Parentage assignment of progeny in mixed milt fertilization of Caspian brown trout Salmo trutta caspius using microsatellite DNA markers: Implications for conservation

Parentage of a stock of mixed milt produced progeny in current artificial breeding protocol of endangered Caspian brown trout, Salmo trutta caspius, was determined using three microsatellite loci chosen after a primary analysis of genetic diversity at nine microsatellite loci in the eight used breeder individuals. Overall, 98.8% of progeny were assigned to their parents using Family Assignment Program (FAP). Selection of hyper-variable microsatellites in Caspian brown trout to identify unique alleles was effective for unambiguous parentage determination and estimation of genetic diversity in this study. Effective population size of breeder individuals (Ne) was lower than the number of breeder individuals used (Nb) indicating unbalanced contribution of breeder individuals to progeny. Indeed, one of the four male breeder individuals produced about 70 % and the other three produced only from 4.86 % to 18.83 % of progeny. The average observed and expected heterozygosity of progeny (0.723 ± 0.011 and 0.684 ± 0.009, respectively) was significantly lower than that of their parents (0.833 and 0.800, respectively). Our data indicate that the current breeding protocol of Caspian brown trout may not provide equal opportunity for all the breeder individuals to contribute equally to progeny. Therefore, appropriate fertilization designs in the hatchery should be established in order to equalize the genetic contribution of different breeder individuals.Key words: Parentage assignment, effective population size, genetic diversity, Salmo trutta caspius

Island survivors: population genetic structure and demography of the critically endangered giant lizard of La Gomera, Gallotia bravoana

Background: The giant lizard of La Gomera (Gallotia bravoana), is an endemic lacertid of this Canary Island that lives confined to a very restricted area of occupancy in a steep cliff, and is catalogued as Critically Endangered by IUCN. We present the first population genetic analysis of the wild population as well as of captive-born individuals (for which paternity data are available) from a recovery center. Current genetic variability, and inferred past demographic changes were determined in order to discern the relative contribution of natural versus human-mediated effects on the observed decline in population size. Results: Genetic analyses indicate that the only known natural population of the species shows low genetic diversity and acts as a single evolutionary unit. Demographic analyses inferred a prolonged decline of the species for at least 230 generations. Depending on the assumed generation time, the onset of the decline was dated between 1200-13000 years ago. Pedigree analyses of captive individuals suggest that reproductive behavior of the giant lizard of La Gomera may include polyandry, multiple paternity and female long-term sperm retention. Conclusions: The current low genetic diversity of G. bravoana is the result of a long-term gradual decline. Because generation time is unknown in this lizard and estimates had large credibility intervals, it is not possible to determine the relative contribution of humans in the collapse of the population. Shorter generation times would favor a stronger influence of human pressure whereas longer generation times would favor a climate-induced origin of the decline. In any case, our analyses show that the wild population has survived for a long period of time with low levels of genetic diversity and a small effective population size. Reproductive behavior may have acted as an important inbreeding avoidance mechanism allowing the species to elude extinction. Overall, our results suggest that the species retains its adaptive potential and could restore its ancient genetic diversity under favorable conditions. Therefore, management of the giant lizard of La Gomera should concentrate efforts on enhancing population growth rates through captive breeding of the species as well as on restoring the carrying capacity of its natural habitat.Spanish Ministry of Education; European Life Project [LIFE 02 NAT-E-008614]; Ministerio de Ciencia e Innovacion [REN 2001- 1514/GLO, CGL 2010-18216]info:eu-repo/semantics/publishedVersio

Heritability of skeleton abnormalities (lordosis, lack of operculum) in gilthead seabream (Sparus aurata) supported by microsatellite family data

Body abnormalities constitute an important problem to fish aquaculture. Pigmentation, scale and skeleton abnormalities have been reported in different cultured fish species. Environmental and genetic factors or their interaction have been suggested to explain their origin. Gilthead seabream (Sparus aurata) is one of the most important European cultured species. Lordosis and lack of operculum are common abnormalities in this species, and can reach frequency of 80% or more of commercial batches. In spite of an important body of data suggesting environmental factors related with their origin, scarce and not conclusive genetic data have been reported to date. In our study, a large number of families originating from the usual production process of a commercial farm were used to estimate the heritabilities of both characters. Two independent experiments were carried out for each abnormality. Nine hundred and ninety four (5.6% lordotics; 157 full-sib families; 6.3 offspring/family) and 808 (7.9% lacking operculum; 83 families; 9.7 offspring/family) individuals were used for lordosis and lacking operculum heritability estimations, respectively. The results obtained adjusting an animal model indicated non significant heritabilities for both characters (0.021 (s.e. 0.019) and 0.032 (s.e. 0.023) for lordosis and lack of operculum, respectively). The corresponding values when a threshold model was used, though higher (0.152 and 0.203, respectively), evidenced large standard errors (0.119 and 0.146, respectively) suggesting h2 = 0 as the most confident hypothesis (P = 0.838 and 0.766, respectively). A non-parametric permutation test was also applied to evaluate if more related individuals had a higher phenotypic resemblance. The results obtained suggested only a slight familiar association (P < 0.05) when comparing individuals lacking operculum, but neither between lordotics nor between normal ones. These results suggest that most phenotypic variation observed for lordosis and lack of operculum in gilthead seabream is due to environmental factors. © 2008 Elsevier B.V. All rights reserved

Development and validation of molecular tool for assesing triploidy in turbot (Scophthalmus maximus)

Production of triploid individuals is a relevant goal for the aquaculture industry due to the benefits associated with their sterility and growth. Thus, methods for assessing triploidy have been developed based on genome, chromosome or gene triploid-associated properties. In this study, we developed a new cheap, technically simple and accurate method to validate triploidy in turbot (Scophthalmus maximus) based on microsatellite markers. Five crosses were performed to produce diploid and triploid progenies that were used to validate this molecular tool. Flow cytometry, one of the most widely used and accurate techniques for ploidy determination, was used as reference to contrast results. A set of four highly polymorphic and largely distant to centromere microsatellites was selected for this purpose. Ploidy was easily evaluated according to the maximum number of alleles at the microsatellite loci tested, diploids showing two and triploids three. These microsatellites were combined in a single multiplex and were able to identify triploids with 100% accuracy in all analyzed crosses.Publicado

Permanent genetic resources added to Molecular Ecology Resources Database 1 December 2010-31 January 2011.

This article documents the addition of 238 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Alytes dickhilleni, Arapaima gigas, Austropotamobius italicus, Blumeria graminis f. sp. tritici, Cobitis lutheri, Dendroctonus ponderosae, Glossina morsitans morsitans, Haplophilus subterraneus, Kirengeshoma palmata, Lysimachia japonica, Macrolophus pygmaeus, Microtus cabrerae, Mytilus galloprovincialis, Pallisentis (Neosentis) celatus, Pulmonaria officinalis, Salminus franciscanus, Thais chocolata and Zootoca vivipara. These loci were cross-tested on the following species: Acanthina monodon, Alytes cisternasii, Alytes maurus, Alytes muletensis, Alytes obstetricans almogavarii, Alytes obstetricans boscai, Alytes obstetricans obstetricans, Alytes obstetricans pertinax, Cambarellus montezumae, Cambarellus zempoalensis, Chorus giganteus, Cobitis tetralineata, Glossina fuscipes fuscipes, Glossina pallidipes, Lysimachia japonica var. japonica, Lysimachia japonica var. minutissima, Orconectes virilis, Pacifastacus leniusculus, Procambarus clarkii, Salminus brasiliensis and Salminus hilarii