Abyssal hydrothermal springs-Cryptic incubators for brooding octopus.

Does warmth from hydrothermal springs play a vital role in the biology and ecology of abyssal animals? Deep off central California, thousands of octopus (Muusoctopus robustus) migrate through cold dark waters to hydrothermal springs near an extinct volcano to mate, nest, and die, forming the largest known aggregation of octopus on Earth. Warmth from the springs plays a key role by raising metabolic rates, speeding embryonic development, and presumably increasing reproductive success; we show that brood times for females are ~1.8 years, far faster than expected for abyssal octopods. Using a high-resolution subsea mapping system, we created landscape-scale maps and image mosaics that reveal 6000 octopus in a 2.5-ha area. Because octopuses die after reproducing, hydrothermal springs indirectly provide a food supplement to the local energy budget. Although localized deep-sea heat sources may be essential to octopuses and other warm-tolerant species, most of these unique and often cryptic habitats remain undiscovered and unexplored.

Effects of Phenanthrene Exposure on the B-esterases Activities of Octopus maya (Voss and Solís Ramírez, 1996) Embryos.

No ecotoxicological information exists on phenanthrene (Phe) exposure in cephalopods, animals of commercial and ecological importance. This study investigated the effect of Phe on two B-esterases, Acetylcholinesterase (AChE) and Carboxylesterases (CbE), in Octopus maya embryos. Octopus embryos were exposed to different treatments: control (seawater), solvent control (seawater and DMSO 0.01%), 10 and 100 µg/L of Phe. AChE and CbE activities were measured at different developmental stages (blastula, organogenesis, and growth). B-esterase activities increased in control and solvent control as the embryos developed, showing no statistically significant differences between them. On the other hand, the embryos exposed to Phe had significant differences from controls, and between the high and low concentrations. Our results indicate that B-esterases are sensitive biomarkers of exposure to Phe in O. maya. Still, complementary studies are needed to unravel the toxicodynamics of Phe and the implications of the found inhibitory effect in hatched organisms.

Elemental and spectral chemometric analyses of Octopus vulgaris beaks as reliable markers of capture location.

The high demand and economic relevance of cephalopods make them prone to food fraud, including related to harvest location. Therefore, there is a growing need to develop tools to unequivocally confirm their capture location. Cephalopod beaks are nonedible, making this material ideal for traceability studies as it can also be removed without a loss of commodity economic value. Within this context, common octopus (Octopus vulgaris) specimens were captured in five fishing areas along the Portuguese coast. Untargeted multi-elemental total X-ray fluorescence analysis of the octopus beaks revealed a high abundance of Ca, Cl, K, Na, S, and P, concomitant with the keratin and calcium phosphate nature of the material. We tested a suite of discrimination models on both elemental and spectral data, where the elements contributing most to discriminate capture location were typically associated with diet (As), human-related pressures (Zn, Se, and Mn), or geological features (P, S, Mn, and Zn). Among the six different chemometrics approaches used to classify individuals to their capture location according to their beaks' element concentration, classification trees attained a classification accuracy of 76.7%, whilst reducing the number of explanatory variables for sample classification and highlighting variable importance for group discrimination. However, using X-ray spectral features of the octopus beaks further improved classification accuracy, with the highest classification of 87.3% found with partial least-squares discriminant analysis. Ultimately, element and spectral analyses of nonedible structures such as octopus beaks can provide an important, complementary, and easily accessible means to support seafood provenance and traceability, whilst integrating anthropogenic and/or geological gradients.

Inhibition of Pathogenic Bacteria and Fungi by Natural Phenoxazinone from Octopus Ommochrome Pigments.

Cephalopods, in particular octopus (Octopus vulgaris), have the ability to alter their appearance or body pattern by showing a wide range of camouflage by virtue of their chromatophores, which contain nanostructured granules of ommochrome pigments. Recently, the antioxidant and antimicrobial activities of ommochromes have become of great interest; therefore, in this study, the pH-dependent redox effect of the extraction solvent on the antioxidant potential and the structural characterization of the pigments were evaluated. Cell viability was determined by the microdilution method in broth by turbidity, MTT, resazurin, as well as fluorescence microscopy kit assays. A Live/Dead Double Staining Kit and an ROS Kit were used to elucidate the possible inhibitory mechanisms of ommochromes against bacterial and fungal strains. The results obtained revealed that the redox state alters the color changes of the ommochromes and is dependent on the pH in the extraction solvent. Natural phenoxazinone (ommochromes) is moderately toxic to the pathogens Staphylococcus aureus, Bacillus subtilis, Salmonella Typhimurium and Candida albicans, while the species Pseudomonas aeruginosa and Pseudomonas fluorescens, and the filamentous fungi Aspergillus parasiticus, Alternaria spp. and Fusarium verticillioides, were tolerant to these pigments. UV/visible spectral scanning and Fourier- transform infrared spectroscopy (FTIR) suggest the presence of reduced ommatin in methanol/ HCl extract with high intrinsic fluorescence.

Octopuses and lobsters are sentient, says review.

Cephalopods and decapods are to be recognised as sentient beings by the government, as Georgina Mills reports.

The potential impact of Octopus cyanea extracts to improve eggplant resistance against Fusarium-wilt disease: in vivo and in vitro studies.

The novelty of the present research is conducting a new method in the systemic resistance of plant diseases by using distinct marine extracts. The ability of two octopus extracts to reduce the wilt disease caused by Fusarium oxysporum was observed. The applied methods are soaked roots (SR) and foliar shoots (FS). The antioxidant enzyme activities, percent disease index (PDI), and growth parameters were measured. In vitro antifungal potential of the octopus extracts against F. oxysporum was examined. The obtained result shows that SR extracts reduced PDI. Additionally, all the tested treatments promoted the growth and photosynthetic pigments of the infected plants. SR (in ethanolic extracts) was the most prominent inducer which offered a high advancement in the total soluble protein contents. Also, SR (in methanolic extracts) was the most suitable inducer which provided a very necessary development not only in the total phenol but also in the peroxidase (POD) and polyphenol oxidase (PPO) activities. GC-MS investigation of the octopus extracts exhibited that the compounds which possess antifungal activity were furoscrobiculin B and/or eugenol. They demonstrated a notable antifungal potential against F. oxysporum with a maximum activity of 38.5 and 12.7 mm ZOI after the treatment with the ethanolic and methanolic extract, respectively. FTIR results illustrated the functional group of the compound responsible for the antifungal activity. Additionally, an atomic absorption result reveals that there are traces of metals detected such as Pb, Ag, Cu, Zn, and Mg. The antifungal activity was decreased as the concentrations were reduced. Accordingly, the present extracts may be used as the vital agents in the agricultural field to restrain the plant pathogenic fungi, especially F. oxysporum from a proliferation.

Comportamento alimentar de paralarvas do polvo Octopus vulgaris Tipo II (Cuvier, 1797) alimentadas com artêmia enriquecida com microalgas e suplementada com DHA / Feeding behavior of octopus Octopus vulgaris Type II (Cuvier, 1797) paralarvae fed on Artemia enriched with microalgae and supplemented with DHA

The inexistence of nutritionally adequate diets in paralarval rearing is the main bottleneck for commercial production of the common octopus Octopus cf. vulgaris. We report the feeding behavior of O. vulgaris Type II paralarvae fed on Artemia sp (0.1 individual. mL-1) nauplii enriched with microalgae Isocrysis galbana and Pavlova lutheri microalgae from 0 to 7 Day After Hatching (DAH).; metanauplii enriched with microalgae and DHA SELCO® lipid emulsion from the 8 DAH. The paralarvae showed active swimming and predation by the 14 DAH, feeding in the most superficial portion of the water column. From the 15 DAH, the paralarvae remained near the bottom and there a decrease in the consumption of artemia was observed. The mortality observed from the 18 DAH and mass mortality of paralarvae on 20 DAH can be attributed mainly to the nutritional composition of the diet. Studies analyzing the biochemical composition and ontogeny of the digestive system during the early life stages should shed some light on the running for an appropriate feeding protocol to the paralarval rearing.(AU)

Lactogenic Activity of an Enzymatic Hydrolysate from Octopus vulgaris and Carica papaya in SD Rats.

The traditional Chinese medicine theory believes that octopus papaya soup can stimulate milk production in lactating women. The objective of this study was to determine whether dietary supplementation with an enzymatic hydrolysate of Octopus vulgaris and Carica papaya (EHOC) could increase milk production and nutritional indexes in Sprague Dawley (SD) rats. Female SD rats (n = 24) were fed a control diet (n = 8), EHOC-supplemented diet, or a positive control diet (Shengruzhi) from day 10 of pregnancy to day 10 of lactation. Maternal serum, mammary gland (day 10 of lactation), milk, and pup weight (daily) were collected for analysis. Results showed that the EHOC diet obviously elevated daily milk yield and pup weight compared to the control group (P < .05). The EHOC diet was found to increase the concentration of prolactin (PRL), progesterone (P), estradiol (E2), and growth hormone (GH) significantly in the circulation and mammary gland. Mammary glands of EHOC-treated dams showed clear lobuloalveolar development and proliferation of myoepithelial cells, but no striking variations were observed among the groups. Furthermore, the nutrition content and immune globulin concentration in the milk of EHOC-supplemented dams were higher than those of the control group, especially the cholesterol, glucose, and IgG were higher by 44.98% (P < .001), 42.76% (P < .01), and 42.23% (P < .01), respectively. In conclusion, this article demonstrates that EHOC administration has beneficial effects on milk production in the dams and on performance of the dam and pup. These results indicate that EHOC could be explored as a potentially lactogenic nutriment for lactating women.

Ethnoecological knowledge of the artisan fishermen of octopi in the community of Coroa Vermelha.

Coral reefs are quite diverse ecosystems that carry out several ecological functions and plays a relevant socioeconomic role. The artisan fishing of octopi (Octopus spp.) is practiced for the survival of part of the inhabitants of Coroa Vermelha community, in the south of the state of Bahia. We intended to study the knowledge of the octopi fishermen of Coroa Vermelha using the comprehensive ethnoecological proposal of Marques. The data were collected between July, 2006 and April, 2008 through direct observation and from interviews with fishermen met by chance and through the "native specialists" criterion. Twenty semi-structured interviews were carried out following an itinerary of pre-established questions about the activity of octopi capture, and the biological and ecological aspects of the resource. The data showed that the fishermen have knowledge about biological and ecological aspects of the octopi. Two capture techniques are used: octopus fishing (polvejamento) in the reefs and through diving. Two specific folk are recognized: the "normal octopus" (Octopus insularis) and the "east octopus" (Octopus macropus (?)). The intervieews demonstrated ecological knowledge sometimes compatible with the scientific literature, mainly in which concerns the trophic ecology and behavior of the octopi.

Ethnoecological knowledge of the artisan fishermen of octopi (Octopus spp.) in the community of Coroa Vermelha (Santa Cruz Cabrália, Bahia)

Coral reefs are quite diverse ecosystems that carry out several ecological functions and plays a relevant socioeconomic role. The artisan fishing of octopi (Octopus spp.) is practiced for the survival of part of the inhabitants of Coroa Vermelha community, in the south of the state of Bahia. We intended to study the knowledge of the octopi fishermen of Coroa Vermelha using the comprehensive ethnoecological proposal of Marques. The data were collected between July, 2006 and April, 2008 through direct observation and from interviews with fishermen met by chance and through the "native specialists" criterion. Twenty semi-structured interviews were carried out following an itinerary of pre-established questions about the activity of octopi capture, and the biological and ecological aspects of the resource. The data showed that the fishermen have knowledge about biological and ecological aspects of the octopi. Two capture techniques are used: octopus fishing (polvejamento) in the reefs and through diving. Two specific folk are recognized: the "normal octopus" (Octopus insularis) and the "east octopus" (Octopus macropus (?)). The intervieews demonstrated ecological knowledge sometimes compatible with the scientific literature, mainly in which concerns the trophic ecology and behavior of the octopi.

Os recifes de coral são ecossistemas muito diversos que realizam várias funções ecológicas e possuem um relevante papel socioeconômico. A pesca artesanal de polvo (Octopus spp.) é realizada para a sobrevivência de uma parte da população da comunidade de Coroa Vermelha, no Sul do Estado da Bahia. A intenção deste estudo foi avaliar o conhecimento dos pescadores de polvos de Coroa Vermelha, usando a proposta da etnoecologia abrangente de Marques. Os dados foram coletados entre julho de 2006 e abril de 2008 através da observação direta e entrevistas com pescadores encontrados oportunisticamente e com os especialistas "nativos". Vinte entrevistas semi-estruturadas foram realizadas seguindo um roteiro de perguntas pré-estabelecidas sobre a atividade de captura polvos, e os aspectos biológicos e ecológicos do recurso capturado. Os dados demonstraram que os pescadores têm conhecimento sobre alguns aspectos biológicos e ecológicos dos polvos. Duas técnicas de captura são utilizadas: pesca sobre o recife (polvejamento) e através de mergulho. Dois específicos do povos são reconhecidos: o polvo "normal" (Octopus insularis) e o polvo "de leste" (Octopus macropus (?)). Os entrevistados demonstraram conhecimento ecológico, por vezes, compatíveis com a literatura científica, principalmente no que diz respeito à ecologia trófica e comportamento dos polvos.

Determination of biogenic amines and endotoxin in squid, musky octopus, Norway lobster, and mussel stored at room temperature.

Little research has been published on the indicators of spoilage in Mediterranean molluscan shellfish and crustaceans. Thus is why we studied changes in the concentrations of endotoxin and four biogenic amines (histamine, putrescine, tyramine and cadaverine) in European common squid (Loligo subulata, Lamarck, 1798), musky octopus (Eledone moschata, Lamarck, 1798), Norway lobster (Nephrops norvegicus, Linnaeus, 1758), and mussel (Mytilus galloprovincialis, Lamarck, 1819) from the Adriatic Sea stored at room temperature for 24 h. Endotoxin load in fresh squid, Norway lobster, and mussel (<1 EU mg-1) indicated good microbiological quality of raw samples. Biogenic amine index (as the sum of histamine, putrescine, tyramine, and cadaverine) correlated well with endotoxin load in squid (r=0.978, p<0.001) and musky octopus (r=0.874, p<0.01). A good correlation was also found between endotoxin and putrescine in Norway lobster (r=0.777, p<0.05). The highest endotoxin load was found in decomposed mussels and was associated with histamine alone. In conclusion, increase in biogenic amine levels is species-specific. Endotoxin analysis could be used for rapid assessment of microbiological quality of cephalopods and shellfish.

Relations between mercury, methyl-mercury and selenium in tissues of Octopus vulgaris from the Portuguese coast.

Mercury, methyl-mercury (MeHg) and selenium were determined in digestive gland and mantle of Octopus vulgaris, from three areas of the Portuguese coast. To our knowledge these are the first data on MeHg in cephalopods. Concentrations were higher in the digestive gland and percentage of MeHg in mantle. Enhanced Hg and MeHg levels were obtained in digestive gland of specimens from Olhão (3.1-7.4 and 2.0-5.0 microg g(-1), respectively). Differences between areas may be partially related to Hg availability. Relationships between concentrations in mantle and digestive gland pointed to proportional increases of Hg and MeHg in tissues of specimens from Matosinhos and Cascais, but relatively constant values in mantle of individuals from Olhão (higher contamination). Se:Hg molar ratio in digestive gland was 32 and 30 in octopus from Matosinhos and Cascais, respectively, and 5.4 from Olhão. The proximity to the unit suggests demethylation as response to elevated MeHg levels in digestive gland.

Intake of essential and non-essential elements from consumption of octopus, cuttlefish and squid.

Total concentrations of essential (Cu, Zn, Se and Cr) and non-essential (Hg, Cd, Pb and As) trace elements were measured in the flesh and hepatopancreas of Octopodidae (Eledone moschata, Eledone cirrhosa, Octopus salutii), Sepiidae (Sepia elegans, Sepia orbignyana) and Loliginidae (Illex coindeti, Loligo vulgaris) from the Mediterranean Sea. As expected, the hepatopancreas showed higher metal concentrations than flesh; the only exceptions were Hg and As, which were equally distributed in the two tissues. Regarding the edible portion, the highest toxic metal concentrations were in Octopodidae (Hg: 0.44, Cd: 0.49, Pb: 0.10 µg g(-1) wet weight) and Sepiidae (Hg: 0.27, Cd: 0.50, Pb: 0.12 µg g(-1) wet weight), while Loliginidae tended to accumulate less metal, especially Hg (Hg: 0.11, Cd: 0.30, Pb: 0.05 µg g(-1) wet weight). The other elements showed a heterogeneous distribution among the different cephalopod families. Loliginidae showed the highest Se concentrations (1.18 µg g(-1) wet weight), Octopodidae of Cu (37.37 µg g(-1) wet weight) and Zn (42.00 µg g(-1) wet weight) and Sepiidae of As (61.43 µg g(-1) wet weight), while Cr was uniformly distributed among the various families (0.38-0.43 µg g(-1) wet weight). In these seafoods, the concentrations of essential and non-essential elements were within the prescribed limits set by various authorities, except for Cu and As. Health risks posed by toxic elements to humans via dietary intake of these mollusks were assessed on the basis on Provisional Tolerable Weekly Intake (PTWI), while the estimated intakes of essential elements were compared to Dietary Reference Intakes (RDIs). A 70-g serving of these mollusks was shown to provide a large contribution to Cd intake (0.89 µg kg(-1) body weight), corresponding to 35.6% of PTWI. Concerning the essential elements, the consumption of these mollusks made an important contribution to daily dietary intake of Se, Cu and Zn.

Acoustically evoked potentials in two cephalopods inferred using the auditory brainstem response (ABR) approach.

It is still a matter of debate whether cephalopods can detect sound frequencies above 400 Hz. So far there is no proof for the detection of underwater sound above 400 Hz via a physiological approach. The controversy of whether cephalopods have a sound detection ability above 400 Hz was tested using the auditory brainstem response (ABR) approach, which has been successfully applied in fish, crustaceans, amphibians, reptiles and birds. Using ABR we found that auditory evoked potentials can be obtained in the frequency range 400 to 1500 Hz (Sepiotheutis lessoniana) and 400 to 1000 Hz (Octopus vulgaris), respectively. The thresholds of S. lessoniana were generally lower than those of O. vulgaris.

Combined effect of vacuum-packaging and oregano essential oil on the shelf-life of Mediterranean octopus (Octopus vulgaris) from the Aegean Sea stored at 4 degrees C.

The present study evaluated the use of vacuum packaging (alone) or with addition of oregano essential oil (EO), as an antimicrobial treatment for shelf-life extension of fresh Mediterranean octopus stored under refrigeration for a period of 23 days. Four different treatments were tested: A, control sample; under aerobic storage in the absence of oregano essential oil; VP, under vacuum packaging in the absence of oregano essential oil; and VO1, VO2, treated samples with oregano essential oil 0.2 and 0.4% (v/w), respectively, under VP. Of all the microorganisms enumerated, Pseudomonas spp., H2S-producing bacteria and lactic acid bacteria (LAB) were the groups that prevailed in octopus samples, irrespective of antimicrobial treatment. With regard to the chemical freshness indices determined, thiobarbituric acid (TBA) values were low in all octopus samples, as could have been expected from the low fat content of the product. Both trimethylamine nitrogen (TMA-N) and total volatile basic nitrogen (TVB-N) values of oregano treated under VP octopus samples were significantly lower compared to control samples during the entire refrigerated storage period. Based primarily on sensory evaluation (odor), the use of VP, VO1 and VO2 extended the shelf-life of fresh Mediterranean octopus by ca. 3, 11 and 20 days, respectively.

[Immunoloregulation effect of polysaccharide isolated from octopus dollfusi on mouse].

OBJECTIVE: To study the immunoloregulation effect of three polysaccharides OGI, OG2and OG3 extracted from Octopus dollfusi muscle, gonad, digestive gland, respectively. METHODS: Spleen cell proliferation was measured by MTT assay and index of immune organs were weighed and calculated. RESULTS: OGI and OG2 could increase the proliferation of mouse spleen cell of normal mice, and significantly increased the proliferation of the spleen of immunosuppression mice caused by sccyclophosphamide, while showed no cooperation with ConA; OG3 appeared suppression for the two spleens. The three polysaccharides could increase index of immune organs of normal mice and remarkably increased the indexof immunosuppression mice caused by sccyclophosphamide ; while showed obvious function on thymus index. CONCLUSION: These results suggest that OG1 and OG2 have enhancement immunity for normal and immunosuppression mice, and have better effect for the latter;OG3 has suppression proliferation the spleen cell in vitro, however it has better effect for increase index of immune organs in vivo.

Quantification and speciation of mercury and selenium in fish samples of high consumption in Spain and Portugal.

Mercury (Hg) and selenium (Se) determinations were carried out to evaluate human exposure to those elements through fish consumption in Spain and Portugal. Atomic fluorescence spectroscopy (AFS) was applied in a cold vapor mode for total mercury quantification and was also hyphenated to gas chromatography (GC) to achieve the speciation of organomercurial species in fish samples. The results obtained show the highest concentration of Hg in swordfish and tuna (0.47+/-0.02 and 0.31+/-0.01 microg g-1, respectively), and a much lower concentration in sardine, mackerel shad, and octopus (0.048+/-0.002, 0.033+/-0.001, and 0.024+/-0.001 microg g-1, respectively). The determination of alkyl mercury compounds revealed that 93-98% of mercury in the fish samples was in the organic form. Methylmercury (MeHg) was the only species found in the three fish species with higher mercury content. Total selenium concentration was high in sardine, swordfish, and tuna (0.43+/-0.02, 0.47+/-0.02, and 0.92+/-0.01 microg g-1, respectively), but low in mackerel shad and octopus (0.26+/-0.01 and 0.13+/-0.01 microg g-1, respectively). Speciation of selenium compounds was done by high-performance liquid chromatography in conjunction with inductively coupled plasma mass spectrometry (LC-ICP-MS). Selenomethionine (SeMet) was the only selenium compound identified in the fish samples with higher selenium content. Among the fish species studied, sardine had the most favourable Se:Hg and SeMet:MeHg molar ratios; therefore, its consumption seems to be preferable.

The Study of Normal Visual Fields Using Goldmann Module in OCTOPUS 101 Automated Perimetery in Koreans

PURPOSE: Kinetic automated perimetric tests were performed with OCTOPUS 101 perimeter using Goldmann module. Normal isopter positions in the peripheral visual field were visualized by the average position +/- 2 standard deviations. METHODS: We examined 102 eyes of 51 normal healthy Koreans who had no family history of glaucoma, no specific ophthalmologic disease, best corrected visual acuity more than 1.0 and normal intraocular pressure less than 21 mmHg with OCTOPUS 101 perimeter using Goldmann module in 5 isopters (I1e, I2e, I3e, I4e, II4e) at 8 meridians (0degree, 45degrees, 90degrees, 135degrees, 180degrees, 225degrees, 270degrees, 315degrees). RESULTS: The visual field was oval shape, and widest at the inferotemporal area, followed by temporal, and inferior. CONCLUSIONS: The normal position of 4 isopters can be used as a reference index for the peripheral visual field test.

Cloning of Octopus cephalotocin receptor, a member of the oxytocin/vasopressin superfamily.

We reported that the common octopus, Octopus vulgaris, in common with vertebrates, possesses two members of the oxytocin/vasopressin superfamily: octopressin (OP) and cephalotocin (CT). This was the first observation of its kind in invertebrates. As OP and CT have different biological activities, the presence of specific receptors has been proposed. We cloned the cDNA of an orphan receptor from Octopus brain and found it to encode a polypeptide of 397 amino acids that displays sequences characteristic of G-protein coupled receptors. The orphan receptor showed high homology to receptors of the oxytocin/vasopressin superfamily and seemed to conserve the agonist-binding pocket common to the oxytocin and vasopressin receptors. Xenopus oocytes that express the orphan receptor responded to the application of CT by an induction of membrane Cl(-) currents coupled to the inositol phosphate/Ca(2+) pathway. OP and the other members of the oxytocin/vasopressin superfamily did not activate this receptor. HPLC fractionation of the Octopus brain extract combined with an oocyte assay yielded a single substance that was identical to CT. On the basis of these results, we conclude that the cloned receptor is the CT receptor (CTR). Expression of CTR mRNA in Octopus was detected in the central and the peripheral nervous systems, the pancreas, the oviduct and the ovary. This receptor may mediate physiological functions of CT in Octopus such as neurotransmission, reproduction and metabolism.

[Urinary and blood markers of internal mercury dose in workers from a chlorakali plant and in subjects not occupationally exposed: relation to dental amalgam and fish consumption]. / Indicatori urinari ed ematici di dose interna di mercurio in lavoratori di un impianto clorosoda e in soggetti non professionalmente esposti: relazione con gli amalgami dentali ed il consumo di pesce di mare.

OBJECTIVES: The aim of this paper was both to evaluate the internal dose of Hg in occupationally exposed workers (35 Chloralkali workers) compared to that of non occupationally exposed controls (40 workers of the same plant of Portotorres and 22 residents on the island of Carloforte, usual consumers of local fish, mostly tuna fish with relatively high Hg levels) and to assess the relevance of environmental and individual exposure factors linked to lifestyle, sea fish consumption and amalgam fillings. METHODS: All subjects filled out a questionnaire concerning the working history and lifestyle. The amalgam fillings area was measured by medical inspection using a standardised schedule attached to the questionnaire. Mercury in urine (HgU) was measured in all cases, while in a subgroup of our study total blood mercury (HgB) and its organic and inorganic component were also assessed. Furthermore, for 8 of the Carloforte group mercury in hair was also available. RESULTS: Values of urinary mercury excretion of the Chloralkali workers were significantly higher (median value of 15.4, range 4.8-35.0 micrograms/g creatinine, 94.3% of the cases having values > 5 micrograms/g creatinine) than those observed both among the reference group (median value of 1.9, range 0.4-5.6 micrograms/g creatinine, 12.5% of the cases having values a little greater than 5 micrograms/g creatinine) and among the residents in Carloforte (median value of 6.5, range 1.8-21.5 micrograms/g creatinine, 59.1% of the cases having values > 5 mcg/g creatinine). The HgU values observed in this group were in turn significantly higher than those of the non occupationally exposed workers living near Sassari (p = 0.03). Only in this last group were the HgU concentrations statistically significantly related to the extension of the amalgam fillings area (Pearson r = 0.53, p < 0.01). In the Carloforte group HgU was significantly related to the number of fish meal consumed per week (Pearson r = 0.48, p < 0.02). HgB (median value of 5.9, range 3.4-21.6 micrograms/l) as well as its inorganic component (median value of 2.4, range 1.8-4.6 micrograms/l) were significantly higher in the Chloralkali group compared to the other two groups. In all cases of the Carloforte group the ratio between the organic component and the total HgB was higher than 85%, while this ratio was significantly lower in the other two groups. The relationship between HgU and HgB was statistically significant, considering both total blood mercury and the inorganic and the organic components separately. A statistically significant relationship between the sea fish consumption per week and both total HgB (Pearson r = 0.82) and the organic component in this matrix (Pearson r = 0.84, p < 0.001) was observed among 16 non-occupationally exposed subjects. However, the significant relationship between organic blood mercury and sea fish consumption was almost entirely supported by the data observed in the Carloforte group. Total hair mercury levels analysed in 8 subjects of the Carloforte group were high (median value of 9.6, range 1.4-34.5 micrograms/g) and significantly related to sea fish consumption, and to both the individual Hg urinary excretion (Pearson r = 0.83) and to the organic component of blood mercury (Pearson r = 0.87). CONCLUSIONS: According to several experimental human and animal trials and to some recent studies on methylmercury toxicokinetic models, our results suggest that the organic compounds absorbed by usual sea fish consumption may be partially demethylated, increasing the inorganic Hg concentration in the kidney and consequently its urinary excretion, as was observed in the Carloforte group.