The use of a non-lethal tool for evaluating toxicological hazard of organochlorine contaminants in Mediterranean cetaceans: new data 10 years after the first paper published in MPB

In the Mediterranean Sea, top predators, and particularly cetacean odontocetes, accumulate high concentrations of organochlorine contaminants and toxic metals, incurring high toxicological risk. In this paper we investigate the use of the skin biopsies as a non-lethal tool for evaluating toxicological hazard of organochlorines in Mediterranean cetaceans, presenting new data 10 years after the paper published by Fossi and co-workers [Mar. Poll. Bull. 24 (9) (1992) 459] in which this new methodology was first presented. Some organochlorine compounds, now with worldwide distribution, are known as endocrine disrupting chemicals (EDCs). Here the unexplored hypothesis that Mediterranean cetaceans are potentially at risk due to organochlorines with endocrine disrupting capacity is investigated. High concentrations of DDT metabolites and PCB congeners (known as EDCs) were found in the different Mediterranean species (Stenella coeruleoalba, Delphinus delphis, Tursiops truncatus and Balaenoptera physalus). In this paper we also propose benzo(a)pyrene monooxygenase (BPMO) activity in marine mammal skin biopsies (non-lethal biomarker) as a potential indicator of exposure to organochlorines, with special reference to the compounds with endocrine disrupting capacity. A statistically significant correlation was found between BPMO activity and organochlorine levels (DDTs, pp(')DDT, op(')DDT, PCBs and PCB99) in skin biopsies of males of B. physalus. Moreover a statistical correlation was also found between BPMO activity and DDT levels in skin biopsies of the endangered Mediterranean population of D. delphis. These results suggest that BPMO induction may be an early sign of exposure to organochlorine EDCs and can be used for periodic monitoring of Mediterranean marine mammal toxicological status.     

Ecological Significance of Endocrine Disruption in Marine Invertebrates

Anthropogenic chemicals which can disrupt the hormonal (endocrine) systems of wildlife species are currently a major cause for concern. Reproductive hormone-receptor systems appear to be especially vulnerable. In the past few years, numerous effects of endocrine disrupting chemicals on wildlife have emerged including changes in the sex of riverine fish, reproductive failure in birds and abnormalities in the reproductive organs of alligators and polar bears. Much less is known regarding endocrine disruption in marine invertebrates, the key structural and functional components of marine ecosystems.

In this paper, potential effects of different classes of endocrine disrupting chemicals are reviewed. The endocrinology of several major invertebrate groups is briefly examined to identify which phyla are most likely to be at risk. Gaps in our knowledge concerning the availability and uptake of endocrine disruptors are identified. For example, the relative importance of different routes of chemical uptake (from seawater vs food) is considered. Feeding strategies (herbivores, carnivores, deposit feeders, suspension feeders) in relation to uptake of endocrine disruptors are also discussed.

Examples of endocrine disruption in marine invertebrates in situ are provided, including imposex in gastropod molluscs exposed to organotin compounds and intersex in crustaceans exposed to sewage discharges. Laboratory data are presented concerning the effects of endocrine disruptors on the growth and reproductive output of the deposit feeding amphipod Corophium volutator and the polychaete worm Dinophilus gyrociliatus. Recent findings are reported which demonstrate reductions in settlement following exposure of barnacle larvae to the xeno-estrogen, 4 nonyl phenol. The potential use of cyprid major protein as a biomarker of oestrogenicity is explored. The ecological significance of endocrine disruption in marine invertebrates is discussed. With regard to environmental management action, an evidence-based approach is advocated. A protocol for collecting evidence of ecologically significant endocrine disruption is outlined.     

Endocrine Disrupters in the Aquatic Environment: An Overview

Although a relatively new area of environmental research, the field of endocrine disruption has grown very rapidly, and currently many hundreds, perhaps even a few thousand, papers are published annually on the many different aspects covered by the field. As far as endocrine disruption in wildlife is concerned, most attention has been focused on aquatic organisms, for two reasons. Firstly, the aquatic environment receives most of the pollutants intentionally released into the environment, through effluents from wastewater treatment plants, and secondly because many of the best documented examples of endocrine disruption in wildlife are of partially or completely aquatic species. These two reasons are probably not unconnected, of course. Hence, aquatic organisms can receive continuous exposure to endocrine‐disrupting chemicals throughout their lives, albeit usually to low concentrations of these chemicals. Analysis of effluents has identified many of the endocrine‐disrupting chemicals present, and shown that these are both natural and man‐made, and vary greatly in potency. Most attention has been directed to identifying the main estrogenic chemicals, because many of the effects reported in wildlife appear to be a consequence of ‘feminization’ of males. However, chemical analysis of effluents has also demonstrated that chemicals with other types of endocrine activity are present, such as androgens, anti‐androgens, progestagens, etc. The effects (if any, of course) of such chemicals on aquatic organisms are unknown, and largely uninvestigated, presently. Much of the biological research has centred on the effects of estrogenic chemicals, especially to fish. These effects, such as elevated vitellogenin concentrations and intersexuality, have to date been studied almost exclusively at the level of the individual, and hence whether endocrine‐disrupting chemicals cause population‐level consequences is largely unknown (the undeniable effects of TBT on molluscs, leading to local extinctions, being the exception). It is my opinion that rather too much of the recent research has not advanced our understanding of endocrine disruption a great deal, and we are probably not much further forward now than we were five years or so ago. It is surely time to tackle some of the outstanding, unresolved issues, such as the impact of endocrine disruption at the population level, and the issue of how organisms respond when exposed to complex mixtures of endocrine active chemicals. Such research will not be easy, and will require multidisciplinary teams, including people with expertise in areas not yet involved in the field of endocrine disruption, such as mathematical modellers. However, until such research is done, it will not be possible to decide how important an issue endocrine disruption is to wildlife, and how that importance compares to the other factors adversely affecting wildlife, such as habitat loss, climate change, and the introduction of exotic species and novel diseases.     

Gonadal histology and serum vitellogenin levels of bigeye tuna Thunnus obesus from the Northern Pacific Ocean--absence of endocrine disruption bio-indicators

Endocrine disrupting chemicals such as organochlorines have been detected in a large number of marine fish. Histological observation of the gonads, measurement of serum vitellogenin (VTG) level and of liver polychlorinated biphenyl (PCB) content were performed to evaluate the reproductive health and the contamination with endocrine disruptors in bigeye tuna Thunnus obesus, collected in the northern Pacific Ocean in 1999 and 2000. Abnormalities commonly found in species affected by endocrine disruptors such as the presence of oocytes in the testis or elevated serum VTG levels were not found in any of males examined. Both males and females had only small amounts of liver PCB content. The results suggest that currently there is little if any risk of organochlorine contamination or endocrine disruption of gonadal function in bigeye tuna from the northern Pacific Ocean. However, further studies are necessary to evaluate the health status of the open sea fishery resources.     

Butyltin Contamination in Marine Mammals – A Review

In order to understand organotin accumulation in marine mammals, the present study determined the concentrations of butyltins (BTs) in various tissues and organs and described their distribution patterns relative to sex, age and geographical factors. Both cetaceans and pinnipeds showed higher BTs concentrations in the liver amongst various tissues and organs. In addition, noticeably high concentrations were found in the hair of pinnipeds, indicating possible excretion of BTs through shedding. BTs composition in mammals and their prey organisms suggested that pinnipeds have a stronger capacity to degrade BTs as compared to cetaceans. No age trends of BTs concentrations were observed in pinnipeds, while cetaceans showed increasing levels in immature growth stages. Comparing butyltin concentrations in various marine mammals, cetaceans retained higher butyltin concentrations than pinnipeds. These specific accumulation patterns found in marine mammals are probably attributable to the lower breakdown capacity of BTs in cetaceans and the significant excretion of BTs through shedding in pinnipeds. Unlike organochlorines, comparable residue levels of butyltins were found in male and female marine mammals. Such a trend suggests that butyltins are less transferable through gestation and lactation from mother to fetus/pup. On a global perspective, residue levels were found to be prominent in the coastal waters of developed nations. The present contamination by BTs may pose a considerable toxic threat to some coastal species of cetaceans. A relatively high percentage of BTs in total tin was found in the liver of cetaceans. This finding suggests that the hepatic tin in cetaceans exists predominantly in organic forms such as butyltin compounds, implying further that tin residues in marine mammals mostly reflects input from anthropogenic sources.     

Accumulation of organochlorines and brominated flame retardants in estuarine and marine food chains: field measurements and model calculations

Food chain accumulation of organochlorines and brominated flame retardants in estuarine and marine environments is compared to model estimations and fresh water field data. The food chain consists of herbivores, detritivores and primary and secondary carnivores i.e. fish, fish-eating birds and marine mammals. Accumulation of polychlorinated biphenyls is predicted well by OMEGA for herbi-detritivores and primary and secondary carnivorous fish. Ratios are similar to those found for fresh water species. Accumulation ratios for fish-eating birds and mammals are overestimated by the model, which is attributed partly to biotransformation of meta-para unsubstituted congeners. Additionally, birds may feed in other less polluted areas. For brominated diphenylethers (BDE) accumulation patterns are highly species and congener specific. Accumulation depends on both K(ow) and metabolization capacities. BDE47 is the predominant congener in lower trophic levels. For marine birds and mammals accumulation ratios of BDE99 and 100 are similar to or higher than ratios of persistent PCBs.     

Inter-population variability in the reproductive morphology of the shore crab (Carcinus maenas): evidence of endocrine disruption in a marine crustacean?

Environmental contaminants that are capable of causing endocrine disrupting effects are currently a major cause for concern. These chemicals are known to influence the reproductive development of vertebrates by mimicking or antagonising the actions of endogenous hormones. However, little is known regarding their potential effects on invertebrates. Here we examine variations in the reproductive morphology of the shore crab (Carcinus maenas) for evidence of endocrine disruption. Crabs were collected from a number of sites comprising a putative gradient of exposure to endocrine disrupting chemicals. Patterns of inter-population variability in the expression of sexually dimorphic traits were then examined for evidence of hormone disruption. Extensive variability was detected and patterns of chelal morphology were consistent with the gradient of endocrine disruption. However, overall, the patterns of morphological variability were not consistent with hormonally-mediated effects. This suggests that shore crabs are not susceptible to the same type of endocrine disrupting effects that have been detected in vertebrates, which are most commonly mediated via the oestrogen receptor. However, the potential for androgenic effects on crustacean morphology are discussed.     

An assessment of marine pollution in Puget Sound

The Marine Ecosystems Analysis (MESA) Puget Sound Project was initiated to determine the existing levels, sources, fates and effects of toxic chemicals in Puget Sound, Washington. An approach was developed to assess the concentrations of a suite of contaminants and to measure a suite of biological parameters known to be responsive to contaminants. Metals, aromatic hydrocarbons and synthetic organics were found throughout the Sound. Highest concentrations were in bays near urban areas, particularly Elliott Bay near Seattle and Commencement Bay near Tacoma. A variety of biological disorders, including liver tumors in flatfish, and benthic community parameters were correlated with elevated contaminant levels. Because of the high contaminant concentrations there, and the high potential for exposure of humans to them, Commencement Bay has been identified by the US Environmental Protection Agency as a high priority toxic dump site in the United States, and remedial action is planned.     

Toxic chemicals in Canadian fish-eating birds

Cross-country comparison of DDE and PCB residue levels in cormorant, gull and tern eggs in Canada reveal that bird populations at the Great Lakes are most contaminated with those pollutants. DDE levels have been correlated with reproductive failure in Double-crested Cormorants in the Great Lakes with eggshell thinning as a major factor. Low reproductive success in Herring Gull colonies at Lake Ontario is associated with high chlorinated hydrocarbon levels in eggs. Fish-eating birds in the Wabigoon River system, northwestern Ontario, are among the most known mercury contaminated birds. It is suggested that the effects of mercury on the reproduction of fish-eating birds should be further examined there.Fish-eating birds occupy the highest levels of the food web and magnification of toxic chemicals through prey organisms in this web makes those birds vulnerable to the effects of environmental contaminants. Since fish-eating birds are present everywhere in Canada's freshwater and marine habitats and occupy various niches there, they may serve as pollution indicators in various food chains of our aquatic environment. Colonial birds are especially valuable indicators as pollution effects on total bird populations can be studied. Baseline information on fish-eating bird populations should now be collected everywhere in Canada for measuring present and future effects of environmental pollutants, as well as other man-made disturbances on their populations.     

Comparative study of 17 β-estradiol on endocrine disruption and biotransformation in fingerlings and juveniles of Japanese sea bass Lateolabrax japonicus

Estrogenic contaminants in the aquatic environment are associated with endocrine disruption and feminization in fish. The effects of endocrine disrupting chemicals (EDCs) on fish have been well documented. However, very few studies have focused on 17 β-estradiol (E2) and its effects on endocrine system and biotransformation in a single prolonged exposure. This study investigated changes in the levels of serotonin (5-hydroxytryptamine) and acetyl choline esterase (AchE) in brain, cortisol in plasma and Ethoxyresorufin-O-deethylase (EROD) activity in gill of two different size groups (fingerlings and juveniles) of Japanese sea bass (Lateolabrax japonicus) upon exposure to two sub-lethal concentrations (200 and 2000 ng L⁻¹) of E2 for 30 d. The results indicate that cortisol level and EROD activity significantly increased in both groups, whereas serotonin level increased in juveniles and decreased in fingerlings due to E2 exposure. The correlation analysis revealed that E2 significantly affected the endocrine and biotransformation systems in both age groups.     

Perfluorinated compounds in blood of Caretta caretta from the Mediterranean Sea

Perfluorinated compounds (PFCs), widely used for their hydro-oil repellent properties, are almost non-degradable in the environment; there is scientific evidence that indicate bioaccumulation. They represent a threat to many organisms, because they are toxic and are endocrine disruptors. Scientific studies have demonstrated the presence of PFCs in blood and liver samples of fish, turtles, birds and mammals of marine ecosystems in different geographical areas. The aim of this study was to determine the distribution of PFOS and PFOA in blood samples of the marine turtle Caretta caretta, using a minimally invasive sampling procedure. 49 blood samples of marine turtle, taken from several Italian marine turtle rescue centers, were analyzed. While PFOA was never detected, measurable concentrations of PFOS were found in 15 blood samples; the values show a range from 1.14 ng/g to 28.51 ng/g (wet wt.). No differences between groups of samples taken from different areas were found.     

Evaluating legacy contaminants and emerging chemicals in marine environments using adverse outcome pathways and biological effects-directed analysis

Natural and synthetic chemicals are essential to our daily lives, food supplies, health care, industries and safe sanitation. At the same time protecting marine ecosystems and seafood resources from the adverse effects of chemical contaminants remains an important issue. Since the 1970s, monitoring of persistent, bioaccumulative and toxic (PBT) chemicals using analytical chemistry has provided important spatial and temporal trend data in three important contexts; relating to human health protection from seafood contamination, addressing threats to marine top predators and finally providing essential evidence to better protect the biodiversity of commercial and non-commercial marine species. A number of regional conventions have led to controls on certain PBT chemicals over several years (termed ‘legacy contaminants’; e.g. cadmium, lindane, polycyclic aromatic hydrocarbons [PAHs] and polychlorinated biphenyls [PCBs]). Analytical chemistry plays a key role in evaluating to what extent such regulatory steps have been effective in leading to reduced emissions of these legacy contaminants into marine environments. In parallel, the application of biomarkers (e.g. DNA adducts, CYP1A-EROD, vitellogenin) and bioassays integrated with analytical chemistry has strengthened the evidence base to support an ecosystem approach to manage marine pollution problems. In recent years, however, the increased sensitivity of analytical chemistry, toxicity alerts and wider environmental awareness has led to a focus on emerging chemical contaminants (defined as chemicals that have been detected in the environment, but which are currently not included in regulatory monitoring programmes and whose fate and biological impacts are poorly understood). It is also known that natural chemicals (e.g. algal biotoxins) may also pose a threat to marine species and seafood quality. Hence complex mixtures of legacy contaminants, emerging chemicals and natural biotoxins in marine ecosystems represent important scientific, economic and health challenges. In order to meet these challenges and pursue cost-effective scientific approaches that can provide evidence necessary to support policy needs (e.g. the European Marine Strategy Framework Directive), it is widely recognised that there is a need to (i) provide marine exposure assessments for priority contaminants using a range of validated models, passive samplers and biomarkers; (ii) integrate chemical monitoring data with biological effects data across spatial and temporal scales (including quality controls); and (iii) strengthen the evidence base to understand the relationship between exposure to complex chemical mixtures, biological and ecological impacts through integrated approaches and molecular data (e.g. genomics, proteomics and metabolomics). Additionally, we support the widely held view that (iv) that rather than increasing the analytical chemistry monitoring of large number of emerging contaminants, it will be important to target analytical chemistry towards key groups of chemicals of concern using effects-directed analysis. It is also important to evaluate to what extent existing biomarkers and bioassays can address various classes of emerging chemicals using the adverse outcome pathway (AOP) approach now being developed by the Organization for Economic Cooperation and Development (OECD) with respect to human toxicology and ecotoxicology.     

The Pelagos Sanctuary for Mediterranean marine mammals: Marine Protected Area (MPA) or marine polluted area? The case study of the striped dolphin (Stenella coeruleoalba)

The concurrence of man-made pressures on cetaceans in the Mediterranean Sea is potentially affecting population stability and marine biodiversity. This needs to be proven for the only pelagic marine protected area in the Mediterranean Sea: the Pelagos Sanctuary for Mediterranean Marine Mammals. Here we applied a multidisciplinary tool, using diagnostic markers elaborated in a statistical model to rank toxicological stress in Mediterranean cetaceans. As a case study we analyzed persistent, bioaccumulative and toxic chemicals combined with a wide range of diagnostic markers of exposure to anthropogenic contaminants and genetic variation as marker of genetic erosion in striped dolphin (Stenella coeruleoalba) skin biopsies. Finally, a statistical model was applied to obtain a complete toxicological profile of the striped dolphin in the Pelagos Sanctuary and other Mediterranean areas (Ionian Sea and Strait of Gibraltar). Here we provide the first complete evidence of the toxicological stress in cetaceans living in Pelagos Sanctuary.     

Contaminants still high in top-level carnivores in the Southern California Bight: levels of DDT and PCBs in resident and transient pinnipeds

Highly industrialized areas, such as the Southern California Bight, often have high levels of contaminants in marine sediments, which can cause chronic exposure to organisms long after their use has ceased. tDDT and tPCB were analyzed in the blubber of 145 stranded pinnipeds that died at local marine mammal centers between 1994 and 2006. Resident species (California sea lion and Pacific harbor seal) had significantly higher concentrations of tDDT and tPCB than the transient species (northern elephant seal). Adult female California sea lions had significantly lower concentrations of tDDT and tPCB than pups, yearlings, and adult males. Concentrations of both tDDT and tPCB in California sea lions significantly declined over time, but did not change in northern elephant seals. Current concentrations of tDDT and tPCB in California sea lions and harbor seals are among the highest values reported worldwide for marine mammals and exceed those reported to cause adverse health effects.     

Cellular and molecular responses to endocrine-modulators and the impact on fish reproduction

Anthropogenic chemicals in the aquatic environment are known to cause reproductive disturbances in vertebrate and invertebrate organisms, by interfering with the endocrine systems. Laboratory-based in vivo and in vitro studies have indicated that several of the anthropogenic and other naturally occurring chemicals in the environment can cause adverse reproductive effects. Various definite or possible reproductive abnormalities caused by endocrine disruption have been identified, but in majority of the reported cases, it is not known whether adverse effects have occurred in the population level of biological organization. Disruption of the hormonal functions in fish may have effects on a number of events, including sexual maturation, gamete production and transport, sexual behaviour, fertility, gestation, lactation or modifications in other functions that are dependent on the integrity of the reproductive system. Although several reproductive effects have been reported, but the degree of causality established between the abnormalities observed and exposure to particular chemicals is variable, and understanding of the mechanism(s) is limited. Fishes are a vital source of proteins and lipids for humans and domestic animals, forming the basis for economically important fisheries and aquaculture. Large efforts have recently been denoted to dissect the mechanisms of action of xenobiotics in aquatic species, with the ultimate aim of detecting, controlling and possibly intervening in chemical exposure and its effects on the aquatic ecosystem and humans. In this context, we ought to be concerned with the health and safety of aquatic species per se, as well as a resource for human needs.     

Impacts of marine debris on wild animals in the coastal area of Korea

Over the last decade, marine debris has become a major factor affecting the coastal ecosystem of Korea. This study compiled information regarding how marine debris impacts wildlife in Korea. Cases of marine debris impacting wildlife were collected from experts of various fields and from local participants through an open access website from February 2010 to March 2012. A total of 21 species were affected by marine debris: 18 species of birds, 2 species of mammals, and 1 species of crustacean. Five threatened or protected species were identified: black-faced spoonbill, finless porpoise, water deer, whooper swan, and greater painted snipe. Recreational fishing gears were the types of debris that most frequently impacted wildlife, especially birds. Black tailed gulls were the most vulnerable species to recreational fishing hooks and lines. Although it was preliminary, this study revealed that recreational fishing activities should be prioritized when managing marine debris in Korea.     

Derelict fishing nets in Puget Sound and the Northwest Straits: Patterns and threats to marine fauna

Derelict fishing gear remains in the marine environment for years, entangling, and killing marine organisms worldwide. Since 2002, hundreds of derelict nets containing over 32,000 marine animals have been recovered from Washington’s inland waters. Analysis of 870 gillnets found many were derelict for years; most were recovered from northern Puget Sound and high-relief rocky habitats and were relatively small, of recent construction, in good condition, stretched open, and in relatively shallow water. Marine organisms documented in recovered gillnets included 31,278 invertebrates (76 species), 1036 fishes (22 species), 514 birds (16 species), and 23 mammals (4 species); 56% of invertebrates, 93% of fish, and 100% of birds and mammals were dead when recovered. For all taxa, mortality was generally associated with gillnet effectiveness (total area, age and condition, and suspension in the water). Mortality from derelict fishing gear is underestimated at recovery and may be important for species of economic and conservation concern.     

An Asian quandary: where have all of the PBDEs gone?

Polybrominated diphenyl ethers (PBDEs) are widely used fire retardant compounds which, like polychlorinated biphenyls (PCBs), have become ubiquitous in environmental media. Time trend analyses of PBDE levels in sediments and biota generally show an increasing trend. The lower (tetra- and penta-) PBDEs accumulate and predominate in biota, whilst the higher congeners (deca-) are prevalent in aquatic environments and sediments. PBDEs also biomagnify, with highest concentrations of the lower brominated congeners being seen in top predators including birds, marine mammals and humans. Toxicological studies of PBDEs have been limited, although effects mediated via the thyroid gland may be extremely important. Disturbingly, PBDEs may be sequestering in certain Asian environments to a higher degree than in other parts of the world, due to the unregulated disposal of computers and electronic equipment. However, this potential problem remains poorly investigated, to date.