The Effect of Longterm Exposure to Mercury on the Bacterial Community in Marine Sediment

The aim of this study was to investigate the effect of mercury contamination on bacterial community structure and function. Bacterial communities from two sites, a mercury-contaminated site inside the harbor of Copenhagen, Denmark (CH) and a unpolluted control site, K?ge Buge (KB), were compared with respect to diversity indices, of antibiotic- and heavy metal-resistance patterns, abundance and self transmissibility of plasmids in resistant isolates (endogenous isolation). Furthermore, the potential for gene transfer between indigenous bacteria was assessed by the exogenous plasmid isolation approach.  It was found that resistance to all the tested compounds was higher in the mercury-polluted sediment than the control sediment. The abundance of plasmids was higher at the polluted site, where 62% of the isolates contained plasmids, whereas only 29% of the isolates from the control sediment contained plasmids. Furthermore, the frequencies of large plasmids and plasmids per isolates were found to be higher in the contaminated sediment. Exogenous plasmid isolations revealed high occurrence of Hg and tetracycline resistance, self-transmissible plasmids in CH sediment (1.8 × 10⁻⁵ transconjugants per recipients) relative to KB sediment (3.0 × 10⁻⁸ T/R). Shannon-Weaver diversity indices showed no difference in the diversity of the isolates from the two sites, and Hg-resistant isolates from CH were found to be as diverse as the CH isolates in total. This may be owing to high level of self-transmissible Hg resistance plasmids found in CH. Received: 25 September 1997 / Accepted: 3 November 1997     

Distribution of plasmids in groundwater bacteria

Summary Bacteria isolated from groundwater aquifer core materials of pristine aquifers at Lula and Pickett, OK, and from a site with a history of aromatic hydrocarbon contamination and natural renovation located at Conroe, TX, were screened for the presence of plasmid DNA by alkaline or enzyme lysis and agarose gel techniques. Some of the isolates were also subjected to taxonomic tests in addition to screening for resistance to antibiotics, tolerance to heavy metal salts, and bacteriocin production. There was no significant difference in the distribution of the traits usually associated with plasmid occurrence in isolates from the three sites. These traits, which occurred at low frequencies, were not restricted to plasmid-bearing strains of the communities. Plasmids were found in isolates from all three sites, but on the average there was a significantly higher percentage of isolates containing plasmids in the samples from Conroe (19.4%) than from either Lula (1.8%) or Pickett (7.7%). The sizes of the plasmids found ranged between 3.5 and 202 kilobases but, for the Conroe samples, many more isolates (67%) contained smaller plasmids (<10 kb) rather than larger ones. No plasmids were found in bacteria recovered from naturally renovated aquifer material at the Conroe site.     

Resistance to broad-spectrum antibiotics in aquatic systems: anthropogenic activities modulate the dissemination of bla(CTX-M)-like genes

We compared the resistomes within polluted and unpolluted rivers, focusing on extended-spectrum beta-lactamase (ESBL) genes, in particular bla(CTX-M). Twelve rivers from a Portuguese hydrographic basin were sampled. Physicochemical and microbiological parameters of water quality were determined, and the results showed that 9 rivers were classified as unpolluted (UP) and that 3 were classified as polluted (P). Of the 225 cefotaxime-resistant strains isolated, 39 were identified as ESBL-producing strains, with 18 carrying a bla(CTX-M) gene (15 from P and 3 from UP rivers). Analysis of CTX-M nucleotide sequences showed that 17 isolates produced CTX-M from group 1 (CTX-M-1, -3, -15, and -32) and 1 CTX-M that belonged to group 9 (CTX-M-14). A genetic environment study revealed the presence of different genetic elements previously described for clinical strains. ISEcp1 was found in the upstream regions of all isolates examined. Culture-independent bla(CTX-M)-like libraries were comprised of 16 CTX-M gene variants, with 14 types in the P library and 4 types in UP library, varying from 68% to 99% similarity between them. Besides the much lower level of diversity among CTX-M-like genes from UP sites, the majority were similar to chromosomal ESBLs such as bla(RAHN-1). The results demonstrate that the occurrence and diversity of bla(CTX-M) genes are clearly different between polluted and unpolluted lotic ecosystems; these findings favor the hypothesis that natural environments are reservoirs of resistant bacteria and resistance genes, where anthropogenic-driven selective pressures may be contributing to the persistence and dissemination of genes usually relevant in clinical environments.     

Genetic variation and heavy metal tolerance in the ectomycorrhizal basidiomycete Suillus luteus

Twenty-one isolates of the ectomycorrhizal fungus Suillus luteus were screened for their tolerance to the heavy metals Zn, Cd, Cu and Ni, measured as inhibition of radial growth and biomass production. Two populations from even-aged pine stands were investigated: 10 isolates were obtained from an area polluted with high levels of Zn, Cd and Cu, and 11 isolates were obtained from a control population located in a nearby unpolluted area. RFLP patterns of the internal transcribed spacer region of the isolates confirmed the morphological identification of the carpophores. All isolates were maintained on basic medium without elevated metals to avoid phenotypically acquired metal tolerance. The in vitro Zn and Cd tolerance of the S. luteus isolates from the polluted habitat were significantly higher than the tolerances measured in the isolates from the nonpolluted site. This observation suggests that the elevated soil metal concentrations might be responsible for the evolution of adaptive Zn and Cd tolerance. Tolerance was maintained in an isolate not exposed to elevated metals for 3 yr. The two S. luteus populations did not differ in tolerance to Cu and Ni. The mechanisms for the adaptive Zn and Cd tolerance are not identical as there was no correlation between response to the two metals; the most Zn-tolerant isolate was the most sensitive for Cd in the metal-tolerant population. Zinc did not accumulate in basidiocarp tissue, whereas Cd levels in basidiocarps were significantly higher in the population on the polluted site. Inter-simple sequence-repeat fingerprints showed that 90% of the isolates were from different individuals. The genetic variation in the population from the unpolluted site was considerably larger than that observed at the polluted site.     

Impact of wastewater discharge on the plant diversity,community structure and heavy metal pollution of range plants in eastern Saudi Arabia

The main objectives of this study were to determine the floristic composition of the vegetation cover and to find the effects of wastewater pollution on the plant community structure in eastern Saudi Arabia. 28 stands which were distributed among polluted and unpolluted sites, were chosen for this study. A total of 42 plant species were recorded (14 in the polluted and 28 in the unpolluted sites). The recorded plants comprised of 13 perennial plant species and 29 annual plant species. Seven vegetation communities were determined using TWINSPAN and DCA classification and ordination techniques. Three in the polluted sites, two in the unpolluted sites and the remaining two were mixed communities. The (Sarcopoterium spinosum – Pistacia len‐tiscus) community in the polluted sites, had the highest values of soil moisture, salinity, sulphate, calcium and potassium. On the other hand, Juniperus phoenicea – Olea europaea community in the unpolluted site, had the lowest value of organic matter, salinity and magnesium. In respect of genetic diversity, the community of Foeniculum vulgare - Nicotiana glauca in the unpolluted site, was the most diverse, while Ricinus communis - Chrysanthemum coronarium in the polluted sites was the least one. Both soil and wastewater heavy metal analysis indicated that Pb, Cd, Cr, Cu and Ni concentrations in the polluted sites were significantly higher than those in the unpolluted ones. The impact of wastewater discharge led to the appearance of new invasive plant species that may significantly affect plant diversity and community structure in eastern Saudi Arabia. Finally wastewater discharge in open rangelands could adversely affect the growth of plant species in the rangelands and thus adversely affect plant community structure and diversity.     

Productivity differences between dandelion (Taraxacum officinale; Asteraceae) clones from pollution impacted versus non-impacted soils

Common dandelions (Taraxacum officinale Weber, sensu lato; Asteraceae) introduced to North America form an assemblage of asexual (agamospermous), clonal lineages derived from Eurasian mixed sexual and asexual populations. We investigated whether selection for more pollution tolerant clonal lineages occurs at polluted sites and selection for more pollution intolerant lineages occurs at unpolluted sites. We tested the above hypothesis by performing reciprocal greenhouse productivity experiments in which unique dandelion clones (12 clones, identified by DNA fingerprinting, from each site type) sampled from two unpolluted and two polluted (moderately enhanced Cu, Pb and Zn soil concentrations) sites were grown pairwise in both unpolluted (nutrient solution only) and polluted (nutrient solution + Cu, Pb and Zn) media (n?=?48 paired tests for each media type). Dandelion clones from polluted sites produced fewer and smaller leaves, shorter roots and smaller root diameters, reduced shoot and root dry weights, and reduced total biomass compared to clones from unpolluted sites when clones were grown in unpolluted-media (P?≤?0.05). In contrast, clones taken from unpolluted sites were shown to produce significantly fewer and shorter leaves, shorter roots and smaller root diameters, reduced shoot and root dry weights, reduced total biomass, a reduced shoot : root biomass ratio, and have much lower survival compared to clones from polluted sites when both were grown in polluted-media (P?≤?0.05). These results reveal that there was increased selection against unpolluted-site clonal lineages in polluted-media and against polluted-site clonal lineages in unpolluted-media. Across all treatments, clones from unpolluted sites growing in unpolluted-media had the highest proximate measures of fitness. Overall, these findings provide insight into the relationships among anthropogenic environmental contamination and the consequent effects of selective forces acting on dandelion clones and their population genetic architecture.     

Yeasts from Marine and Estuarine Waters with Different Levels of Pollution in the State of Rio de Janeiro, Brazil

Yeast counts were made at 24 marine and estuarine sites in the vicinity of Rio de Janeiro, Brazil. Mean salinities of estuarine sites ranged from 14.2 to 27.4‰, and mean temperatures ranged from 25 to 28°C. Total coliform counts varied from 80% above 100,000 colony-forming units (CFU)/100 ml at heavily polluted sites to 100% below 100 CFU/100 ml at unpolluted sites. Total yeast counts above 100 CFU/100 ml were typical of heavily and moderately polluted water but atypical of lightly polluted and unpolluted water. Mean total yeast counts were 2,880 CFU/100 ml for heavily polluted sites, 202 CFU/100 ml for moderately polluted sites, and 3 CFU/100 ml for lightly polluted and unpolluted sites. Total yeast counts had a positive response to increased pollution levels, and Candida krusei and phenotypically similar yeasts as a group were prevalent in polluted estuarine water but rare in unpolluted seawater. The 549 strains of yeasts and yeast-like organisms isolated were grouped into 67 species, of which the 21 most prevalent made up 86% of the total yeast population. The prevalent genera in the polluted estuary were Candida, Rhodotorula, Torulopsis, Hanseniaspora, Debaryomyces, and Trichosporon.     

Heavy metals in water,sediments and invertebrates from a metal-contaminated river free of organic pollution

A study was made of water, sediments and invertebrates in the R. Derwent, North-East England, at one site above and three sites below a stream bringing in high concentrations of Zn, Cd and Pb derived from an active fluorspar mine. The mean concentrations of these metals in filtrable water at the unpolluted site were 0.020, <0.0003, 0.005 mg l^-1, respectively, while those at the first polluted site were 0.29, 0.0006, 0.016 mg l^-1. The benthic macroinvertebrate fauna was dominated by insects; all taxa present at the unpolluted site were represented at one or more of the polluted sites. In almost all cases the elevated concentrations of metals in water and sediments at polluted sites were paralleled by higher concentrations in animals. Significant positive correlations were demonstrated between metal concentrations in certain taxa and those in their environment e.g. Ecdyonurus venosus and aqueous Pb. Mayflies tended to concentrate Zn, Cd and Pb to higher levels than other groups. Comparison of carnivorous species with other taxa revealed no indication of increased metal concentrations at higher trophic levels.     

Superficial and hyporheic oligochaete communities as indicators of pollution and water exchange in the River Moselle, France

Benthic oligochaetes were sampled on three occasions (June, August and October 1992) in the upper (0–10 cm) and hyporheic (35–45 cm depths) sediments at five sites of the River Moselle, from upstream of the town of Epinal to Velle-sur-Moselle. The first site (upstream from Epinal) is considered as unpolluted and the four remaining sites are polluted by industrial effluents. The most polluted stations were generally dominated by the pollution tolerant taxon Limnodrilus. Numbers of individuals of this taxon decreased at the less polluted last site in recovery zone, and were also scarce in the first unpolluted site. It is noteworthy that these tendencies were observed in both superficial and hyporheic substrates and to the greatest degree in hyporheic ones. At the unpolluted site, the hyporheic habitat is dominated by the groundwater species Propappus volki, Pristina spp., Pristinella spp. At the less polluted site (last site), the deep sediments are dominated by groundwater species and the Tubificidae without hair setae decrease from June to October. As a result of water exchange between superficial and subterranean waters, superficial substrates of the first and the last stations tend to be colonised by a high proportion of hyporheic species that suggests that flow is primarily from subterranean to superficial waters. The contrary is the case at other polluted stations which are characterised by the invasion of hyporheic substrates by the pollution tolerant superficial taxa Limnodrilus. This suggests that water flows from the river to the deeper groundwater. These two stations are located near drinking water plants which utilise groundwater, thus increasing the vulnerability of groundwater to surface contaminants.     

Activity and heavy metal resistance of non-specific esterases in leaf beetle Chrysomela lapponica from polluted and unpolluted habitats

We compared the general activity and heavy metal resistance of non-specific esterases in two populations of the leaf beetle Chrysomela lapponica from habitats severely contaminated by heavy metals (mostly Ni and Cu) and two populations from unpolluted habitats. Concentrations of Ni and Cu in adult beetles from the most polluted site were 7.7 and 3.6 times higher that in beetles from unpolluted habitats. Larval esterases showed higher activity and lower susceptibility to heavy metals than esterases of adults. Larval esterase activity did not differ between populations from polluted and unpolluted sites, but adult beetles from polluted localities had lower esterase activity than beetles from unpolluted habitats. Both Cu and Ni sulfates in millimolar concentrations in vitro suppressed esterase activity of larvae from unpolluted habitats, but caused no negative effect on esterases of larvae from polluted sites. Similarly, inhibition of adult esterase activity by Ni was stronger in beetles from unpolluted localities than in beetles from polluted localities. This indicates that resistance of non-specific esterases to heavy metals is higher in leaf beetle populations from contaminated environment.     

基于树叶凋落物分解速率的溪流生态系统健康评价——以广东横石水河为例

用Gessner等提出的树叶凋落物分解模式对横石水河的功能完整性和生态系统健康进行评价,对比了藜蒴和荷木2个树种的树叶凋落物在横石水河各污染样点和未受污染样点的分解速率.结果表明：藜蒴和荷木树叶凋落物在研究区污染样点的分解速率均显著低于清洁样点（P<0.05）;根据Gessner模式,各污染样点的环境得分均为0,而各清洁样点的环境得分均为2,表明横石水河污染河段的健康状态已受到严重破坏.研究区生态系统健康的生物学评价与水质分析结果一致,表明树叶凋落物分解模式是一个合适的评价河流状态的指数.     

Responses and structural recovery of periphytic diatom communities after short-term disturbance in some rivers (Hanoi, Vietnam)

Field transfer experiments of periphytic diatom assemblages developed on artificial substrates were set up to assess the responses of those communities to environmental disturbances. The glass slides were positioned for colonization at the relatively unpolluted site (Red, in the Red River) and at the heavily polluted site (TL, in the To Lich River) in the beginning of the experiment. After a period of 2?weeks, the colonized glass slides were concomitantly transferred from the unpolluted Red site to the heavily polluted TL site and to the moderate polluted site (NT₂, in the Nhue River) and, conversely, from the TL site to the Red site, and then to the NT₂ site. The responses and the adapting capacity of periphytic diatom communities to environmental changes were assessed through the cell density, diversity index, species richness, taxonomic composition, and diatom indices after 2 and 4?weeks of transfer periods. For all transfers except for the transfer from the Red to the TL site in which the growth inhibition of diatom cells was found, the diatom density significantly increased until the end of the experiment. Thus, the diatom communities have expressed their pollution tolerance or sensitivities by changing their composition to adapt themselves to environmental changes. Characteristic species of the Red site (Gyrosigma scalproides, Navicula recens) were replaced by Nitzschia palea, Nitzschia umbonata, Aulacoseira granulate typical species of the NT₂ site, in the biofilm transferred from the Red site to the NT₂ site. The relative abundances of typical diatom species of the Red site proliferated in the biofilm transferred from the TL site to the Red site. The replacement of periphytic diatom communities appeared after the transfer from the second week at the different sites. The slow shift of the species towards the typical species at the TL site could result from the organized structure of diatoms within biofilm before the transfer from the Red site to the TL site. The shifts in values of the Index of Specific Polluosensitivity and Diatom Assemblage Index to organic pollution throughout the experiment indicated the clear sensitivity of these indices to water quality changes.     

Calcium oxalate and sulphate-containing structures on the thallial surface of the lichen Ramalina lacera: response to polluted air and simulated acid rain

The formation of calcium‐containing structures on the thallial surface of the lichen Ramalina lacera (With.) J.R. Laund. in response to air pollution and to simulated acid rain, was studied in in situ and transplanted thalli. In situ thalli were collected from an unpolluted site and transplanted to heavily polluted and less polluted sites for a 10 month period. Additional thalli were treated either with double distilled water or with simulated acid rain. Scanning electron microscopy and infrared spectrometry revealed that thallial surfaces of in situ R. lacera samples collected in unpolluted sites were covered with two kinds of calcium oxalate crystals: whewellite and weddellite. These aggregates of calcium oxalate crystals appear to disintegrate and provide a crystal layer on the thallial surface. Infrared spectroscopy of powder scraped from thallial surfaces of transplants, retrieved from non‐polluted sites, showed the presence of whewellite and weddellite, whereas powders obtained from thalli retrieved from polluted sites contained whewellite, weddellite and gypsum. It is suggested that a certain fraction of the gypsum detected in crater‐like structures in transplants from polluted sites and in thalli treated with simulated acid rain is endogenous and should be considered a biomineral.     

Flow cytometric analysis of bacterial respiratory and enzymatic activity in the natural aquatic environment

The respiratory and enzymatic activity of bacteria in polluted and unpolluted river water was investigated by flow cytometry. Double staining with 6CFDA (6-carboxy fluorescein diacetate) and PI (propidium iodide) was used to examine bacterial esterase activity. CTC (5-cyano-2,3-ditolyl tetrazolium chloride) was employed as the indicator of bacterial respiration. The ratios of colony-forming units to total bacterial number were less than 2%, except highly polluted sites. The number of respiring bacteria determined by flow cytometry amounted to 10–15% of the total bacterial number at both unpolluted and polluted sampling stations, while it was 30% at the highly polluted station. Almost 50% of total bacteria demonstrated esterase activity in the unpolluted areas, whereas 70–90% of total bacteria retained this enzymatic activity in the polluted areas. Thus, many of the non-culturable bacteria in the natural aquatic environment have enzymatic activity regardless of the pollution level, and 6CFDA was more superior in the sensitivity for the evaluation of physiological activity of bacteria in the natural environment. The ratio of bacteria with esterase activity increased in direct proportion to the pollution level of river water and suggested that this ratio would be a useful indicator in evaluating the pollution levels in an aquatic environment.     

Distribution and assessment of heavy metals in the aquatic environment of Lake Manzala,Egypt

Analysis of trace metals from sediment and fish collected from eight locations in Lake Manzala, Egypt, during May, June and July of 2011, indicated anthropogenic impacts. The highest concentrations of metals, except Mn and Sr, were detected at Bahr Al-Baqar (S1) which drains from Cairo. At the other sampling locations, the concentrations of selected trace metals fall below the levels of concern while the geo-accumulation index (I_geo) suggests unpolluted conditions for the majority of the studied metals at most sites. However, near the Bahr Al-Baqar (S1) drain concentrations of Ag, Hg and Zn fall above the Long and Morgan's (1990) effects range low (ERL) while Pb was above their effects range median (ERM). I_geo values suggest unpolluted to moderately polluted condition for As and Sn, moderately polluted conditions for Zn, moderate to extremely polluted conditions for Ag, and severely to extremely polluted conditions for Pb at Bahr Al-Baqar. Two sites (Legan, S5, and Al-Ginka, S8) were indicated by I_geo as moderately polluted for Ag while two other sites (West of Bashar, S3, and Al Dehdy, S6) were indicated as unpolluted to moderately polluted for Ag. The Legan and Al-Ginka sites were also indicated as being moderately polluted for Zn. Similar to the sediments, the highest concentrations of metals in fish tissue were from Bahr Al-Baqar (S1). Al concentrations at all sites were comparable to concentrations known to cause lesions in fish. At all sites concentrations of selected trace metals in fish tissue were below the limits set by the FAO.     

Specific regions in the Sod1 locus of the ericoid mycorrhizal fungus Oidiodendron maius from metal-enriched soils show a different sequence polymorphism

The huge diversity of fungi may reflect both the heterogeneity of the niches they occupy and the diverse stresses they must cope with. In order to investigate the genetic and functional diversity in the ericoid mycorrhizal fungus Oidiodendron maius subjected to heavy metal stress, we isolated O. maius strains from a serpentine site naturally enriched by heavy metals. Despite the high Cr and Ni soil concentrations, a high level of diversity was found in the serpentine fungal community. The growth of these isolates in the presence of different metal contaminants identified some tolerant strains, suggesting a site-specific adaptation. To investigate within-species gene divergence in stressful environments, we then compared the sequence polymorphism of a neutral (internal transcribed spacer) and a functional (Cu,ZnSOD) gene in O. maius isolates derived from the serpentine site, from a site heavily polluted with industrial wastes and from unpolluted sites. For all isolates tested, the polymorphism was higher in the nucleotide sequence of the functional gene. However, when compared with isolates from the serpentine area, isolates from industrially polluted sites showed a significantly higher polymorphism in the Cu,ZnSOD promoter region, suggesting that environmental stress may influence the rate of mutations in specific regions of the Sod1 locus.     

Water quality as a determinant of the composition of fish parasite communities

To investigate the relationship between fish parasite communities and water quality level, metazoan parasites were examined in 157 specimens of chub (Leuciscus cephalus L.), sampled in four lowland water courses (northern Italy): Ticino river (unpolluted), Naviglio Pavese Canal (slightly polluted), Lambro river near the Merone village (polluted) and near Monza (severely polluted). Dactylogyrus vistulae, Paradiplozoon ergensi, Bucephalus polymorphus, Acanthocephalus anguillae and larval stages of Tylodelphys clavata were found in all the sampled sites. The distribution of Lamproglena pulchella and Pomphorhynchus laevis was limited to the unpolluted and slightly polluted river sectors, while Asymphylodora tincae, glochidia, along with larval stages of Diplostomum spathaceum, were absent in the severely polluted site. The variability of the calculated infection indices (prevalence, mean abundance and mean intensity), the degree of interactivity among parasites, as well as parameters of species richness and diversity suggest that the structure of parasite communities are affected by the water contamination level.     

Soil Bacterial Biomass, Activity, Phospholipid Fatty Acid Pattern, and pH Tolerance in an Area Polluted with Alkaline Dust Deposition

Soil bacterial biomass, phospholipid fatty acid pattern, pH tolerance, and growth rate were studied in a forest area in Finland that is polluted with alkaline dust from an iron and steel works. The pollution raised the pH of the humus layer from 4.1 to 6.6. Total bacterial numbers and the total amounts of bacterial phospholipid fatty acids in the humus layer did not differ between the unpolluted control sites and the polluted ones. The number of CFU increased by a factor of 6.4 in the polluted sites compared with the controls, while the bacterial growth rate, measured by the thymidine incorporation technique, increased about 1.8-fold in the polluted sites. A shift in the pattern of phospholipid fatty acids indicated a shift in the bacterial species composition. The largest proportional increase was found for the fatty acid 10Me18:0, which indicated an increase in the number of actinomycetes in the polluted sites. The levels of the fatty acids i14:0, 16:1ω5, cy17:0, 18:1ω7, and 19:1 also increased in the polluted sites while those of fatty acids 15:0, i15:0, 10Me16:0, 16:1ω7t, 18:1ω9, and cy19:0 decreased compared with the unpolluted sites. An altered pH tolerance of the bacterial assemblage was detected either as a decrease in acid-tolerant CFU in the polluted sites or as altered bacterial growth rates at different pHs. The latter was estimated by measuring the thymidine incorporation rate of bacteria extracted from soil by homogenization-centrifugation at different pHs.