Gilthead seabream (Sparus aurata) carrying antibiotic resistant enterococci. A potential bioindicator of marine contamination?

Antibiotic resistance in bacteria is a growing problem that is not only restricted to the clinical setting but also to other environments such as marine species that harbor antibiotic resistant bacteria and therefore may serve as reservoirs for antibiotic-resistance genetic determinants. The aim of this study was to evaluate antibiotic resistance phenotypes in enterococci isolated from fecal samples of gilthead seabream and the associated mechanisms of resistance. A collection of 118 samples were analyzed and 73 enterococci were recovered. The strains showed high percentages of resistance to erythromycin and tetracycline (58.9% and 17.8%, respectively). Lower level of resistance (<13%) was detected for quinupristin-dalfopristin, ampicillin, high-level-gentamicin, high-level-streptomycin, high-level-kanamycin, ciprofloxacin and chloramphenicol. The erm(B), tet(L) or tet(M), aac(6′)-aph(2″) and aph(3′)-IIIa genes were shown in isolates resistant to erythromycin, tetracycline, high-level gentamicin and high-level kanamycin, respectively. Antibiotic resistance in natural microbiota is becoming a concern of human and environmental health.     

Echinoderms from Azores islands: An unexpected source of antibiotic resistant Enterococcus spp. and Escherichia coli isolates

The prevalence of antibiotic resistance and the implicated mechanisms of resistance were evaluated in Enterococcus spp. and Escherichia coli, isolated from a total of 250 faecal samples of echinoderms collected from Azorean waters (Portugal). A total of 144 enterococci (120 Enterococcus faecium, 14 E. hirae, 8 E. faecalis, 2 E. gallinarum) and 10 E. coli were recovered. High percentages of resistance in enterococci were found for erythromycin, ampicillin, tetracyclin and ciprofloxacin. The erm(A) or erm(B), tet(M) and/or tet(L), vat(D), aac(6′)-aph(2″) and aph(3′)-IIIa genes were found in isolates resistant to erythromycin, tetracycline, quinupristin/dalfopristin, high-level gentamicin and high-level kanamycin, respectively. Resistance in E. coli isolates was detected for streptomycin, amikacin, tetracycline and tobramycin. The aadA gene was found in streptomycin-resistant isolates and tet(A) + tet(B) genes in tetracycline-resistant isolates. The data recovered are essential to improve knowledge about the dissemination of resistant strains through marine ecosystems and the possible implications involved in transferring these resistances either to other animals or to humans.     

Significance of <Emphasis Type="Italic">Vibrio</Emphasis> species in the marine organic carbon cycle—A review

The genus Vibrio, belonging to Gammaproteobacteria of the phylum Proteobacteria, is a genetically and ecologically diverse group of heterotrophic bacteria, that are ubiquitous in marine environments, especially in coastal areas. In particular, vibrios dominate, i.e. up to 10% of the readily culturable marine bacteria in these habitats. The distribution of Vibrio spp. is shaped by various environmental parameters, notably temperature, salinity and dissolved organic carbon. Vibriospp. may utilize a wide range of organic carbon compounds, including chitin (this may be metabolized by most Vibrio spp.), alginic acid and agar. Many Vibrio spp. have very short replication times (as short as ~10 min), which could facilitate them developing into high biomass content albeit for relatively short durations. Although Vibriospp. usually comprise a minor portion (typically ~1% of the total bacterioplankton in coastal waters) of the total microbial population, they have been shown to proliferate explosively in response to various nutrient pulses, e.g., organic nutrients from algae blooms and iron (Fe⁺) from Saharan dust. Thus, Vibrio spp. may exert large impacts on marine organic carbon cycling especially in marginal seas. Genomics and related areas of investigation will reveal more about the molecular components and mechanisms involved in Vibrio-mediated biotransformation and remineralization processes.     

Molecular characterizations of oxytetracycline resistant bacteria and their resistance genes from mariculture waters of China

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.     

Impact of milk fish farming in the tropics on potentially pathogenic vibrios

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.     

The occurrence, spatial and temporal distribution, and environmental routes of tetracycline resistance and integrase genes in Crassostrea virginica beds

Rapid spread of antibiotic resistance genes has placed them under consideration as emerging environmental contaminants. The occurrence and distribution of tetracycline resistance genes (TRG) and integrons (INT) have been monitored for 2 years in seven oyster beds located in the Altamaha River (Georgia, USA) estuary. TRG/INT profiles varied both temporally (p < 0.025) and spatially between oysters and their environment (p < 0.005) suggesting shifts in sources of contamination and selection of TRG/INT carriers by oysters. The latter was confirmed via analysis of tet(D) and tet(G) distribution between oysters and their environment. Uncommon class 3 integrons dominated the integrons’ profile in oysters. The frequency of TRG/INT incidents correlated (R² ? 0.9) to (i) dissolved solids, (ii) potential water density, and (iii) turbidity but not to the amount of local precipitation suggesting the watershed as the dominant source of TRG/INT contamination and other than directly rainfall-caused run-offs as its environmental route.     

Role of shellfish hatchery as a reservoir of antimicrobial resistant bacteria

The main aim of this study was to determine the occurrence of resistant bacteria in florfenicol-treated and untreated scallop larval cultures from a commercial hatchery and to characterize some selected florfenicol-resistant strains. Larval cultures from untreated and treated rearing tanks exhibited percentages of copiotrophic bacteria resistant to florfenicol ranging from 0.03% to 10.67% and 0.49–18.34%, respectively, whereas florfenicol resistance among oligotrophic bacteria varied from 1.44% to 35.50% and 3.62–95.71%, from untreated and treated larvae, respectively. Florfenicol resistant microbiota from reared scallop larvae mainly belonged to the Pseudomonas and Pseudoalteromonas genus and were mainly resistant to florfenicol, chloramphenicol, streptomycin and co-trimoxazole. This is the first study reporting antimicrobial resistant bacteria associated to a shellfish hatchery and the results suggest that a continuous surveillance of antimicrobial resistance even in absence of antibacterial therapy is urgently required to evaluate potential undesirable consequences on the surrounding environments.     

Microbial water quality of the Georgian coastal zone of the Black Sea

From 2006 to 2008, microbial water quality was monitored along the Georgian coast of the Black Sea. Temperature, pH, salinity, and dissolved oxygen were measured, along with a variety of aquatic microbial parameters, including heterotrophic plate count (HPC), total culturable bacterial count (TCBC), and chlorophyll a (Chl-a) concentration. Total and fecal coliforms and total enterococci counts were recorded as indicators of fecal pollution. Vibrio bacteria, and Escherichia coli- and Vibrio-specific bacteriophages were isolated and enumerated to determine their relationships to standard marine pollution indicators.Persistent microbial pollution was observed, particularly in the summer months, with a higher rate of contamination in estuaries. Microbial indicators generally showed seasonal dependence, suggesting that temperature may influence bacterial dynamics in this environment. No correlation was apparent between fecal pollution indicators and physical-chemical and aquatic microbial parameters, although there were significant relationships amongst the indicators themselves, as well as with the prevalence of Vibrio bacteria and phage.     

Microbiological composition of native and exotic clams from Tagus estuary: Effect of season and environmental parameters

The influence of seasonal and environmental parameters on the occurrence of bacteria was investigated in two clam species (Venerupis pullastra and Ruditapes philippinarum), water and sediment from the Tagus estuary. Total viable counts (TVC), Escherichia coli, Salmonella spp. and Vibrio spp. were evaluated during one-year. Overall, significant seasonal variations were found in both sampling sites, especially for E. coli and Vibrio spp. levels. In summer, significantly higher Vibrio spp. levels were found in R. philippinarum and sediment samples, but not in V. pullastra clams and water samples. In contrast, significantly higher TVC and E. coli levels were observed in winter months in water and sediment samples. Salmonella spp. was generally isolated when higher levels of E. coli were detected, particularly in R. philippinarum. This study is useful for authorities to develop monitoring strategies for coastal contamination and to estimate human health risks associated with the consumption of bivalves.     

Bacterial flora and antibiotic resistance from eggs of green turtles Chelonia mydas: An indication of polluted effluents

Sea turtles migrate to various habitats where they can be exposed to different pollutants. Bacteria were collected from turtle eggs and their resistance to antibiotics was used as pollutant bio-indicators of contaminated effluents. Eggs were collected randomly from turtles when they were laying their eggs. A total of 90 eggs were collected and placed into sterile plastic bags (3 eggs/turtle) during June-December of 2003. The bacteria located in the eggshell, albumen and yolk were examined, and 42% of the eggs were contaminated with 10 genera of bacteria. Pseudomonas spp. were the most frequent isolates. The albumen was found to be the part of the egg to be the least contaminated by bacterial infection. Bacterial isolates tested with 14 antibiotics showed variations in resistance. Resistance to ampicillin was the highest. The presence of antibiotic resistant bacteria in eggs indicates that the green turtle populations were subjected to polluted effluents during some of their migratory routes and feeding habitats. Scanning electron microscopy revealed that Salmonella typhimurium penetrated all eggshell layers.     

Oxygen and hydrogen isotope studies of serpentinization in the Troodos ophiolite complex,Cyprus

Oxygen and hydrogen isotopic compositions were measured on 12 serpentine and 2 actinolite samples from the Troodos ophiolite complex, Cyprus. The single analyzed antigorite(δD= ?60, δ¹⁸O= 7.1) is isotopically similar to all previously analyzed antigorites from ultramafic bodies. However, although their D/H ratios are relatively “normal”(δD= ?70to?92), the δ¹⁸O values of most of the Troodos lizardite-chrysotile serpentines (+12.6 to +14.1) are much higher than the 2.0–9.3‰ range typically found in such serpentines. Such high δ¹⁸O values have previously been found only in the serpentine-like mineraloids termed “deweylites”, which apparently formed at Earth-surface temperatures, and in a single sample from Vourinos, Greece that is in contact with high-¹⁸O limestone. The Troodos lizardite-chrysotile samplescannot have formed by reaction with heated ocean waters, but instead must have formed in contact with large amounts of some type of meteoric, metamorphic, or formation water, either (1) at very low temperatures in a near-surface environment, or (2) at about 100°C from waters that were abnormally enriched in¹⁸O(δ¹⁸O ≈ +4 to +8). The latter possibility seems most plausible inasmuch as extensive evaporites were deposited throughout the Mediterranean Sea during the late Miocene, and this would have been accompanied by strong¹⁸O enrichments of the local meteoric waters. Heated ocean waters, however, probably were responsible for the formation of the actinolitic amphiboles(δ¹⁸O= 4.6 to 5.5; δD= ?51to?46) from the gabbro and ultramafic zones in the Troodos complex. The amphiboles must have formed at considerably higher temperatures and at an earlier stage than the lizardite-chrysotile serpentinization.     

Resistance to low oxygen in the Ponto–Caspian amphipod Pontogammarus robustoides varies among lentic habitats of its northern invaded range

The Ponto–Caspian amphipod Pontogammarus robustoides was introduced into Lithuanian inland waters more than 50 years ago and is now among the most successful local crustacean invaders. Existing as an oxyphilic species in its native range, in Lithuania it managed to establish in some lentic eutrophic waters facing long-term hypoxic conditions under winter ice cover, or shorter periods of hypoxia during the warm season. Recently, it has been observed to be further expanding in such waters. The aim of this study was to explore the possible divergence in anaerobic metabolism among introduced populations facing different selective pressures. A closed-bottle experiment was conducted using individuals from three Lithuanian lentic water bodies of different trophic status: mesotrophic, eutrophic and hypertrophic. Severe hypoxia was gradually reached, after which lactate dehydrogenase activity was significantly higher in amphipods from eutrophic and hypertrophic, than from mesotrophic environments, and was well correlated with trophic status of inhabited environments as assessed by chlorophyll a concentration. These findings suggest a physiological acclimation response to oxygen deficiency faced by the species in some environments in the northern invaded range, which may have a genetic background. Such resistance to oxygen deficiency may expand the environmental niche and promote species’ spread into previously unsuitable habitats, which may pose a threat to native species residing in refuges that are currently free of the invader.     

Comparisons of picophytoplankton abundance, size, and fluorescence between summer and winter in northern South China Sea

The abundance, size, and fluorescence of picophytoplankton cells were investigated during the summer (July-August of 2009) and winter (January of 2010) extending from near-shore coastal waters to oligotrophic open waters in northern South China Sea, under the influence of contrasting seasonal monsoons. We found that the median abundance of Prochlorococcus averaged over top 150 m decreased nearly 10 times in the winter compared to the summer in the whole survey area, while median abundance of Synechococcus and picoeukaryotes increased 2.6 and 2.4 folds, respectively. Vertical abundance profiles of picoeukaryotes usually formed a subsurface maximum during the summer with the depth of maximal abundances tracking the depth of nutricline, whereas their vertical distributions were more uniform during the winter. Size and cellular fluorescence of Prochlorococcus and Synechococcus usually increased with depth in the summer, while the size of picoeukaryotes was smallest at the depth of maximal abundances. Size, cellular fluorescence, and chlorophyll-to-carbon ratio of Prochlorococcus and Synechococcus in surface waters were generally higher in the winter than in the summer and onshore than offshore, probably resulting from different temperature, nutrient, and light environments as well as different ecotype compositions. Prochlorococcus cells were most abundant in warm and oligotrophic environments, while the abundance of Synechococcus and picoeukaryotes was the highest in waters with intermediate chlorophyll and nutrient concentrations. The distributional patterns of picophytoplankton groups are consistent with their specific physiology documented in previous studies and can be possibly predicted by environmental physical and chemical variables.     

The plankton community in Norwegian coastal waters—abundance, composition, spatial distribution and diel variation

The purpose of the present study was to explore the composition and variation of the pico-, nano- and micro-plankton communities in Norwegian coastal waters and Skagerrak, and the co-occurrence of bacteria and viruses. Samples were collected along three cruise transects from Jæren, Lista and Oksøy on the south coast of Norway and into the North Sea and Skagerrak. We also followed a drifting buoy for 55 h in Skagerrak in order to observe diel variations. Satellite ocean color images (SeaWiFS) of the chlorophyll a (chl a) distribution compared favorably to in situ measurements in open waters, while closer to the shore remote sensing chl a data was overestimated compared to the in situ data. Using light microscopy, we identified 49 micro- and 15 nanoplankton sized phototrophic forms as well as 40 micro- and 12 nanoplankton sized heterotrophic forms. The only picoeukaryote (0.2-2.0 μm) we identified was Resultor micron (Pedinophyceae). Along the transects a significant variation in the distribution and abundance of different plankton forms were observed, with Synechococcus spp and autotrophic picoeukaryotes as the most notable examples. There was no correlation between viruses and chl a, but between viruses and bacteria, and between viruses and some of the phytoplankton groups, especially the picoeukaryotes. Moreover, there was a negative correlation between nutrients and small viruses (Low Fluorescent Viruses) but a positive correlation between nutrients and large viruses (High Fluorescent Viruses). The abundance of autotrophic picoplankton, bacteria and viruses showed a diel variation in surface waters with higher values around noon and late at night and lower values in the evening. Synechococcus spp were found at 20 m depth 25-45 nautical miles from shore apparently forming a bloom that stretched out for more than 100 nautical miles from Skagerrak and up the south west coast of Norway. The different methods used for assessing abundance, distribution and diversity of microorganisms yielded complementary information about the plankton community. Flow cytometry enabled us to map the distribution of the smaller phytoplankton forms, bacteria and viruses in more detail than has been possible before but detection and quantification of specific forms (genus or species) still requires taxonomic skills, molecular analysis or both.     

Distribution of Sargassum muticum on the North West coast of Spain: Relationships with urbanization and community diversity

Invasions are common in coastal marine environments where proximity to urban areas should influence the number of non-native organisms due to the likelihood of unintentional introductions from urban centers. Invasive species are widely recognised as important agents of global change, and can colonize new habitats and even cause local extinction of native species. This study was focused on the invasive marine macroalga Sargassum muticum, a problematic invasive species on the West coast of America and in European waters. An intensive survey was carried out along the Galician coast (North West of Spain) from March to August 2008 to determine the current distribution of this species, and its relationship with level of urbanization and native macroalgal diversity. We sampled the presence of S. muticum and native macroalgae in the mid and low intertidal zones of twenty rocky shores distributed along ten rias. Rias were grouped into high and scarcely urbanized based on their population density. Results indicated that S. muticum was present in nine out of the ten rias, confirming its presence in most of the intertidal rocky shores of Galicia twenty years after it was first observed. The presence of S. muticum was not related to the degree of urbanization of the rias, and its distribution and abundance did not seem to be influenced by diversity or functional groups of native algae. In addition, there was a great variability in the percentage cover, number and length of individuals amongst localities and rias suggesting that both large-scale and local processes may play a role in the distribution of this species. This study highlights the importance of future research into the distribution and impact of introduced algae and proves the urgent need for monitoring programmes and increasing efforts to prevent and control new introductions.     

Benthic foraminiferal assemblages of the South Brazil: Relationship to water masses and nutrient distributions

Using distributions of benthic Foraminifera and bottom-water variables (depth, salinity, temperature, oxygen, suspended matter, organic matter, phosphate, silicate, nitrite, and nitrate), we investigated movements of water masses on the South Brazilian Shelf (27–30°S) and assessed the seasonality of continental runoff on the distribution of shelf water masses. The data were obtained from water and sediment samples collected in the austral winter of 2003 and austral summer of 2004 in three transects. The terrestrial nutrient input was significantly reduced at stations away from the coast, but high values of nutrients were maintained in subsurface waters due the presence of South Atlantic Central Water (SACW) at greater depths. At shallow sampling stations the influence of freshwater runoff was related to (1) the dominance of calcareous benthic Foraminifera, such as lagoon-related Pseudononion atlanticum, Hanzawaia boueana, Bulimina marginata, Bolivina striatula, Elphidium poeyanum, together with several agglutinated species, including Arenoparrella mexicana, Gaudryina exilis, and Trochammina spp., common in coastal environments subject to wide salinity fluctuations. In contrast, smaller forms and higher species diversity characterized the assemblage at offshore stations. In winter, the presence of Buccella peruviana and Uvigerina peregrina at Santa Marta Cape suggest the possible transport of those species of Subantarctic Shelf Waters (SASW) origin. Foraminifera associated to Subtropical Shelf Water (STSW) were dominated by Globocassidulina subglobosa in both seasons. In summer, the occurrence of U. peregrina in the shallower stations suggested the influence of SACW nutrients brought up by upwelling of deeper waters.     

Abilities and genes for PAH biodegradation of bacteria isolated from mangrove sediments from the central of Thailand

PAH-degrading bacteria, including Novosphingobium sp. PCY, Microbacterium sp. BPW, Ralstonia sp. BPH, Alcaligenes sp. SSK1B, and Achromobacter sp. SSK4, were isolated from mangrove sediments. These isolates degraded 50–76% of 100 mg/l phenanthrene within 2 weeks. Strains PCY and BPW also degraded pyrene at 98% and 71%, respectively. Furthermore, all of them probably produced biosurfactants in the presence of hydrocarbons. Interestingly, PCY has a versatility to degrade various PAHs. Molecular techniques and plasmid curing remarkably revealed the presence of the alpha subunit of pyrene dioxygenase gene (nidA), involving in its pyrene/phenanthrene degrading ability, located on megaplasmid of PCY which has never before been reported in sphingomonads. Moreover, genes encoding ferredoxin, reductase, extradiol dioxygenase (bphA3A4C) and exopolysaccharide biosynthetase, which may be involved in PAH degradation and biosurfactant production, were also found in PCY. Therefore, we conclude that these isolates, especially PCY, can be the candidates for use as inoculums in the bioremediation.     

Detection of spatial fluctuations of non-point source fecal pollution in coral reef surrounding waters in southwestern Puerto Rico using PCR-based assays

Human fecal contamination of coral reefs is a major cause of concern. Conventional methods used to monitor microbial water quality cannot be used to discriminate between different fecal pollution sources. Fecal coliforms, enterococci, and human-specific Bacteroides (HF183, HF134), general Bacteroides-Prevotella (GB32), and Clostridium coccoides group (CP) 16S rDNA PCR assays were used to test for the presence of non-point source fecal contamination across the southwestern Puerto Rico shelf. Inshore waters were highly turbid, consistently receiving fecal pollution from variable sources, and showing the highest frequency of positive molecular marker signals. Signals were also detected at offshore waters in compliance with existing microbiological quality regulations. Phylogenetic analysis showed that most isolates were of human fecal origin. The geographic extent of non-point source fecal pollution was large and impacted extensive coral reef systems. This could have deleterious long-term impacts on public health, local fisheries and in tourism potential if not adequately addressed.     

青藏高原陆相介形类的分布特征及其对生态环境的响应

统计分析了青藏高原不同区域水体环境表层沉积物陆相介形类的属种分布特征,探讨了介形类在不同水环境下（盐度、pH值及水深）对生态环境的响应.结果显示,青藏高原现生介形类共计21属67种,其中Candona candida、Ilyocypris bradyi、Eucypris inflata、Limnocythere dubiosa、Limnocythere inopinata、Paracypricerus angulata、Leucocytherella sinensis和Leucocythere mirabilis为青藏高原地区的常见种.湖泊、河流、洼地和湿地4类水体环境中,湖泊中介形类最为丰富,达19属62种;青藏高原东北部（祁连山和柴达木盆地）、北部（昆仑山）、西部和南部不同区域的介形类常见种存在较大差异,可能是区域海拔、pH值和盐度综合作用的结果.淡水和微咸水环境介形类属种数量较咸水及盐湖中丰富,分别有17属41种和13属42种,Limnocythere dubiosa （0.52~90.6 g/L）和Leucocythere mirabilis（0.51~174.63 g/L）在淡水、咸水及盐湖中均有出现,适应盐度范围较广;pH值在8.0~10.0范围内介形类属种多样性最丰富,表明大部分介形类具有嗜碱性的特征;青藏高原陆相介形类属种多样性随水深的增加而降低,浅湖（0~15 m）中介形类属种最为丰富,达到17属52种,其中Candona candida（0.2~80 m）和Leucocythere dorsotuberosa（0.3~110 m）从滨湖至深湖区均有分布,二者均具有较大的水深适应范围.     

Some effects of sewage discharge upon phytoplankton in Hong Kong

The coastal waters of Hong Kong constitute a transition from estuarine conditions in the west to more oceanic conditions in the east, with a major discharge of untreated sewage located at the mid-point. Chlorophyll a was determined and net phytoplankton was sampled at 45 stations throughout this transition. Over a period of 20 months, chlorophyll a values rarely exceeded 2 μg l.^?1 in unpolluted coastal waters. Estuarine waters generally contained 2–6 μg l.^?1 and, in waters influenced by sewage discharge, values sometimes exceeded 20 μg l.^?1. There was no evidence of a reduction in taxonomic diversity in polluted areas except in summer, when the net phytoplankton was dominated by Chaetoceros spp. In the autumn and early winter, Skeletonema costatum was abundant in the central polluted areas.