Genetic ecotoxicology: an overview

The techniques currently available for detecting genotoxin exposure are briefly described and evaluated with regard to the goals of genetic ecotoxicology. The occurrence and significance of genotoxin-induced neoplasia in marine organisms is described. Although there are numerous examples of hotspots where tumour incidences in fish and shellfish have been correlated with raised concentrations of anthropogenic chemicals, causal mechanisms are seldom established. Insufficient information is available to gauge the seriousness of genotoxicity for marine organisms on regional or global scales. The possibility of using marine organisms as sentinels to provide early warning of potential threats to Man is examined. Recognition of the genotoxic disease syndrome in lower animals highlights the need to explore the relationships between DNA damage (adduct formation, gene mutations, etc.) and its phenotypic consequences. Within a given population, not all individuals are equally susceptible to pollutant toxicity (including genotoxicity). The potential for using similarities in phenotypic traits to recognise subsets of individuals within populations possessing similar genotypes is discussed. Changes in heterozygosity and the evolution of genetically resistant populations following exposure to pollution are evaluated in the context of genetic ecotoxicology. Risk assessment procedures are required which enable genotoxin exposure to be related to specific consequences at the community and ecosystem levels. This necessitates both a sound scientific understanding of the mechanisms involved and the development of pragmatic ecotoxicological tools that can be employed by environmental managers.     

Ecological consequences of photomovement and photobleaching in the marine flagellate Cryptomonas maculata

Abstract The marine flagellate Cryptomonas maculata is bleached and eventually killed by exposure to even moderate white-light fluence rates. Bleaching affects all of its photosynthetic pigments and the kinetics depend on the fluence rate of the radiation the organisms are exposed to. Nitrogen-deficient cells which show a reduced pigment concentration and impaired photosynthetic efficiency tolerate bleaching white-light exposure far better than the normally colored cells. In their natural environment the organisms escape this situation by a pronounced negative phototaxis at fluence rates above 3.6 klx (= 15 W.m⁻²), while they show positive phototaxis at lower fluence rates. In nitrogen-deficient cells, however, though being less prone to photobleaching, negative phototaxis commences even at a fluence rate of about 830 lx (= 3.5 W.m⁻²). The ecological consequences of the remarkable light sensitivity and the phototactic orientation are being discussed.     

Can EMF exposure during development leave an imprint later in life?

People in industrialized nations live in an environment of ubiquitous electromagnetic field (EMF) exposure, both natural and anthropogenic. The intensity, variety, and geographic distribution of anthropogenic EMF exposures have grown dramatically since the mid 20th century, with many uses serving, and in close proximity to, human populations, such as electric power distribution, radio and television transmission, and more recently, personal cell phone communication units and transmitting towers. Thus, it is reasonable to ask if this EMF exposure could cause alterations in the physiology of developing organisms, since they are generally assumed to be the most sensitive to chemical stressors. In this report, we review work published beginning in the late 1980s. Initial reports indicated that exposure of chicken eggs during embryonic development to power-line electric fields of 50 and 60 Hz, at 10 V/m in air (which is frequently in locations inhabited by humans), could cause the brain tissues of the hatched chickens to respond differently in a particular test. More recently, an anecdotal report of human sensitivity to EMF has appeared that shows a health-related influence of prior exposure history to particular power-line frequencies in chemically sensitized individuals. These reports open the question of whether the ambient electromagnetic environment can leave an imprint on developing organisms and if such imprint changes have the potential for health consequences.     

A note on the relevance of human population genetic variation and molecular epidemiology to assessing radiation health risk for space travellers

We discuss the relevance to space medicine of studies concerning human genetic variation and consequent variable disease susceptibility or sensitivity between individuals. The size of astronaut and cosmonaut populations is both presently and cumulatively small, and despite the launch of the International Space Station, unlikely to increase by orders of magnitude within the foreseeable future. In addition, astronauts–cosmonauts constitute unrepresentative samples of their national populations. While the context of exposure for the astronaut–cosmonaut group is one unlikely to be replicated elsewhere than in space, aspects of specific exposures may be simulated by events such as occupational radiation exposure or radiation therapy. Hence, population-based studies of genetic susceptibility or sensitivity to disease, especially where it is precipitated by events that may simulate consequences of the space environment, likely will prove of value in assessing long-term health risks.     

Reduced vitamin A (retinol) levels indicate radionuclide exposure in Streaked Shearwaters (Calonectris leucomelas) following the 2011 Fukushima nuclear accident

The Fukushima nuclear accident in 2011 released significant amounts of radionuclides into the marine environment. Exposure to radiation reduces levels of antioxidants such as carotenoids and vitamins A and E within exposed individuals. Such reductions can cause teratogenic or mutagenetic effects leading to reduced reproductive viability and fitness. Reduced antioxidant levels therefore may be used as an indicator of radionuclide contamination and to infer individual or population level impacts; however, the taxa-specific responses of marine organisms, such as seabirds, are poorly understood. As top predators, seabirds are ideal bio-indicators of the prevalence of contaminants and pollutants in marine ecosystems. At-sea foraging distributions of Streaked Shearwaters (Calonectris leucomelas) from Mikura Island (MKR), Japan during the post egg-laying period coincide with the Fukushima nuclear plume while the breeding colony on Birou Island (BRU) lies outside the affected zone. We examined the physiological responses of Streaked Shearwater chicks at MKR and BRU to possible radiation exposure during the 2011 breeding season, four to seven months after the Fukushima nuclear accident. Fledging mass did not differ between islands but fledglings from MKR displayed significantly reduced vitamin A levels. Available information suggests these depletions most likely result from radiation exposure due to the Fukushima nuclear accident, implying that the risk of radionuclide contamination is considerably elevated for Streaked Shearwaters on MKR, where more than 60% of the world's population breeds. While additional negative impacts are expected due to delayed effects of radionuclide transport via biomagnification in the food chain, this study highlights the potential immediate and worrisome consequences of the Fukushima nuclear accident for marine wildlife.     

Salmonellae in the Environment Around a Chicken Processing Plant

Studies have been conducted over a 2-year period to determine the extent to which a poultry processing plant served as a reservoir of salmonellae reaching the external environment, to examine the question of the importance of salmonellae in the environment, and to consider how best to control the spread of the organisms. The studies have been undertaken at a chicken processing plant handling between 75,000 and 80,000 birds per day. Populations of salmonellae and indicator bacteria were estimated in the raw wastes, through the waste treatment plant, and in the receiving stream waters. The results demonstrate that salmonellae are present in poultry processing wastes in a surprisingly constant relation to fecal coliforms (in excess of 1 Salmonella per 500 fecal coliforms), that serotypes in the environment are constantly changing, and that they may reflect unusual conditions in the processing plant, or a possible source of infection among human and animal residents of the environment. Disinfection of poultry processing wastes is recommended.     

Control of marine bacterioplankton populations: Measurement and significance of grazing

A variety of methods have been used to estimate the degree of control exercised upon marine bacterioplankton by grazing organisms. These include filtration or dilution of samples to reduce grazers, the use of specific inhibitors to prevent growth or grazing, and the use of artificial particles or radio-labelled bacteria as tracers for the natural bacterioplankton. Each of these techniques has drawbacks which may lead to under- or overestimates of grazing. In addition, they tell us little about which organisms are doing the grazing or the degree to which viruses or lytic bacteria compete with grazers for bacterial production. Because measurements of grazing and bacterioplankton growth rates are uncertain, exact comparisons are not presently possible. Thus measurements of bacterial and bacterivore abundance, concentrated on comparisons between seasons, on diel cycles and on spatial variations, have been used to evaluate mechanisms controlling bacterial populations. These give an idea of the degree of coupling between bacterial growth and bacterivore activity and of the time scales over which growth and grazing balance. Combined with laboratory studies of grazing, they currently provide the best insight into what controls populations of bacteria in the sea.     

Effects of Ultraviolet Radiation on Locomotion and Orientation in Roughskin Newts (Taricha granulosa)

Environmental changes, including those associated with the atmosphere may significantly affect individual animals and ultimately populations. Ultraviolet (UV) radiation, perhaps increasing due to stratospheric ozone depletion, has been linked to mortality in a number of organisms, including amphibians. The eggs and larvae of certain amphibian species hatch at significantly lower rates when exposed to ambient ultraviolet light. Yet little is known about the sublethal effects of UV radiation. For example, UV radiation may affect specific behaviors of an animal that could alter its ability to survive. To examine if UV radiation affects amphibian behavior, we used roughskin newts ( Taricha granulosa ) as a model. Newts were exposed to low-level doses of UV in the laboratory and then tested in the field to examine if UV-exposed and control (no UV) newts differed in orientation towards water or in locomotor activity levels. UV-exposed and control newts both exhibited a significant orientation towards water in field tests but there was no significant difference in orientation between treatments. However, UV-exposed newts were significantly more active than control newts. Our results suggest that exposure to short-term low levels of UV radiation alters certain behaviors. Environmentally induced changes in behavior may have significant ecological and evolutionary consequences.     

Morphometric Indices of Scots Pine Needle under Chronic Radiation Exposure

We have studied the morphometric indices of needles in Scots pine populations that grow in Bryansk oblast in sites with radioactive contamination long after the Chernobyl accident. The variability in needle weight and length, as well as the fluctuating asymmetry indices and occurrence of necroses and morphoses, were studied in four contaminated and two reference populations of Scots pine in 2011, 2013, and 2014. The exposure of needles in radioactively contaminated sites varied from 7 to 130 mGy/yr. We found brachyblasts with three needles in contaminated Scots pine populations; this morphosis was absent in the reference populations. A dependence of the occurrence of needles damaged by necrosis on the levels of radiation exposure in 2011 was found. However, it was statistically insignificant in other years. The length and weight of needles in contaminated populations differed from the control values; however, the dependence of these indices on the level of radiation exposure was not revealed in the studied range of doses. In 2011 and 2013, the index of fluctuating asymmetry in needle length exceeded the control levels in sites where the absorbed doses were 90 and 130 mGy; this index also tended to grow (statistically significantly in 2011) with an increase in the characteristics of radioactive contamination of the studied plots: the exposure dose rate and the specific activity of ⁹⁰Sr and ¹³⁷Cs in cones and of ¹³⁷Cs in the soil. Therefore, one can observe the consequences of chronic radiation exposure at the organismic level in populations of Scots pine (one of the most radiosensitive plant species) even 25 years after the accident. The data that we obtained in natural habitats confirm the international estimates, according to which the annual chronic radiation exposure of 100 mGy can be considered the limit of safe radiation exposure of natural populations according to morphological and ontogenetic parameters.     

Field bioassays for early detection of chronic impacts of chemical wastes upon marine organisms

A major problem facing those who must assess the environmental effects of the disposal in the ocean of industrial and municipal wastes, including dredged materials, is determining whether given wastes elicit chronic deteriorative responses in important species of organisms. The full importance of such low-level, nonlethal effects is not known, but it is suspected that repeated elicitations may result in ecosystem changes as important as those caused by more easily determinable acute effects. Such considerations are important to the marine environment, where dumped pollutants may be quickly diluted to legal nonlethal concentrations, but may still bring forth cumulative chronic response patterns. One objective of this study has been to develop a field method of assessing the impacts of the disposal of various industrial and municipal wastes. The measure of the impact is not mortality measured against time, but the increase or decrease in activity of certain metabolic enzymes that signal whether an organism is under stress from a class of wastes. Also, by analysing tissues of test and indigenous species for the accumulation of metals, PCBs, and high molecular weight hydrocarbons as well as for the enzyme activity, one gains an insight into the actual effect, if any, of the accumulation upon the whole organism. The test organisms are exposed for selected periods of time in the field in devices called Biotal Ocean Monitors (BOMs); they are then assayed for enzyme induction. At present the following enzymes are used: mitochondrial ATPase, which responds particularly to excess biphenyls in the environment; catalase that is dissolved in the cytosol and responds to excesses of toxic metals; and cytochrome P-420 and P-450, which respond to cyclic and long-chain hydrocarbons. The applicability of the adenylate energy charge system to this problem is also studied.     

RecA Expression in Response to Solar UVR in the Marine Bacterium Vibrio natriegens

Solar ultraviolet radiation may produce daily stress on marine and estuarine communities as cells are damaged and repair that damage. Reduction in the earth's stratospheric ozone layer has increased awareness of the potential effects that ultraviolet radiation may have in the environment, including how marine bacteria respond to changes in solar radiation. We examined the use of the bacterial RecA protein as an indicator of the potential of bacteria to repair DNA damage caused by solar UV irradiation using the marine bacterium Vibrio natriegens as a model. RecA is universally present in bacteria and is a regulator protein for the so-called Dark Repair Systems, which include excision repair, postreplication recombinational repair, and mutagenic or SOS repair. Solar UVB and UVA both reduced V. natriegens viability in seawater microcosms. After exposure to unfiltered solar radiation or radiation in which UVB was blocked, survival dropped below 1%, whereas visible light from which UVA and UVB had been filtered had no effect on survival. Using a RecA-specific antibody for detection, RecA protein was induced by solar radiation in a diel pattern in marine microcosms conducted in the Gulf of Mexico. Peak induction was observed at dusk each day. Although RecA expression was correlated with the formation of UVB-induced cyclobutyl pyrimidine dimers, longer wavelength UVA radiation also induced recA gene expression. Our results demonstrate that RecA-regulated, light-independent repair is an important component in the ability of marine bacteria to survive exposure to solar ultraviolet radiation and that RecA expression is a useful monitor of bacterial repair after exposure to solar UVR.     

Marine Bacterial Isolates Display Diverse Responses to UV-B Radiation

The molecular and biological consequences of UV-B radiation were investigated by studying five species of marine bacteria and one enteric bacterium. Laboratory cultures were exposed to an artificial UV-B source and subjected to various post-UV irradiation treatments. Significant differences in survival subsequent to UV-B radiation were observed among the isolates, as measured by culturable counts. UV-B-induced DNA photodamage was investigated by using a highly specific radioimmunoassay to measure cyclobutane pyrimidine dimers (CPDs). The CPDs determined following UV-B exposure were comparable for all of the organisms except Sphingomonas sp. strain RB2256, a facultatively oligotrophic ultramicrobacterium. This organism exhibited little DNA damage and a high level of UV-B resistance. Physiological conditioning by growth phase and starvation did not change the UV-B sensitivity of marine bacteria. The rates of photoreactivation following exposure to UV-B were investigated by using different light sources (UV-A and cool white light). The rates of photoreactivation were greatest during UV-A exposure, although diverse responses were observed. The differences in sensitivity to UV-B radiation between strains were reduced after photoreactivation. The survival and CPD data obtained for Vibrio natriegens when we used two UV-B exposure periods interrupted by a repair period (photoreactivation plus dark repair) suggested that photoadaptation could occur. Our results revealed that there are wide variations in marine bacteria in their responses to UV radiation and subsequent repair strategies, suggesting that UV-B radiation may affect the microbial community structure in surface water.     

Fish introduction and parasites in marine ecosystems: a need for information

Invasive species provide unique and useful systems by which to examine various ecological and evolutionary issues, both in terms of the effects on native environments and the subsequent evolutionary impacts. While biological invasions are an increasing agent of change in aquatic systems, alien species also act as vectors for new parasites and diseases. To date, colonizations by hosts and parasites have not been treated and reviewed together, although both are usually interwoven in various ways and may have unpredictable negative consequences. Fish are widely introduced worldwide and are convenient organisms to study parasites and diseases. We report a global overview of fish invasions with associated parasitological data. Data available on marine and freshwater are in sharp contrast. While parasites and diseases of inland freshwater fish, ornamental, reared and anadromous fish species are well documented, leading to the emergence of several evolutionary hypotheses in freshwater ecosystems during the last decade, the transfer of such organisms are virtually unexplored in marine ecosystems. The paucity of information available on the parasites of introduced marine fish reflects the paucity of information currently available on parasites of non-indigenous species in marine ecosystems. However, such information is crucial as it can allow estimations of the extent to which freshwater epidemiology/evolution can be directly transferred to marine systems, providing guidelines for adapting freshwater control to the marine environment.     

Genotoxic, carcinogenic, and teratogenic hazards in the marine environment, with special reference to the Mediterranean Sea.

Genotoxic, carcinogenic, and teratogenic hazards arising out of pollution in the marine environment are discussed in this article, with special reference to the situation in the Mediterranean area. A number of chemical compounds or complex mixtures relevant to marine pollution, either natural or of anthropogenic origin, are tentatively listed, along with protective factors which may play a counteracting role in the same environment. Harmful substances tend to undergo interactions and transformations in seawater, sediments, and marine biota, due to physical, chemical, microbial, or light-mediated mechanisms. Bioaccumulation phenomena in marine organisms may result from food-chain biomagnification processes or from concentration of pollutants by filter feeders. A variety of sources can account for marine pollution by genotoxic, carcinogenic, and teratogenic compounds, but there is a relative paucity of analytical data concerning the Mediterranean. Metabolic transformations of xenobiotics occur in all marine organisms, the biochemical mechanisms in fish being comparable to those which have been extensively investigated in mammals. Induction of metabolic pathways, and especially of the mixed-function oxygenase system, represents the earliest warning signal of exposure to pollutants. Occurrence of neoplastic diseases is documented by experimental and field studies in marine vertebrates as well as in invertebrates. The association with local pollution phenomena has been recognized in several studies, but other etiopathogenetic factors may be also involved, and in some cases tumors have been reported to be unrelated to chemical pollution. Genotoxic agents have been detected by means of suitable techniques in seawater, sediments, and marine organisms. Several studies have investigated the presence of carcinogen-DNA adducts, DNA damage and repair processes, and cytogenetic alterations, such as chromosomal aberrations, sister-chromatid exchanges, and micronuclei, in tissues of marine organisms. However, monitoring of these end-points under field conditions encounters some limitations and problems. Even more fragmentary is the information on teratogenic effects in marine organisms, although interesting test systems have been set up. On the whole, a quite extensive database on all these toxicological issues is already available in the literature, but further studies are warranted for an adequate assessment of genotoxic, carcinogenic, and teratogenic hazards, and possibly counteracting factors in the marine environment, and specifically in the Mediterranean Sea.     

A new look at evolution in the Galápagos: evidence from the late Cenozoic marine molluscan fauna

Endemism is not as common in the marine invertebrate fauna of the Galápagos Islands region as in the adjacent terrestrial biota. Marine invertebrates in the Galápagos are largely cosmopolitan species from the Panamic, Indo-Pacific, Californian, or Peruvian faunal provinces. However, an endemic component is also present in the fauna. The observed pattern among marine invertebrate organisms can be accounted for by at least two processes: (1) genetic continuity between mainland and island populations mediated through planktonic larvae; and (2) lower rates of intrinsic evolutionary change. The evolutionary scenario standardly applied to terrestrial organisms in the Galápagos, namely, adaptive radiation and speciation in reproductive isolation from mainland source populations, does not apply to all marine invertebrates. Evidence in support of the alternative scenario for marine invertebrates comes from both published records of species occurring in the islands and recent studies of fossil-bearing deposits on several islands in the archipelago. Two misconceptions–considering the islands and sedimentary deposits to be older than now thought, and equating the rate of evolution of the terrestrial biota with the marine biota–can lead to an incorrect interpretation of evolution in the Galápagos Contrasts between marine invertebrate and terrestrial organisms serve to illustrate some fundamental differences which have important evolutionary implications. Some of these are: endemism; dispersal; taxonomic relationships; island definitions; rates of evolutionary change; and age of fossils. In terms of Darwin's evolutionary scenario, terrestrial organisms represent the paradigm and marine organisms represent the paradox.     

Dissecting the Rhizosphere complexity: The truffle-ground study case

The word Rhizosphere describes the part of the soil which is immediately adjacent to and affected by plant roots. This is a very dynamic environment where plants, soil and microorganisms interact. The plant releases, in addition to biologically active substances, nutritive substances (exudates), which create a privileged habitat for many microbial populations. The same microbes that live in the rhizosphere may be useful for the plant. Interest in this fascinating environment has increased over the years. However, our knowledge of the biology and diversity of microbial populations in the rhizosphere is still limited, because it has always been linked to traditional culture-based techniques. These methods, which only allow the study of cultured microorganisms, do not allow the majority of the organisms existing in nature to be characterized. Over the last few years, this limitation has been overcome through the introduction of methodologies that are independent of culture techniques. This different approach, which has revolutionized scientific research, is known as metagenomics. In this review, the rhizosphere environment is considered with particular attention to the fungal and symbiotic organisms, which populate it. The new environmental genomic techniques and how these have been applied to the study of the various environments and the rhizosphere are described. Finally, a specific rhizosphere, a truffle-ground, is described as our study case.      

Phenotype–environment mismatches reduce connectivity in the sea

The connectivity of marine populations is often surprisingly lower than predicted by the dispersal capabilities of propagules alone. Estimates of connectivity, moreover, do not always scale with distance and are sometimes counterintuitive. Population connectivity requires more than just the simple exchange of settlers among populations: it also requires the successful establishment and reproduction of exogenous colonizers. Marine organisms often disperse over large spatial scales, encountering very different environments and suffering extremely high levels of post-colonization mortality. Given the growing evidence that such selection pressures often vary over spatial scales that are much smaller than those of dispersal, we argue that selection will bias survival against exogenous colonizers. We call this selection against exogenous colonizers a phenotype–environment mismatch and argue that phenotype–environment mismatches represent an important barrier to connectivity in the sea. Crucially, these mismatches may operate independently of distance and thereby have the potential to explain the counterintuitive patterns of connectivity often seen in marine environments. We discuss how such mismatches might alter our understanding and management of marine populations.
Ecology Letters (2010) 13: 128–140     

Nuclear waste management activities in the pacific basin and regional cooperation on the nuclear fuel cycle

Abstract

Pacific Ocean and island sites have been used since World War II for nuclear activities, including effluent discharges from nuclear facilities, sea dumping of packaged radioactive wastes, and testing of nuclear explosives. In the future, the amounts of radioactive wastes deliberately released into the Pacific Ocean may increase in connection with planned commercial‐scale nuclear fuel reprocessing operations, recommencement of plutonium production for weapons purposes, and resumption of sea dumping of low‐level wastes. Proposed storage of spent nuclear fuel on Pacific island sites or disposal of high‐level wastes in the deep seabed of the Pacific could also expose the ocean to a risk of contamination by long‐lived radio‐nuclides. The consequences of all these activities should be assessed in practical terms—their likely effects on the living marine resources of the Pacific and the economic development of the societies benefited by them; in terms of the legal principles which govern activities such as marine radioactive waste disposal that could pollute the marine environment; and in relation to current and future organizational arrangements that could achieve political resolution of outstanding international nuclear energy issues. Despite the prospective dangers of marine nuclear activities, the use of relatively remote or extraterritorial marine locations including those in the Pacific basin for nuclear operations could provide a basis for international cooperation on management of the “back end”; of the nuclear fuel cycle, including storage and reprocessing of spent fuel and high‐level waste disposal. A broadly recognized international regime for the nuclear fuel cycle could be based on regional organization of such back‐end operations, provided local acceptance could be obtained.     

Population genomic evidence for adaptive differentiation in the Baltic Sea herring

Detecting and estimating the degree of genetic differentiation among populations of highly mobile marine fish having pelagic larval stages is challenging because their effective population sizes can be large, and thus, little genetic drift and differentiation is expected in neutral genomic sites. However, genomic sites subject to directional selection stemming from variation in local environmental conditions can still show substantial genetic differentiation, yet these signatures can be hard to detect with low‐throughput approaches. Using a pooled RAD‐seq approach, we investigated genomewide patterns of genetic variability and differentiation within and among 20 populations of Atlantic herring in the Baltic Sea (and adjacent Atlantic sites), where previous low‐throughput studies and/or studies based on few populations have found limited evidence for genetic differentiation. Stringent quality control was applied in the filtering of 1 791 254 SNPs, resulting in a final data set of 68 182 polymorphic loci. Clear differentiation was identified between Atlantic and Baltic populations in many genomic sites, while differentiation within the Baltic Sea area was weaker and geographically less structured. However, outlier analyses – whether including all populations or only those within the Baltic Sea – uncovered hundreds of directionally selected loci in which variability was associated with either salinity, temperature or both. Hence, our results support the view that although the degree of genetic differentiation among Baltic Sea herring populations is low, there are many genomic regions showing elevated divergence, apparently as a response to temperature‐ and salinity‐related natural selection. As such, the results add to the increasing evidence of local adaptation in highly mobile marine organisms, and those in the young Baltic Sea in particular.