RESUMO
Two metallothionein (low-molecular-weight, metal-binding proteins) preparations, MT-1 and MT-2, have been isolated from the digestive gland of American lobster (Homarus americanus) contaminated with Cd. MT-1 contains Cd- and Cu-binding proteins, whereas MT-2 is a reasonably pure Cd-binding protein. The properties of MT-1 and MT-2 with respect to amino acid and elemental compositions, heat stabilities, polarographic, high-performance liquid chromatography (HPLC), and isoelectric focussing behaviors are reported. Lobster metallothioneins share a number of similarities with mammalian metallothioneins with respect to the presence of Cd and Cu, apparent molecular weights, amino acid compositions, UV absorption spectra at various pH, and polarographic behavior, but differ substantially in their electrophoretic behavior.
Assuntos
Cádmio/análise , Sistema Digestório/química , Metalotioneína/isolamento & purificação , Aminoácidos/análise , Animais , Braquiúros , Cromatografia em Gel , Cromatografia Líquida de Alta Pressão , Cobre/análise , Cavalos , Focalização Isoelétrica , Rim/química , Fígado/química , Metalotioneína/química , Peso Molecular , Nephropidae , Ostreidae , Zinco/análiseRESUMO
The reactivity of EDTA, Cu2+, and copper citrate with two metallothionein preparations (MT-1 and MT-2) isolated from the digestive gland of Cd-contaminated American lobster (Homarus americanus) was studied. Under pseudo-first-order conditions, metallothioneins reacted with EDTA for removal of Cd2+ in a multiphasic manner. Cadmium(+II) removal by Cu2+ was complex and non-stoichiometric, suggesting different binding sites. Rabbit liver metallothionein reacted similarly. Cadmium removal from lobster metallothionein by copper citrate was slow and triphasic in nature. EDTA removed Cu2+ from lobster metallothionein very slowly.
Assuntos
Cádmio/metabolismo , Citratos/farmacologia , Cobre/farmacologia , Sistema Digestório/metabolismo , Ácido Edético/farmacologia , Metalotioneína/metabolismo , Animais , Cádmio/farmacologia , Ácido Cítrico , Cinética , Metalotioneína/isolamento & purificação , Nephropidae , Ligação ProteicaRESUMO
Samples of blubber, liver, kidney and brain, obtained from 10 male, 6 female neonatal, and 4 lactating female harp seals (Pagophilus groenlandicus), were analysed for DDT, dieldrin, PCB, and total mercury. Methyl mercury levels in blood were also determined. Biocide deposition was not significantly different in female and male ten day old pups. There were no significant differences in biocide levels in the liver of the 14/+ day old males, but in blubber there were significant differences in dieldrin and DDT. There was no clear relationship between biocide levels in the 6-18 year old lactating adults and their pups. Younger adult seals (6 and 7 years) were found to have higher levels of PCB and sigmaDDT levels in their blubber than did older females (10 and 18 years). Wide intraspecific variation was noted in organochlorine and mercury residue levels. Pups taken in 1973 were found to have lower organochlorine residues than pups taken in the same area in 1971. Preliminary investigation indicates that detectable amounts of organochlorine and mercury residues are capable of crossing the placenta in the harp seal.
Assuntos
Caniformia/metabolismo , Hidrocarbonetos Clorados , Inseticidas/metabolismo , Mercúrio/metabolismo , Resíduos de Praguicidas/análise , Focas Verdadeiras/metabolismo , Tecido Adiposo/metabolismo , Animais , Animais Recém-Nascidos , Encéfalo/metabolismo , Canadá , Poluentes Ambientais/análise , Feminino , Rim/metabolismo , Lactação , Fígado/metabolismo , Masculino , GravidezRESUMO
Hematological and blood chemistry values were examined in harp seals (Pagophilus groenlandicus) exposed to daily oral dosages of methylmercuric chloride (MMC). Two seals, exposed to 0.25 mg MMC/kg body weight/day for 60 and 90 days, respectively, did not show abnormal blood values. Two other seals exposed to 25.0 mg MMC/kg body weight/day died on day 20 and 26 of exposure. Blood parameters indicated toxic hepatitis, uremia and renal failure. Total mercury and methylmercury values in the tissues of the experimental animals indicated that harp seals can tolerate high levels of mercury in the brain and that the observed renal and hepatic dysfunction were related to the high accumulation of mercury in these tissues. Tests of renal function are useful in cases of severe methylmercury poisoning.
Assuntos
Caniformia/fisiologia , Compostos de Metilmercúrio/intoxicação , Focas Verdadeiras/fisiologia , Animais , Comportamento Animal/efeitos dos fármacos , Peso Corporal/efeitos dos fármacos , Feminino , Masculino , Mercúrio/metabolismo , Compostos de Metilmercúrio/sangue , Atividade Motora/efeitos dos fármacos , Fatores de TempoRESUMO
A method is presented for estimating replicate polycyclic aromatic hydrocarbon (PAH) concentrations in American lobster (Homarus americanus) digestive gland tissue based on recoveries of added perdeuterated surrogates from a single satisfactory analysis. PAH concentrations demonstrated a large interanimal variance, even in specimens captured at the same time in the same place. Principal component analysis showed that the variability of the total system of biological variables (carapace length, lobster weight, and digestive gland weight) could be adequately summarized by the first principal component alone in each data set. Ranks provide ordered classification of individuals, allowing data analysis by statistical methods for continuous variables (i.e., analysis of variance). PAH concentrations in individual lobsters were generally highly sensitive to animal size, sex, and fishing area. Efficient monitoring would result from analyzing individual animals of a single sex from a study area, using as small a geographical study area as possible, measuring a single biological variable, and using individual specimens of as narrow a size range as possible.
Assuntos
Nephropidae/química , Hidrocarbonetos Policíclicos Aromáticos/análise , Poluentes Químicos da Água/análise , Análise de Variância , Animais , Feminino , MasculinoRESUMO
Twenty-five laboratories were sent 2 materials, one an acetone powder of lobster digestive gland, the other, the oil which had been extracted during preparation of the powder. Each laboratory was requested to measure the levels of a suite of polycyclic aromatic hydrocarbons in both materials. The response was poor with only 10 laboratories submitting results. Both intra- and interlaboratory precisions were poor; the interlaboratory error was so great as to preclude statistical analysis of the error. Relative standard deviations for oil results determined by liquid chromatography ranged from 39 to 96%.
Assuntos
Compostos Policíclicos/análise , Frutos do Mar/análise , Poluentes Químicos da Água/análise , Poluentes da Água/análise , Animais , Bivalves/metabolismo , Cromatografia Gasosa-Espectrometria de Massas , Nephropidae/metabolismo , Espectrofotometria UltravioletaRESUMO
Fresh interrenal tissue of Atlantic salmon (Salmo salar) parr transformed [3H]pregnenolone and [14C]progesterone, in vitro, to double-labeled cortisol and corticosterone with yields of less than 3% 3H and 5% 14C at 0-5 degrees, 8-10 degrees, and 22-24 degrees over various incubation periods. Incubations at 22-24 degrees gave the highest yields. A time-yield study at 22-24 degrees for 0.5 to 3 hr in late March showed that metabolic patterns did not change with time as metabolic yields increased. Double-labeled metabolites, 11-deoxycortisol, 21-deoxycortisol, and 17 alpha-hydroxyprogesterone, were all detected in hatchery-reared Annapolis River stock in April, but not in November. The 3H:14C ratio of cortisol was significantly lower (P < 0.05) in late April than in early November. Isotopic ratios of corticosterone, but not cortisol, differed significantly (P < 0.05) between immature and precocious parr in November. The isolation of double-labeled corticosterone from all incubations, except immature parr in November, may indicate a metabolic role for the 17-deoxycorticosteroids. No differences in the histology of interrenal tissues were found between Spring (April-May) and Autumn (October-November) parr. Sulfuric acid chromogens on TLC typical of cortisol, corticosterone, and 11-deoxycortisol and other, less polar delta-5-steroids, but not cortisone, in amounts estimated to be > or = 0.1 microgram, were detected in head kidney tissues (0.5-4.5 g) and blood plasma (7-18 ml) of parr in July, late November, and early December.
Assuntos
Corticosteroides/metabolismo , Rim/metabolismo , Salmão/metabolismo , Animais , Cromatografia em Camada Fina , Congelamento , Histocitoquímica , Rim/anatomia & histologia , Pregnenolona/farmacologia , Progesterona/farmacologia , Estações do Ano , Esteroides/sangue , Esteroides/metabolismo , Ácidos Sulfúricos/química , TemperaturaRESUMO
A simple, rapid, easily automated method is described for the determination of polycyclic aromatic hydrocarbons (PAHs) in shellfish such as American lobster (Homarus americanus) and blue mussel (Mytilus edulis). PAHs are extracted from small amounts (1-8 g) of tissue by saponification in 1N ethanolic potassium hydroxide followed by partitioning into 2,2,4-trimethylpentane. This solution is evaporated just to dryness by rotary evaporation and the residue is dissolved in cyclohexane-dichloromethane (1 + 1) for gel permeation chromatography (GPC) on Bio-Beads SX-3. The GPC procedure is ideal as a screening method in the range 25-18 000 ng PAHs/g tissue. If individual PAH measurements are required, the appropriate GPC fraction is collected and PAHs are separated by reverse phase liquid chromatography (LC) with fluorometric detection. Individual PAHs at concentrations as low as 0.25-10 ng/g can be determined. Recoveries of added fluoranthene, pyrene, benz[a]anthracene, chrysene, benzo[e]pyrene, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[a]pyrene, dibenz[a,h]anthracene, benzo[ghi]perylene, and indeno[1,2,3-cd]pyrene were quantitative, with relative standard deviations ranging from 0.0 to 16.9%.
Assuntos
Compostos Policíclicos/análise , Frutos do Mar/análise , Poluentes Químicos da Água/análise , Poluentes da Água/análise , Animais , Autoanálise , Cromatografia em Gel , Cromatografia LíquidaRESUMO
The reduction of arsenate ion to arsenite ion by a variety of tissues was widely distributed, and not a property of any particular tissue. The nature of the reduction was investigated by using various chemical inducers and inhibitors and is believed to be chemical in nature.
Assuntos
Arseniatos/metabolismo , Arsênio/metabolismo , Animais , Oxirredução , Fatores de TempoRESUMO
Twenty-three laboratories from 13 countries in continental Europe, Scandinavia, the British Isles, and North America participated in a polychlorinated biphenyl (PCB) Check Sample Program conducted under the auspices of the International Council for Exploration of the Sea (ICES). PCBs were determined in unspiked and spiked (1.00 mg Aroclor 1254/kg oil) herring (Clupea harengus harengus) oil by the participants, each of which used his or her own cleanup and quantitation techniques; a common Aroclor 1254 mixture was used as a standard and a common quantitation technique was used for comparative purposes. Results for the unspiked oil ranged from 0.48 to 3.416 mg PCB/kg oil, while spiked oil results ranged from 0.70 to 3.891 mg/kg. Calculated spike recoveries ranged from 22 to 136%. Serious deficiencies were found in most steps in the procedures. No evidence was found to support the use of a common PCB standard or a common method of calculation using packed column chromatography. The chromatographic stationary phase used appeared to affect the PCB levels obtained. Florisil cleanup adsorbent yielded higher mean results for both unspiked and spiked oils than did alumina. Large coefficients of variation were found (25-50%), the principal source of which was systematic error (interlaboratory).
Assuntos
Óleos de Peixe/análise , Bifenilos Policlorados/análise , Animais , Técnicas de Química Analítica/normasRESUMO
Chromatographic and chemical confirmatory evidence is presented for the presence of residues of toxaphene, a polychlorinated camphene pesticide, in herring (Clupea harengus harengus) and cod (Gadus morhua) from widely-separated areas of the Canadian east coast. Toxaphene residues were not detected in a sample of deep-sea scallops (Placopecten magellanicus). Toxaphene was determined by capillary gas chromatography following a combination of chromatography and fuming nitric-concentrated sulfuric acid cleanup, a procedure which greatly simplified the capillary gas chromatograms and eliminated many co-extractives. Concentrations in the fish tissues ranged from 0.4 to 1.1 micrograms/g on a net weight basis and from 2.4 to 12 micrograms/g on a fat weight bais. These data indicate widespread contamination of the marine environment by chlorinated camphenes.
Assuntos
Peixes/metabolismo , Inseticidas/análise , Resíduos de Praguicidas/análise , Toxafeno/análise , Animais , Canadá , Cromatografia Gasosa , Fígado/análiseRESUMO
Analysts participating in a multi-laboratory comparative study were asked to identify 4 chlorobiphenyls (CBs) supplied in neat form, to measure the amounts of these present in spiked and unspiked fish oil, to measure other CBs and organochlorines in the unspiked fish oil, and to compare results for their own standard solution and those for standard solutions prepared from the supplied CB compounds. Comparisons were done for a common supplied method and the individual methods used in each laboratory. Participants had no trouble identifying 2,2',5,5'-tetrachlorobiphenyl, 2,2',4,5,5'-pentachlorobiphenyl, and 2,2',4,4',5,5'-hexachlorobiphenyl. Most misidentified 2,2',3,4,5-pentachlorobiphenyl. Approximately one-half of the quantitative comparisons between participants' standards and those prepared from the supplied CBs differed by more than 10%. Two standards prepared from one of the CBs differed by an average of 6.6% (range 0.0-24.5%). Recoveries of added CBs from the fish oil ranged from 24 to 294% for spikes of 63-85 ng/g with no clear distinction between results for the common method vs individual laboratory methods. The common methodology gave a lower coefficient of variation (CV) for most other CBs and organochlorines, but in most cases the CVs for the individual CBs were not smaller than that for Aroclor 1254 equivalents.
Assuntos
Óleos de Peixe/análise , Contaminação de Alimentos/análise , Hidrocarbonetos Clorados/análise , Bifenilos Policlorados/análise , AnimaisRESUMO
A rapid, simple method for screening polychlorinated dibenzo-p-dioxin and dibenzofuran classes in shellfish tissue at pg g-1 wet mass concentrations is described. The method does not require a clean room facility and is based on saponification followed by extraction into hexane, clean-up using gel-permeation chromatography and sulfuric acid treatment, and measurement using capillary gas chromatography-low resolution mass spectrometry with selected ion monitoring. Selected ion monitoring using multiple ions eliminates interferences not removed by clean-up or chromatography. The detector response factor is constant for isomers within a class, e.g., tetrachlorodibenzo-p-dioxin isomers gave a mean response of 0.977 +/- 0.075 area counts fg-1, but varied significantly between classes. Thus one isomer serves as a 'standard' for all members of its class. Recoveries of added polychlorinated dibenzo-p-dioxins and dibenzofurans (20-500 pg g-1 wet mass) averaged 95.6 +/- 6.9 and 99.0 +/- 5.7%, respectively. The limits of detection (five times the noise level) are 20 pg g-1 tetrachlorodibenzo-p-dioxins and dibenzofurans, 20 pg g-1 pentachlorodibenzo-p-dioxins and dibenzofurans, 40 pg g-1 hexachlorodibenzo-p-dioxins and dibenzofurans, 40 pg g-1 heptachlorodibenzo-p-dioxins and dibenzofurans, and 100 pg g-1 octachlorodibenzo-p-dioxin and dibenzofuran. Above the limits of detection, the method gave results for polychlorinated dibenzo-p-dioxin and dibenzofuran classes in shellfish tissue comparable to those obtained by gas chromatography-high resolution mass spectrometry.
Assuntos
Benzofuranos/análise , Dioxinas/análise , Hidrocarbonetos Clorados/análise , Frutos do Mar/análise , AnimaisRESUMO
Differences observed in autoradiogram profiles and isotope ratios of corticosteroid metabolites (such as cortisol), in vitro, in interrenal activity between and within hatchery reared salmon (Salmo salar) parr stocks were not observed in corresponding wild stocks. Wild fish from nine Nova Scotian rivers (Annapolis, Black, East, Gold, LaHave, Medway. Mushamush, Salmon, and Stewiacke Rivers), sampled during May to December all showed similar corticosteroid profiles. Differences were observed in corticosteroid profiles in December between: (1) hatchery stock held in acidic and circumneutral rivers; and (2) hatchery stock held in cages in a limited river, wild stock from this river, and hatchery stock maintained at the hatchery.
Assuntos
Corticosteroides/metabolismo , Rim/metabolismo , Salmão/metabolismo , Animais , Autorradiografia , Cromatografia em Camada Fina , Histocitoquímica , Concentração de Íons de Hidrogênio , Rim/anatomia & histologia , Nova Escócia , Especificidade da Espécie , Esteroides/sangue , Esteroides/metabolismoRESUMO
A method is described for determining selenium in fish tissues, meat, cereals, milk powder, and other materials by flameless atomic absorption spectrophotometry. Samples are solubilized in HNO3 and atomized in a graphite furnace in the presence of nickel nitrate. Recoveries of 0.500 and 1.000 microgram selenium added to several fish samples averaged 99.0 and 98.3%, respectively, with standard deviations of 5.3 and 4.0. Results agreed with those obtained for samples previously analyzed by fluorometry, and with results for NBS Standard Reference Material. The detection limit was 3 ng/ml solution and 50 ng/g sample.
Assuntos
Produtos Pesqueiros/análise , Análise de Alimentos , Selênio/análise , Temperatura Alta , Espectrofotometria AtômicaRESUMO
A new, simple, and rapid cleanup procedure is described for di-2-ethylhexyl phthalate (DEHP) residues in fish lipids. Extracted are chromatographed on small alumina:sulfuric acid-impregnated alumina (layered) columns after gel permeation chromatography of the fish lipid extracts on BioBeads SX-3. Quantitative analyses were carried out by electron capture gas chromatography using 2 columns. Spiked DEHP recoveries varied from 79.3% in mackerel to 86.3% in herring. DEHP concentrations were determined in plaice, eel, redfish, herring, cod, and mackerel tissues and ranged from trace (less than 0.001 microgram/g) to approximately 10 microgram/g (wet wt basis). Ethanolic KOH saponification of the purified DEHP fraction, resulting in the disappearance of the DEHP peak, was used as a confirmatory test. Limited studies with dibutyl, di-n-heptyl, and diethyl phthalates suggest that the method can be made more versatile.
Assuntos
Dietilexilftalato/análise , Peixes/metabolismo , Lipídeos/análise , Ácidos Ftálicos/análise , Animais , Cromatografia Gasosa/métodos , Cromatografia em Gel/métodosRESUMO
The mean levels ± s.d., (ranges) of organochlorines, pesticides, chlorobenzenes and polychlorinated biphenyl were determined in the livers of 100 Atlantic cod (Gadus morhua) caught off the east coast of Canada in 1980 and were as follows: PCB 1.71±0.90 (0.24-4.33); p,p'-DDE 0.28±0.15 (0.01-0.84); p,p'-DDD 0.10±0.09 (trace-0.84); p,p'-DDT 0.15±0.09 (ND-0.58); α-HCH 0.06±0.02 (trace-0.10); α-HCH (ND-trace); heptachlor 0.02±0.01 (ND-0.04); heptachlor epoxide 0.15±0.37 (ND-2.16); dieldrin 0.06±0.11 (ND-0.71); hexachlorobenzene 0.02±0.01 (trace-0.05) mg kg(-1) (wet wt.). The levels of these compounds in 1980 have not changed over the previous eight years with the exception of PCB and the DDT group where there appears to be a general decline between 1972 and 1975 with no significant change thereafter.
RESUMO
Tissue from a harp seal given methyl mercury at a concentration of 0.25 mg/kg in its diet for 61 days, was highly contaminated with mercury. Over 70% of the mercury in the seal's liver (64.0 p.p.m.) was in the inorganic form indicating a demethylating system in this organ. Most of the mercury in the liver, spleen and kidney of an untreated seal was also in the inorganic form. In contrast, over 75% of the mercury in the adrenals and gonads (14.2 and 13.0 p.p.m., respectively) of the treated seal was methyl mercury. Mercury was not detectable in the gonads and not analyzed in the adrenals of the untreated seal. Biosynthesized (in vitro) cortisol, corticosterone, cortisone, and 11-ketotestosterone were isolated and identified from the adrenal incubations, and delta4-androstene-3,17-dione and testosterone were isolated and identified from ovarian incubations from both untreated and methyl mercury (in vivo) treated seals. The ovaries and adrenals from both seals appeared to be normal under the light microscope. The ovaries from both seals were in the same follicular phase, but in vitro incubations of tissue from these organs indicated that the methyl mercury and treatment caused a marked alteration of steroid biosynthesis in tissue from the treated seal. The altered pattern of steroid biosynthesis was also demonstrated by autoradiography, and it is suggested that this technique could be used as an indicator of incipient contamination by a pollutant.
Assuntos
Caniformia/metabolismo , Compostos de Metilmercúrio/toxicidade , Focas Verdadeiras/metabolismo , Esteroides/biossíntese , Poluentes Químicos da Água/toxicidade , Poluentes da Água/toxicidade , Glândulas Suprarrenais/metabolismo , Androstenodiona/biossíntese , Animais , Corticosterona/biossíntese , Cortisona/biossíntese , Feminino , Hidrocortisona/biossíntese , Cetosteroides/biossíntese , Rim/metabolismo , Fígado/metabolismo , Masculino , Mercúrio/metabolismo , Intoxicação por Mercúrio/metabolismo , Ovário/metabolismo , Baço/metabolismo , Testosterona/biossínteseRESUMO
Arsenite ion, as arsenic trichloride was extracted into benzene from strongly acidified tissue homogenates. Following this, arsenite was extracted from the benzene into water, made up in 1N HCl and analyzed polarographically. Arsenate ion, left in the homogenate after arsenite extraction is isolated in exactly the same way following treatment of the homogenate with cuprous ion to reduce arsenate to arsenite ion. Treatment of the acidified homogenate with cuprous ion prior to extraction gives a homogenate which is readily analyzed for total "inorganic" arsenic. The method was efficient to a maximum level of about 20 mug inorganic arsenic since at higher levels lower recoveries were found. Analysis of a variety of marine biological specimens, with levels of total arsenic up to 40.5 ppm, indicated little of this arsenic was present in an inorganic form. Post mortem reduction of arsenate to arsenite was found to occur rapidly in fish tissue.