Fernando Ayllon

Induction of micronuclei and other nuclear abnormalities in European minnow Phoxinus phoxinus and mollie Poecilia latipinna: an assessment of the fish micronucleus test.

In this work, we have measured both micronuclei and other nuclear abnormalities in renal erythrocytes from European minnow Phoxinus phoxinus and mollie Poecilia latipinna, with the aim to contribute to the standardisation of the micronucleus test for fish species. Intraperitoneal injections of colchicine (10 mg/kg), cyclophosphamide (40 mg/kg), or mitomycin C (20 mg/kg) for 24 h induced diverse nuclear abnormalities in minnow erythrocytes, therefore nuclear abnormalities should be added to micronuclei as genotoxicity indicators in fish micronucleus tests. The adequacy of administration protocols based on intraperitoneal injections has been evaluated by injecting saline solution to both species: single or double injections have not induced neither micronuclei nor other nuclear abnormalities in any case. Finally, the differential sensitivity of both species to toxic heavy metals was evaluated by exposing individuals of both species to different doses (0.17, 1.7, 2x1.7, and 3.4 mg/kg) of cadmium and mercury for 24 h; we concluded that the mollie is sensitive to both metals whereas the minnow is not sensitive to mercury.

MICRONUCLEUS TEST IN FRESHWATER FISH SPECIES: AN EVALUATION OF ITS SENSITIVITY FOR APPLICATION IN FIELD SURVEYS

Brown trout, Salmo trutta, European eel, Anguilla anguilla, and European minnow, Phoxinus phoxinus, three fish species inhabiting European freshwater ecosystems, were evaluated for their use as in situ pollution biomarkers using the micronucleus test in renal erythrocytes. Experimental exposure (by immersion) to different concentrations of cyclophosphamide, colchicine, and cadmium showed that brown trout are more sensitive to the three compounds than minnows and eels. In situ surveys of wild freshwater ecosystems with different levels of pollution showed that minnows and eels living in polluted sites do not present higher micronuclei averages than those caught in clean rivers systems, whereas micronuclei are induced in brown trout inhabiting polluted sites. Our results demonstrated the suitability of brown trout for in situ biomonitoring of freshwater ecosystems as well as for laboratory tests using the micronucleus test.

Assessing pollution in coastal ecosystems: a preliminary survey using the micronucleus test in the mussel Mytilus edulis

Mussels Mytilus edulis were sampled at increasing distances from urban effluents in two very different locations, Gijon (northern Spain, Europe, 43 degrees N) and Puerto Madryn (Argentina, South America, 43 degrees S), and from an industry effluent in Puerto Madryn. The micronucleus test was performed on branchial cells. For the three situations, a statistically significant negative association was found between the distance of sampling site from the effluent and the mean number of micronuclei per 1000 cell counts, in a range of distances as short as 300m. The micronucleus test in Mytilidae, here revealed to be sensitive enough to monitor urban pollution, is proposed for routine surveys of pollution as a bioindicator of choice for coastal ecosystems.

Interspecific hybridization between Atlantic salmon and brown trout introduced in the subantarctic Kerguelen Islands

In the subantarctic French territory of the Kerguelen Islands, Atlantic salmon Salmo salar was stocked to create a new population in the Korrigans drainage system. The analysis of historical scale collections showed that hybrids were inadvertently introduced with the original Atlantic salmon stocks. Later colonization of the Korrigans system by brown trout Salmo trutta was followed by interspecific hybridization between the two species. Our results highlight the importance of both routine genetic monitoring of hatchery stocks and considering the possible influence of other species when stocking is envisaged.

Introgression in the genus Salmo via allotriploids

Hybridization between sympatric species is not uncommon in the wild. Wild allotriploids (individuals with two chromosome sets from a species + one chromosome set from another species) are generally the result of a backcross between interspecific hybrids that produce unreduced gametes and one of the parental species. In animals, allotriploids are commonly sterile, except for some vertebrate species complexes in which allotriploids reproduce by parthenogenesis, gynogenesis and/or hybridogenesis, producing generally clonal or hemiclonal gametes; nuclear DNA introgression between hybridizing species is considered to be extremely rare. Employing species‐specific molecular markers, we show genetic introgression between the chromosomally well‐differentiated salmonids Atlantic salmon (2n = 58) and brown trout (2n = 80) through spontaneous bisexual reproduction of allotriploids leading to salmon‐like offspring bearing some brown trout genes. Although introgression between these Salmo species can occur via allotriploids, we hypothesize that extinction of parental species can be discarded based on very low survival of allotriploid offspring.

Dispersal and rapid evolution in brown trout colonizing virgin Subantarctic ecosystems

Two brown trout Salmo trutta stocks of different origin (wild Polish, domestic commercial) came into secondary contact after deliberate releases conducted in virgin rivers systems of the Subantarctic Kerguelen Islands (70° E 49° S). Samples obtained in 2001–2003 and a historical sample from 1993 were analysed for genetic variation at seven microsatellite loci and one allozyme locus (LDH‐C1*). Bayesian clustering analysis demonstrated that rapid genetic differentiation formed separate genetic units in neighbouring rivers in less than 20 years. These genetic units were characterized by a large proportion of Polish genotypes mixed with some genomes of domestic origin (up to 30%). A different colonization strategy of the naturalized stocks, likely related with differential performance, was identified as a cause of rapid population differentiation in this area.

Conventional armament wastes induce micronuclei in wild brown trout Salmo trutta

We analysed micronuclei in brown trout Salmo trutta specimens sampled in the Trubia River, upstream and downstream of the emissions from a Spanish military factory to assess genotoxicity risks derived from military wastes. A significant exponential increase in micronuclei counts was found in fish living downstream of the military wastes with respect to fishes inhabiting upstream areas of the same river. In comparison, we only found a linear increase in micronuclei counts in a control stream where an old military factory had been demolished 6 months before sampling. This difference suggests that active discharge of armament factory wastes can directly induce micronuclei and therefore represents a genotoxic risk for the ecosystem.