Molecular detection of the swim bladder parasite Anguillicola crassus (Nematoda) in fecal samples of the endangered European eel Anguilla anguilla.

The European eel Anguilla anguilla is listed as critically endangered by the IUCN. Among many threats, the introduced parasitic nematode Anguillicola crassus is suspected to alter the eels' swim bladder and jeopardize their reproductive oceanic migration. To date, gaining knowledge about the distribution and prevalence of A. crassus requires individual sacrifice (over 50,000 eels were sacrificed for epidemiology studies since 2010). This paper describes a non-lethal molecular protocol for identifying prevalence of A. crassus in A. anguilla, based on searching for A. crassus DNA in the feces of eels. Tests using three DNA microsatellite markers specific to the nematode showed that molecular detection provided similar results to visual examination of the swim bladder in up to 80% of the cases, and allowed for comparison of prevalence among sites. Easy to implement, this non-lethal protocol for detecting A. crassus could be valuable for management plans of this endangered species.

Tolerance of young allis shad Alosa alosa (Clupeidae) to oxy-thermic stress.

The ecology of the young stages of allis shad Alosa alosa is poorly documented, although they can be exposed to many pressures during their freshwater phase and their downstream migration. When passing through systems such as the Gironde-Garonne-Dordogne watershed (GGD, SW France), they can be subjected to high temperatures and low levels of oxygen (hypoxia). The aim of this work is to assess the tolerance of young Alosa alosa at four ages (c. 10, 30, 60 and 85 days old) by challenging them to different temperatures (18, 22, 26 and 28°C) together with decreasing oxygen saturation levels (from 100% to 30%). Survival of the 10-day-old individuals was not influenced by oxy-thermic conditions, but high stress levels were detected and perhaps this age class was too fragile regarding the constraint of the experimental design. Survival at 30 and at 60 days old was negatively influenced by the highest temperatures tested alone (from 26°C and from 28°C, respectively) but no effect was detected at 85 days old up to 28°C. A combined effect of temperature and oxygen level was highlighted, with heat accelerating survival decrease when associated with oxygen level depletion: essentially, survival was critical (<50%) at 30 days old at temperature ≥22°C together with 30% O2 ; at 60 days old, at temperature = 28°C with 30% O2 ; at 85 days old, at temperature ≥26°C with ≤40% O2 . Tolerance to oxy-thermic pressures appeared to be greater among the migratory ages (60 and 85 days old) than among the 30-day-old group. Based on environmental data recorded in the GGD system and on our experimental results, an exploratory analysis allowed a discussion of the possible impact of past oxy-thermic conditions on the local population dynamics between 2005 and 2018. The oxy-thermic conditions that may affect Alosa alosa at ages when they migrate downstream (60 and 85 days old) were not frequently recorded in this period, except in cases of extreme episodes of heat together with hypoxia that occurred in some years, in summertime in the turbidity maximum zone of the Gironde estuary (particularly in the year 2006). Interestingly, oxy-thermic conditions that are likely to threaten the 30-day-old individuals occurred more frequently in the lower freshwater parts of the GGD system between the years 2005 and 2018. In the context of climate change, a general increase in temperature is predicted, as well as more frequent and severe hypoxic events, therefore we suggest that local Alosa alosa population recruitment could encounter critical oxy-thermic conditions more frequently in the future if no adaptive management of water resources occurs.

New insights regarding gonad development in European eel: evidence for a direct ovarian differentiation.

In European eel, it has been proposed that the undifferentiated gonad would develop into either an intersexual stage (Syrski organ) or directly into an ovary. The Syrski organ could then develop into either an ovary or a testis. In the present study, glass eels were raised until they reached a minimum size of 29 cm for histological sex assessment. In addition, some undifferentiated individuals with size encompassing 15-28 cm were sampled in a female-biased population (Oir River). We also investigated aromatase gene expression, which is known to be involved in the process of fish sex differentiation. Gonad histology revealed that intersexual eels were characterized by a small number of oocytes within a predominant testis-like structure. Males were significantly smaller than intersexual eels, which suggests that all males do not necessarily pass through an intermediate intersexual stage. Aromatase transcript levels in intersexual eels gonads and testes were similar but significantly lower than in ovaries and were comparable between ovaries and undifferentiated gonads from the females-biased population. In addition, condition factor was lower in female than in intersexual individuals. Together, these results provide evidence that ovaries would not develop from the Syrski organ.

A complete check-up of European eel after eight years of restocking in an upland river: Trends in growth, lipid content, sex ratio and health status.

By combining field research and careful laboratory analysis of samples over the course of an eight-year study, we met the challenge of assessing the life history traits and health status of eels restocked in freshwater ecosystems. We found that restocked eels exhibited good growth performance; moreover, the stocks were female-dominated, showed a good Fulton's condition factor (K) and lipid stores and had high survival probability estimated using the best model of Jolly-Seber stock assessment method for open populations. A necropsy revealed the absence of internal lesions. A swim bladder examination revealed the absence of the parasite Anguillicola crassus. Polymerase chain reaction (PCR) analyses revealed an increase of Anguillid herpesvirus-1 (AngHV-1) prevalence throughout the study. Most positive subjects expressed viral loads compatible with a latent infection and correlated positively with K. All restocked eels were contaminated by at least one of the organic pollutant congeners studied, but the pollution loads corresponded to the lowest range of pollutant concentrations reported in the available literature for European eels and did not exceed the maximum residue and contaminant limits in food and feed of several national and international regulations. Pollutant loads were negatively correlated with K, lipid content and eel density for polychlorinated biphenyls PCB 138, 153 and 180 and K for pesticides p.p'-DDE, p.p'-DDD, p.p'-DDT and PBDE47. This study highlights the potential role played by upland aquatic ecosystems in enhancing riverine silver eel production from the perspective of species conservation. To be successful, restocking must be accompanied by improved ecosystem quality and migration routes for eels in inland freshwaters. We also provide some recommendations for future research to improve the management of restocking programmes.

Video tracking in the extreme: a new possibility for tracking nocturnal underwater transparent animals with fluorescent elastomer tags.

Initially developed so that an individual could be recognized in mark-recapture studies of aquatic animals, fluorescent visible implant elastomer (VIE) tags are used here for a new application in ethometry: the study of the behavior of transparent animals in dim light or in darkness using automatic tracking technology. The application and validation of this multitracking method is tested in the context of research on the estuarine migratory behavior of the glass eel (Anguilla anguilla), a crucial point to better understand the dynamics of this endangered species. The method makes it possible to measure the activity (notably the distance and speed) of four individuals as a function of tidal and nycthemeral rhythms in the same flume (a circular aquarium simulating river or estuarine conditions) across a wide time scale (from seconds to weeks).

Genetic architecture of threshold reaction norms for male alternative reproductive tactics in Atlantic salmon (Salmo salar L.).

Alternative mating tactics have important ecological and evolutionary implications and are determined by complex interactions between environmental and genetic factors. Here, we study the genetic effect and architecture of the variability in reproductive tactics among Atlantic salmon males which can either mature sexually early in life in freshwater or more commonly only after completing a migration at sea. We applied the latent environmental threshold model (LETM), which provides a conceptual framework linking individual status to a threshold controlling the decision to develop alternative traits, in an innovative experimental design using a semi-natural river which allowed for ecologically relevant phenotypic expression. Early male parr maturation rates varied greatly across families (10 to 93%) which translated into 90% [64-100%] of the phenotypic variation explained by genetic variation. Three significant QTLs were found for the maturation status, however only one collocated with a highly significant QTL explaining 20.6% of the variability of the maturation threshold located on chromosome 25 and encompassing a locus previously shown to be linked to sea age at maturity in anadromous Atlantic salmon. These results provide new empirical illustration of the relevance of the LETM for a better understanding of alternative mating tactics evolution in natural populations.

Kleptoparasitism and aggressiveness are influenced by standard metabolic rate in eels.

Kleptoparasitism refers to either interspecific or intraspecific stealing of food already procured by other species or individuals. Within a given species, individuals might differ in their propensity to use such a tactic, in a similar manner to which they differ in their general level of aggressiveness. Standard metabolic rate is often viewed as a proxy for energy requirements. For this reason, it should directly impact on both kleptoparasitism and aggressiveness when individuals have to share the same food source. In the present study we first assessed the standard metabolic rate (SMR) of 128 juvenile European eels (Anguilla anguilla) by the determination of oxygen consumption. We then tested how the SMR could influence agonistic behavior of individuals competing for food in three distinct trials evenly distributed over three months. We demonstrate that SMR positively correlates with attacks (sum of bite and push events) in all trials. Similarly SMR correlated positively with kleptoparasitism (food theft), but this was significant only for the third trial (month 3). To our knowledge, the present study is the first reporting a link between kleptoparasitism and SMR in a fish species. This has ecological implications owing to the fact that this species is characterized by an environmental sex determination linked to early growth rate. We discuss theses findings in the light of the producer-scrounger foraging game.

Sexually dimorphic gene expressions in eels: useful markers for early sex assessment in a conservation context.

Environmental sex determination (ESD) has been detected in a range of vertebrate reptile and fish species. Eels are characterized by an ESD that occurs relatively late, since sex cannot be histologically determined before individuals reach 28 cm. Because several eel species are at risk of extinction, assessing sex at the earliest stage is a crucial management issue. Based on preliminary results of RNA sequencing, we targeted genes susceptible to be differentially expressed between ovaries and testis at different stages of development. Using qPCR, we detected testis-specific expressions of dmrt1, amh, gsdf and pre-miR202 and ovary-specific expressions were obtained for zar1, zp3 and foxn5. We showed that gene expressions in the gonad of intersexual eels were quite similar to those of males, supporting the idea that intersexual eels represent a transitional stage towards testicular differentiation. To assess whether these genes would be effective early molecular markers, we sampled juvenile eels in two locations with highly skewed sex ratios. The combined expression of six of these genes allowed the discrimination of groups according to their potential future sex and thus this appears to be a useful tool to estimate sex ratios of undifferentiated juvenile eels.

Spatial and seasonal variations of methylmercury in European glass eels (Anguilla anguilla) in the Adour estuary (France) and relation to their migratory behaviour.

Overall recruitment of European glass eels (Anguilla anguilla) has decreased significantly since the early 1980s. Due to their long life cycle, benthic/demersal habits and high lipid content, eels might accumulate high concentrations of contaminants, but data concerning glass eels are still scarce. This study provides original data on methylmercury (MeHg) concentrations in glass eels at spatial (marine and estuarine), annual and seasonal scales. The relationship between MeHg concentrations in glass eels and their propensity to migrate up estuaries was also investigated. MeHg data were individually related to the eels' energetic condition which was estimated by dry weight. Glass eel migratory behaviour was investigated in an experimental flume and related to the MeHg concentration and dry weight at the individual scale. Marine and estuarine glass eels were caught from 2004 to 2011. There was a strong inverse correlation between MeHg concentrations and dry weight. MeHg concentrations increased in marine and estuarine glass eels from 2004 to 2009 and from 2004 to 2010, respectively, and then, both groups decreased in 2011. On a seasonal time scale, MeHg concentrations were higher at the end of the fishing season (April). MeHg bioaccumulation is likely to result from different sources, but the lack of significant differences between marine and estuarine glass eels suggests that direct contamination during estuarine migration is low. Other sources such as maternal transfer or oceanic contamination are discussed.

Endocrine control of Anguilla anguilla glass eel dispersal: effect of thyroid hormones on locomotor activity and rheotactic behavior.

Dispersal, one of the most important processes in population ecology, is an issue linking physiological and behavioral features. However, the endocrine control of animal dispersal remains poorly understood. Here, we tested whether and how thyroid hormones may influence dispersal in glass eels of Anguilla anguilla, by testing their influence on locomotor activity and rheotactic behavior. Glass eels were caught during their estuarine migration and treated by immersion in either a l-thyroxine (T(4)) or a thiourea (TU) solution. As measured by radioimmunoassay, T(4) and TU treatments induced, respectively, increased and decreased whole-body thyroid hormone levels relative to untreated controls. We tested a total of 960 glass eels distributed into control, and T(4) and TU treatment groups, on their swimming behavior in experimental flume tanks equipped with upstream and downstream traps that allowed us to concurrently measure both the locomotor activity and the rheotactic behavior. Compared to controls, locomotor activity significantly increased among the hyperthyroid, T(4)-treated eels, but significantly decreased among the hypothyroid, TU-treated eels. The results on rheotactic behavior suggested a more complex regulatory mechanism, since TU but not T(4) treatment significantly affected rheotactic behavior. The influence of thyroid hormones on locomotor activity suggests a central role for these hormones in the regulation of mechanisms leading to the colonization of continental habitats by glass eels. Thyroid hormones are also implicated in the control of locomotor activity in mammals and migratory behavior in birds, suggesting that these hormones represent conserved, proximate mediators of dispersal in vertebrates.