Growth potential can affect timing of maturity in a long-lived semelparous fish.

Many diadromous fishes such as salmon and eels that move between freshwater and the ocean have evolved semelparous reproductive strategies, but both groups display considerable plasticity in characteristics. Factors such as population density and growth, predation risk or reproduction cost have been found to influence timing of maturation. We investigated the relationship between female size at maturity and individual growth trajectories of the long-lived semelparous European eel, Anguilla anguilla A Bayesian model was applied to 338 individual growth trajectories of maturing migration-stage female silver eels from France, Ireland, the Netherlands and Hungary. The results clearly showed that when growth rates declined, the onset of maturation was triggered, and the eels left their growth habitats and migrated to the spawning area. Therefore, female eels tended to attain larger body size when the growth conditions were good enough to risk spending extra time in their growth habitats. This flexible maturation strategy is likely related to the ability to use diverse habitats with widely ranging growth and survival potentials in the catadromous life-history across its wide species range.

Transcriptome profile analysis reveals specific signatures of pollutants in Atlantic eels.

Identifying specific effects of contaminants in a multi-stress field context remain a challenge in ecotoxicology. In this context, "omics" technologies, by allowing the simultaneous measurement of numerous biological endpoints, could help unravel the in situ toxicity of contaminants. In this study, wild Atlantic eels were sampled in 8 sites presenting a broad contamination gradient in France and Canada. The global hepatic transcriptome of animals was determined by RNA-Seq. In parallel, the contamination level of fish to 8 metals and 25 organic pollutants was determined. Factor analysis for multiple testing was used to identify genes that are most likely to be related to a single factor. Among the variables analyzed, arsenic (As), cadmium (Cd), lindane (γ-HCH) and the hepato-somatic index (HSI) were found to be the main factors affecting eel's transcriptome. Genes associated with As exposure were involved in the mechanisms that have been described during As vasculotoxicity in mammals. Genes correlated with Cd were involved in cell cycle and energy metabolism. For γ-HCH, genes were involved in lipolysis and cell growth. Genes associated with HSI were involved in protein, lipid and iron metabolisms. Our study proposes specific gene signatures of pollutants and their impacts in fish exposed to multi-stress conditions.

Abnormal ovarian DNA methylation programming during gonad maturation in wild contaminated fish.

There is increasing evidence that pollutants may cause diseases via epigenetic modifications. Epigenetic mechanisms such as DNA methylation participate in the regulation of gene transcription. Surprisingly, epigenetics research is still limited in ecotoxicology. In this study, we investigated whether chronic exposure to contaminants experienced by wild female fish (Anguilla anguilla) throughout their juvenile phase can affect the DNA methylation status of their oocytes during gonad maturation. Thus, fish were sampled in two locations presenting a low or a high contamination level. Then, fish were transferred to the laboratory and artificially matured. Before hormonal treatment, the DNA methylation levels of the genes encoding for the aromatase and the receptor of the follicle stimulating hormone were higher in contaminated fish than in fish from the clean site. For the hormone receptor, this hypermethylation was positively correlated with the contamination level of fish and was associated with a decrease in its transcription level. In addition, whereas gonad growth was associated with an increase in DNA methylation in fish from the clean site, no changes were observed in contaminated fish in response to hormonal treatment. Finally, a higher gonad growth was observed in fish from the reference site in comparison to contaminated fish.

The use of rapid coverage monitoring in the national rubella vaccination campaign, Haiti 2007-2008.

BACKGROUND: Prior to introduction of rubella vaccine in Haiti's national immunization program, the Haitian government conducted a nationwide rubella-measles immunization campaign targeting persons 1-19 years of age to accelerate elimination of rubella and congenital rubella syndrome, while strengthening measles elimination. The national immunization campaign was conducted in phases by geographic region and combined multiple interventions to reach high coverage in all districts. METHODS: We analyzed reported data on number of doses administered and results of rapid monitoring by "commune" (district) to evaluate coverage for each vaccine and intervention in target populations. We reviewed measles and rubella surveillance data from Haiti's national surveillance system. RESULTS: Immunization registers recorded 4.7 million doses of measles-rubella (MR) vaccine administered to persons 1-19 years of age, reaching 80.2% of the estimated population of 1-4 year-olds and surpassing the target among 5-19 year-olds. In addition, 1 million children under 5 years of age received oral polio vaccine and vitamin A supplements, 1.5 million school children received deworming treatment nationwide, and over 500000 women 15-49 years old in 2 major population centers received diphtheria-tetanus vaccine. Based on administrative data, 102 (76.7%) of 133 communes attained 95% or greater coverage with MR vaccine among persons 1-19 years of age. Rapid monitoring in 118 communes indicated that coverage targets were reached in 52.5%. From 2007 to 2010, no confirmed cases of measles or rubella were reported from Haiti. CONCLUSIONS: The experience in Haiti suggests that rubella and congenital rubella syndrome can be eliminated through mass vaccination in countries with weak national immunization programs. However, high routine immunization coverage and improved surveillance are urgently needed to maintain measles and rubella elimination.

PBDE and PCB contamination of eels from the Gironde estuary: from glass eels to silver eels.

Since the 1980s, the eel population has been decreasing dangerously. Persistent Organic Pollutants (POPs) such as Polychlorinated Biphenyls (PCBs) are one of the suspected causes of this decline. A preliminary study of PCB contamination carried out on different fish from the Gironde estuary (southwest of France, Europe) has shown a relatively high level of contamination of eel muscles. In order to characterize the contamination level of PCBs and PBDEs (PolyBrominated Diphenyl-Ethers) in eels from this estuary more than 240 eels were collected during the years 2004-2005 in the Gironde estuarine system, from glass eels to silver eels. Individual European eels were grouped according to length and localization sites. The results have shown a low contamination level of glass eels: respectively 28±11 ng g(-1)dw for PCBs and 5±3 ng g(-1)dw for PBDEs. The contamination level in eels (expressed in ng g(-1)dw) increases from glass eels to silver eels up to 3399 ng g(-1)dw of PCBs for the most contaminated silver eel. Such levels of PCBs similar to those observed in Northern Europe, could raise sanitary problems connected with the World Health Organization (WHO) recommendations. These results are worrying for the local people who regularly eat eels caught in the Gironde estuary.

Exoskeletal deformities in Palaemonidae: are they a threat to survival?

Shrimps of the Palaemon genus have been affected for over 15 years by a problem of exoskeletal deformities, particularly in the Gironde estuary (France). Given the large numbers of individuals affected, this study focus on a better description of the phenomenon to estimate its impact on shrimps. This study, on Palaemon longirostris and Palaemon macrodactylus, is based on samples collected from 1992 to 2007 in the Gironde estuary with particular focus on the year 2007, and on a 4-month rearing of adult shrimps in experimental system. The different approaches reveal the relative persistence of the phenomenon since 1992 (with on average more than 58% of ovigerous females affected) and its persistence during shrimp life cycle, even after several moults. Important consequences associated with deformities at the individual level were demonstrated: a higher mortality, a lighter egg mass and a lower weight for a given size. The presence of exoskeletal deformities is now a major phenomenon, with profound effects on individual shrimps, and which may in turn have an important impact on the survival of the P. longirostris population in particular.

Ovarian gene transcription and effect of cadmium pre-exposure during artificial sexual maturation of the European eel (Anguilla anguilla).

European eels are dangerously threatened with extinction. Recent advances tend to show that pollution could, in addition to other already identified factors, contribute to this drama. In a previous report, cadmium (Cd) pre-exposure was found to strongly stimulate the pituitary-liver-gonad axis of maturing female silver eels, leading, lastly, to oocytes atresia and eels mortality. The present work was performed to get more insights into the effects of Cd preexposure on eels' ovaries. The transcription levels of various genes involved in mitochondrial metabolism, in the cellular response to metal (metallothioneins, MTs) and oxidative stress (catalase, CAT) were investigated. Our results show that ovarian growth is associated with an up-regulation of mitochondrial genes. However, Cd pre-exposure was found to significantly impair this up-regulation. Such findings could explain, at least in part, why oocytes of Cd pre-contaminated eels could not reach final maturation. Concerning MTs, despite the end of the experiment was marked by a strong increase in their gene transcription levels in both eel groups, MTs protein content was found to increase only in the case of Cd pre-contaminated eels. Since this increase in MTs protein content was associated with a massive entry of Cd in gonads, our findings suggest that MTs mRNA, that are normally accumulated in oocytes to cope with the future needs, can be activated and translated in response to Cd exposure.

How cadmium could compromise the completion of the European eel's reproductive migration.

The European eel (Anguilla anguilla L.) is severely threatened with extinction. Surprisingly, even though their unusual life cycle makes them particularly vulnerable to pollution, the possible contribution of contamination remains especially poorly known. Here we have investigated the possible effect of cadmium (Cd), a widespread nonessential metal, on eel reproductive capacities. Both control and Cd precontaminated female silver eels were experimentally matured and forced to swim in metal-free conditions to mimic their reproductive migration. Cd pre-exposure was found to strongly stimulate the pituitary-gonad-liver axis of maturing female silver eels leading to early and enhanced vitellogenesis. This was followed by a strong phenomenon of oocyte atresia and eel mortality. These phenomena occurred before oocytes could reach full maturation and were associated with a large entry of both vitellogenin and Cd into the ovaries. Indeed, a redistribution of previously stored cadmium, even from the low Cd levels of control eels, was observed during sexual maturation. Atresia and mortality phenomena were also associated with an overexpression of the pituitary gene encoding the growth hormone, a marker of physiological stress and energy reserves exhaustion. Significantly, these devastating effects of Cd were observed in organisms that presented liver and kidney Cd concentrations still below those observed in eels from Cd contaminated hydrosystems. Our research shows how common levels of cadmium contamination could disrupt endocrine pathways implicated in gonad maturation and subsequently impair reproductive capacity of eel future genitors.

Cadmium uptake by the European eel: trophic transfer in field and experimental investigations.

Due to its status of threatened species and being heavily contaminated by metals, the European eel (Anguilla anguilla) was selected to investigate cadmium contamination levels of fish settled along a historically cadmium-contaminated hydrosystem, the Garonne-Gironde continuum (France), according to its various location sites and fish length. Results have shown an important site effect on cadmium concentrations in liver but not in gills, highlighting the possible predominance of the trophic exposure route. Subsequently, uncontaminated eels were experimentally exposed to cadmium by water uptake and/or trophic route(s). Eels were fed with different preys: white shrimps collected in an unpolluted area in the Gironde estuary, and cadmium-enriched shrimps. Data obtained tend to show that the use of cadmium-enriched food during experimental investigations triggers an underestimation of the metal trophic transfer rate. These two complementary approaches provide some elements to suggest that the trophic route plays an important role in cadmium contamination of wild eels.

Relationships between locomotor behavior, morphometric characters and thyroid hormone levels give evidence of stage-dependent mechanisms in European eel upstream migration.

In order to decipher movements during freshwater eel colonization, we experimentally characterized individual locomotor behavior of two eel life history stages: elvers and yellow eels. A ramp located at the flume tank upstream side required a specific locomotor behavior to be ascended. Placing individually tagged eels in the middle of the tank three times successively tested behavioral consistency. Eels climbing the ramp on each trial were classified as "upstream climbers" whereas eels settling in the tank middle were classified as "inactive". Both stages exhibited these two opposite consistent behaviors. However, elvers were predominantly "upstream climbers" (58.1%) whereas yellow eels were predominantly "inactive" (79.6%). We measured morphometric characters and thyroid hormones to determine if upstream activity was related to body condition and thyroid status. Elver upstream climbers had higher body condition as well as higher thyroxine (T(4)) and triiodothyronine (T(3)) levels compared with inactive elvers. Yellow eel upstream climbers had lower body length as well as higher T(3) and (T(3):T(4)) ratio compared with inactive yellow eels. This indicated that the physiological release factors for eel upstream migration may be stage dependent. For elvers, high thyroid gland activity, together with high body condition, may be the physiological release factors for migration. In contrast, for yellow eels, physiological stress may be the release factor with an increase in T(4) deiodination activity in the smallest eels. Our study revealed inter-stage and intra-stage locomotor behavior plasticity and suggested stage-dependent opposite impacts of physiological condition on eel upstream migration.

Common pattern of gene expression in response to hypoxia or cadmium in the gills of the European glass eel (Anguilla anguilla).

European eel (Anguilla anguilla) populations are in decline. Glass eel recruitment has fallen 10-fold since the early 1980s. Estuaries play a fundamental role in the life history of eels because glass eels must pass through them to reach freshwater ecosystems. Unfortunately, because of their geographical position at the upstream basin slopes, estuaries accumulate metals like cadmium and are important sites of hypoxia events. In this context, we studied the effect of the oxygen level on the ventilation of the glass eel. In parallel, glass eels were submitted to different dissolved cadmium concentrations (0, 2, and 10 microg L(-1)) under two oxygen levels (normoxia PO2 = 21 kPa and Hypoxia PO2 = 6 kPa). The expression level of various genes involved in the mitochondrial respiratory chain, in the cellular response to metal and oxidative stresses, was investigated. Our results showed that hypoxia enhances (1) ventilation of the postlarval stage and (2) Cd accumulation in gills only at the lowest metal water concentration tested (2 microg Cd L(-1)). At the gene level, Cd exposure mimics the effect of hypoxia since we observed a decrease in expression of genes involved in the respiratory chain and in the defense against oxidative stress.

Impairment of lipid storage by cadmium in the European eel (Anguilla anguilla).

Because European silver eels (Anguilla anguilla) fast during their reproductive migration to the Sargasso Sea, the successful completion of their unusual life cycle depends on quantity of lipids stored beforehand. These lipids are mainly accumulated during the growth phase stage of the animals, called yellow eel, as triglycerides in muscle. They are then catabolized to provide sufficient energy to enable migration, gonad maturation and spawning. In the laboratory, we investigated the possible impact of cadmium on the lipid storage efficiency of yellow eels in order to evaluate the possible contribution of this pollutant to the reported decline of European eel populations. Eels were exposed to dissolved cadmium at nominal concentrations of 0 and 5 microgL(-1) for 1 month. Cd toxicity was then examined by studying the activity and expression level of several enzymes involved in liver lipolysis and lipogenesis and by determining lipid content in muscle. Contaminated eels showed a lower body weight growth with a lower efficiency of lipid storage compared to controls. Using two complementary approaches, genetic and enzymatic, it was possible to conclude that this impairment is mainly explained by an increased utilisation of triglycerides since cadmium contamination did not trigger a reduced fatty acid synthesis. These observations suggest an increased fat consumption in presence of cadmium, which could compromise successful reproduction.

Differential regulation of luteinizing hormone and follicle-stimulating hormone expression during ovarian development and under sexual steroid feedback in the European eel.

Pituitary gonadotropins, luteinizing hormone (LH) and follicle-stimulating hormone (FSH) are, in teleosts as in mammals, under the control of hypothalamic factors and steroid feedbacks. In teleosts, feedback regulations largely vary depending on species and physiological stage. In the present study the regulation of FSH and LH expression was investigated in the European eel, a fish of biological and phylogenetical interest as a representative of an early group of teleosts. The eel FSHbeta subunit was cloned, sequenced and together with earlier isolated eel LHbeta and glycoprotein hormone alpha (GPalpha) subunits used to study the differential regulation of LH and FSH. In situ hybridization indicated that FSHbeta and LHbeta are expressed by separate cells of the proximal pars distalis of the adenohypophysis, differently from the situation in mammals. The profiles of LHbeta and FSHbeta subunit expression were compared during experimental ovarian maturation, using dot-blot assays. Expression levels for LHbeta and GPalpha increased throughout ovarian development with a positive correlation between these two subunits. Conversely, FSHbeta mRNA levels decreased. To understand the role of sex steroids in these opposite variations, immature eels were treated with estradiol (E2)and testosterone (T), both steroids being produced in eel ovaries during gonadal development. E2 treatment induced increases in both LHbeta and GPalpha mRNA levels, without any significant effect on FSHbeta. In contrast, T treatment induced a decrease in FSHbeta mRNA levels, without any significant effect on the other subunits. These data demonstrate that steroids exert a differential feedback on eel gonadotropin expression, with an E2-specific positive feedback on LH and a T-specific negative feedback on FSH, leading to an opposite regulation of LH and FSH during ovarian development.

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.