Arsenic-contaminated freshwater: assessing arsenate and arsenite toxicity and low-dose genotoxicity in Gammarus elvirae (Crustacea; Amphipoda).

Arsenic (As) contamination of freshwater is largely due to geogenic processes, but As is also released into the environment because of improper anthropic activities. The European regulatory limits in drinking water are of 10 µg L-1 As. However, knowledge of the genotoxic effects induced by low doses of As in freshwater environments is still scanty. This study was designed to investigate arsenate (As(V)) and arsenite (As(III)) toxicity and low-dose genotoxicity in Gammarus elvirae, which has proved to be a useful organism for genotoxicity assays in freshwater. As(V) and As(III) toxicity was assessed on the basis of the median lethal concentration, LC(50), while estimates of DNA damage were based on the Comet assay. The G. elvirae LC (50-240 h) value we calculated was 1.55 mg L-1 for As(V) and 1.72 mg L-1 for As(III). Arsenic exposure (240 h) at 5, 10, and 50 µg L-1 of As in assays with either arsenate or arsenite-induced DNA damage in hemocytes of G. elvirae in a concentration-dependent manner. Our study provides a basis for future genotoxic research on exposure to freshwater that contains low levels of arsenic.

Evaluation of Genotoxic Potential of Waters from Two Italian Rivers in Gammarus elvirae (Amphipoda).

The aim of the present work is to evaluate the genotoxic impact of contaminants along the whole course of Ninfa-Sisto and Amaseno (Latium, Italy) rivers. The authors performed the alkaline Comet assay to assess DNA damage in the freshwater amphipod Gammarus elvirae, exposed ex situ for 24 hours and 7 days to water collected at different sites. The assay, applied on haemocytes, provides a sensitive tool to reveal effects even at low concentrations of pollutants. The results indicate significant increase of DNA damage along the course of the two rivers, compared to the unpolluted upstream sites, even if the analytes do not exceed the permissible limits. Moreover, the results show that there is not a linear correlation between the concentration of analytes and DNA damage. Based on this study's results, it would be desirable to use Comet assay, on proposed test species, as an early warning method to detect genotoxic potential of waters.

Genetic connectivity between land and sea: the case of the beachflea Orchestia montagui (Crustacea, Amphipoda, Talitridae) in the Mediterranean Sea.

INTRODUCTION: We examined patterns of genetic divergence in 26 Mediterranean populations of the semi-terrestrial beachflea Orchestia montagui using mitochondrial (cytochrome oxidase subunit I), microsatellite (eight loci) and allozymic data. The species typically forms large populations within heaps of dead seagrass leaves stranded on beaches at the waterfront. We adopted a hierarchical geographic sampling to unravel population structure in a species living at the sea-land transition and, hence, likely subjected to dramatically contrasting forces. RESULTS: Mitochondrial DNA showed historical phylogeographic breaks among Adriatic, Ionian and the remaining basins (Tyrrhenian, Western and Eastern Mediterranean Sea) likely caused by the geological and climatic changes of the Pleistocene. Microsatellites (and to a lesser extent allozymes) detected a further subdivision between and within the Western Mediterranean and the Tyrrhenian Sea due to present-day processes. A pattern of isolation by distance was not detected in any of the analyzed data set. CONCLUSIONS: We conclude that the population structure of O. montagui is the result of the interplay of two contrasting forces that act on the species population genetic structure. On one hand, the species semi-terrestrial life style would tend to determine the onset of local differences. On the other hand, these differences are partially counter-balanced by passive movements of migrants via rafting on heaps of dead seagrass leaves across sites by sea surface currents. Approximate Bayesian Computations support dispersal at sea as prevalent over terrestrial regionalism.

In-vivo assesment of the genotoxic and oxidative stress effects of particulate matter on Echinogammarus veneris.

Seven types of atmospheric dusts (road dust, soil dust, brake dust, desert dust, pellet ash and coke and certified material NIST1648a - urban dust) have been tested for their genotoxicity on specimens of Echinogammarus veneris, a small aquatic amphipod. Experiments were carried out in vivo, by exposing the animals for 24 h to water containing 25 mg/L of dust. Each dust has been chemically analyzed for ions, elemental carbon, organic carbon and for the soluble and insoluble fractions of elements. Non-specific damages to DNA have been evaluated by the comet test, while oxidative damages have been estimated by coupling the comet test with formamido pyrimidine DNA glycosylase reaction. The animal tissues have been acid digested and analyzed for their elemental content to evaluate the bioaccumulation. All the considered dusts have caused a significant non-specific DNA damage, while the oxidative stress was shown only by dust types containing high concentration of elements. Furthermore, the oxidative damage has shown a positive correlation with the total bio-accumulated elemental concentration. For all the dust samples, the correlation with bio-accumulation in the tissues was more satisfactory for the insoluble fraction than for the soluble fraction of elements. Elements contained in solid particles seem then to be the main responsible bioaccumulation and for the oxidative stress.

An integrated study on Gammarus elvirae (Crustacea, Amphipoda): perspectives for toxicology of arsenic-contaminated freshwater.

The Italian region Latium is characterized by extensive quaternary volcanic systems that contribute greatly to arsenic (As) contamination of freshwater, including drinking water supplies. However, knowledge of the possible toxic effects in these aquatic environments is, despite being highly relevant to public health, still limited. In this paper, we approach this issue using Gammarus elvirae, an amphipod species that inhabits rivers and streams in central Italy, including Latium. We explored the possibility of using G. elvirae in the toxicology of freshwater by addressing the most relevant issues. First, we tested the usefulness of hemocytes from G. elvirae in determining non-specific DNA damage by means of the Comet assay after exposure (24 h and 7 days) to different river water samples in Latium; second, we provided an interpretative overview of the usefulness of hepatopancreatic epithelial cells of G. elvirae as a means of assessing toxicity after long-term exposure to As and other pollutants; third, the LC (50-240 h) value for G. elvirae was estimated for arsenate, which is usually the dominant arsenic species in surface waters. Our study sheds light on G. elvirae at different levels, providing a background for future toxicological research of freshwater.

Status of insecticide resistance in Culex pipiens field populations from north-eastern areas of Italy before the withdrawal of OP compounds.

BACKGROUND: Heavy and constant use of organophosphorus (OP) larvicides selected Culex pipiens L. resistant populations through two main mechanisms of genetic resistance, the increased activity of detoxifying esterase and the production of alterate acetylcholinesterase-1 (AChE1) by G119S mutation. The aim of this study was the assessment of the distribution of Cx. pipiens populations resistant to temephos and chlorpyrifos in the north-eastern regions of Italy and the occurrence of the insensitive AChE in these populations. Data describe the situation in the last years before European legislation prohibited the use of OP larvicides in mosquito control, up until 2007. RESULTS: For the first time a high level of OP resistance in the samples from Ravenna (182-fold, 80% A4/B4 or A5/B5 esterases and 38.3% Ester(5)), Emilia Romagna region, was detected; therefore, new data from the Veneto and Friuli Venezia Giulia regions were obtained and reinforced existing knowledge about resistance previously studied along the Adriatic coast. Nearby, in the Villa Verucchio locality, the highest (87.5%) AChE1R was found. CONCLUSION: Cx. pipiens resistance esterases A5/B5 and A4/B4 spread southward along the Adriatic coastal plain while OPs were being used in mosquito control, as confirmed by the first molecular screening of the AChE1 gene in these populations.

Molecular phylogeny of two lineages of Leuciscinae cyprinids (Telestes and Scardinius) from the peri-Mediterranean area based on cytochrome b data.

We examined phylogenetic relationships in two lineages of Leuciscinae cyprinid fishes based on the sequence data of the complete mitochondrial DNA region coding for the cytochrome b gene (1140 bp). Telestes includes obligate riverine, moderately cold water-adapted species whereas Scardinius comprises warm-adapted species living in lowland lakes and still waters of rivers and streams. We also analysed selected representatives of Leuciscus and Phoxinellus because the taxonomic status of some species belonging to these genera is dubious and they could be placed in the genus Telestes. The study includes 18 species, 43 populations, and 111 individuals from 9 of the 14 peri-Mediterranean ichthyogeographic districts. Clades recovered from the phylogenetic analyses do not support previous taxonomic assumptions based on morphology. Telestes, Leuciscus, and Phoxinellus do not form monophyletic assemblages; phylogenetic analyses suggest that L. polylepis, L. turskyi, P. croaticus, and P. metohiensis should be included in Telestes. Similarly, populations of Scardinius erythrophthalmus do not cluster together and the endangered S. scardafa, endemic to central Italy and surviving in a single locality, is nested within them. The radiations of Telestes and Scardinius occurred in different time periods. A major diversification of Telestes is consistent with a sea dispersal during the freshwater Messinian "Lago Mare" phase of the Mediterranean Sea. Cladogenetic events within Scardinius are likely related to the extension and confluence of river drainages in lowlands following multiple lowering of the sea level during the Quaternary glaciations.