Andrea Sabatini

Endangered trout populations can be monitored with a variety of methods, the selection of which should consider social constraints and environmental variables known to affect method effectiveness. Here, we confront the effectiveness of four monitoring methods (removal with electrofishing, ELE; underwater camera survey, UCS; streamside visual survey, SVS; visual surveys with angling, VSA) to estimate the relative abundance of three populations of the endangered Mediterranean brown trout. The trout counts obtained via different methods were well correlated (r = 0.65–0.72), providing a coherent description of the relative pool abundance across the methods. However, the methods were differently affected by environmental variables, depending on the age classes of trout. Specifically, the adult and subadult counts provided by ELE and VSA were negatively and positively affected by the maximum pool depth, respectively; adult and subadult counts of VSA and the SVS were positively affected by pool area; the juvenile counts provided by the UCS were positively affected by pool shade and negatively affected by water turbidity; juvenile counts provided by VSA were positively affected by shade. Variables such as pool depth, area, shading, water turbidity and proportion of age classes can be hardly controlled in monitoring programs, and their bias could be modelled. Different sampling methods provided similar information about relative abundance and appeared equivalent. While ELE could be selected to collect samples and biometric data, monitoring relative abundance with the UCS, VSA, and SVS appears more suited and can also involve citizen scientists.

An otolith shape and morphometric analysis was performed on European eel (Anguilla anguilla) subpopulations from five rivers and three coastal lagoons of Sardinia (central-western Mediterranean) to assess the role of different habitats on otolith development. Sagittal otolith shape was described by 11 harmonics from elliptic Fourier descriptors. Comparisons among the harmonics were run through canonical discriminant analyses (CDAs). The CDA reclassification rate (75.7%) demonstrated a spatial environmental discrimination among local eel subpopulations of Sardinia. The Euclidean distance values demonstrated a dissimilarity between the river and lagoon groups. The form factor and roundness shape indices were significantly higher in the river group than in the lagoon group. The distances of the first three rings to the otolith core revealed site-specific otolith development. Moreover, the annual otolith growth rate was faster in the lagoon group than in the river group. The differences among the studied sites in terms of sagittal otolith shape could relate to changes in different local stocks potentially related to environmental peculiarities. Establishing a direct correlation between otolith morphology and environmental factors is challenging, and further studies are needed to investigate the relationship between habitat type/environmental variation and growth/body characteristics of eels. Nevertheless, the achieved results suggest that this method can be considered to be a valuable tool for studying the ontogeny of the European eel.

Monitoring the conservation status of endangered freshwater fish using less invasive methods poses challenges for ecologists and conservationists. Visual surveys have been proposed as an alternative to electrofishing, which is a standard methodology that can cause injuries, physiological stress and post-release mortality in organisms. To test the efficacy of visual methods, a study was conducted in an intermittent stream of Sardinia (Italy). Two visual methods were employed: a visual survey from streambanks (VSS) and an underwater visual survey (UVS) using cameras. The aims of this study were (1) to compare the effectiveness of these methods in detecting patch occupancy patterns and (2) to investigate the effect of environmental variables on the detection probability of Mediterranean native trout. Environmental variables characterizing pool habitats were recorded, and generalized linear models (GLMs) were employed to assess the correlation between these variables and trout presence/absence. GLM analysis revealed that UVS had higher detection probability with larger pool volume, whereas submerged macrophytes negatively affected detection probability. Detection from streambanks (VVS) was negatively affected by a high turbulence rate. In conclusion, our study suggests the utility of visual methods to describe patterns of patch occupancy of Mediterranean trout. However, methods can be differently affected by environmental variables. Therefore, monitoring programs using these methods should consider these factors to ensure a reliable description of within-stream trout distribution in intermittent streams.

Olfaction could represent a pivotal process involved in fish orientation and migration. The olfactory bulb can manage olfactive signals at the granular cell (GC) and dendritic spine levels for their synaptic plasticity properties and changing their morphology and structural stability after environmental odour cues. The GCs’ dendritic spine density and morphology were analysed across the life stages of the catadromous Anguilla anguilla. According to the head and neck morphology, spines were classified as mushroom (M), long thin (LT), stubby (S), and filopodia (F). Total spines’ density decreased from juvenile migrants to no-migrant stages, to increase again in the adult migrant stage. Mean spines’ density was comparable between glass and silver eels as an adaptation to migration. At non-migrating phases, spines’ density decreased for M and LT, while M, LT, and S density increased in silver eels. A great dendritic spine development was found in the two migratory phases, regressing in trophic phases, but that could be recreated in adults, tracing the migratory memory of the routes travelled in juvenile phases. For its phylogenetic Elopomorph attribution and its complex life cycle, A. anguilla could be recommended as a model species to study the development of dendritic spines in GCs of the olfactory bulb as an index of synaptic plasticity involved in the modulation of olfactory stimuli. If olfaction is involved in the orientation and migration of A. anguilla and if eels possess a memory, these processes could be influenced by the modification of environmental stimuli (ocean alterations and rapid climate change) contributing to threatening this critically endangered species.

This paper reports heavy metal and metalloid accumulation in wild brown trout (Salmo trutta L., 1758 complex) raised in freshwater and uncontaminated Sardinia system (Italy). Metals are widespread pollutants of aquatic systems, and their contamination can originate from anthropogenic activities such as industrial waste, agricultural and domestic environments, and geochemical release. Fish has a relevant position within the human diet; moreover, fishes can accumulate metals, making them a valuable tool as biomarkers for risk assessment studies. The concentration of 22 metals and metalloids after chemical digestion was assessed by inductively coupled plasma-optic emission spectroscopy (ICP-OES) in both the guts and the edible part (EP, muscle + skin) of brown trout. The results, expressed as μg g⁻¹, showed different levels of accumulation in the EP and guts, following the series Cu > Zn > Ba > Al > Sr > Fe > Pb and Fe > Al > Hg > As > Mn > Cu > Ba > B > Zn > Pb, respectively. PCA analysis showed a fairly good correlation between the total lipid and SAFA content and Cd, Hg, and Pb accumulation in the gut. Non-carcinogenic risk assessment, expressed as THQ (target hazard quotient), showed values far below 1 for all metals in muscles, while high As and Hg contamination of the gut draws attention to possible health risks which should be discarded from the fish before consumption. TR (target cancer risk) values showed alarmingly high values for As and Cd when the fish were consumed entirely (gut + EP), while Pb levels were far below the safety levels.

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.

Both morphological and molecular data are presented and discussed for indigenous Salmo sp. from Corsica and Sardinia, here called Tyrrhenian trout. For comparison, morphological data obtained from museum specimens, including the Algerian S. macrostigma, are discussed in the light of recent and new molecular findings. In total, 29 measurements and 20 meristic characters were taken from each specimen. Out of the meristic characters, 12 were obtained by means of X-ray. One important morphometric character in the present study is the size of the head measured from premaxilla to posterior margin of preoperculum. This character was particularly stable in all Tyrrhenian trout, showing relatively large head compared to Atlantic trout and to S. macrostigma. On the contrary, other characters like body punctuations, black and white edges of fins, body depth or number of epurals in the caudal skeleton are quite polymorphic. In certain meristic characters, range of variation of Tyrrhenian trout even exceeds that of the extensive comparative material. Each trout has been genetically characterized. New haplotypes from Tyrrhenian trout were discovered, belonging to three mitochondrial lineages viz. Adriatic, marble and Mediterranean, however, Adriatic haplotypes are dominant. Comparing morphological and genetic data, observed morphology lacks any obvious correlation to mitochondrial lineages and it is concluded that Tyrrhenian trout show no particular affinity to S. macrostigma from Algeria.

The systematic status of the Italian trout in the Salmo trutta L., 1758 complex (including marble, Mediterranean and lacustrine trout), has long been - and is still today – subject of controversies among ichthyologists. The specific name and the taxonomic rank changed several times in the last years, and the natural occurrence of this salmonid fish in some Italian areas was debated due to spread of alien strains. The main difficulty with the taxonomy of the Italian trout stems from the impossibility of disentangling it “on paper” or, even worse, trying to face this systematic issue considering only a very limited (local/national scale) part of the brown trout range. The taxonomy of the Italian trout population is inextricably linked to the necessity of clarifying first phylogeny and phylogeography in an overall Mediterranean context. The opportunity of a non “self-referential” taxonomy is even more fundamental for a vulnerable salmonid like the Italian brown trout, for which there is a very conflicting management problem related to sport fishing and, at the same time, the urgent need for effective conservation measures. It is however necessary to emphasize that conservation is independent from taxonomy but must start from the level of the local population. In fact, management units need stability and they cannot, therefore, coincide with entities – the Linnean species – requiring continuous taxonomic revisions. Modern molecular methods are the best tools for defining these units of management and conservation in an evolutionary perspective.

Invasive alien species (IAS) are considered one of the major threats to biodiversity worldwide, thus requiring severe control strategies. Based on the promising results obtained in the field of insect pest management with polyvinyl chloride (PVC) for long-lasting release of attractants, the main aim of this study was to ascertain the efficacy of PVC/attractant dispensers also in the aquatic environment. Therefore, we developed PVC/food dispensers and evaluated their attractiveness, by means of behavioural bioassays on whole animals, over a 60-day period of continuous use towards the red swamp crayfish Procambarus clarkii, one of the top 100 worst IAS. The attractiveness in PVC of trehalose, leucine and taurocholic acid was also tested. Our results show that the PVC dispensers release the food and are strongly attractive for crayfish over a prolonged time, even if their effectiveness depends on the storage conditions: From 18 days when stored underwater at 23 °C up to 50 days when stored out of the water at −20 °C. Besides, trehalose, leucine and taurocholic acid in PVC resulted in reliable attractants for P. clarkii. The development of PVC dispensers for long-lasting release of attractants may help improve the efficiency of mass trapping strategies in the management and control of invasive crayfish.

The common dentex, Dentex dentex, is a fish species which inhabits marine environments in the Mediterranean and Northeast Atlantic regions. This is an important species from an ecological, economic and conservation perspective, however critical information on its population genetic structure is lacking. Most samples were obtained from the Mediterranean Sea (17 sites) with an emphasis around Corsica (5 sites), plus one Atlantic Ocean site. This provided an opportunity to examine genetic structuring at local and broader scales to provide science based data for the management of fishing stocks in the region. Two mitochondrial regions were examined (D-loop and COI) along with eight microsatellite loci. The COI data was combined with publicly available sequences and demonstrated past misidentification of common dentex. All markers indicated the absence of population genetic structure from the Bay of Biscay to the eastern Mediterranean Sea. Bayesian approaches, as well as the statistical tests performed on the allelic frequencies from microsatellite loci, indicated low differentiation between samples; there was only a slight (p = 0.05) indication of isolation by distance. Common dentex is a marine fish species with a unique panmictic population in the Mediterranean and likely in the Atlantic Ocean as well.