Biomarkers of PAHs exposure in crabs Ucides cordatus: laboratory assay and field study.

Pyrene metabolites in urine and micronucleus in haemocytes of crabs (Ucides cordatus) were tested as biomarkers of exposure to oil derived PAHs in mangrove sediments. The goal was to verify how well pyrene metabolites in urine represent levels of oil contamination in mangroves and whether the micronuclei assay indicates exposure. For this, bioassays were performed using crabs from clean and contaminated areas, and field studies were conducted in four mangroves. Results of the bioassay show that U. cordatus assimilates, metabolises, and excretes pyrene in urine as pyrene-1-glucoside, pyrene-sulphate and pyrene-conjugate. OH-pyrene-sulphate was the major metabolite produced/excreted over 120 h of observation by crabs from the clean mangrove. The production/excretion of pyrene-1-glucoside in this case increased linearly with time at a rate of 2.3 x 10(-10)mol L(-1)day(-1). The number of micronuclei in haemocytes also increased with the time after pyrene inoculation, indicating that exposure to pyrene triggers genotoxic and mutagenic response. In crabs from a heavily oil-contaminated mangrove pyrene-1-glucoside was the major metabolite, an indication that production/excretion of a certain metabolite varies depending on adaptation of the animal to the environment. A highly significant correlation was found between the concentration of pyrene metabolites in urine of field crabs expressed as OH-pyrene equivalents and the sum of 38 PAHs determined in hepatopancreas/sediments (r=0.825, n=23, p<0.05). The response of these crabs to the micronuclei assay was not significantly related to concentration of individual or total PAHs. Nevertheless, metabolite results prove U. cordatus as excellent bioindicator for evaluating environmental quality in mangrove areas as related to PAHs and oil contamination.

Validation of Ucides cordatus as a bioindicator of oil contamination and bioavailability in mangroves by evaluating sediment and crab PAH records.

This study is aimed at verifying the relevance of Ucides cordatus as a bioindicator of oil contamination and PAH bioavailability in mangrove sediments. For this, crabs and sediment cores were sampled from five mangroves, including an area suspected of contamination derived from an MF380 oil spillage, and analyzed for the 16 PAH in the USEPA priority list as well as for the five series of alkylated homologues. Concentrations in sediments varied from 35 microg kg-1 in the lower core layer of the control area to 33,000 microg kg-1 in the upper layer of the most contaminated area. Total PAH contents in crabs varied from 206 to 62,000 microg kg-1 and were closely correlated to that in sediments. In general, individual PAH profiles in both matrices were in good agreement. Phenanthrenes, however, were more predominant in crabs making up to 30-46% of the Total PAH. Accumulation factors found in the range of 0.7 to 35 were highly variable even after normalizing concentrations for organic carbon and lipid content. Survival in highly contaminated environment and reliable record of environmental contamination in the tissue provide evidence that U. cordatus is an excellent bioindicator for oil in mangroves.

Formation and identification of PAHs metabolites in marine organisms.

In this study, the development of the technique APCI(+) LC/MS/MS allowed the detection of phenanthrene, pyrene and metabolites of alkyl homologs in fish bile (in situ) and in urine of crabs. Laboratory experiments were carried out exposing crabs from an unpolluted mangrove (Barra de Guaratiba) to phenanthrene, and to the alkylated homologs 1-methyl phenanthrene and 2,6,9-trimethyl phenanthrene. Urine samples were collected at 0, 24, 48, 72, and 96 h. Fishes were captured from strategic sites from Guanabara Bay. Hydroxylated metabolites of phenanthrene, epoxides, orthoquinone and glucoside conjugates were identified in both samples. The method APCI(+) LC/MS/MS showed to be effective in a preliminary assessment of phenanthrene metabolite formation, although the low concentrations of 1-methyl phenanthene and 2,6,9-trimethyl phenanthrene did not allow a systematic evaluation of data. The method, however, proved to be excellent tool for studies of PAHs metabolites due to the high selectivity, sensitivity and separation attained.