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1.
PLoS One ; 15(1): e0214833, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-31899775

RESUMEN

It is a common view that an organism's microbiota has a profound influence on host fitness; however, supporting evidence is lacking in many organisms. We manipulated the gut microbiome of Daphnia magna by chronic exposure to different concentrations of the antibiotic Ciprofloxacin (0.01-1 mg L-1), and evaluated whether this affected the animals fitness and antioxidant capacity. In line with our expectations, antibiotic exposure altered the microbiome in a concentration-dependent manner. However, contrary to these expectations, the reduced diversity of gut bacteria was not associated with any fitness detriment. Moreover, the growth-related parameters correlated negatively with microbial diversity; and, in the daphnids exposed to the lowest Ciprofloxacin concentrations, the antioxidant capacity, growth, and fecundity were even higher than in control animals. These findings suggest that Ciprofloxacin exerts direct stimulatory effects on growth and reproduction in the host, while microbiome- mediated effects are of lesser importance. Thus, although microbiome profiling of Daphnia may be a sensitive tool to identify early effects of antibiotic exposure, disentangling direct and microbiome-mediated effects on the host fitness is not straightforward.


Asunto(s)
Daphnia/microbiología , Microbioma Gastrointestinal/efectos de los fármacos , Aptitud Genética/genética , Reproducción/efectos de los fármacos , Animales , Antibacterianos/farmacología , Antioxidantes/metabolismo , Ciprofloxacina/farmacología , Daphnia/efectos de los fármacos , Daphnia/genética , Microbioma Gastrointestinal/genética
2.
Nanotoxicology ; 12(6): 509-521, 2018 08.
Artículo en Inglés | MEDLINE | ID: mdl-29732936

RESUMEN

Cellulose nanofibril (CNF)-based materials are increasingly used in industrial and commercial applications. However, the impacts of CNF on aquatic life are poorly understood, and there are concerns regarding their potential toxicity. Using a combination of standard ecotoxicological tests and feeding experiments, we assessed the effects of CNF exposure (0.206-20.6 mg/L) on the feeding (food uptake and gut residence time) and life-history traits (growth and reproduction) in the cladoceran Daphnia magna. No mortality was observed in a 48 h acute exposure at 2060 mg/L. Moreover, a 21-day exposure at low food and moderate CNF levels induced a stimulatory effect on growth, likely driven by increased filtration efficiency, and, possibly, partial assimilation of the CNF by the animals. However, at low food levels and the highest CNF concentrations, growth and reproduction were negatively affected. These responses were linked to caloric restriction caused by dilution of the food source, but not an obstruction of the alimentary canal. Finally, no apparent translocation of CNF past the alimentary canal was detected. We conclude that CNF displays a low toxic potential to filter-feeding organisms and the expected environmental risks are low.


Asunto(s)
Celulosa/toxicidad , Daphnia/efectos de los fármacos , Nanofibras/toxicidad , Animales , Daphnia/fisiología
3.
Mar Environ Res ; 125: 10-24, 2017 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-28038348

RESUMEN

The aim of this study was to determine a suitable set of biomarker based methods for environmental monitoring in sub-arctic and temperate offshore areas using scientific knowledge on the sensitivity of fish species to dispersed crude oil. Threshold values for environmental monitoring and risk assessment were obtained based on a quantitative comparison of biomarker responses. Turbot, halibut, salmon and sprat were exposed for up to 8 weeks to five different sub-lethal concentrations of dispersed crude oil. Biomarkers assessing PAH metabolites, oxidative stress, detoxification system I activity, genotoxicity, immunotoxicity, endocrine disruption, general cellular stress and histological changes were measured. Results showed that PAH metabolites, CYP1A/EROD, DNA adducts and histopathology rendered the most robust results across the different fish species, both in terms of sensitivity and dose-responsiveness. The reported results contributed to forming links between biomonitoring and risk assessment procedures by using biomarker species sensitivity distributions.


Asunto(s)
Monitoreo del Ambiente/métodos , Peces/fisiología , Petróleo/toxicidad , Contaminantes Químicos del Agua/toxicidad , Animales , Regiones Árticas , Biomarcadores/metabolismo , Citocromo P-450 CYP1A1/metabolismo , Peces Planos/metabolismo
4.
Environ Sci Technol ; 48(21): 12886-92, 2014 Nov 04.
Artículo en Inglés | MEDLINE | ID: mdl-25247638

RESUMEN

To apply biomarkers of oxidative stress in laboratory and field settings, an understanding of their responses to changes in physiological rates is important. The evidence is accumulating that caloric intake can increase production of reactive oxygen species and thus affect background variability of oxidative stress biomarkers in ecotoxicological testing. This study aimed to delineate effects of food intake and xenobiotics on oxidative biomarkers in Daphnia magna. Antioxidant capacity measured as oxygen radical absorbance capacity (ORAC) and lipid peroxidation assayed as thiobarbituric acid reactive substances (TBARS) were measured. Food intake was manipulated by varying food densities or by exposing the animals to chemicals inhibiting feeding rate (pharmaceutical haloperidol and pesticide lindane). Feeding rate proved to affect both protein, ORAC, and TBARS in unexposed daphnids. However, there was no significant effect of feeding rate on the protein-specific ORAC values. Both substances affected individual protein and ORAC levels and changed their relationship to feeding rate. Our results show that inhibition of feeding rate influenced the interpretation of biomarker response and further emphasize the importance of understanding (1) baseline variability in potential biomarkers due to variations in metabolic state and (2) the contribution of feeding rate on toxic response of biomarkers.


Asunto(s)
Daphnia/fisiología , Ecotoxicología , Conducta Alimentaria/efectos de los fármacos , Pruebas de Toxicidad , Xenobióticos/toxicidad , Animales , Antioxidantes/metabolismo , Biomarcadores/metabolismo , Daphnia/efectos de los fármacos , Análisis de los Mínimos Cuadrados , Modelos Lineales , Oxidación-Reducción/efectos de los fármacos , Especies Reactivas de Oxígeno/farmacología , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismo
5.
Environ Sci Pollut Res Int ; 21(24): 13758-68, 2014 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-24819434

RESUMEN

Oil spills occur commonly, and chemical compounds originating from oil spills are widespread in the aquatic environment. In order to monitor effects of a bunker oil spill on the aquatic environment, biomarker responses were measured in eelpout (Zoarces viviparus) sampled along a gradient in Göteborg harbor where the oil spill occurred and at a reference site, 2 weeks after the oil spill. Eelpout were also exposed to the bunker oil in a laboratory study to validate field data. The results show that eelpout from the Göteborg harbor are influenced by contaminants, especially polycyclic aromatic hydrocarbons (PAHs), also during "normal" conditions. The bunker oil spill strongly enhanced the biomarker responses. Results show elevated ethoxyresorufin-O-deethylase (EROD) activities in all exposed sites, but, closest to the oil spill, the EROD activity was partly inhibited, possibly by PAHs. Elevated DNA adduct levels were also observed after the bunker oil spill. Chemical analyses of bile revealed high concentrations of PAH metabolites in the eelpout exposed to the oil, and the same PAH metabolite profile was evident both in eelpout sampled in the harbor and in the eelpout exposed to the bunker oil in the laboratory study.


Asunto(s)
Perciformes/metabolismo , Contaminación por Petróleo/efectos adversos , Petróleo/efectos adversos , Animales , Bilis/química , Bilis/metabolismo , Biomarcadores/metabolismo , Citocromo P-450 CYP1A1/metabolismo , Aductos de ADN/genética , Monitoreo del Ambiente , Proteínas de Peces/metabolismo , Perciformes/genética , Hidrocarburos Policíclicos Aromáticos/análisis , Hidrocarburos Policíclicos Aromáticos/metabolismo , Contaminantes Químicos del Agua/análisis
6.
PLoS One ; 9(4): e92771, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24699280

RESUMEN

The intense artificial sweetener sucralose has no bioconcentration properties, and no adverse acute toxic effects have been observed in standard ecotoxicity tests, suggesting negligible environmental risk. However, significant feeding and behavioural alterations have been reported in non-standard tests using aquatic crustaceans, indicating possible sublethal effects. We hypothesized that these effects are related to alterations in acetylcholinesterase (AChE) and oxidative status in the exposed animals and investigated changes in AChE and oxidative biomarkers (oxygen radical absorbing capacity, ORAC, and lipid peroxidation, TBARS) in the crustacean Daphnia magna exposed to sucralose (0.0001-5 mg L(-1)). The sucralose concentration was a significant positive predictor for ORAC, TBARS and AChE in the daphnids. Moreover, the AChE response was linked to both oxidative biomarkers, with positive and negative relationships for TBARS and ORAC, respectively. These joint responses support our hypothesis and suggest that exposure to sucralose may induce neurological and oxidative mechanisms with potentially important consequences for animal behaviour and physiology.


Asunto(s)
Acetilcolinesterasa/metabolismo , Biomarcadores/análisis , Daphnia/efectos de los fármacos , Peroxidación de Lípido/efectos de los fármacos , Sacarosa/análogos & derivados , Edulcorantes/farmacología , Animales , Oxidación-Reducción , Especies Reactivas de Oxígeno/metabolismo , Sacarosa/farmacología , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismo
7.
Sci Total Environ ; 410-411: 248-57, 2011 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-21995877

RESUMEN

We surveyed four Chesapeake Bay tributaries for skin and liver tumors in brown bullhead (Ameiurus nebulosus). We focused on the South River, where the highest skin tumor prevalence (53%) in the Bay watershed had been reported. The objectives were to 1) compare tumor prevalence with nearby rivers (Severn and Rhode) and a more remote river (Choptank); 2) investigate associations between tumor prevalence and polynuclear aromatic hydrocarbons (PAHs) and alkylating agents; and 3) statistically analyze Chesapeake Bay bullhead tumor data from 1992 through 2008. All four South River collections exhibited high skin tumor prevalence (19% to 58%), whereas skin tumor prevalence was 2%, 10%, and 52% in the three Severn collections; 0% and 2% in the Choptank collections; and 5.6% in the Rhode collection. Liver tumor prevalence was 0% to 6% in all but one South River collection (20%) and 0% to 6% in the three other rivers. In a subset of samples, PAH-like biliary metabolites and (32)P-DNA adducts were used as biomarkers of exposure and response to polycyclic aromatic compounds (PACs). Adducts from alkylating agents were detected as O6-methyl-2'-deoxyguanosine (O6Me-dG) and O6-ethyl-2'-deoxyguanosine (O6Et-dG) modified DNA. Bullheads from the contaminated Anacostia River were used as a positive control for DNA adducts. (32)P-DNA adduct concentrations were significantly higher in Anacostia bullhead livers compared with the other rivers. We identified alkyl DNA adducts in bullhead livers from the South and Anacostia, but not the Choptank. Neither the PAH-like bile metabolite data, sediment PAH data, nor the DNA adduct data suggest an association between liver or skin tumor prevalence and exposure to PACs or alkylating agents in the South, Choptank, Severn, or Rhode rivers. Logistic regression analysis of the Chesapeake Bay database revealed that sex and length were significant covariates for liver tumors and length was a significant covariate for skin tumors.


Asunto(s)
Enfermedades de los Peces/inducido químicamente , Ictaluridae , Neoplasias Hepáticas/veterinaria , Neoplasias Cutáneas/veterinaria , Alquilantes/metabolismo , Animales , Biomarcadores/metabolismo , Aductos de ADN/análisis , District of Columbia/epidemiología , Femenino , Enfermedades de los Peces/epidemiología , Enfermedades de los Peces/patología , Hígado/metabolismo , Hígado/patología , Neoplasias Hepáticas/inducido químicamente , Neoplasias Hepáticas/epidemiología , Neoplasias Hepáticas/patología , Modelos Logísticos , Masculino , Maryland/epidemiología , Hidrocarburos Policíclicos Aromáticos/metabolismo , Prevalencia , Ríos/química , Piel/metabolismo , Piel/patología , Neoplasias Cutáneas/inducido químicamente , Neoplasias Cutáneas/epidemiología , Neoplasias Cutáneas/patología , Virginia/epidemiología
8.
J Toxicol Environ Health A ; 74(7-9): 582-604, 2011.
Artículo en Inglés | MEDLINE | ID: mdl-21391100

RESUMEN

The Norwegian water column monitoring program investigates the biological effects of offshore oil and gas activities in Norwegian waters. In three separate surveys in 2006, 2008, and 2009, bioaccumulation and biomarker responses were measured in mussels (Mytilus edulis) and Atlantic cod (Gadus morhua) held in cages at known distances from the produced water (PW) discharge at the Ekofisk oil field. Identical monitoring studies performed in all three years have allowed the biological effects and bioaccumulation data to be compared, and in addition, enabled the potential environmental benefits of a PW treatment system (CTour), implemented in 2008, to be evaluated. The results of the 2009 survey showed that caged animals were exposed to low levels of PW components, with highest tissue concentrations in mussels located closest to the PW discharge. Mussels located approximately 1-2 km away demonstrated only background concentrations of target compounds. Concentrations of polycyclic aromatic hydrocarbons (PAH) and alkyl phenol (AP) metabolites in the bile of caged cod were elevated at stations 200-250 m from the discharge. There was also a signal of exposure relative to discharge for the biomarkers CYP1A in fish and micronuclei in mussels. All other fish and mussel biomarkers showed no significant exposure effects in 2009. The mussel bioaccumulation data in 2009 indicated a lower exposure to the PW effluent than seen previously in 2008 and 2006, resulting in an associated general improvement in the health of the caged mussels. This was due to the reduction in overall discharge of PW components (measured as oil in water) into the area in 2009 compared to previous years as a result of the improved PW treatment system.


Asunto(s)
Monitoreo del Ambiente/métodos , Industria Procesadora y de Extracción , Gadus morhua/metabolismo , Mytilus edulis/efectos de los fármacos , Petróleo , Contaminantes Químicos del Agua/toxicidad , Animales , Biomarcadores/sangre , Biomarcadores/metabolismo , Carga Corporal (Radioterapia) , Citocromo P-450 CYP1A1/metabolismo , Proteínas del Huevo/sangre , Femenino , Gadus morhua/sangre , Glutatión Transferasa/metabolismo , Hígado/metabolismo , Masculino , Mytilus edulis/metabolismo , Noruega , Hidrocarburos Policíclicos Aromáticos/metabolismo , Agua de Mar/química , Vitelogeninas/sangre , Contaminantes Químicos del Agua/análisis
9.
Mutat Res ; 702(1): 24-31, 2010 Sep 30.
Artículo en Inglés | MEDLINE | ID: mdl-20643223

RESUMEN

Adult and young herring gulls (Larus argentatus) in Sweden and Iceland were investigated with respect to DNA adducts, analysed with the nuclease-P1 version of the (32)P-postlabelling method, and micronucleated erythrocytes. Three important aims were: (1) to estimate the degree of exposure to genotoxic environmental pollutants in the Baltic Sea area and Iceland, (2) to evaluate the utility of the investigated biomarkers in birds, and (3) to investigate if there was any relationship between genotoxic effects and thiamine deficiency. The results demonstrate that both Swedish and Icelandic herring gulls are exposed to genotoxic pollution. Urban specimens have higher levels of DNA adducts than rural specimens, but background exposure to genotoxic environmental pollutants, such as PAHs, is also significant. In the herring gull the general level of DNA adducts in the liver seems to be higher than in fish. DNA adducts were most abundant in the liver, followed by the kidney, intestinal mucosa, and whole blood, in decreasing order. The frequency of micronucleated erythrocytes was probably slightly elevated in all the investigated sites, reflecting a significant background exposure. The level of DNA adducts was unrelated to the frequency of micronucleated erythrocytes, and both these variables were unrelated to symptoms of thiamine deficiency. The investigation confirmed the utility of DNA adducts, and probably also micronucleated erythrocytes, as biomarkers of genotoxicity in birds.


Asunto(s)
Charadriiformes/genética , Aductos de ADN , Contaminantes Ambientales/toxicidad , Micronúcleos con Defecto Cromosómico , Animales , Biomarcadores , Eritrocitos/ultraestructura , Peces/genética , Islandia , Suecia
10.
Environ Sci Technol ; 43(9): 3329-34, 2009 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-19534154

RESUMEN

Large discharges from oil and gas production platforms (produced water) have led to concerns for adverse biological effects in marine areas. The aim of this study was to investigate the development of DNA adductformation and related biomarkers in fish after chronic exposure to water-soluble components of oil. Atlantic cod (Gadus morhua) were exposed for up to 44 weeks to three treatments (low, pulsed, high) containing environmentally relevant concentrations of low-molecular-weight polycyclic aromatic hydrocarbons (PAHs) and short-chained alkylphenols (APs). A time- and dose-related pattern of DNA adduct formation (measured using 32P-postlabeling) was observed. The results suggested that an extended exposure period (more than 16 weeks) would be required for the formation of DNA adduct levels above background. Interestingly, fish receiving pulsed high exposure did not develop elevated concentrations of DNA adducts, possibly due to DNA repair processes. No obvious relationship between DNA adduct concentration and cytochrome P4501A activity (EROD) was observed. This study has demonstrated the genotoxic potential of water-soluble oil components, relevant for operational discharges (produced water) and chronic oil spills. The quantification of PAH metabolites in bile and hepatic DNA adduct formation appear to be suitable for environmental monitoring of chronic oil pollution.


Asunto(s)
Ambiente , Gadus morhua/metabolismo , Aceites/toxicidad , Contaminantes Químicos del Agua/toxicidad , Animales , Bilis/efectos de los fármacos , Bilis/metabolismo , Citocromo P-450 CYP1A1/metabolismo , Aductos de ADN/metabolismo , Monitoreo del Ambiente , Femenino , Hidroxilación/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/enzimología , Pruebas de Mutagenicidad , Fenantrenos/metabolismo , Isótopos de Fósforo , Hidrocarburos Policíclicos Aromáticos/metabolismo , Pirenos/metabolismo , Solubilidad/efectos de los fármacos
11.
Environ Sci Technol ; 41(8): 2972-7, 2007 Apr 15.
Artículo en Inglés | MEDLINE | ID: mdl-17533866

RESUMEN

This study reports on the bioavailability, maternal transfer, and genotoxicity in feral fish of organic sediment pollutants. Northern pike (Esox lucius) and perch (Perca fluviatilis) were caught in a polluted bay before and during dredging activities and from reference areas. Polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) were analyzed in ovulating eggs to investigate if the bay sediment posed a threat to early life-stages of fish. On the basis of previous investigations in this area, the level of exposure via maternal transfer and diffusive uptake of water-borne pollutants after hatch is likely sufficient to cause abnormalities in early life-stages of fish. During dredging, hepatic DNA adducts were elevated in adult fish, demonstrating an increased release of genotoxic compounds, which may contribute to adverse effects in aquatic organisms for several years. Although no substantial increase of maternally transferred pollutants were observed during dredging, this is the first time a correlation between hepatic DNA adducts in fish and pollutant burden in their eggs is demonstrated. Our findings underline the importance of combining chemical and toxicological methods as well as a need for greater emphasis on other polycyclic aromatic compounds in environmental risk evaluations.


Asunto(s)
Aductos de ADN/análisis , Daño del ADN , Esocidae/genética , Óvulo/química , Percas/genética , Contaminantes Químicos del Agua/toxicidad , Contaminación Química del Agua/análisis , Animales , Aductos de ADN/efectos de los fármacos , Aductos de ADN/genética , Femenino , Agua Dulce , Sedimentos Geológicos/química , Masculino , Óvulo/efectos de los fármacos , Contaminación Química del Agua/efectos adversos
12.
Mar Pollut Bull ; 53(8-9): 437-50, 2006.
Artículo en Inglés | MEDLINE | ID: mdl-16904705

RESUMEN

By measuring a battery of basic physiological biomarkers and the concentration of SigmaDDT in adult female perch (Perca fluviatilis), an assumed aquatic pollution gradient was confirmed, with the city of Stockholm (Sweden) as a point source of anthropogenic substances. The investigation included an upstream gradient, westwards through Lake Mälaren (46 km), and a downstream gradient, eastwards through the Stockholm archipelago (84 km). The results indicated a severe pollution situation in central Stockholm, with poor health status of the perch: retarded growth, increased frequency of sexually immature females, low gonadosomatic index, and disturbed visceral fat metabolism. SigmaDDT, measured as a pollution indicator, was 10-28 times higher than the background in perch from the Baltic Proper. Besides the main gradient other sources of pollution also influenced the response pattern of the measured biomarkers. In particular, there were strong indications of pollution coming from the Baltic Sea.


Asunto(s)
Biomarcadores/análisis , DDT/análisis , Monitoreo del Ambiente/estadística & datos numéricos , Agua Dulce/química , Percas/metabolismo , Contaminantes Químicos del Agua/análisis , Análisis de Varianza , Animales , Peso Corporal , DDT/toxicidad , Femenino , Gónadas/fisiología , Metabolismo de los Lípidos , Tamaño de los Órganos , Percas/crecimiento & desarrollo , Maduración Sexual/efectos de los fármacos , Suecia , Contaminantes Químicos del Agua/toxicidad
13.
Mar Pollut Bull ; 53(8-9): 451-68, 2006.
Artículo en Inglés | MEDLINE | ID: mdl-16750226

RESUMEN

A battery of biochemical biomarkers and the SigmaPCB concentration in adult female perch (Perca fluviatilis) verified an aquatic pollution gradient with the city of Stockholm (Sweden) as a point source of anthropogenic substances. The investigation included both an upstream gradient, 46 km westwards through Lake Mälaren, and a downstream gradient, 84 km eastwards through the Stockholm archipelago. Besides the main gradient from Stockholm, there were strong indications of pollution coming from the Baltic Sea. The results indicated a severe pollution situation in central Stockholm, with poor health status of the perch, characterised by increased specific EROD activity in the liver, increased liver EROD somatic index, decreased AChE activity in the muscle, increased amount of DNA adducts in the liver, and a high concentration of biliary 1-pyrenol. In addition, laboratory exposure to common EROD inducers elicited an abnormal response, suggestive of chronic intoxication.


Asunto(s)
Biomarcadores/análisis , Monitoreo del Ambiente/estadística & datos numéricos , Agua Dulce/química , Percas/metabolismo , Contaminantes Químicos del Agua/análisis , Acetilcolinesterasa/análisis , Análisis de Varianza , Animales , Citocromo P-450 CYP1A1/análisis , Aductos de ADN/análisis , Femenino , Glutatión Transferasa/análisis , Hígado/metabolismo , Hígado/patología , Metalotioneína/análisis , Músculo Esquelético/metabolismo , Percas/anatomía & histología , Bifenilos Policlorados/análisis , Estaciones del Año , Suecia , Contaminantes Químicos del Agua/toxicidad
14.
Biomarkers ; 8(6): 445-60, 2003.
Artículo en Inglés | MEDLINE | ID: mdl-15195677

RESUMEN

In order to understand and apply DNA adduct formation in fish liver as a biomarker for aquatic pollution, information concerning the natural background levels in noncontaminated organisms, caused by endogenous compounds, is of fundamental importance. In this study, DNA adducts were analysed in liver of 11 fish species from arctic and sub-arctic areas in the northern Atlantic using the nuclease P1 version of the 32P-postlabelling technique. The collected fish were assumed not to have been influenced by anthropogenic pollution apart from possible long-range transported pollutants. As polycyclic aromatic hydrocarbons (PAHs) are thought to be fundamental in forming the type of DNA adducts detected by the method used, biliary PAH metabolite levels were measured in a selection of the investigated species. In all investigated individuals, the levels of PAH metabolites were undetectable. Controlled on-site exposure experiments with benzo[a]pyrene (polar cod) and laboratory experiments with crude oil (polar cod and Atlantic cod) were conducted. DNA adducts were formed in both these species. The field-sampled fish showed undetectable levels of DNA adducts or levels just above the detection limit. The present study supports the assumption that when DNA adducts are detected by the nuclease P1 version of the 32P-postlabelling method in fish liver, it can be interpreted as DNA damage caused by pollutants.


Asunto(s)
Aductos de ADN/análisis , Monitoreo del Ambiente/métodos , Peces/metabolismo , Endonucleasas Específicas del ADN y ARN con un Solo Filamento , Animales , Océano Atlántico , Biomarcadores/análisis , Femenino , Hígado/química , Masculino , Isótopos de Fósforo , Hidrocarburos Policíclicos Aromáticos/análisis , Contaminantes del Agua/metabolismo
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