Alfred J. Vella

Imposex in Hexaplex trunculus (Gastropoda: Muricidae): first results from biomonitoring of tributyltin contamination in the Mediterranean

Imposex (the occurrence of penis and vas deferens in females) in Hexaplex trunculus collected from Malta (Central Mediterranean) in 1992 is reported. This phenomenon is related to the levels of tributyltin (TBT) in the organisms as well as in sediments. Imposex indices (relative penis size and vas deferens sequence) are used to biomonitor coastal contamination by TBT for the first time in the Mediterranean. In its response to TBT, H. trunculus is ranked as one of the most sensitive neogastropod species studied so far. Accumulation of TBT and its metabolites in the digestive glands, gonads and the rest of the body are described, and sex-related differences are shown. No preferential female mortalities are recorded in populations exposed to high levels of TBT. However, a reported shift in the size frequency distribution of H. trunculus in contaminated sites, towards bigger snails, may suggest reduced reproductive potential.

Imposex as a biomonitoring tool for marine pollution by tributyltin: some further observations

Imposex, i.e., the imposition of male sexual characteristics on female neograstropods, has been used worldwide as a bioindicator of pollution by the antifouling agent TBT as well as to assess the related ecological impact. The recent total ban on the use of TBT in the maritime industry has been partly based on the use of such a biomarker. This is mainly based on the assumption that no other pollutant, or environmental stress, is able to induce such a biological response. Nonetheless, several authors had challenged this idea. The present paper will present further field observations on imposex in Hexaplex trunculus from Malta (Central Mediterranean) which may help clarify this issue. Furthermore, the incidence of imposex in this species will be related to body burdens and to the environmental levels of organotins as measured analytically. First histological observations will also be presented which may help clarify the ecological significance of this response in this species. Finally a number of alternative scenarios regarding the relationship between imposex and TBT will be discussed.

Evaluation of environmental levels and biological impact of TBT in Malta (central Mediterranean)

Chemical monitoring for butyltins in bulk seawater, surface microlayer and superficial sediments determined that the two main local sources of marine contamination by tributyltin (TBT) compounds in Malta are ship-repairing dockyards and marinas. In bulk seawater, TBT values ranged from below the detection limit of 5 ng Sn l(-1) to 300 ng Sn l(-1); in sediments derived from the most polluted areas, TBT concentrations as high as 1500 ng Sn g(-1) were measured. At TBT levels found in local harbours, several sublethal biological responses are possible and were observed, including a significant reduction in MFO enzyme system activities of fish; digestive cell atrophy in the oyster Ostrea edulis; and induction of imposex in the snail Hexaplex trunculus. The latter two responses are evident at TBT concentrations below the environmental quality standard (20 ng TBT l(-1)). The ecological implications of these results are discussed and it is concluded that exposure of marine organisms to TBT in local harbours may be expected to lead to a reduction in the long-term survival of a number of such organisms. For these reasons, both the use of TBT-based antifouling paints for pleasure boats as well as drydock practices need to be controlled by appropriate regulations and their enforcement.

Laboratory and field investigations on the effects of organotin (tributyltin) on the oyster, Ostrea edulis

Abstract Specimens of Ostrea edults collected from coastal sites exposed to antifouling tributyltin (TBT) were found to exhibit shell thickening as indicated by a particular shell thickness index. It is suggested that this effect was qualitatively different from that reported for other bivalves and known to be specifically caused by TBT. Nonetheless, laboratory exposure experiments showed that nominal levels of 10 ng/l of TBT in seawater significantly reduce digestive cell volume in this species, and this may lead to reduced somatic growth and thus to shell abnormalities in this species.

Ether-derived alkanes from sedimentary organic matter

Abstract Glycerol ether lipids have been detected in the bitumen of DSDP sediments from Sites 467, 440B and 380 and from the Green River Shale. The alkyl side groups of these ethers were determined by conversion into deuteroalkanes. The presence of glycerol ethers produced by methanogenic bacteria was indicated in the DSDP bitumens by the formation of monodeuterated phytane and dideuterated biphytane. Other ethers were found with novel non-isoprenoidal side groups which may belong to sulfate-reducing or other, probably anaerobic, bacteria. Kerogen-bound alkoxy groups were determined using hydrogen iodide cleavage of the ether link followed by conversion of the iodoalkanes into corresponding deuteroalkanes. For this reaction, the kerogen was not isolated from the rock matrix. The structures so produced were found to include alkyl groups which have known bacterial precursors as well as others that are presently unknown in organisms. The Green River ether biomarker profile is interpreted as possibly indicative of bacterial diagenesis exclusive of biomethanogenesis.

Methodology to identify badly weathering limestone using geochemistry: case study on the Lower Globigerina Limestone of the Maltese islands

This paper proposes a new objective method to classify building limestone into types of varying durability working on samples that are visually identical in appearance. Geochemical data were obtained for 90 samples of the Lower Globigerina Limestone of the Maltese islands. Multivariate analyses, including cluster analysis, principal components analysis and discriminant analysis allowed the identification of variables SiO2, Al2O3, K2O, TiO2, Fe2O3 as indicators to distinguish between more durable ‘franka’ limestone and the less durable facies locally called ‘soll’. This hypothesis was confirmed by differences in geochemical data from samples of both weathered and unweathered stone taken from abandoned quarry faces. In the less durable ‘soll’, SiO2 concentrations are >3.4±0.3%, Al2O3 >0.78±0.05%, K2O >0.13±0.01%, TiO2 >0.06±0.002% and Fe2O3 >0.47±0.02%. However, some overlap occurs between the two stone types for all five components, demarcating another stone type we designated as ‘intermediate’. The geochemical differences are reflected in mineralogical data and in acid-insoluble residues. This geochemical method is simple and innovative. It can be used to identify newly extracted limestone for use in building and also for replacement of deteriorated stones in old buildings.

Perchlorate in dust fall and indoor dust in Malta: An effect of fireworks.

We report on the presence of perchlorate in the settleable dust of Malta, a small central Mediterranean island. Both dust fall collected directly as it precipitated from atmosphere over a period of one month and deposited indoor dust from domestic residences were studied. Perchlorate was determined by ion chromatography of water extracts of the collected dusts. Dust fall was collected from 43 towns during 2011 to 2013 and indoor dust was sampled from homes in the same localities. Perchlorate was detected in 108 of 153 samples of dust fall (71%) and in 28 of 37 indoor dust samples (76%). Detectable perchlorate in dust fall ranged from 0.52μgg(-1) to 561μgg(-1) with a median value of 6.2μgg(-1); in indoor dust, levels were from 0.79μgg(-1) to 53μgg(-1) with a median value of 7.8μgg(-1), the highest recorded anywhere to date. Statistical analysis suggested that there was no significant difference in perchlorate content of indoor dust and dust fall. Perchlorate levels in dust fall escalate during the summer in response to numerous religious feasts celebrated with fireworks and perchlorate persists at low μgg(-1) concentrations for several months beyond the summer festive period. In Malta, perchlorate derives exclusively from KClO4, imported for fireworks manufacture. Its residue in dust presents an exposure risk to the population, especially via ingestion by hand to mouth transfer. Our results suggest that wherever intensive burning of fireworks takes place, the environmental impact may be much longer lived than realised, mainly due to re-suspension and deposition of contaminated settled dust in the urban environment.

Organotin pollution in Malta coastal zone

The speciation of organotins in seawater, surface microlayer and sediments in the marine environment of Malta is described. Concentrations of tributyltin (TBT) in bulk seawater inside the harbours were as high as 0.3 μg Sn L‐1 but were below detection limits (5 ng Sn L‐1) in open sea, 1.6 km offshore. In sediments, TBT levels were highest for the yacht marinas and ranged between 0.03 and 1.5 ug Sn g‐1. Dibutyltin is more common in the surface microlayer than TBT. Diphenyl and monophenyltin were found infrequently in bulk seawater and in sediments. Tetrasubstituted organotins, namely, MenBu(4_n)Sn, where n = 1, 2, and 3, were found frequently in TBT‐contaminated sediments (0.1–9μg Sng‐1), in seawater and in the microlayer where concentrations as high as 140 μg Sn L‐1 (Me3BuSn) were measured. Direct environmental methylation of TBT and that of its debutylated analogues may play a significant role in the geochemical cycling of tin under certain environmental conditions.

Analytical aspects of the gas chromatographic determination of tributyltin and metabolites in environmental samples

Tributyltin and its metabolites were determined in environmental samples by gas chromatography using a flame photometric detection method. The analytical method involved the propylation of the organotins and the use of a recovery standard to correct for inefficient recovery. A detection limit of 5 ng Sn l(-1) for seawater, and 5 ng Sn g(-1) for sediments, was obtained when the detector was equipped with an optical filter; however, this concentration limit, which is higher than the environmental quality standard for TBT, could be improved by a factor of 100 on removing the filter. The limitations imposed by the choice of recovery and internal standards were discussed and it was shown that the possible presence of Sn(II) had no effect on the total extractable inorganic tin, as determined by the analytical protocol. The identification of unexpected mixed methylbutyltins in TBT-contaminated sediments was described, and the implications of the existence of such compounds were discussed.

Distribution of isoprenoid hydrocarbons and alkylbenzenes in immature sediments: evidence for direct inheritance from bacterial/algal sources

Abstract Alkanes having unusual saturated isoprenoidal and methyl-branched structures have been isolated from the bitumen of several sediments. The methanogenic biomarkers 2,6,10,15,19-pentamethyleicosane and squalane were found in sediments which also contained bacteriogenic glycerol ethers. However, in one ether-containing sediment, 2,6,10,13,17,21-hexamethyldocosane was tentatively identified and this compound was found in place of the established alkane biomarkers. Other hydrocarbons found were regular C21 and C23 isoprenoid alkanes, compounds which cannot be derived from phytol; two isoprenoids of the type 3,7,11…-polymethylalkane, previously reported only in petroleums; 8-methylheptadecane, a probable biomarker for cyanobacteria and a number of its homologs and other methyl-branched alkanes. Ubiquitous branched-chain alkylbenzenes and a homology of trimethylalkylbenzenes were also isolated. Most, or all, of the compounds reported here are probably not catagenetic products but may represent direct algal or bacterial bioinputs.