Chromosome aberrations in pressure-induced triploid Atlantic salmon.

BACKGROUND: Triploid organisms have three sets of chromosomes. In Atlantic salmon, hydrostatic pressure treatment of newly fertilized eggs has been extensively used to produce triploids which are functionally sterile due to their unpaired chromosomes. These fish often perform poorly on commercial farms, sometimes without explanation. Inheritance patterns in individuals subjected to pressure treatment have not been investigated in Atlantic salmon thus far. However, work on other species suggests that this treatment can result in aberrant inheritance. We therefore studied this in Atlantic salmon by genotyping 16 polymorphic microsatellites in eyed eggs and juveniles which had been subjected to pressure-induction of triploidy. Communally reared juveniles including fish subjected to pressure-induction of triploidy and their diploid siblings were included as a control. RESULTS: No diploid offspring were detected in any of the eggs or juveniles which were subjected to hydrostatic pressure; therefore, the induction of triploidy was highly successful. Aberrant inheritance was nevertheless observed in 0.9% of the eggs and 0.9% of the juveniles that had been subjected to pressure treatment. In the communally reared fish, 0.3% of the fish subjected to pressure treatment displayed aberrant inheritance, while their diploid controls displayed 0% aberrant inheritance. Inheritance errors included two eyed eggs lacking maternal DNA across all microsatellites, and, examples in both eggs and juveniles of either the maternal or paternal allele lacking in one of the microsatellites. All individuals displaying chromosome aberrations were otherwise triploid. CONCLUSIONS: This is the first study to document aberrant inheritance in Atlantic salmon that have been subjected to pressure-induction of triploidy. Our experiments unequivocally demonstrate that even when induction of triploidy is highly successful, this treatment can cause chromosome aberrations in this species. Based upon our novel data, and earlier studies in other organisms, we hypothesize that in batches of Atlantic salmon where low to modest triploid induction rates have been reported, aberrant inheritance is likely to be higher than the rates observed here. Therefore, we tentatively suggest that this could contribute to the unexplained poor performance of triploid salmon that is occasionally reported in commercial aquaculture. These hypotheses require further investigation.

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 degrees E 49 degrees 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.

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.

Micronuclei and other nuclear lesions as genotoxicity indicators in rainbow trout Oncorhynchus mykiss.

The induction of micronuclei and other nuclear abnormalities in renal erythrocytes of rainbow trout Oncorhynchus mykiss by six genotoxic compounds is evaluated. Colchicine, mitomycin, cyclophosphamide, acrylamide, methyl-methanesulfonate, and N-ethyl-N-nitrosourea were intraperitoneally injected in trout. Our results show that cyclophosphamide induces the formation of micronuclei and also the other nuclear abnormalities; N-ethyl-N-nitrosourea, acrylamide, and colchicine induce only micronuclei; mitomycin-C induces only other nuclear abnormalities but not micronuclei. Methyl-methanesulfonate does not induce nuclear abnormalities in rainbow trout at the dose assayed in this work. The possible genotoxic origin for the different nuclear abnormalities is discussed.

Induction of micronuclei in eel (Anguilla anguilla L.) by heavy metals.

Micronucleus test was performed in situ on eels (Anguilla anguilla) from river sites with different levels of heavy metal pollution (cadmium and mercury). Cadmium content in eel liver but not micronuclei averages in kidney were associated with cadmium content in sediments. Mercury content in liver was not significantly associated with mercury content in sediments. Both cadmium and mercury induced micronuclei expression in eels when injected, the concentration tested being 1.7 mg metal/kg body weight and the micronuclei induction being 2.64 and 2.35 micronuclei per 1000 cells for cadmium and mercury respectively. It was concluded that these heavy metals are genotoxic for European eel, that eel liver metal content is a sensitive indicator for environmental monitoring of cadmium pollution, and that the micronuclei scores in eels are not a sensitive method to detect heavy metals pollution in freshwater ecosytems.

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.

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.