Soil: the environmental source of Escherichia coli and Enterococci in Guam's streams

We have previously documented that faecal indicator bacteria (Escherichia coli, faecal coliform, enterococci) recommended by the U.S. Environmental Protection Agency (USEPA) to establish recreational water quality standards are naturally found in high concentrations in the surface and subsurface of soils in Hawaii. Rain, the source of all streams in Hawaii, washes the soil sources of faecal bacteria into all the streams of Hawaii, at concentrations which consistently exceed the USEPA recreational water quality standards. The objective of this study was to test the hypothesis that faecal bacteria are able to establish themselves in the soil environments of tropical islands by conducting the same study in Guam, a tropical pacific island with warmer temperatures and higher humidity than Hawaii. The same methods and study design used in Hawaii was used in Guam. The results of the study conducted in Guam revealed that all streams contain consistently high concentrations of faecal coliform, E. coli, and enterocci which exceeded the old USEPA recreational water quality standard of 200 faecal coliform/100 ml as well as the new water quality standards of 126 E. coli/100 ml or 33 enterococci/100 ml. These same faecal indicator bacteria were recovered in high concentrations in surface and subsurface (18-36 cm depth) soil samples in Guam. Limited coastal water analysis showed that most coastal marine waters contain low concentrations of faecal bacteria but coastal waters impacted by stream run-off showed elevated levels of faecal bacteria. The results of this study support the hypothesis that environmental conditions in the tropical areas of the world can support the growth and establishment of populations of faecal bacteria in the soil. Thus, soil becomes an environmental, non-faecal source of faecal indicator bacteria. These results indicate that USEPA water quality standards may not be directly applicable to tropical island environments.     

Distribution of Synechococcus spp. determined by immunofluorescent assay

By using two polyclonal antisera against WH 7803 strain (Synechococcus sp.) and WH 5701 strain (Synechococcus bacillaris) it is possible to detect and to enumerate cells of the two cyanobacterial serogroups. The immunofluorescence technique was used to study the distribution of the two serogroups in the estuarine, coastal and upwelling waters of the Mediterranean Sea surrounding Messina. In the estuarine waters of the Alcantara River (Ionian Sea), the WH 7803 serogroup was present at a concentration in the order of 10² cells ml⁻¹ and the WH 5701 serogroup at a concentration of 5·5 × 10² cellsml⁻¹. In the coastal waters of Messina, where urban and industrial wastes are usuallydumped, the concentration of total phycoerythrin- Synechococcus ranged from 1·3 × 10² to 4·1 × 10³ cells ml⁻¹; the WH 7803 serogroup accounted for 50–94% of the totalpopulation in Ionian stations, whereas the WH 5701 serogroup ranged from1·4 × 10¹ to6·7 × 102cells ml⁻¹. In the upwelling area (Straits of Messina) bothserogroups were found. Vertical distribution of two Synechococcus strains had anopposite trend and their concentrations were of the order of 10¹–10²cells ml⁻¹. Theuse of the Scan laser system allows both autofluorescent and labelled organismsto be distinguished in a preparation for optical microscopy. It also allows false-positivecells to be distinguished.     

Isolation of faecal coliform bacteria from the American alligator (Alligator mississippiensis)

Aims: To determine whether American alligators (Alligator mississippiensis) are an unrecognized poikilothermic source of faecal coliform and/or potential pathogenic bacteria in South Carolina’s coastal waters. Methods and Results: Bacteria from the cloaca of American alligators, as well as bacteria from surface water samples from their aquatic habitat, were isolated and identified. The predominant enteric bacteria identified from alligator samples using biochemical tests included Aeromonas hydrophila, Citrobacter braakii, Edwardsiella tarda, Escherichia coli, Enterobacter cloacae, Plesiomonas shigelloides and putative Salmonella, and these were similar to bacteria isolated from the surface waters in which the alligators inhabited. Based on most‐probable‐number enumeration estimates from captive alligator faeces, faecal coliform bacteria numbered 8·0 × 10⁹ g^?1 (wet weight) of alligator faecal material, a much higher concentration than many other documented endothermic animal sources. Conclusions: A prevalence of enteric bacteria, both faecal coliforms and potential pathogens, was observed in American alligators. The high faecal coliform bacterial density of alligator faeces may suggest that alligators are a potential source of bacterial contamination in South Carolina coastal waters. Significance and Impact of the Study: These findings help to increase our understanding of faecal coliform and potential pathogenic bacteria from poikilothermic reptilian sources, as there is the potential for these sources to raise bacterial water quality levels above regulatory thresholds.     

Lignicolous marine fungi in the Straits of Messina,Italy

The occurrence and distribution of lignicolous marine fungi in the Straits of Messina has been studied. Using submerged panels of pine, beech and poplar, twenty fungal species were identified. The lignicolous mycoflora of the Messina Straits was not significantly different from that reported in the literature for other temperate marine coastal environments, Ascomycotina being frequent, while Basidiomycotina were rare.     

Detection of the nifH gene of Methanobrevibacter smithii: a potential tool to identify sewage pollution in recreational waters

AIMS: The goal of this study was to develop and test the efficacy of a PCR assay for the environmental detection of the nifH gene of Methanobrevibacter smithii, a methanogen found in human faeces and sewage. METHODS AND RESULTS: PCR primers for the nifH gene of M. smithii were designed, tested and used to detect the presence or absence of this organism in faecal and environmental samples. Specificity analysis showed that the Mnif primers amplified products only in M. smithii pure culture strains (100%), human faeces (29%), human sewage samples (93%) and sewage-contaminated water samples (100%). No amplification was observed when primers were tested against 43 bacterial stock cultures, 204 animal faecal samples, 548 environmental bacterial isolates and water samples from a bovine waste lagoon and adjacent polluted creek. Sequencing of PCR products from sewers demonstrated that a 222-bp product was the nifH gene of M. smithii. The minimal amount of total DNA required for the detection of M. smithii was 10 ng for human faeces, 10 ng for faecally contaminated water and 5 ng for sewage. Recreational water seeded with M. smithii established a lower detection limit of 13 cells ml(-1). CONCLUSIONS: The Mnif assay developed during this investigation showed successful detection of M. smithii in individual human faecal samples, sewage and sewage-contaminated water but not in uncontaminated marine water or bovine-contaminated waters. The Mnif assay appears to be a potentially useful method to detect sewage-polluted coastal waters. SIGNIFICANCE AND IMPACT OF THE STUDY: This study was the first to utilize methanogens as an indicator of sewage pollution. Mnif PCR detection of M. smithii was shown to be a rapid, inexpensive and reliable test for determining the presence or absence of sewage pollution in coastal recreational waters.     

江苏近海生态系统服务价值评估

海洋生态系统服务是人类从海洋生态系统中获得的效益,对沿海地区社会经济具有重要的支撑作用。采用国家标准《海洋生态资本评估技术导则》的方法,评估了江苏省管辖海域的生态系统服务价值,并揭示了其空间分布特征。江苏近海海域面积1.18万km2,2008年提供的生态系统服务总价值为426.02亿元,供给服务、文化服务、调节服务和支持服务价值分别为226.38亿元,186.61亿元,12.68亿元和0.34亿元;3个沿海地市比较,盐城近海生态系统服务价值最高,为184.55亿元;其次是连云港近海,为155.58亿元;南通近海最低,为85.89亿元。江苏近海生态系统服务价值空间分布密度平均为361.91万元/km2,从近岸到远海总体呈逐渐降低趋势,高值区主要分布在滨海旅游区和海洋保护区,中值区出现在养殖区,其它海域的服务价值则较低。江苏近海生态系统服务利用属于供给服务和文化服务共同主导的综合开发利用型,连云港近海属于文化服务主导的开发利用型,盐城近海属于供给服务和文化服务共同主导的综合开发利用型,南通近海则属于供给服务主导的开发利用型。建议在江苏近海开展离岸养殖工程,以提高近海供给服务价值;在盐城,特别是南通近岸海域开发海洋公园、渔家乐和其他亲水性旅游活动以提高文化服务价值。     

Enumeration of faecal coliforms from recreational coastal sites: evaluation of techniques for the separation of bacteria from sediments

AIMS: To identify the most efficient techniques for the separation of micro-organisms from coastal sediments and, using these techniques, to determine the concentration of faecal indicator organisms in recreational coastal water and sediment. METHODS AND RESULTS: Sediment samples were taken from a range of recreational coastal sites and subjected to various physical techniques to separate micro-organisms from sediment particles. Techniques investigated included manual shaking, treatment by sonication bath for 6 and 10 min, respectively, and by sonication probe for 15 s and 1 min, respectively. The use of the sonication bath for 10 min was the most successful method for removing micro-organisms from sediment particles where sediments consisted mainly of sand. When sediments contained considerable proportions of silt and clay, however, manual shaking was most successful. Faecal coliforms were then enumerated by membrane filtration in both water and sediment from three recreational coastal sites, chosen to represent different physical sediment characteristics, over a 12-month period. Faecal coliform concentrations were generally greater in sediment compared with overlying water for all samples. This was most evident in sediment consisting of greater silt/clay and organic carbon content. CONCLUSIONS: This study demonstrated the importance of sediment characteristics in determining the most efficient method for the separation of micro-organisms from coastal sediments. Sediment characteristics were also found to influence the persistence of micro-organisms in coastal areas. SIGNIFICANCE AND IMPACT OF THE STUDY: Recreational coastal sediments can act as a reservoir for faecal coliforms; therefore, sampling only overlying water may greatly underestimate the risk of exposure to potentially pathogenic micro-organisms in recreational waters.     

Distribution of potentially pathogenic bacteria as free living and plankton associated in a marine coastal zone

AIMS: To determine the abundance of faecal and nonfaecal bacteria related to human and animal health, as free living or associated with small (>64 microm) and large (>200 microm) plankton, samples were collected monthly from the coastal zone at Messina (Italy). METHODS AND RESULTS: Different enrichment and selective cultural methods were used to determine the abundance of bacteria in sea water and plankton. The bacteria were more frequently isolated from water and large plankton than from small plankton. Vibrio and Aeromonas spp. showed different distribution patterns in water and plankton. Faecal indicators were always present in water and the large size class plankton samples. Enterococci associated with large plankton were more abundant than E. coli in the winter. Vibrio species distributions were different in water and plankton samples. Among arcobacters only A. butzleri was isolated from water and plankton samples. Campylobacter spp. was always absent in small plankton and more frequent in large plankton than in water. CONCLUSIONS: The colonization of zooplankton by potentially pathogenic bacteria is a widespread phenomenon. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of potentially pathogenic bacteria in sea water and associated with plankton can have ecological and epidemiological implications.     

Genetic structure analysis of mantis shrimp <Emphasis Type="Italic">Oratosquilla oratoria</Emphasis> based on mitochondrial DNA control region sequence

The genetic relationships of mantis shrimp Oratosquilla oratoria between the coastal waters of China and Japan were not well studied. To reveal the genetic differentiation and genetic structure among populations, we collected populations of mantis shrimp O. oratoria from the coastal waters of China and Japan to analyze the mtDNA control region variation. A total of 309 individuals of O. oratoria were collected from 13 localities (11 from China and 2 from Japan) and a segment of mitochondrial DNA control region was sequenced. Three hundred nine haplotypes were defined, yielding a very high haplotype diversity and nucleotide diversity. Two lineages of O. oratoria were revealed and displayed strong differences in the geographical distribution. In the coastal waters of China, the geographic distribution of the two lineages was completely separated by the Yangtze River estuary; however, the lineages showed geographic sympatry in two populations from Japan. Based on the lineage distribution, three groups were defined. There was no significant genetic differentiation among the populations within the three groups, indicating high gene flow within each group. Significant and negative values for Tajima D and Fu’s Fs tests, and mismatch distributions for two lineages indicated population expansion. The present result confirmed that the freshwater outflow from the Yangtze River formed a physical barrier and affected gene exchange. The different distribution patterns of the two lineages in coastal waters of China and Japan indicated that the larvae of O. oratoria were transferred from China to the coastal waters of Japan with a one-way gene flow.     

Contribution of Crenarchaeota and Euryarchaeota to the prokaryotic plankton in the coastal northwestern Black Sea

The NW Black Sea is an area with peculiar hydrological, hydrochemicaland biological features (river plume dynamics, coastal upwelling,intense eutrophication, blooms algae). We determined the abundanceand distribution of the two major groups of Archaea (Crenarchaeotaand Euryarchaeota) in three contrasting sites of the NW BlackSea from May to August 2004 to estimate the contribution ofnon-extremophilic Archaea to the coastal prokaryotic plankton.Using catalyzed reporter deposition-fluorescence in situ hybridization,Euryarchaeota were found to be particularly prominent (11–22%of total prokaryotic plankton) in the low-salinity waters duringlate-spring and summer, whereas Crenarchaeota contributed only5% to the prokaryotic community in these waters. The abundanceof Crenarchaeota was highest in nutrient- and particle-richwaters and lowest in offshore Black Sea surface waters. Theseresults confirm recent studies on the distribution of planktonicArchaea with Euryarchaeota dominating the archaeal communityin surface waters. Statistical analyses indicated significantcorrelations between crenarchaeotal abundance, ammonia and nitrateconcentrations at each sites. Our data show that the archaealplankton constitute an important and dynamic component of theprokaryotic plankton in the coastal oxygenated waters of theNW Black Sea and might play a significant role in the nitrogencycle.     

Relation between Aeromonas and faecal coliforms in fresh waters

A possible correlation between the presence of mesophilic aeromonads and the number of faecal coliforms present in three fresh-water habitats subject to differing levels of faecal pollution was investigated. Concentration of Aeromonas spp. between 10(2)-10(9) cfu/100 ml and faecal coliforms of between 9-10(7) cfu/100 ml were found in the waters. In water free from faecal pollution there was no correlation but in polluted waters there was a significant relationship between the numbers of aeromonads, faecal coliform and the concentration of organic matter measured by biological oxygen demand.     

Virological quality of the Ria de Aveiro: Validity of potential microbial indicators

The summer occurrence of human enteroviruses and rotaviruses in the bacteriologically clean area of the Ria de Aveiro, a coastal marine lagoon, prompted the question of the assessment of the virological quality of recreational waters and shellfish raising beds. Enteroviruses were present in surface water at a density of 3 pfu 10 l^–1 and were accumulated in sediments and, especially, in cockles where they reached concentrations 2 to 3¹⁰log units greater. Rotaviruses were detected at one¹⁰log unit below the density of enteroviruses in sediments and cockles and were not detected in water. Four bacteriophage systems were assayed as indicators of human enteric viruses: somatic coliphages ofE.coli C, sexual and sexual-RNA coliphages plated onSalmonella WG49 and phages againstBacteroides fragilis HSP40. The results obtained from 2 lagoon stations sampled in summer, autumn and winter showed that the four systems failed to indicate the presence of enteroviruses and rotaviruses in water, sediment and shellfish samples. The absence of phages ofB. fragilis HSP40 in all types of samples taken from the lagoon, but not from the residual waters of the treatment station, suggests that they may suffer a strong negative pressure in this ecosystem as their proportion to the coliphages in the cockles deviated strongly from the ratio of 1100 to 11000 observed at the sewage outfall. In fact, no correlation was observed between these phages and enteric viruses or coliphages. Alternatively, it is possible that the importance of diffuse faecal pollution and the interference of faecal pollutants of animal origin, including migratory sea birds which are abundant in winter, can alter the proportions of the faecal bacteriophages beyond recognition. It is apparent that bacteriophage monitoring of the health risk linked to the occurrence of viruses in the marine environment is not yet fully resolved, what may leave viral quality assessment dependent on direct detection of human enteric virus.     

Fauna of anemonefishes (Pomacentridae,Amphiprioninae) and their host sea anemones (Cnidaria,Actiniaria) on reefs of Phu Quy,Con Son,and An Thoi Islands (South China Sea,South Vietnam,and Gulf of Thailand) and a review of these groups from the coastal waters of Vietnam

A full species composition of anemonefishes (Pomacentridae, Amphiprioninae) and their host sea anemones (Cnidaria, Actiniaria) from reefs of Phu Quy and Con Son Islands (off South Vietnam) and An Thoi Islands (northeastern Gulf of Thailand) is presented for the first time. A comparison of the faunas of anemonefishes and their host sea anemones is conducted in the coastal waters of Vietnam and adjacent waters. The presence of intermittent ranges of the anemonefishes in the coastal waters of Vietnam is demonstrated. For example, the ranges of two species, Amphiprion clarkii and A. frenatus, include the coastal waters of Vietnam, excluding the Gulf of Thailand, where they are not found even at the eastern border of the gulf. A. perideraion is absent in the Gulf of Tonkin, but it is distributed to the south along the entire coastal zone of Vietnam, including the Gulf of Thailand. In the coastal waters of Vietnam, A. sandaracinos is distributed only between the Gulf of Tonkin and Gulf of Thailand. A. polymnus is reliably indicated in the coastal waters of Vietnam between 12°50′ and 9°54′ N, but, most likely, its range is continuous along the entire Vietnamese coast. The richness of species composition of host sea anemones decreases (from nine to three species) in the southern direction, from Central Vietnam to the eastern border of the Gulf of Thailand.     

基于突变理论的近岸海域生态风险综合评价方法——以罗源湾为例

基于突变理论建立了“海水水质-沉积物-生物体”三相一体、“富营养化-重金属-有机污染”3种类型的近岸海域生态风险综合评价指标体系和递级突变模型,综合相关质量标准,建立了近岸海域生态风险4级评判标准（无、低、中、高）,采用2007和2009年监测数据对罗源湾进行生态风险综合评价.结果表明： 2007-2009年,罗源湾生态风险空间变化特征表现为从湾顶到湾口呈降低趋势;2009年罗源湾生态风险等级总体小于2007年;罗源湾关键生态风险因素为富营养化.本文所建方法能够识别近岸海域生态风险的时空变化特征,能较好地反映出关键风险因素,为近岸海域生态风险有效预测预警提供了基础方法.     

Distribution of Aeromonas species in waters with different levels of pollution

Strains of Aeromonas spp. (883) were isolated from 10 stations in the north-west of Spain. Biotyping of the strains gave: 55% Aeromonas caviae , 34% A. hydrophila , 6% A. sobria and 5% Aeromonas spp. Phenotypic characters that have been claimed to be related to virulence such as haemolysis and the Voges-Proskauer reaction were detected mostly in A. hydrophila and A. sobria. The distribution of the species was significantly related to levels of faecal pollution in waters. Aeromonas caviae predominated in sewage and waters with a high degree of faecal pollution. In less polluted waters, either fresh or marine, A. caviae and A. hydrophila were almost equally distributed. In waters with low or no faecal pollution, the proportion of A. sobria to other species increased considerably.     

Distribution of Aeromonas species in waters with different levels of pollution

Strains of Aeromonas spp. (883) were isolated from 10 stations in the north-west of Spain. Biotyping of the strains gave: 55% Aeromonas caviae, 34% A. hydrophila, 6% A. sobria and 5% Aeromonas spp. Phenotypic characters that have been claimed to be related to virulence such as haemolysis and the Voges-Proskauer reaction were detected mostly in A. hydrophila and A. sobria. The distribution of the species was significantly related to levels of faecal pollution in waters. Aeromonas caviae predominated in sewage and waters with a high degree of faecal pollution. In less polluted waters, either fresh or marine, A. caviae and A. hydrophila were almost equally distributed. In waters with low or no faecal pollution, the proportion of A. sobria to other species increased considerably.     

Impact of synthetic pyrethroid-sheep dip on the indigenous microflora of animal slurries

The chemical constituents of sheep dip in the UK are currently changing from organophosphate-based to synthetic pyrethroid-based insecticides. As a result, changes are also being made to the methods of disposal of these chemicals in the environment, such that pyrethroid sheep dips must now be diluted in animal slurry or water. To date, there is a lack of quantitative information on the impact of the insecticide on the indigenous microflora of animal slurries. This paper investigated the impact of Bayticol (synthetic pyrethroid sheep dip) over a range of concentrations on selected populations of bacteria within animal slurry. It was found that, with increasing pesticide concentration, there was up to a four orders of magnitude increase in the numbers of faecal coliforms and pathogens, such as putative Salmonella spp. These findings have implications for the disposal of sheep dip-amended animal slurries to land from several aspects: (i) the longevity of putative pathogens in the field may require re-evaluation of the time required before the return of grazing livestock to a slurry-amended field; (ii) the potential for the transfer of pathogenic bacteria and faecal coliforms into human and animal foodchains, and (iii) the increased potential for faecal coliforms being washed into streams, rivers and coastal bathing waters.     

Use of microcosms to determine persistence of Escherichia coli in recreational coastal water and sediment and validation with in situ measurements

AIMS: To determine the persistence of the faecal indicator organism Escherichia coli in recreational coastal water and sediment using laboratory-based microcosms and validation with in situ measurements. METHODS AND RESULTS: Intact sediment cores were taken from three distinct coastal sites. Overlying estuarine water was inoculated with known concentrations of E. coli and decay rates from both overlying water and sediment were determined following enumeration by the membrane filtration method at fixed time intervals over a 28-day period. It was demonstrated that E. coli may persist in coastal sediment for >28 days when incubated at 10 degrees C. Escherichia coli survival was found to have an inverse relationship with temperature in both water and sediment. In general the decay rate for E. coli was greater in water than in sediment. Small particle size and high organic carbon content were found to enhance E. coli survival in coastal sediments in the microcosms. CONCLUSIONS: Results of this microcosm study demonstrated the more prolonged survival of E. coli in coastal sediments compared with overlying water, which may imply an increased risk of exposure because of the possible resuspension of pathogenic micro-organisms during natural turbulence or human recreational activity. SIGNIFICANCE AND IMPACT OF THE STUDY: A more accurate estimate of exposure risk has been described which may subsequently be used in a quantitative microbial risk assessment for recreational coastal waters.     

Relation between Aeromonas and faecal coliforms in fresh waters

A possible correlation between the presence of mesophilic aeromonads and the number of faecal coliforms present in three fresh-water habitats subject to differing levels of faecal pollution was investigated. Concentration of Aeromonas spp. between 10²–10⁹ cfu/100 ml and faecal coliforms of between 9–10⁷ cfu/100 ml were found in the waters. In water free from faecal pollution there was no correlation but in polluted waters there was a significant relationship between the numbers of aeromonads, faecal coliform and the concentration of organic matter measured by biological oxygen demand.