Demonstrating the value and importance of combining DNA barcodes and discriminant morphological characters for polystome taxonomy (Platyhelminthes, Monogenea).

Polystomes are monogenean parasites that infest mainly semi aquatic vertebrates, such as amphibians and chelonians. Owing to the lack of discriminative morphological characters and because polystomes are considered to be strictly host- and site-specific, host identity is often used as an additional character for parasite identification. Recent genetic studies, however, showed that polystomes infecting freshwater turtles in outdoor turtle enclosures and natural environments, were not strictly host-specific. Therefore, we proposed a new procedure for turtle polystome taxonomy based on the combination of Cytochrome c Oxydase I sequences and two discriminant morphological characters, namely the number of genital spines and the testis shape. We tested the validity of this procedure with Polystomoides oris, which was collected from the pharyngeal cavity of the American painted turtle Chrysemys picta and two undescribed species, both collected from the pharyngeal cavity of the American slider Trachemys scripta and two other European turtles, namely the European pond turtle Emys orbicularis and the Mediterranean turtle Mauremys leprosa. A Principal Component Analysis based on both morphological characters allowed the separation of all specimens in three morphological groups, which matched well with the molecular data. As a result, we describe two new polystome species, i.e., Polystomoides soredensis n. sp. and Polystomoides scriptanus n. sp.

Oxidative stress induced by glyphosate-based herbicide on freshwater turtles.

Freshwater ecosystems face very strong anthropogenic pressures, among which overexploitation, habitat degradation, flow modification, species invasion, and water pollution lead to growing threats on biodiversity. Urbanization through wastewater treatment, industry through the release of inorganic and organic chemicals, and agriculture through the use of pesticides and herbicides are the main factors involved in water pollution. In France, more precisely in the Pyrénées-Orientales department, the poor quality of the watercourses is attributable overall to the use of glyphosate-based herbicides in agricultural activities. Because these chemicals can impact individuals, populations, and biodiversity, we investigated, under experimental conditions, the physiological response of animals facing abiotic contaminants. We selected as a model, juveniles of the freshwater turtle Trachemys scripta elegans. We measured the gene expression and activity of the catalase and superoxide dismutase enzymes as well as the levels of lipid peroxidation, which are all oxidative stress biomarkers, in turtles challenged with high concentrations of glyphosate-based herbicides, on the one hand, and with degraded waters collected from a local watercourse, on the other. We also measured the acetylcholinesterase activity across the same animals. We showed through variations in gene expression and enzyme activity that a glyphosate commercial formulation induced a stress in turtles. A similar outcome was obtained when turtles faced degraded waters. The results indicated that the poor quality of regional waters could be a real threat for animal health. Because turtles are globally less sensitive to contaminants than amphibians, which are lacking in the degraded waters of the Pyrénées-Orientales department, they could constitute an excellent model to follow the evolution of water quality through the study of oxidative stress biomarkers. Environ Toxicol Chem 2017;36:3343-3350. © 2017 SETAC.

Oxidative stress biomarkers in the Mediterranean pond turtle (Mauremys leprosa) reveal contrasted aquatic environments in Southern France.

Increasing anthropogenic activities, like agricultural practices, constitute the main causes of the loss of water quality and disruption of freshwater ecosystems. High concentrations of pesticides, as shown under experimental conditions, can indeed impact freshwater animals. In Southern France, especially in the Pyrénées-Orientales department, because agricultural activities are mainly based on fruit crops and vineyards, glyphosate and AMPA were detected in some watercourses. Thereby we investigated the effects of degraded waters on the physiology of the endemic endangered freshwater species, namely the Mediterranean pond turtle Mauremys leprosa, in contrasted environments along the same rivers on the one hand and between different rivers on the other. We measured the activity and gene expression of two enzymes involved in the oxidative detoxification processes, namely the Catalase and the Superoxide dismutase. We showed significant variations in the Catalase gene expression and activity within turtles of the Fosseille River depending of their location, i.e. upstream or downstream of the wastewater treatment plants (WTP). Because agricultural environments are similar all along this river, they can no be longer considered as the unique source of turtle stress. The processed waters discharged by the WTP, which contribute to watercourses degradation, could therefore considerably impact the biodiversity of the freshwater environments.

The high resolution melting analysis (HRM) as a molecular tool for monitoring parasites of the wildlife.

In an interconnected world, the international pet trade on wild animals is becoming increasingly important. As a consequence, non-native parasite species are introduced, which affect the health of wildlife and contribute to the loss of biodiversity. Because the investigation of parasite diversity within vulnerable host species implies the molecular identification of large samples of parasite eggs, the sequencing of DNA barcodes is time-consuming and costly. Thereby, the objectives of our study were to apply the high resolution melting (HRM) approach for species determination from pools of parasite eggs. Molecular assays were validated on flatworm parasites (polystomes) infecting the Mediterranean pond turtle Mauremys leprosa and the invasive red-eared slider Trachemys scripta elegans in French natural environments. HRM analysis results indicated that double or multiple parasitic infections could be detected from wild animal populations. They also showed that the cycle of parasite eggs production was not regular over time and may depend on several factors, among which the ecological niche and the target species. Thereby, monitoring parasites from wild endangered animals implies periodic parasitological surveys to avoid false negative diagnostics, based solely on eggs production.