This study assessed the effectiveness of a novel conservation strategy for the native brown trout populations in Sardinia (Italy). The conservation measures included: (i) installation of an electric fish barrier; (ii) removal of all the resident introgressed brown trout upstream from the barrier; (iii) restocking of the upstream region with native Sardinian trout genetically characterised; and (iv) estimation of native trout population size with prediction of future growth. Introgressed brown trout were removed using multiple-pass electrofishing methods in a 2-km region upstream from the barrier. In two eradication campaigns, conducted in 2010 and 2012, a total of 1801 trout were captured, comprising a total biomass of 55,501 g. The capture efficiency analysis showed total capture rates of over 99% were achieved for parr (0+ age cohort), and 100% for adult trout. After the eradication campaigns we restocked this area with native brown trout (Adriatic lineage, AD, according to the mitochondrial DNA control region nomenclature). The monitoring campaigns showed good survival (S = 60%) and reproduction of 3.1 recruits/adult trout. Successful breeding was recorded starting from the summer of 2015 and a self-sustaining population resulted after only two restocking events. The results of this restoration project allow recommendations to be made for future conservation eradication and repopulation projects for restoration of typical Mediterranean streams.

Skeletal anomalies are commonplace among farmed fish. The pug-headedness anomaly is an osteological condition that results in the deformation of the maxilla, pre-maxilla, and infraorbital bones. Here, we report the first record of pug-headedness in an isolated population of the critically endangered native Mediterranean trout Salmo trutta L., 1758 complex from Sardinia, Italy. Fin clips were collected for the molecular analyses (D-loop, LDH-C1* locus. and 11 microsatellites). A jaw index (JI) was used to classify jaw deformities. Ratios between the values of morphometric measurements of the head and body length were calculated and plotted against values of body length to identify the ratios that best discriminated between malformed and normal trout. Haplotypes belonging to the AD lineage and the genotype LDH-C1*100/100 were observed in all samples, suggesting high genetic integrity of the population. The analysis of 11 microsatellites revealed that observed heterozygosity was similar to the expected one, suggesting the absence of inbreeding or outbreeding depression. The frequency of occurrence of pug-headedness was 12.5% (two out of 16). One specimen had a strongly blunted forehead and an abnormally short upper jaw, while another had a slightly anomaly asymmetrical jaw. Although sample size was limited, variation in environmental factors during larval development seemed to be the most likely factors to trigger the deformities.

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