Morphological variations and demographic responses of the Mediterranean pond turtle Mauremys leprosa to heterogeneous aquatic habitats.

Human activities affect terrestrial and aquatic habitats leading to changes at both individual and population levels in wild animal species. In this study, we investigated the phenotype and demographics of the Mediterranean pond turtle Mauremys leprosa (Schweigger, 1812) in contrasted environments of Southern France: two peri-urban rivers receiving effluents from wastewater treatment plants (WWTP), and another one without sewage treatment plant. Our findings revealed the presence of pesticides and pharmaceuticals in the three rivers of investigation, the highest diversities and concentrations of pollutants being found in the river subsections impacted by WWTP effluents. Principal component analysis and hierarchical clustering identified three levels of habitat quality, with different pollutant concentrations, thermal conditions, nutrient, and organic matter levels. The highest turtle densities, growth rates, and body sizes were estimated in the most disturbed habitats, suggesting potential adult benefits derived from harsh environmental conditions induced by pollution and eutrophication. Conversely, juveniles were the most abundant in the least polluted habitats, suggesting adverse effects of pollution on juvenile survival or adult reproduction. This study suggests that turtles living in polluted habitats may benefit from enhanced growth and body size, at the expense of reproductive success.

Molecular Phylogenies of Leeches and Haemoparasites Infecting Freshwater Turtles in Aquatic Ecosystems of Northern Africa Suggest Phylogenetic Congruence between Placobdella costata Sensu Lato and Haemogregarina stepanowi Sensu Lato.

Haemogregarines are blood parasites with a life-cycle involving a vertebrate as the intermediate host and an invertebrate as the definitive host and vector. Extensive phylogenetic investigations based on 18S-rRNA gene sequences have shown that Haemogregarina stepanowi (Apicomplexa: Haemogregarinidae) is able to infest a large diversity of freshwater turtle species, including the European pond turtle Emys orbicularis, the Sicilian pond turtle Emys trinacris, the Caspian turtle Mauremys caspica, the Mediterranean pond turtle Mauremys leprosa, and the Western Caspian turtle Mauremys rivulata, among others. From the same molecular markers, H. stepanowi is further considered to be a complex of cryptic species predisposed to infect the same host species. While Placobdella costata is known to be the unique vector of H. stepanowi, it is only recently that independent lineages within P. costata have been illustrated-suggesting the presence of at least five unique leech species across Western Europe. The aims of our study were therefore to investigate from mitochondrial markers (COI) the genetic diversity within haemogregarines and leeches infecting freshwater turtles of the Maghreb, in order to identify processes of parasite speciation. We showed that H. stepanowi consists of at least five cryptic species in the Maghreb, while two Placobella species were identified in the same area. Although an Eastern-Western speciation pattern was apparent for both leeches and haemogregarines, we cannot make definitive conclusions regarding co-speciation patterns between parasites and vectors. However, we cannot reject the hypothesis of a very strict host-parasite specificity within leeches.

Population structure and dynamics of the Mediterranean Pond Turtle Mauremys leprosa (Schweigger, 1812) in contrasted polluted aquatic environments.

Pollution contributes to the degraded state of continental aquatic ecosystems and biodiversity. Some species appear to be tolerant to aquatic pollution, yet little is known about the effects of such pollution on population structure and dynamics. Here, we investigated how wastewater treatment plant (WWTP) effluents of the Cabestany City, in southern France, contribute to the pollution levels of the Fosseille River, and we tested how they could affect population structure and medium-term dynamics of the native freshwater turtle, the Mediterranean Pond Turtle Mauremys leprosa (Schweigger, 1812). Amongst the 68 pesticides surveyed from water samples collected along the river in 2018 and 2021, a total of 16 pesticides were detected, among which eight were found in the upstream section of the river, 15 in the river section located downstream of the WWTP, and 14 in the outfall of the WWTP, exhibiting the contribution of effluents to the river pollution. From 2013 to 2018 and in 2021, capture-mark-recapture protocols were carried out on the freshwater turtle population living in the river. Using robust design and multi-state models, we showed a stable population throughout the study period, with high year-dependent seniority, and a bidirectional transition occurring primarily from the upstream to the downstream river sections of the WWTP. The freshwater turtle population consisted mostly of adults, with a male biased sex ratio detected downstream of the WWTP neither related to sex-dependent survival, recruitment, nor transition, suggesting a male bias in the hatchlings or primary sex ratio. Also, the largest immatures and females were captured downstream of the WWTP, with females having the highest body condition, whereas no such differences were observed in males. This study highlights that population functioning of M. leprosa is driven primarily by effluents induced resources, at least over the medium-term.

A quantum leap in the evolution of platyhelminths: Host-switching from turtles to hippopotamuses illustrated from a phylogenetic meta-analysis of polystomes (Monogenea, Polystomatidae).

While monogenean worms are mainly parasites of the gills and skin of fish, and to a lesser extent parasites of the oral cavity, urinary bladder, and/or conjunctival sacs of amphibians and freshwater turtles, Oculotrema hippopotamiStunkard, 1924 is the single monogenean polystome reported from a mammal, the common hippopotamus (Hippopotamus amphibius Linnaeus). Several hypotheses have been suggested in the last decade to explain the origin of this enigmatic parasite which infects the conjunctival sacs of H. amphibius. Based on a molecular phylogeny inferred from nuclear (28S and 18S) and mitochondrial (12S and COI) sequences of O. hippopotami and chelonian polystomes, we found a sister group relationship between O. hippopotami and Apaloneotrema moleri (Du Preez & Morrison, 2012). This result suggests lateral parasite transfer between freshwater turtles and hippopotamuses, thus likely reflecting one of the most exceptional known examples of host-switching in the course of vertebrate evolution. It also demonstrates that the proximity in the ecological habitat of parasites within host species is an important feature for their speciation and diversification. Because A. moleri and its host, the Florida softshell turtle (Apalone ferox (Schneider)), are restricted to the USA, we suggest that an ancestral stock of parasites may have been isolated on primitive African trionychids after they diverged from their American relatives, and then switched to hippopotamuses or anthracotheres in Africa.

Metapolystoma ohlerianum n. sp. (Monogenea: Polystomatidae) from Aglyptodactylus madagascariensis (Anura: Mantellidae).

PURPOSE: Despite Madagascar's high amphibian diversity of more than 400 species, only a few polystome species are known from the island. The dissection of frogs from museum collections, together with amphibian and parasite surveys conducted in Madagascar led to the discovery of an undescribed polystome infecting Aglyptodactylus madagascariensis. The purpose of this study is to formally describe this species. METHODS: Polystomes recovered from A. madagascariensis were stained (Acetocarmine) and mounted (Canada balsam) to facilitate morphometrics and taxonomic drawings. Some specimens were fixed in absolute alcohol, a Bayesian tree inferred from the analysis of concatenated 18S, 28S and COI gene sequences was constructed and pairwise distances were calculated. Parasites collected from archived hosts in museums were used for histology and scanning electron microscopy (SEM). RESULTS: Polystomes recovered from A. madagascariensis display characteristics of the genus Metapolystoma and morphologically differed from all other known metapolystomes. The Bayesian phylogeny shows that Metapolystoma n. sp. ex. A. madagascariensis and M. falcatum are sister species with high Bayesian posterior probability. Histological and SEM investigations contributed to morphological descriptions. CONCLUSIONS: Morphological examination supported by phylogenetic analysis and genetic divergences revealed distinct differences from all known metapolystome species, supporting the description of a new species. Differences between the life cycles of Metapolystoma and Polystoma provided additional evidence for the validity of that genus as taxon. Whereas Polystoma may display ovoviviparity on rare occasions after incomplete egg expulsion towards the end of the breeding season, Metapolystoma displays true ovoviviparity. We emphasize the need for parasite surveys in Madagascar and recommended for museum material to be examined for polystomes to provide supplementary material and localities for further field investigation.

Insights on the particle-attached riverine archaeal community shifts linked to seasons and to multipollution during a Mediterranean extreme storm event.

Even if Archaea deliver important ecosystem services and are major players in global biogeochemical cycles, they remain poorly understood in freshwater ecosystems. To our knowledge, no studies specifically address the direct impact of xenobiotics on the riverine archaeome. Using environmental DNA metabarcoding of the 16S ribosomal gene, we previously demonstrated bacterial communities significant shifts linked to pollutant mixtures during an extreme flood in a typical Mediterranean coastal watercourse. Here, using the same methodology, we sought to determine whether archaeal community shifts coincided with the delivery of environmental stressors during the same flood. Further, we wanted to determine how archaea taxa compared at different seasons. In contrast to the bacteriome, the archaeome showed a specific community in summer compared to winter and autumn. We also identified a significant relationship between in situ archaeome shifts and changes in physicochemical parameters along the flood, but a less marked link to those parameters correlated to river hydrodynamics than bacteria. New urban-specific archaeal taxa significantly related to multiple stressors were identified. Through statistical modeling of both domains, our results demonstrate that Archaea, seldom considered as bioindicators of water quality, have the potential to improve monitoring methods of watersheds.

Morphology and molecular characterization of Polystoma goeldii n. sp. (Monogenea, Polystomatidae) parasite from the urinary bladder of Physalaemus ephippifer (Steindachner) (Anura, Leptodactylidae).

Polystomatid flatworms of amphibians are represented in the Neotropical realm by species of Mesopolystoma, Nanopolystoma, Parapseudopolystoma, Polystoma, Riojatrema and Wetapolystoma but only species of Polystoma are known from Brazil, namely Polystoma cuvieri, P. knoffi, P. lopezromani and P. travassosi. During a survey of monogeneans infecting amphibians in the north-eastern region of Pará State, the Cayenne Caecilian Typhlonectes compressicauda was found to be infected with Nanopolystoma tinsleyi and the Veined Tree Frog Trachycephalus typhonius was found to harbor Polystoma lopezromani. A yet unknown species of Polystoma was also encountered in the urinary bladder of the Steindachner's Dwarf Frog, Physalaemus ephippifer. This new species, which is the second species reported from Physalaemus spp., is described herein as Polystoma goeldii n. sp. and its life cycle is also illustrated. The new species can be distinguished from Polystoma spp. from other neotropical realm by a combination of characteristics, including hamuli morphology, outer/inner hamuli length ratio, haptor/total body length ratio, genital bulb/total body length ratio, genital spine number and COI molecular characters.

Revision of the systematics of the Polystomoidinae (Platyhelminthes, Monogenea, Polystomatidae) with redefinition of Polystomoides Ward, 1917 and Uteropolystomoides Tinsley, 2017.

Polystomatids are platyhelminth parasites that infect mainly amphibians and freshwater turtles. For more than seven decades, chelonian polystomes were classified into three genera according to the number of hamuli, i.e. absent for Neopolystoma, one pair for Polystomoidella and two pairs for Polystomoides. Following re-examination of morphological characters, seven new genera were erected the past six years, namely Apaloneotrema, Aussietrema, Fornixtrema, Manotrema, Pleurodirotrema, Uropolystomoides and Uteropolystomoides. However, the polyphyly of Neopolystoma and Polystomoides on the one hand, and the nested position of Uteropolystomoides within a clade encompassing all Neopolystoma and Polystomoides spp. on the other, still raised questions about the validity of these genera. We therefore re-examined several types, paratypes and voucher specimens, and investigated the molecular phylogeny of polystomes sampled from the oral cavity of North American turtles to re-evaluate their systematic status. We show that all Polystomoides Ward, 1917, sensu Du Preez et al., 2022, Neopolystoma Price, 1939, sensu Du Preez et al., 2022 and Uteropolystomoides Tinsley, 2017 species, display vaginae that are peripheral and extend well beyond the intestine. We thus reassign all species of the clade to Polystomoides and propose nine new combinations; however, although Uteropolystomoides is nested within this clade, based on its unique morphological features, we propose to keep it as a valid taxon. Polystomoides as redefined herein groups all polystome species infecting either the oral cavity or the urinary bladder of cryptodires, with peripheral vaginae and with or without two pairs of small hamuli. Uteropolystomoides nelsoni (Du Preez & Van Rooyen 2015), originally described from Pseudemys nelsoni Carr is now regarded as Uteropolystomoides multifalx (Stunkard, 1924) n. comb. infecting three distinct Pseudemys species of North America.

Title: Révision de la systématique des Polystomoidinae (Plathelminthes, Monogenea, Polystomatidae) avec redéfinition des genres Polystomoides Ward, 1917 et Uteropolystomoides Tinsley, 2017. Abstract: Les Polystomatidés sont des plathelminthes parasites qui infestent principalement les amphibiens et les tortues d'eau douce. Pendant plus de sept décennies, les polystomes de chéloniens ont été classés en trois genres selon le nombre d'hamuli, absents pour Neopolystoma, une paire pour Polystomoidella et deux paires pour Polystomoides. Suite au réexamen des caractères morphologiques, sept nouveaux genres ont été érigés ces six dernières années, à savoir Apaloneotrema, Aussietrema, Fornixtrema, Manotrema, Pleurodirotrema, Uropolystomoides et Uteropolystomoides. Cependant, la polyphylie de Neopolystoma et Polystomoides d'une part, et la position imbriquée d'Uteropolystomoides au sein d'un clade englobant toutes les espèces de Neopolystoma et Polystomoides d'autre part, soulèvent encore des questions sur la validité de ces trois genres. Nous avons donc réexaminé plusieurs types, paratypes et vouchers et étudié la phylogénie moléculaire de polystomes prélevés dans la cavité buccale de tortues d'Amérique du Nord pour réévaluer leur statut systématique. Nous montrons que toutes les espèces de Polystomoides Ward, 1917, sensu Du Preez et al., 2022, Neopolystoma Price, 1939, sensu Du Preez et al., 2022 et Uteropolystomoides Tinsley, 2017, présentent des vagins périphériques qui s'étendent bien au-delà de l'intestin. Nous réattribuons ainsi toutes les espèces du clade à Polystomoides et proposons neuf nouvelles combinaisons; cependant, nous proposons de conserver Uteropolystomoides sur la base de ses caractéristiques morphologiques exceptionnelles, bien que son espèce soit imbriquée au sein de ce clade. Polystomoides tel que redéfini ici regroupe toutes les espèces de polystomes infectant soit la cavité buccale, soit la vessie des cryptodires, avec des vagins périphériques, et deux paires de petits hamuli ou sans hamuli. Uteropolystomoides nelsoni (Du Preez & Van Rooyen 2015), l'unique espèce décrite à l'origine à partir de Pseudemys nelsoni Carr est maintenant considérée comme Uteropolystomoides multifalx (Stunkard, 1924) n. comb., qui infecte trois espèces distinctes de Pseudemys d'Amérique du Nord.

Classification of pleurodire polystomes (Platyhelminthes, Monogenea, Polystomatidae) revisited with the description of two new genera from the Australian and Neotropical Realms.

Polystomatids are platyhelminth parasites that infect mostly amphibian and chelonian hosts. Polystomatid of testudines were, for more than seven decades, classified in the three genera - Neopolystoma Price, 1939, Polystomoides Ward, 1917 and Polystomoidella Price, 1939. The genus delimitation was primarily based on the absence of hamuli in Neopolystoma, the presence of one pair of hamuli in Polystomoidella, and two pairs in Polystomoides. From 2016 to 2020, five new genera were erected - namely Uropolystomoides Tinsley and Tinsley, 2016, Uteropolystomoides Tinsley, 2017, Apaloneotrema Du Preez and Verneau, 2020, Aussietrema Du Preez and Verneau, 2020 and Fornixtrema Du Preez and Verneau, 2020. The generic diagnosis was based not only on the size and shape of morphological characters such as hamulus 1, uterus and eggs, but also on the site of infestation (i.e. urinary bladder, oral cavity or conjunctival sacs). Despite large advancements in polystome classification over the last decade, Neopolystoma was still polyphyletic with some species nested within Polystomoides and others being closely related to the Australian Aussietrema. Regarding the distribution of freshwater turtles of the two suborders Pleurodira (Southern continents) and Cryptodira (distributed worldwide except in Australia), one may wonder whether Australian chelonian polystomes of the genus Neopolystoma may have diverged from species infecting other pleurodires of South America. In the present study based on the analysis of several species selected among all genera, we reveal striking morphological differences within polystomes infecting pleurodiran turtles, which herein led to the proposal of two new chelonian polystome genera, Pleurodirotrema n. g. and Manotrema n. g. Pleurodirotrema n. g. is characterized by the absence of hamuli, presence of latero-ventral vaginae and includes species that infect either the oral region or the urinary bladder of Australian hosts of the Pleurodira. Manotrema n. g. is characterized by the presence of small hamuli, latero-ventral vaginae, deep incisions between suckers, a low genital spine number (<10) and includes species that infect the oral region of South American Pleurodira.

First record of Metapolystoma (Monogenea: Polystomatidae) from Boophis tree frogs in Madagascar, with the description of five new species.

Although Madagascar has more than 350 frog species of which all but two are endemic to the island, the known polystome (Monogenea: Polystomatidae) diversity parasitizing Malagasy frogs is low, encompassing five species of Madapolystoma, one species of Kankana and one Metapolystoma. Investigating the parasite diversity of frog parasites at selected Malagasy localities led to the discovery of undescribed polystomes. Five treefrogs, Boophis albilabris, Boophis doulioti, Boophis luteus, Boophis madagascariensis and Boophis occidentalis were found to be infected and are reported here as hosts for new Metapolystoma species. Morphological investigation, combining examination of body length, haptor length, genital bulb width, genital crown diameter, genital spine number, genital spine length, ovary length, egg length, hamulus length, hamulus guard length and hamulus hook length, revealed five distinct morphotypes. Phylogenetic analysis and genetic divergences obtained for three of the five morphotypes, support the distinction of new species. Metapolystoma ansuanum n. sp. is described from B. luteus, Metapolystoma falcatum n. sp. from B. doulioti, Metapolystoma multiova n. sp. from B. occidentalis, Metapolystoma theroni n. sp. from B. madagascareniensis and Metapolystoma vencesi n. sp. from B. albilabris. Finally, although the validity of Metapolystoma as taxon is not fully resolved yet, the phylogenetic position of the described species and their morphology provide clear evidence for new metapolystome taxa.

Pseudocapillaria (Ichthyocapillaria) bumpi n. sp. (Nematoda: Capillariidae) Parasitising West African Lungfish Protopterus annectens (Owen, 1839) (Lepidosireniformes: Protopteridae) in Mozambique and Its Phylogenetic Position Within Capillariid Nematodes.

INTRODUCTION: Pseudocapillaria (Ichthyocapillaria) bumpi n. sp. (Nematoda: Capillariidae) is described from specimens infecting the digestive tract of the West African lungfish, Protopterus annectens (Owen) (Lepidosireniformes: Protopteridae). METHODS: Nematodes were collected in Karingani Game Reserve, Southern Mozambique and studied using light microscopy; obtained sequences of the 18S ribosomal (18S rRNA) gene used for phylogenetic studies. RESULTS: The new species is assigned to Pseudocapillaria Freitas, 1959 by having a stichosome consisting of a single row of stichocytes and ventrolateral lobes on the male tail as well as by lacking spines on the specular sheath, caudal alae, a membranous caudal bursa, and a lateral expansion of the caudal end. It is also assigned to the sub-genus Ichthyocapillaria Moravec, 1982 by having a membrane between the ventrolateral lobes and by lacking a vulvar appendage. Pseudocapillaria (I.) bumpi n. sp. differs from its nominal congeners by having a comparatively large body size with relatively long spicule, bearing the thin membrane non-extending further than ventrolateral lobes in males and relatively smaller eggs without protruding polar plugs. It is the 20th species of the genus, the first from the Afrotropical Realm, and the first from any lungfish species. Phylogenetic analyses suggested that P. (I.) bumpi is deeply nested in a clade that associates species of the genera Pearsonema, Aonchotheca and Baruscapillaria.

Cytochrome c oxydase I phylogenetic analysis of Haemogregarina parasites (Apicomplexa, Coccidia, Eucoccidiorida, Haemogregarinidae) confirms the presence of three distinct species within the freshwater turtles of Tunisia.

Species of Haemogregarina are apicomplexan blood parasites that use vertebrates as intermediate hosts. Due to limited interspecific morphological characters within the genus during the last decade, 18S rRNA gene sequences were widely used for species identification. As coinfection patterns were recently reported from nuclear molecular data for two sympatric freshwater turtles Mauremys leprosa and Emys orbicularis from Tunisia, our objectives were to design COI specific primers to confirm the presence of three distinct species in both host species. Blood samples were collected from 22 turtles, from which DNAs were extracted and used as templates for amplification. Following different rounds of PCR and nested PCR, we designed specific Haemogregarina COI primers that allowed the sequencing of nine distinct haplotypes. Phylogenetic Bayesian analysis revealed the occurrence of three well-differentiated sublineages that clustered together into a single clade. Based on pairwise genetic distances (p-distance), we confirmed the occurrence of three distinct but phylogenetically closely related species coinfecting M. leprosa and E. orbicularis in the same aquatic environments. Our results demonstrate that the use of fast evolving genes within Haemogregarina will help to investigate the parasite diversity within both intermediate vertebrate and definitive invertebrate hosts, and to assess the evolution, historical biogeography and specificity of haemogregarines.

First Record of a Polystome from Alligator Snapping Turtle, Macrochelys temminckii (Cryptodira: Chelydridae) or Mississippi; with Comments on "Neopolystoma orbiculare (Stunkard, 1916)" and Its Junior Subjective Synonyms.

Herein, we describe several newly-collected specimens of Neopolystoma cf. orbiculare from the urinary bladder of 2 alligator snapping turtles, Macrochelys temminckii (Troost in Harland, 1835) (Cryptodira: Chelydridae Gray, 1831) from Comet Lake (30°35'46.94″N, 88°36'3.12″W), Pascagoula River, Mississippi. Our specimens differed from all previous descriptions of N. orbiculare and its junior subjective synonyms by the combination of having intestinal ceca adorned with triangular pockets and that terminate dorsal to the haptor, distinctive hooklets each having a handle and guard of approximately equal length and having a much longer and curved blade, 16 genital coronet spines that each possess 1-2 flanges per spine, pre-testicular vaginal pores, and vaginal ducts that are anterior to the junction of the oviduct and genito-intestinal canal. Some of our specimens were enantiomorphic (4 and 3 had a dextral and sinistral ovary, respectively). Nucleotide sequences (large subunit ribosomal DNA [28S], small subunit ribosomal DNA [18S], and cytochrome oxidase subunit 1 mitochondrial gene [COI]) for our specimens were most similar to GenBank sequences ascribed to N. orbiculare. Single-gene and concatenated phylogenetic analyses confirmed that NeopolystomaPrice, 1939 is polyphyletic and that our isolates share a recent common ancestor with those ascribed to N. orbiculare. This is the first record of a polystomatid from Mississippi, from the Pascagoula River, and from the alligator snapping turtle (and only the second species of Neopolystoma reported from any snapping turtle).

Eye to eye: classification of conjunctival sac polystomes (Monogenea: Polystomatidae) revisited with the description of three new genera Apaloneotrema n. g., Aussietrema n. g. and Fornixtrema n. g.

Polystomes (Monogenea: Polystomatidae) of freshwater turtles are currently represented by five genera, namely Neopolystoma, Polystomoides, Polystomoidella, Uropolystomoides and Uteropolystomoides. These parasites can infect the urinary, oral and/or the conjunctival sac systems of their hosts, showing strict site specificity. A recent phylogenetic study showed that the two most diverse genera within chelonian polystomes, i.e. Neopolystoma and Polystomoides, are not monophyletic. Furthermore, polystomes infecting the conjunctival sacs of their host, except for one species, formed a robust lineage. A fusiform egg shape has been reported for conjunctival sac polystomes and it was assumed that this characteristic could be a good character for the systematics of polystomes. Our objective in the present work was, therefore, to study more in depth the morphology of polystomes collected from the conjunctival sacs of chelonians to find characters defining a putative new genus. To achieve this objective, more specimens were collected in 2018 and 2019 from turtles sampled in North Carolina and Florida (USA) to extend taxon sampling for the phylogenetic analysis. Morphological characters of relevant polystome specimens were re-examined from several collections from Asia, Australia, Europe, South Africa, South America and North America. Based on a Bayesian tree inferred from the analysis of four concatenated genes, namely 12S, 18S, 28S and COI, polystomes found in the conjunctival sacs were grouped in three distinct lineages, the first one including a single species infecting an Australian pleurodire turtle; the second one including eleven species infecting cryptodire turtles of South America, North America and Asia; and the last one including a single species infecting a softshell cryptodire turtle of North America. Based on observations of live specimens by Dr. Sylvie Pichelin and our morphological analysis, the conjunctival sac polystomes from Australian turtles are small, cannot extend their body significantly, have a spherical ovary and egg, have a large genital bulb and possess latero-ventral vaginae at the level of the testis. Based on observations of live specimens and morphological analysis of whole mounted specimens, polystomes of the second lineage share the following morphological characteristics: the ability to stretch out and double their length, a long oval ovary, a separate egg-cell-maturation-chamber, fusiform to diamond-shaped eggs with acute tips, small genital bulb and vaginae peripheral on the side of the body at the level of the testis. The polystome species of the third lineage occupies a basal position, has the ability to stretch out and possess an elongated ovary, a large fusiform egg with rounded tips, a small genital bulb and small latero-ventral vaginae at the level of the ovary. These three distinct conjunctival sac polystome lineages are herein described as separate new genera, Aussietrema, Fornixtrema and Apaloneotrema, respectively.

The genetic diversity of blood parasites within the freshwater turtles Mauremys leprosa and Emys orbicularis in Tunisia reveals coinfection with Haemogregarina spp.

Haemogregarina species are apicomplexan blood parasites infecting vertebrates such as fish, lizards, and turtles. Due to the high morphological similarity of the erythrocytic stages infecting host species, it has always been a challenge to identify the true diversity of these parasites. Therefore, taxonomic studies are presently based on the combination of morphological and molecular data. In Tunisia, two species of Haemogregarina have been reported within the freshwater turtle Mauremys leprosa (Geoemydidae) for more than 40 years. Since M. leprosa occurs in the same aquatic environments as Emys orbicularis (Emydidae) in Tunisia, our objectives were to assess parasite diversity and specificity on the basis of both morphological and molecular approaches. The turtles were surveyed and sampled across six aquatic areas of Tunisia. Among the 39 specimens of M. leprosa and seven of E. orbicularis that were trapped and investigated, the presence of haemogregarines was detected in the blood of turtles only at sites where leeches were observed. Three 18S variants were identified, which corresponded to three distinct Haemogregarina species, among which one was identified as Haemogregarina stepanowi. The two other species that were detected are likely new to science. Because we show the occurrence of more than one blood parasite species within a single host specimen, our study provides the first report of coinfection with molecularly distinct Haemogregarina spp.

Particle-attached riverine bacteriome shifts in a pollutant-resistant and pathogenic community during a Mediterranean extreme storm event.

Rivers are representative of the overall contamination found in their catchment area. Contaminant concentrations in watercourses depend on numerous factors including land use and rainfall events. Globally, in Mediterranean regions, rainstorms are at the origin of fluvial multipollution phenomena as a result of Combined Sewer Overflows (CSOs) and floods. Large loads of urban-associated microorganisms, including faecal bacteria, are released from CSOs which place public health - as well as ecosystems - at risk. The impacts of freshwater contamination on river ecosystems have not yet been adequately addressed, as is the case for the release of pollutant mixtures linked to extreme weather events. In this context, microbial communities provide critical ecosystem services as they are the only biological compartment capable of degrading or transforming pollutants. Through the use of 16S rRNA gene metabarcoding of environmental DNA at different seasons and during a flood event in a typical Mediterranean coastal river, we show that the impacts of multipollution phenomena on structural shifts in the particle-attached riverine bacteriome were greater than those of seasonality. Key players were identified via multivariate statistical modelling combined with network module eigengene analysis. These included species highly resistant to pollutants as well as pathogens. Their rapid response to contaminant mixtures makes them ideal candidates as potential early biosignatures of multipollution stress. Multiple resistance gene transfer is likely enhanced with drastic consequences for the environment and human-health, particularly in a scenario of intensification of extreme hydrological events.

Indopolystoma n. gen. (Monogenea, Polystomatidae) with the description of three new species and reassignment of eight known Polystoma species from Asian frogs (Anura, Rhacophoridae).

The polystomes (Monogenea, Polystomatidae) radiated across semi-aquatic tetrapods including all three amphibian orders, freshwater turtles and the hippopotamus. Prior to this study, phylogenetic analyses revealed that the most diverse and widespread genus, Polystoma, was not monophyletic; a lineage comprising four undescribed species from the bladder of Zhangixalus spp. (Rhacophoridae) in Asia occupied a deep phylogenetic position. Regarding vicariance biogeography and molecular dating, the origin of this lineage is correlated with the breakup of Gondwanaland in the Mesozoic period. Based on a Bayesian analysis of four concatenated genes (18S, 28S, COI and 12S) and morphological evidence, one new genus, Indopolystoma n. gen., and three new species, sampled in Japan and China, are described here: Indopolystoma viridi n. sp. from Z. viridis of Japan, Indopolystoma elongatum n. sp. from Z. arboreus of Japan, and Indopolystoma parvum n. sp. from Z. omeimontis of China. Indopolystoma is unique amongst polystome genera infecting anurans by possessing a small haptor relative to the body size, posteriormost marginal hooklet C1 much bigger than hooklets C2-C8 with conspicuous broad blade and guard and a pair of hamuli lacking a deep notch. Eight species of Asian Polystoma, all from rhacophorids, are transferred as Indopolystoma carvirostris (Fan, Li & He, 2008) n. comb., I. hakgalense (Crusz & Ching, 1975) n. comb., I. indicum (Diengdoh & Tandon, 1991) n. comb., I. leucomystax (Zhang & Long, 1987) n. comb., I. mutus (Meng, Song & Ding, 2010) n. comb., I. pingbianensis (Fan, Wang & Li, 2004) n. comb., I. rhacophori (Yamaguti, 1936) n. comb., and I. zuoi (Shen, Wang & Fan, 2013) n. comb.

TITLE: Indopolystoma n. gen. (Monogenea, Polystomatidae) avec description de trois nouvelles espèces et réaffectation de huit espèces connues de Polystoma parasites de grenouilles asiatiques (Anura, Rhacophoridae). ABSTRACT: Les polystomes (Monogenea, Polystomatidae) se sont diversifiés sur des tétrapodes semi-aquatiques, notamment les trois ordres d'amphibiens, les tortues d'eau douce et l'hippopotame. Avant cette étude, des analyses phylogénétiques avaient révélé que le genre le plus diversifié et le plus répandu, Polystoma, n'était pas monophylétique ; une lignée comprenant quatre espèces non décrites de la vessie de Zhangixalus spp. (Rhacophoridae) en Asie occupait une position phylogénétique profonde. En ce qui concerne la biogéographie de vicariance et la datation moléculaire, l'origine de cette lignée est corrélée à l'éclatement du Gondwana au Mésozoïque. D'après une analyse bayésienne de quatre gènes concaténés (18S, 28S, COI et 12S) et des preuves morphologiques, un nouveau genre, Indopolystoma n. gen. et trois nouvelles espèces échantillonnées au Japon et en Chine sont décrites ici : Indopolystoma viridi n. sp. de Z. viridis du Japon, Indopolystoma elongatum n. sp. de Z. arboreus du Japon et Indopolystoma parvum n. sp. de Z. omeimontis de Chine. Indopolystoma est unique parmi les genres de polystomes infectant les anoures. Il possède un hapteur petit par rapport à la taille du corps, un crocheton le plus postérieur C1 beaucoup plus gros que les crochetons C2 à C8, avec lame et garde bien visibles, ainsi qu'une paire d'hamuli dépourvus d'encoche profonde. Huit espèces de polystomes asiatiques, toutes issues de Rhacophoridae, sont transférées comme Indopolystoma carvirostris (Fan, Li & He, 2008) n. comb., I. hakgalense (Crusz et Ching, 1975) n. comb., I. indicum (Diengdoh & Tandon, 1991) n. comb., I. leucomystax (Zhang & Long, 1987) n. comb., I. mutus (Meng, Song & Ding, 2010) n. comb., I. pingbianensis (Fan, Wang et Li, 2004) n. comb., I. rhacophori (Yamaguti, 1936) n. comb. et I. zuoi (Shen, Wang et Fan, 2013) n. comb.

First record of viviparity in polystomatid flatworms (Monogenea: Polystomatidae) with the description of two new species of Madapolystoma from the Madagascan anuran hosts Blommersia domerguei and Mantella expectata.

Two frog species, Blommersia domerguei and Mantella expectata, are reported as hosts for new species of Madapolystoma. Phylogenetic analyses and genetic divergences observed in the genus supported the distinction of two morphotypes infesting selectively each host species and morphological investigation combining marginal hooklet morphometrics, genital spine number and measurements further showed that polystomes from the two host species differed from each other and from all other known polystomes. Madapolystoma magnahami n. sp. and Madapolystoma isaloensis n. sp. are therefore described as two new species. Advanced in utero development was illustrated in both polystome species following the observation of well developed hamuli and two pairs of haptoral suckers in developing embryos. Inside some of these in utero embryos a F2 generation embryo was also observed. This is the first report of true viviparity among polystomatid flatworms.

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.