Antagonistic effects in zebrafish (Danio rerio) behavior and oxidative stress induced by toxic metals and deltamethrin acute exposure.

In natural environments, the aquatic organisms are exposed to complex mixtures of chemicals which may originate from natural sources or from anthropogenic activities. In this context, the aim of the study was to assess the potential effects that might occur when aquatic organisms are simultaneously exposed to multiple chemicals. For that, we have studied the acute effects of cadmium (0.2 µg L-1), nickel (10 µg L-1) and deltamethrin (2 µg L-1) as individual toxicants and as mixture on the behavioral responses, oxidative stress (SOD and GPx), body electrolytes and trace metals profiles of zebrafish (Danio rerio). So far the scientific literature did not report about the combined effects of pesticides and toxic metals on zebrafish behavior using a 3D tracking system. Compared with other studies, in the present paper we investigated the acute effects of two heavy metals associated with a pesticide on zebrafish, in the range of environmentally relevant concentrations. Thus, the environmental concentrations of cadmium and nickel in three rivers affected by urban activities and one river with protected areas as background control were measured. The observations that resulted in our study demonstrated that deltamethrin toxicity was significantly decreased in some of the behavioral variables and oxidative stress when combined with CdNi mixture. Consequently, our study supports previous works concerning the combined toxicity of environmental chemicals since their simultaneous presence in the aqueous environment may lead to higher or lower toxicological effects on biota than those reported from a single pollutant. Therefore, the evaluation of toxic effects of a single contaminant does not offer a realistic estimate of its impact against aqueous ecosystems. This study also supports the idea that the interactions between different chemical compounds which do not exceed the maximum permitted limits in environment may have benefits for aquatic life forms or be more toxic.

Chaotic genetic structure and past demographic expansion of the invasive gastropod Tritia neritea in its native range, the Mediterranean Sea.

To better predict population evolution of invasive species in introduced areas it is critical to identify and understand the mechanisms driving genetic diversity and structure in their native range. Here, we combined analyses of the mitochondrial COI gene and 11 microsatellite markers to investigate both past demographic history and contemporaneous genetic structure in the native area of the gastropod Tritia neritea, using Bayesian skyline plots (BSP), multivariate analyses and Bayesian clustering. The BSP framework revealed population expansions, dated after the last glacial maximum. The haplotype network revealed a strong geographic clustering. Multivariate analyses and Bayesian clustering highlighted the strong genetic structure at all scales, between the Black Sea and the Adriatic Sea, but also within basins. Within basins, a random pattern of genetic patchiness was observed, suggesting a superimposition of processes involving natural biological effects (no larval phase and thus limited larval dispersal) and putative anthropogenic transport of specimens. Contrary to the introduced area, no isolation-by-distance patterns were recovered in the Mediterranean or the Black Seas, highlighting different mechanisms at play on both native and introduced areas, triggering unknown consequences for species' evolutionary trajectories. These results of Tritia neritea populations on its native range highlight a mixture of ancient and recent processes, with the effects of paleoclimates and life history traits likely tangled with the effects of human-mediated dispersal.

Connectivity Among Populations of the Top Shell Gibbula divaricata in the Adriatic Sea.

Genetic connectivity studies are essential to understand species diversity and genetic structure and to assess the role of potential factors affecting connectivity, thus enabling sound management and conservation strategies. Here, we analyzed the patterns of genetic variability in the marine snail Gibbula divaricata from five coastal locations in the central-south Adriatic Sea (central Mediterranean) and one in the adjacent northern Ionian Sea, using 21 described polymorphic microsatellite loci. Observed and expected heterozygosity varied from 0.582 to 0.635 and 0.684 to 0.780, respectively. AMOVA analyses showed that 97% of genetic variation was observed within populations. Nevertheless, significant, although small, genetic differentiation was found among nearly all of the pairwise F ST comparisons. Over a general pattern of panmixia, three groups of populations were identified: eastern Adriatic populations, western Adriatic populations, and a third group represented by the single northern Ionian Sea population. Nonetheless, migration and gene flow were significant between these groups. Gibbula divaricata is thought to have a limited dispersal capacity related to its lecithotrophic trochophore larval stage. Our results indicated high levels of self-recruitment and gene flow that is mainly driven through coastline dispersion, with populations separated by the lack of suitable habitats or deep waters. This stepping-stone mode of dispersion together with the high levels of self-recruitment could lead to higher levels of population structuring and differentiation along the Adriatic Sea. Large effective population sizes and episodic events of long-distance dispersal might be responsible for the weak differentiation observed in the analyzed populations. In summary, the circulation system operating in this region creates natural barriers for dispersion that, together with life-history traits and habitat requirements, certainly affect connectivity in G. divaricata. However, this scenario of potential differentiation seems to be overridden by sporadic events of long-distance dispersal across barriers and large effective population sizes.

Toxic metals biomonitoring based on prey-predator interactions and environmental forensics techniques: A study at the Romanian-Ukraine cross border of the Black Sea.

Marine cross-border areas are ideal for monitoring pollutants so as to increase ecosystems protection. This study was conducted at the Romanian-Ukraine border of the Black Sea to reveal evidence of contamination with toxic metals based on biomonitoring of: cadmium, lead, total chromium, nickel and copper at different water depths and prey-predator interactions, combined with environmental forensics techniques of biological sampling and separation in witnesses size groups. The species used were Mytilus galloprovincialis L. and Rapana venosa V. collected at 17.5m, 28m and 35m depth. An atomic absorption spectrometer with a high-resolution continuum source and graphite furnace was used for toxic metals quantification in various samples: sediments, soft tissue, stomach content, muscular leg, hepatopancreas. The best sample type, based on the pathology of metal location and bioaccumulation, is the hepatopancreas from R. venosa that proved a significant decrease of cadmium and lead at lower depths.

Toxic metals in tissues of fishes from the Black Sea and associated human health risk exposure.

The anthropogenic activities in the Black Sea area are responsible for toxic metal contamination of sea food products. In this study, several toxic metals: cadmium, lead, nickel, chromium, and copper were quantified in different tissues (digestive tract, muscle, skeleton, skin) of nine fish species (Neogobius melanostomus, Belone belone, Solea solea, Trachurus mediterraneus ponticus, Sardina pilchardus, Engraulis encrasicolus, Pomatomus saltatrix, Sprattus sprattus, Scorpaena porcus) by using atomic absorption spectrometer with a high-resolution continuum source and graphite furnace technique (HR-CS GF-AAS), and the risk of fish meat consumption by the young human population was evaluated. These metals are used in high amounts in industries located near the coastline such as shipyard construction and industrial plants. Toxic metal accumulation depends on fish feeding behavior, abiotic conditions, metal chemistry, and animal physiology. For instance, cadmium was measured in the muscle of the investigated species and average values of 0.0008-0.0338 mg kg-1 were obtained. The lowest average value of this metal was measured at benthic species N. melanostomus and the highest at the pelagic predator T. mediterraneus ponticus. Generally, the highest metal concentration was measured in the digestive tract that has the role of biofilter for these contaminants. The risk of contamination is significantly reduced by avoiding the consumption of certain fish tissues (digestive tract and skin for copper and skeleton for nickel). An estimation of the dietary metal intake to young consumers was realized for each of the studied species of fish from Romanian, Bulgarian, and Turkish waters, during the period 2001-2014 in order to evaluate the risks of chronic exposure in time due to metal toxicity. This estimation is important for the prevention of chronic exposure due to metal toxicity. Food exposure to studied metals showed a negative trend for Romania, Turkey, and Bulgaria based on the data provided by this study. The young consumers were highly exposed to these elements during the 2001-2006 period as proven by the results from this study.

Patterns and mechanisms of dispersal in a keystone seagrass species.

Mechanisms and vectors of long-distance dispersal remain unknown for many coastal benthic species, including plants. Indications for the possibility for long-distance dispersal come from dispersal modelling and from genetic assessments, but have rarely been assessed with both methods. To this end, we assessed dispersal of the seagrass Zostera noltei, an important foundation species of the coastal zone. We investigate whether small scale seed dispersal and long-distance propagule dispersal do play a role for meta-population dynamics, using both genetic assessments based on eight microsatellite markers and physical modelling of ocean currents. Such assessments enhance our understanding of the biology and population dynamics of an important coastal foundation species. They are relevant for large scale conservation strategies as they give insights in the maintenance of genetic diversity and connectivity that may enhance resilience and resistance to stresses associated with seagrass loss.