Ultrastructural,biochemical, and immunological characterization of two populations of the mytilid mussel <Emphasis Type="Italic">Bathymodiolus</Emphasis><Emphasis Type="Italic">azoricus</Emphasis> from the Mid-Atlantic Ridge: evidence for a dual symbiosis

A recently described species of mytilid mussel, Bathymodiolus azoricus Von Cosel et al., 1999, was observed to be the dominant organism at the hydrothermal vents off the Azores, at both the Lucky Strike and Menez Gwen sites. Evidence suggests this species of Bathymodiolus represents yet another example of the intriguing dual symbiosis known in three other species of deep-sea mytilid mussels. Transmission electron micrographs (TEM) show the majority of gill bacteriocytes in mussels sampled from both populations to contain two distinct symbiont morphotypes. One morphotype is characterized by large size (mean diameter, 1.25 µm), coccoid shape, and stacked intracytoplasmic membranes that are consistent with the morphology of type I methanotrophs. The second morphotype is smaller (mean diameter, 0.35 µm) and was observed in coccoid or rod shapes. Immunoblots revealed the presence of ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) and methanol dehydrogenase (MeDH) in both populations of mussels. Activities of these enzymes, as well as sulfate adenylyl transferase (ATP sulfurylase) and adenylyl sulfate reductase (APS reductase), were detected in gill extracts. The activities measured for the two populations were highly variable, though the population sampled from Lucky Strike showed higher RubisCO activity. Stable carbon isotope values (Lucky Strike, '¹³C=-32.6ǂ.3‰; Menez Gwen, '¹³C=-22.8ǂ.4‰) are in the range of previously reported stable carbon isotope measurements for mytilid mussels hosting a dual symbiosis. Collectively, these results provide evidence for the activity of both sulfur-oxidizing and methane-oxidizing metabolic pathways in B. azoricus. Furthermore, evidence for a greater dependence on methanotrophy in the Menez Gwen mussel population is offered by analysis of cell counts from TEMs. Higher methanotroph numbers, and putatively activity, in this population of mussels are further supported by published geochemical data indicating higher methane concentrations in the vent fluids at Menez Gwen. This finding suggests that environmental conditions may regulate a balance between the physiological activities of different symbiont populations associated with these mussels. The existence of a dual symbiosis could thus confer greater environmental tolerance and increased niche space to the mytilid host in the stochastic hydrothermal vent habitat.     

Environmental and human exposure assessment monitoring of communities near an abandoned mercury mine in the Philippines: a toxic legacy

Abandoned mines are an important global concern and continue to pose real or potential threats to human safety and health including environmental damage/s. Very few countries had government mine regulation and reclamation policies until the latter part of the century where legal, financial and technical procedures were required for existing mining operations. Major reasons for mine closure may be mainly due to poor economies of the commodity making mining unprofitable, technical difficulties and national security. If the mine is abandoned, more often than not it is the government that shoulders the burden of clean-up, monitoring and remediation. The topic of abandoned mines is complex because of the associated financial and legal liability implications. Abandoned mercury mines have been identified as one of the major concerns because of their significant long-term environmental problems. Primary mercury production is still ongoing in Spain, Kyrgzystan, China, Algeria, Russia and Slovakia while world production declined substantially in the late 1980s. In the Philippines, the mercury mine located southeast of Manila was in operation from 1955 to 1976, before ceasing operation because of the decline in world market price for the commodity. During this time, annual production of mercury was estimated to be about 140,000 kg of mercury yearly. Approximately 2,000,000 t of mine-waste calcines (retorted ore) were produced during mining and roughly 1,000,000 t of these calcines were dumped into nearby Honda Bay to construct a jetty to facilitate mine operations where about 2000 people reside in the nearby three barangays. In October, 1994 the Department of Health received a request from the Provincial Health Office for technical assistance relative to the investigation of increasing complaints of unusual symptoms (e.g. miscarriages, tooth loss, muscle weakness, paralysis, anemia, tremors, etc.) among residents of three barangays. Initial health reports revealed significant elevation of blood mercury levels exceeding the then recommended exposure level of 20ppb in 12 out of the 43 (27.9%) residents examined. The majority of the volunteers were former mine workers. In this study the abnormal findings included gingivitis, mercury lines, gum bleeding and pterydium. The most common neurologic complaints were numbness, weakness, tremors and incoordination. Anemia and elevated liver function tests were also seen in a majority of those examined. The assessment also revealed a probable association between blood mercury level and eosinophilia. The same association was also seen between high mercury levels and the presence of tremors and working in the mercury mine. To date, there are very limited environmental and health studies on the impact of both total and methylmercury that have been undertaken in the Philippines. Thus, this area of study was selected primarily because of its importance as an emerging issue in the country, especially regarding the combined effects of total and methylmercury low-dose and continuous uptake from environmental sources. At present the effects of total mercury exposure combined with MeHg consumption remain an important issue, especially those of low-dose and continuous uptake. Results of the study showed that four (4) species of fish, namely ibis, tabas, lapu-lapu and torsillo, had exceeded the recommended total mercury and methylmercury levels in fish (NV>0.5 microg/gf.w., NV>0.3 microg/gf.w., respectively). Saging and kanuping also exceeded the permissible levels for methylmercury. Total and methylmercury in canned fish, and total mercury in rice, ambient air and drinking water were within the recommended levels, however, additional mercury load from these sources may contribute to the over-all body burden of mercury among residents in the area. Surface water quality at the mining area, Honda Bay and during some monitoring periods at Palawan Bay exceeded total mercury standards (NV>0.002 ng/mL). Soil samples in two sites, namely Tagburos and Honda Bay, exceeded the EPA Region 9 Primary Remediation Goal recommended values for total mercury for residential purposes (NV>23 mg/kg). The hand to mouth activity among infants and children is another significant route for mercury exposure. Statistically significant results were obtained for infants when comparing the results after one year of monitoring for methylmercury levels in hair for both exposed and control sub-groups. Likewise, comparing the initial and final hair methylmercury levels among pregnant women/mothers in the exposed group showed statistically significant (p<0.05) results. Comparing the exposed and control sub-groups' mercury hair levels per sub-group showed statistically significant results among the following: (a) initial and final total mercury hair levels among children, (b) initial and final methylmercury hair levels among children, (c) final total mercury hair levels among pregnant women, (d) initial and final total mercury hair levels among mothers, and (e) initial and final methyl hair levels among mothers.     

Comparison of alkaline industrial wastes for aqueous mineral carbon sequestration through a parallel reactivity study

Thirty-one alkaline industrial wastes from a wide range of industrial processes were acquired and screened for application in an aqueous carbon sequestration process. The wastes were evaluated for their potential to leach polyvalent cations and base species. Following mixing with a simple sodium bicarbonate solution, chemistries of the aqueous and solid phases were analyzed. Experimental results indicated that the most reactive materials were capable of sequestering between 77% and 93% of the available carbon under experimental conditions in four hours. These materials – cement kiln dust, spray dryer absorber ash, and circulating dry scrubber ash – are thus good candidates for detailed, process-oriented studies. Chemical equilibrium modeling indicated that amorphous calcium carbonate is likely responsible for the observed sequestration. High variability and low reactive fractions render many other materials less attractive for further pursuit without considering preprocessing or activation techniques.     

Biodiversity of marine sessile epifauna at an Aegean island subject to hydrothermal activity: Milos, eastern Mediterranean Sea

Sessile macroepifauna was sampled at six rocky sites between 2 and 90 m depth with a number of different methods involving both underwater photography and collection of specimens. A total of 212 species (or varieties) were identified, belonging to seven higher taxa: poriferans (24 species), cnidarians (32), molluscs (8), serpuloidean polychaetes (33), bryozoans (90), brachiopods (4) and ascidians (21). The combined use of a varied array of sampling methods was effective in obtaining a rich faunal inventory. Deep and offshore sites tended to be richer in species than shallow and inshore sites. In all cases species richness was higher at sites closest to hydrothermal vents on the sea floor. Although there are no comparable inventories of marine sessile epifauna in the Aegean, the high number of species found, with a relatively low sampling effort in a restricted area, indicates that the marine biodiversity of this sea is not as low as traditionally believed. Received: 20 January 1999 / Accepted: 25 August 1999     

Intra-laboratory evaluation of Microbial Assay for Risk Assessment (MARA) for potential application in the implementation of the Water Framework Directive (WFD)

The Microbial Assay for Risk Assessment (MARA) is an innovative system based on an array of 11 different microbial species freeze-dried in a 96-well micro-titre plate format. Developed for testing the toxicity of chemicals, mixtures and environmental samples, the assay employs species of a taxonomically diverse range. In addition to ten prokaryotic species, a eukaryote (yeast) is included in the range. The MARA's innate scope of a multi-dimensional test allows determination of toxicity based on a unique assay fingerprint or index, numerically expressed as the mean Microbial Toxic Concentration (MTC). The most significant potential of the test is in the additional inference that can be conveyed to the toxicity evaluation because of the presence of each of the constituent species. In view of the fact that conventional aquatic bioassays, like fish or cladoceran tests, are expensive and impractical, the MARA could provide a cost-effective solution for routine ecotoxicological testing. The performance of the MARA was evaluated to ascertain its capability and potential scope. Sensitivity to toxicants and different environmental samples was assessed. Evaluation included comparison with other tests: namely Microtox, invertebrate (Daphnia magna and Thamnocephalus platyurus) microbiotests, and respiration-inhibition and nitrification-inhibition tests. The most sensitive invertebrate test was found to be the T. platyurus microbiotest for three of the four metals tested. The LC(50) values for this test for Cd(ii), Cr(vi) and As(iii) were 0.2, 0.018 and 0.3 mg l(-1), respectively; and the corresponding most sensitive MARA species MTC values were 4.4, 2.8 and 17 mg l(-1), respectively.     

Uptake and Metabolism of TNT and GTN by Plants Expressing Bacterial Pentaerythritol Tetranitrate Reductase

The manufacture and improper disposal of explosives has resulted in a significant amount of land requiring remediation. The compound 2,4,6-trinitrotoluene (TNT) is the most persistent and toxic of the explosive pollutants with current treatment methods being energy intensive and costly. Bacterial enzymes such as pentaerythritol tetranitrate reductase (PETNR) from Enterobacter cloacae PB2 have been found to have activity against TNT; however, microbes often lack the biomassnecessary for remediation applications. The PETNRgene (onr1) was transformed into tobacco plants in an attempt to combine the metabolic diversity of microbes with the sequestering properties of plants. The resulting transgenic plants were shown to have enhanced tolerance to TNT during germination and as seedlings. Phytoremediation applications with these plants may provide an alternative treatment of TNT contamination.