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1.
In the marine environment, vibrios adhere to a number of substrates including chitin-rich organisms such as crustaceans. Their wide diffusion in coastal waters and pathogenic potential require knowledge of the lifestyle and environmental reservoirs of these bacteria. To test the presence of culturable vibrios in coastal areas and their association with benthic crustaceans, vibrios were isolated from water, sediments and crustaceans (copepods and anphipods) at three stations placed in front of heavily used tourist beaches of the Adriatic Sea. We observed significant correlations between vibrios and temperature. Benthic and planktonic copepods harboured vibrios in summer, while benthic amphipods harboured these bacteria in spring and autumn. Vibrio alginolyticus and Vibrio parahaemolyticus strains gave positive results using primers for Vibrio cholerae toxR and toxS. Sedimentary crustaceans may extend Vibrio persistence in seawater and may represent an additional aquatic reservoir of these bacteria.  相似文献   

2.
A total of 51 potential pathogenic vibrios were isolated from moribund silver sea bream (Sparus sarba) collected from fish farms in Hong Kong. Using the API 20E system and the scheme of Alsina and Blanch (1994), 7 species were identified from all isolates. These species were Vibrio alginolyticus (24 strains), Vibrio vulnificus (12 strains), Vibrio parahaemolyticus (7 strains), Vibrio logei (4 strains), Vibrio pelagius II (2 strains), Vibrio fluvialis (1 strain) and Vibrio meditterranei (1 strain). The three dominant species (V. alginolyticus, V. vulnificus and V. parahaemolyticus) were confirmed to be virulent to sea bream by experimental challenge.

All isolates were screened for plasmid DNA by agarose gel electrophoresis and tested for susceptibility to 16 antibiotics by the agar dilution method. Of the 51 isolates examined, all strains were sensitive to ceftriaxone, streptomycin, nalidixic acid and rifampicin, and almost all were sensitive to ceftazidime, netilimicin, chloramphenicol and sulfamethoxazole. Most isolates were resistant to ampicillin (60.8%), cefuroxime (66.7%), amikacin (55%), kanamycin (58.8%) and trimethoprim (76.5%). Fifteen of the 51 isolates harboured 1–4 plasmids, with sizes ranging from 9 to 123 kb. Both the plasmids and the associated antibiotic resistance (ampicillin, cefuroxime and trimethoprim) of 9 isolates could be transferred to a recipient by single-step conjugation. However, the frequencies were very low, ranging from 10−11 to 10−9. The present results indicate that resistance to these antibiotics is chromosomal.  相似文献   


3.
This study reports molecular markers potentially associated with resistance or sensitivity to the impact of copper in juvenile red abalone, Haliotis rufescens, in the north of Chile under experimental conditions. Genomic analysis was made applying subtractive hybridization libraries (SSH) to identify genes up-and down regulated during cooper exposure in abalone over periods of 12 and 168 h exposed to 2.5 and 10 μg/L of Cu+2. Results obtained from the SSH library revealed 368 different sequences regulated by copper, that correspond to eight major physiological functions. The validation of these sequences obtained by SSH as well as their expression kinetics were made by PCR in real time on 14 potential genes regulated by metal stress. This study provides information for the characterization of potential genomic markers that may be used in future environmental monitoring and to investigate new mechanisms of stress to copper in this commercially important marine species.  相似文献   

4.
《Marine pollution bulletin》2014,78(1-2):325-332
Ratios of sucrose-negative to sucrose-positive vibrios on TCBS agar (suc−/suc+) indicate the abundance of potential human pathogenic non-cholera vibrios in coastal mariculture environments of the Lingayen Gulf (Philippines. In guts of adult maricultured milkfish (Chanos chanos) of suc− vibrios reached extreme peak values ranging between 2 and 545 million per g wet weight. Suc− vibrios outnumbered suc+ vibrios in anoxic sediments, too, and were rarely predominant in coastal waters or in oxidized sediments. Suc−/suc+ ratios in sediments increased toward the mariculture areas with distance from the open sea at decreasing redox potentials. There is circumstantial evidence that suc− vibrios can be dispersed from mariculture areas to adjacent environments including coral reefs. An immediate human health risk by pathogenic Vibrio species is discounted, since milkfish guts contained mainly members of the Enterovibrio group. A representative isolate of these contained proteolytic and other virulence factors, but no genes encoding toxins characteristic of clinical Vibrio species.  相似文献   

5.
基于环境DNA(eDNA)技术的鱼类多样性监测方法因具有灵敏度高、对目标生物无伤害以及成本低等特点,近年来在国内外得到了广泛应用。eDNA方法的有效性往往取决于宏条形码引物的选择,尽管目前已经有一些鱼类环境DNA宏条形码引物,但较少有研究综合评估这些引物的检出效果。本研究评估了来自COI、Cytb、12S rRNA和16S rRNA基因的29对鱼类eDNA宏条形码引物(包括本研究设计的2对和从国内外文献中引用的27对引物),首先基于计算机模拟PCR(in silico PCR)进行了初步分析,随后通过高通量测序对其中效果较好的17对引物开展了更进一步的验证,结果显示:计算机模拟PCR结果良好的引物在进行高通量测序时并非全部表现良好,表明引物的筛选不能仅仅依靠计算机模拟PCR;具有更长扩增片段长度的宏条形码引物并没有获得理想中更好的扩增效果,表现效果好的引物大多是扩增片段长度处于200~300 bp之间的引物;相对于COI和Cytb而言,12S rRNA引物与16S rRNA引物均具有良好的扩增效果,适宜作为鱼类环境DNA宏条形码而用于鱼类多样性研究;同时,鉴于当前的鱼类DNA条形码数据库尚不完备以及不同引物的特性不同,使用多对引物将大大增加物种的检出概率和eDNA研究的可信性。本研究展示了eDNA在评估生物多样性方面的潜力,有助于将来的鱼类eDNA研究,从而为鱼类多样性的保护提供参考。  相似文献   

6.
Black band disease(BBD),characterized by the Cyanobacterial dominated pathogenic consortium,is thought to play a key role in the global decline of the coral reef ecosystems.The present paper originally documents a case of BBD from Yongxing Island(Xisha Islands,South China Sea),and further probes the reasons of this abnormal phenomenon.Prior to 2007,corals at northern reef-flat of Yongxing Isand were in healthy growth.Catastrophic coral mortality occurred between 2007 and 2008.The 16S rRNA gene sequencing and PCR amplification,with universally conserved primers,were applied to detect the contagious bacterial community of the microbial mat.The results demonstrated that six bacterial divisions constituted the clone libraries derived from the BBD mat,and that Cyanobacteria are the most diversely represented group that inhabit BBD bacterial mats,despite the fact that species in five others divisions(α-Proteobacteria,γ-Proteobacteria,Bacteroidetes,Verrucomicrobia and Actinobacteria) are also consistently diverse within the BBD mats of diseased coral.Other factors such as coral bleaching,typhoons,ocean acidification and crown-of-thorns starfish outbreaks,are not primarily responsible for the coral mortality within such a short time interval.The disaster expansion of BBD associated with Cyanobacterial blooms is a more likely mechanism impacting these coral reefs.Excessive human activity enhances the eutrophication of the marine water of the reefal region and may result in occurrence of the BBD.  相似文献   

7.
Transport of ballast water is one major factor in the transmission of aquatic organisms, including pathogenic bacteria. The IMO-guidelines of the Convention for the Control and Management of Ships' Ballast Water and Sediments, states that ships are to discharge <1 CFU per 100 ml ballast water of toxigenic Vibrio cholerae, emphasizing the need to establish test methods. To our knowledge, there are no methods sensitive and rapid enough available for cholera surveillance of ballast water. In this study real-time PCR and NASBA methods have been evaluated to specifically detect 1 CFU/100ml of V. cholerae in ballast water. Ballast water samples spiked with V. cholerae cells were filtered and enriched in alkaline peptone water before PCR or NASBA detection. The entire method, including sample preparation and analysis was performed within 7 h, and has the potential to be used for analysis of ballast water for inspection and enforcement control.  相似文献   

8.
Oxytetracycline-resistant bacteria were isolated from a mariculture farm in China, and accounted for 32.23% and 5.63% of the total culturable microbes of the sea cucumber and the sea urchin rearing waters respectively. Marine vibrios, especially strains related to Vibrio splendidus or V. tasmaniensis, were the most abundant resistant isolates. For oxytetracycline resistance, tet(A), tet(B) and tet(D) genes were detected in both sea cucumber and sea urchin rearing ponds. The dominant resistance type for V. tasmaniensis-like strains was the combination of both tet(A) and tet(B) genes, while the major resistance type for V. splendidus-like strains was a single tet(D) gene. Most of the sea cucumber tet-positive isolates harbored a chloramphenicol-resistance gene, either cat IV or cat II, while only a few sea urchin tet-positive isolates harbored a cat gene, actually cat IV. The coexistence of tet and cat genes in the strains isolated from the mariculture farm studied was helpful in explaining some of the multi-resistance mechanisms.  相似文献   

9.
鄱阳湖流域蚌类环境DNA宏条形码引物的筛选验证   总被引：1，自引：0，他引：1  
环境DNA技术是一种非侵入、高灵敏、高效率且对环境无破坏性,对生物体无损伤的调查工具.为了筛选适合于蚌类环境DNA生物多样性研究的宏条形码引物,本研究通过对鄱阳湖流域24种常见蚌的基因组DNA进行普通扩增和高通量测序筛选了11对引物(设计了9对通用引物及从相关文献中引用了2对引物),结果显示引物cyt b和16S rRNA具有良好的扩增效果和高辨别度.进一步用环境样本(n=6)并结合传统采样技术对这2对引物进行验证,结果表明:使用引物16S rRNA共注释到蚌科物种6属8种,蚌科物种序列占总序列数的26.69%;而使用引物cyt b共注释到蚌科物种4属6种,蚌科物种序列占总序列数的6.60%.引物16S rRNA更适合用于蚌类环境DNA生物多样性研究的宏条形码引物.在生物多样性监测中环境DNA技术可以作为传统方法的有效补充,且同时使用多对引物,可增加可信度和检测率.  相似文献   

10.
In this study, DNAs were extracted from sediment samples at depths of 5, 35, and 69 cm from eutrophic Guanting reservoir, China. 16S rDNAs were amplified by PCR and clone libraries were constructed. The depth-related distribution of bacterial community in the sediment was characterized by using amplified 16S rDNA restriction analysis (ARDRA) and sequencing of the dominant clones. The results indicated that species diversity in the sediment of Guanting reservoir was rather high with the Shannon-Wiener index about 5.8. Bacterial richness varied in different depths: the highest in the sample of 35 cm in depth; followed by the sample of 5 cm in depth; and the lowest bacterial richness in the sample of 69 cm. Dominant species from the three samples were different although there were some common clones. PhyIogenetic analysis showed that all of the dominant clones in the three layers were uncultured bacteria and distantly related to the previously reported species in beta or gamma subclass of proteobacteria, including bacterial groups that have the ability to degrade aromatic hydrocarbons, n-all:anes, chlorinated organic compounds, or to accumulate polyphosphate, etc. Changes of deptt-related bacterial community in the Guanting reservoir sediment might reflect the pollution history and the water quality of the reservoir. In addition, the cloned sequences from the Guanting reservoir sediment were all different from the presently reported ones, indicating that there were some particular bacteria in that environment.  相似文献   

11.
Although Escherichia coli is widely distributed in the marine environment, only a small percentage are pathogenic to humans. Nonetheless, the widespread occurrence of waterborne infections of E. coli origin in humans has become one of the major health problems worldwide. To date, several types of enterovirulent E. coli have been recognized as the aetiologic agents of various gastrointestinal infections in humans. The most commonly encountered are those belonging to the enterotoxigenic (ETEC), enteroinvasive (EIEC), enterohaemorrhagic (EHEC) and enteropathogenic (EPEC) subtypes. In order to better determine the health risks that are associated with exposure to some of these specific subtypes, we have developed a very sensitive multiplex PCR system for the rapid detection and typing of ETEC, EHEC and possibly EPEC strains of E. coli in the aquatic environment. The target genes chosen for this investigation included: the PHO-A housekeeping gene (present in all E. coli); the LT1, LT2 and ST1 genes of ETEC; the VT1 and VT2 verotoxin, and EAE virulence genes of EHEC and EPEC, respectively. Six pairs of oligonucleotide primers were designed to simultaneously amplify internal fragments of these genes by multiplex PCR to generate PCR products that could be analysed and confirmed with relative ease by gel electrophoresis and HincII enzyme digestion. The results showed that the six sets of PCR primers were highly specific for their target genes and produced specific amplimers of the expected size from several control strains of E. coli – ATCC 35401 (LT1+/ST1+); SA53 (LT2+/VT2+); and O157 (VT1+/VT2+/EAE+). The detection sensitivity of the multiplex PCR system for the six target genes in an E. coli cell mixture was optimized and enhanced by preincubating serially diluted cells in Luria-Bertani broth for 6 h prior to PCR analysis. The results obtained indicated a detection sensitivity of 10° CFU (of each strain) per 100 μl reaction. Multiplex PCR analysis of seawater samples collected from four sewage-polluted sites in Hong Kong indicated the presence in all four samples of E. coli bacteria that were positive for LT1, ST1, VT1 and EAE virulence genes. Overall, the data indicated that the multiplex PCR system described in this study is a potentially very useful and powerful method for routine monitoring and risk assessment of water quality.  相似文献   

12.
Water quality in Singapore's coastal area was evaluated with microbial indicators, pathogenic vibrios, chemical tracers and physico-chemical parameters. Sampling sites were grouped into two clusters (coastal sites at (i) northern and (ii) southern part of Singapore). The coastal sites located at northern part of Singapore along the Johor Straits exhibited greater pollution. Principal component analysis revealed that sampling sites at Johor Straits have greater loading on carbamazepine, while turbidity poses greater influence on sampling sites at Singapore Straits. Detection of pathogenic vibrios was also more prominent at Johor Straits than the Singapore Straits. This study examined the spatial variations in Singapore's coastal water quality and provided the baseline information for health risk assessment and future pollution management.  相似文献   

13.
杨海乐  张辉  杜浩 《湖泊科学》2023,35(1):12-31
eDNA (environmental DNA)是指从环境样品（水体、土壤、沉积物、空气、混合物等）中提取的DNA,是各种生物的DNA混合物,区别于传统从单物种样品中提取的DNA。eDNA监测是指从环境样品中提取DNA,用物种特异性引物或特定DNA metabarcoding引物对其进行扩增测序、分类学分析、相对丰度分析、功能预测等,以监测环境中是否存在某特定物种,或者获得环境中物种组成、群落结构、生态功能等相关信息。eDNA监测主要应用在特定物种监测检出、生物多样性调查评估、群落结构和功能分析、生态系统过程分析评估等领域。eDNA监测可以应用在陆地、水体、空气、器物表面、生物（内）表面等任何有可能存在不确定DNA的场景。因为eDNA监测的非侵入性、灵敏检出、大类群检出、易标准化方法与结果、低人力物力时间成本、结果可核查等特点,未来很有希望发展成为一种常规的物种监测、群落功能预测、生态过程分析的方法,对象可涵盖所有生境条件下的所有生物类群。但要实现这个图景,需要从普遍性和特殊性层面完成eDNA监测技术链条中10个关键环节的技术标准化：样品重复数设计、采样时间设计、采样点设计、采样方法设...  相似文献   

14.
Polybrominated diphenyl ethers (PBDEs) are used as flame retardants in numerous products. These compounds have been found to enter the marine environment where they have the potential to bioaccumulate in biota. Limited information is currently available concerning the levels of PBDEs in Australian marine wildlife. This study presents baseline information on PBDE levels in a variety of marine species from Queensland, Australia and considers the influence of species-specific factors on contaminant levels and tissue distribution in marine turtles. Overall, the PBDE levels measured in this study are relatively low compared to marine biota from the northern hemisphere, indicating low level input into the marine system of Queensland. This is in general agreement with global estimates which suggest low PBDE usage in Australia. Previous studies, however, have found relatively high PBDE levels in Australian human milk and sera. This discrepancy in contamination trends between terrestrial and marine biota suggests that future transport of PBDEs may occur to the marine system in Australia.  相似文献   

15.
《Marine pollution bulletin》2009,58(6-12):409-418
Polybrominated diphenyl ethers (PBDEs) are used as flame retardants in numerous products. These compounds have been found to enter the marine environment where they have the potential to bioaccumulate in biota. Limited information is currently available concerning the levels of PBDEs in Australian marine wildlife. This study presents baseline information on PBDE levels in a variety of marine species from Queensland, Australia and considers the influence of species-specific factors on contaminant levels and tissue distribution in marine turtles. Overall, the PBDE levels measured in this study are relatively low compared to marine biota from the northern hemisphere, indicating low level input into the marine system of Queensland. This is in general agreement with global estimates which suggest low PBDE usage in Australia. Previous studies, however, have found relatively high PBDE levels in Australian human milk and sera. This discrepancy in contamination trends between terrestrial and marine biota suggests that future transport of PBDEs may occur to the marine system in Australia.  相似文献   

16.
A molecular PCR-based assay was developed and applied to macrophyte and seawater samples containing mixed microphytobenthic and phytoplanktonic assemblages, respectively, in order to detect toxic Ostreopsis species in Mediterranean Sea. The specificity and sensitivity of the molecular PCR assay were assessed with both plasmidic and genomic DNA of the target genus or species using taxon-specific primers in the presence of background macrophyte DNA. The PCR molecular technique allowed rapid detection of the Ostreopsis cells, even at abundances undetectable within the resolution limit of the microscopy technique. Species-specific identification of Ostreopsis was determined only by PCR-based assay, due to the inherent difficulty of morphological identification in field samples. In the monitoring of the toxic Ostreopsis blooms PCR-based methods proved to be effective tools complementary to microscopy for rapid and specific detection of Ostreopsis and other toxic dinoflagellates in marine coastal environments.  相似文献   

17.
为了检验已记录的3种裸腹溞(发头裸腹溞(Moina irrasa)、短型裸腹溞(Moina brachiata)、微型裸腹溞(Moina micrura))的系统分类,用试剂盒法分别提取3种裸腹溞的基因组DNA.利用特异性引物,通过PCR扩增了3种裸腹溞的16S r DNA部分序列,并与来自Gen Bank中每个种类相似度较高的裸腹溞属序列进行分析.结果表明,3种裸腹溞的平均种间相似度为88.7%,碱基中A+T含量均明显高于G+C含量.本研究的发头裸腹溞的16S r DNA序列与Gen Bank所下载的多刺裸腹溞(Moina macrocopa)的16S r DNA序列相似度为99%,遗传距离(K2P双参考模型)为0.5%,属种内范围;两个地区的短型裸腹溞测得的16S r DNA序列与Gen Bank下载的欧洲短型裸腹溞的16S r DNA序列序列相似度相对较低(88%~90%),遗传距离较大(13.2%~13.5%左右),已达到属内种间分化水平.基于16S r DNA构建的NJ树和贝叶斯树也支持以上结论.结果表明,本研究的发头裸腹溞可能为多刺裸腹溞,本研究用的短型裸腹溞与Gen Bank下载的欧洲短型裸腹溞已经达到种间分化的标准.由于缺乏物种形态资料和其他分子标记的对比,3种裸腹溞的分类地位还需进行更深入的探讨.  相似文献   

18.
Tang  Kaihao  Zhan  Waner  Zhou  Yiqing  Xu  Tao  Chen  Xiaoqing  Wang  Weiquan  Zeng  Zhenshun  Wang  Yan  Wang  Xiaoxue 《中国科学:地球科学(英文版)》2020,63(1):157-166
Scleractinian corals host numerous microbial symbionts with different types of interactions. The gastric cavity of scleractinian coral, as a semiclosed subenvironment with distinct chemical characteristics(e.g., dissolved O2, pH, alkalinity, and nutrients), harbors a distinct microbial community and a diverse array of bacteria that can be pathogenic or beneficial. Galaxea fascicularis is one of the dominant massive scleractinian coral species on inshore fringing reefs in the northern South China Sea.Although the abundance of coral-associated bacteria has been investigated in G. fascicularis, less is known about the microorganisms in the gastric cavity. In this study, we specially isolated cultivable bacterial strains from the gastric cavity of G.fascicularis collected from Hainan Island using a noninvasive sampling approach. Among the 101 representative bacterial strains, one Vibrio coralliilyticus strain, SCSIO 43001, was found to be a temperature-dependent opportunistic pathogen of G.fascicularis. The antagonistic activity between the 100 strains and V. coralliilyticus SCSIO 43001 was tested using a modified Burkholder diffusion assay. Our results showed that V. coralliilyticus SCSIO 43001 inhibits the growth of Erythrobacter flavus and Sphingomonas yabuuchiae. Additionally, we found that three Pseudoalteromonas strains showed moderate to high antibacterial activity against V. coralliilyticus SCSIO 43001 and several other coral-associated Gram-negative bacterial strains.These results suggest that competition between the coral pathogen and other bacteria also occurs in the gastric cavity of coral, and Pseudoalteromonas strains in the gastric cavity of G. fascicularis may provide a protective role in the defense against coinhabiting coral pathogens at elevated temperature.  相似文献   

19.
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20.
A bacterial copper-resistant strain was isolated from a hatchery-conditioned adult of the scallop Argopecten purpuratus and was identified as Vibrio sp. according to its physiological characteristics. The lowest concentration of Cu2+ required for its complete inhibition in VNSS medium was 50 microg ml(-1). The Vibrio strain was found to accumulate copper, exhibiting cellular and loosely bound copper levels of 201.14 and 493.21 microg g(-1) dry weight, respectively, after 24 h of incubation in VNSS medium supplemented with 15 microg ml(-1) of Cu2+, with cellular concentration factors of 10.17 and 14.35 after 12 and 24 h of exposure. When a scallop larvae culture was exposed to a concentration of 6.46 x 10(2) CFU ml(-1) of Cu-enriched Vibrio, they accumulated 20.42 +/- 1.12 and 30.96 +/- 1.85 microg Cu2+ g(-1) dry weight after 12 and 24 h, respectively. This study suggests that bacterial copper accumulation could be very active in marine environments increasing the occurrence of copper transfer to marine food chains.  相似文献   

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