fungal infections. (c) 2007 Elsevier GmbH. All rights reserved.

Phosphorylation of plant actin-depolymerising factor by calmodulin-like domain protein kinase

actin-depolymerising factor (ADF)/cofilin group of proteins are stimulus-responsive actin-severing proteins, members of which are regulated by reversible phosphorylation. The phosphorylation site on the maize ADF, ZmADF3, is Ser-6 but the kinase responsible is unknown [Smertenko et al,, Plant J. 14 (1998) 187-193]. We have partially purified the ADF kinase(s) and found it to be calcium-regulated and inhibited by N-(6-aminohesyl)-[H-3]5-chloro-1-naphthalenesulphonamide. Immunoblotting reveals that calmodulin-like domain protein kinase(s) (CDPK) are enriched in the purified preparation and addition of anti-CDPK to in vitro phosphorylation assays results in the inhibition of ADF phosphorylation, These data strongly suggest that plant ADP is phosphorylation by CDPK(s), a class of protein kinases unique to plants and protozoa. (C) 2001 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.

Organophosphonates revealed: new insights into the microbial metabolism of ancient molecules.

Organophosphonates are ancient molecules that contain the chemically stable C–P bond, which is considered a relic of the reducing atmosphere on primitive earth. Synthetic phosphonates now have a wide range of applications in the agricultural, chemical and pharmaceutical industries. However, the existence of C–P compounds as contemporary biogenic molecules was not discovered until 1959, with the identification of 2-aminoethylphosphonic acid in rumen protozoa. Here, we review advances in our understanding of the biochemistry and genetics of microbial phosphonate metabolism, and discuss the role of these compounds and of the organisms engaged in their turnover within the P cycle.

Prévalence d’excrétion de Giardia et Cryptosporidium chez les humains, les animaux domestiques et les lémuriens de l’écosystème du Parc National de Ranomafana, Madagascar

L’augmentation des interactions entre humains et animaux sauvages en lisière des habitats naturels pourrait faciliter la transmission d’agents pathogènes entre les humains et les différentes espèces animales d’un écosystème et ainsi favoriser l’émergence de maladies. Nous avons effectué une étude transversale portant sur l’infection par Giardia et Cryptosporidium chez les humains, les animaux domestiques, les rongeurs et les lémuriens au sein de l’écosystème de Ranomafana, Madagascar. Des échantillons de fèces ont étés collectés de manière non invasive chez des personnes volontaires, des mammifères domestiques et des rongeurs introduits habitant trois villages situés en lisière du Parc National de Ranomafana (PNR) ainsi que quatre espèces de lémuriens (Propithecus edwardsii, Prolemur simus, Eulemur rubriventer et Microcebus rufus) du PNR. Des analyses coproscopiques par la technique d’immunofluorescence directe ont été réalisées afin de détecter la présence de Cryptosporidium et Giardia. Leur prévalence a été estimée et certaines variables reliées à l’infection par les parasites ont été identifiées. Cryptosporidium et Giardia ont été détectés avec une prévalence estimée à 22,9 % et 13,6 % respectivement chez les humains. La prévalence de ces deux parasites variait de 0 % à 60 % chez les animaux domestiques et les rongeurs au sein des villages. L’espèce hôte, l’âge ainsi que la co-infection par un autre protozoaire sont les seules variables associées à l’infection par Cryptosporidium et Giardia dans cet écosystème tandis qu’aucune association avec une coinfection par un ordre de nématode n’a été détecté. De plus, Cryptosporidium a été détecté chez 10,5 % des lémuriens du PNR. Cette étude documente pour la première fois la présence de Cryptosporidium chez deux espèces de lémuriens du PNR. Par contre, Giardia n’a pas été détecté dans les échantillons issus de lémuriens du PNR.

Interaction d'Escherichia coli entérohémorragique (EHEC) avec Acanthamoeba castellanii et rôle du régulon Pho chez les EHEC

Les EHEC de sérotype O157:H7 sont des agents zoonotiques d’origine alimentaire ou hydrique. Ce sont des pathogènes émergeants qui causent chez l’humain des épidémies de gastro-entérite aiguë et parfois un syndrome hémolytique-urémique. Les EHEC réussissent leur transmission à l’humain à partir de leur portage commensal chez l’animal en passant par l’étape de survie dans l’environnement. L’endosymbiose microbienne est une des stratégies utilisées par les bactéries pathogènes pour survivre dans les environnements aquatiques. Les amibes sont des protozoaires vivants dans divers écosystèmes et connus pour abriter plusieurs agents pathogènes. Ainsi, les amibes contribueraient à transmettre les EHEC à l'humain. La première partie de mon projet de thèse est centrée sur l'interaction de l’amibe Acanthamoeba castellanii avec les EHEC. Les résultats montrent que la présence de cette amibe prolonge la persistance des EHEC, et ces dernières survivent à leur phagocytose par les amibes. Ces résultats démontrent le potentiel réel des amibes à héberger les EHEC et à contribuer à leur transmission. Cependant, l’absence de Shiga toxines améliore leur taux de survie intra-amibe. Par ailleurs, les Shiga toxines sont partiellement responsables de l’intoxication des amibes par les EHEC. Cette implication des Shiga toxines dans le taux de survie intracellulaire et dans la mortalité des amibes démontre l’intérêt d’utiliser les amibes comme modèle d'interaction hôte/pathogène pour étudier la pathogénicité des EHEC. Durant leur cycle de transmission, les EHEC rencontrent des carences en phosphate inorganique (Pi) dans l’environnement. En utilisant conjointement le système à deux composantes (TCS) PhoB-R et le système Pst (transport spécifique de Pi), les EHEC détectent et répondent à cette variation en Pi en activant le régulon Pho. La relation entre la virulence des EHEC, le PhoB-R-Pst et/ou le Pi environnemental demeure inconnue. La seconde partie de mon projet explore le rôle du régulon Pho (répondant à un stress nutritif de limitation en Pi) dans la virulence des EHEC. L’analyse transcriptomique montre que les EHEC répondent à la carence de Pi par une réaction complexe impliquant non seulement un remodelage du métabolisme général, qui est critique pour sa survie, mais aussi en coordonnant sa réponse de virulence. Dans ces conditions le régulateur PhoB contrôle directement l’expression des gènes du LEE et de l’opéron stx2AB. Ceci est confirmé par l’augmentation de la sécrétion de l’effecteur EspB et de la production et sécrétion de Stx2 en carence en Pi. Par ailleurs, l’activation du régulon Pho augmente la formation de biofilm et réduit la motilité chez les EHEC. Ceci corrèle avec l’induction des gènes régulant la production de curli et la répression de la voie de production d’indole et de biosynthèse du flagelle et du PGA (Polymère β-1,6-N-acétyle-D-glucosamine).

Survival of Escherichia coli in a tropical estuary

The survival of Escherichia coli in tropical estuarine water has been studied under controlled laboratory conditions using microcosms. The survival has been assessed in terms of various self purifying factors of the natural waters such as biological, chemical and physical factors. The biological factors considered included competition from other microorganisms, predation by protozoa and coliphages. The suitability of the chemical composition of estuarine water has been studied under chemical factors and negative impact of sunlight has been studied under physical factors. The results revealed that sunlight exerted maximum negative impact, followed by biotic factors contained in the estuarine water. However, the chemical composition of the estuarine water is found to be suitable for the growth and survival of E. coli. The injury exerted by each of the above factors was also evaluated by using a selective and non-selective medium in conjunction. It was found that sunlight resulted in 100% injury of the cells as the cells failed to develop in a selective medium. While, sunlight resulted in the extinction of 90% of the E. coli cells within the first two hours of exposure, biotic factors took nearly 24 hours to remove the same amount of population.