Unregulated introduced fish (Perca fluviatilis Linnaeus, 1758) is host to zoonotic parasites in a small Mediterranean island.

Introduced species have a major impact on freshwater ecosystems, particularly on islands. Numerous fish species have been introduced in Corsica (Mediterranean island, southern France) as part of planned programs or clandestinely. The introduction of non-native freshwater fish species can have a range of impacts on the recipient ecosystem, including through the co-introduction of its pathogens. A sample of introduced perch Perca fluviatilis Linnaeus, 1758 from the artificial reservoir of Padula was examined following a report of parasites by an angler. The analyses revealed the occurrence of Eustrongylides sp. (Nematoda) and Clinostomum complanatum (Digenea), two zoonotic parasites in P. fluviatilis. Both parasites are reported for the first time in France. Eustrongylides sp. and C. complanatum may have been introduced with their fish intermediate hosts or through their final bird hosts. The occurrence of the two parasites raises concerns from both a veterinary and human health perspective as they can use a wide range of amphibians as intermediate hosts and can be acquired in humans through the consumption of raw or undercooked fish.

Unclear host taxonomy hinders parasite studies: An up-to-date checklist of the protozoan and metazoan parasites of Phoxinus minnows (Teleostei: Leuciscidae).

Phoxinus minnows are small freshwater fish found throughout Eurasia. The taxonomy of this genus is still under investigation, and new species are regularly described. They are frequently introduced outside their native range due to their use as live bait for angling, and such fish introductions can have diverse impacts on the recipient areas. Before the rise in interest regarding the diversity of this genus in the past 15 years, studies carried out on their biology and ecology reported their findings for the Eurasian minnow Phoxinus phoxinus Linnaeus, 1758, which was then considered ubiquitous. A parasites checklist for Phoxinus minnows has yet to be published, and the recent progress on the taxonomy of this genus has enabled us to propose a reassignment of parasite records to their proper host. The most thoroughly studied potential species were the North European species Phoxinus sp. 7 sensu Palandacic et al. 2017, Phoxinus isetensis, and P. phoxinus. We were able to highlight a gap in knowledge for numerous species that have received little-to-no attention in terms of parasitology. Systematic molecular identification of Phoxinus minnows should be conducted when studying their parasites, as a reliable identification of the host is vital in parasitology research. Future research will certainly reveal a species-specific composition of the parasitofauna occurring in Phoxinus, especially among widespread parasites such as Gyrodactylus and Diplostomum. Such specificities could even become tools for assessing the introduction routes of Phoxinus species.

Influence of season and site location on European cultured sea bass parasites in Corsican fish farms using indicator species analysis (IndVal).

The parasites of 536 European sea bass, Dicentrarchus labrax, were studied between January 2012 and December 2013 in six Corsican fish farms. The indicator value (IndVal) method, which combines measures of fidelity and specificity, has been used in this study. Because of its resilience to changes in abundance, IndVal is a particularly effective tool for ecological bioindicator. The IndVal method showed how season can influence the occurrence of parasite species in cultured sea bass and also identified parasites as bioindicators relative to fish farm location. The combination of specificity and fidelity highlighted several parasite species as significant indicators. A randomization test identified five parasite species as having a significant indicator value for season (the monogenean Diplectanum aequans; the copepods Lernanthropus kroyeri and Caligus minimus; the isopod Ceratothoa oestroides, and the myxosporidian Ceratomyxa labracis). If gills parasites are compared, they can be seen to be indicator species for two different seasons. The only Monogenea species D. aequans had fidelity and specificity more pronounced in winter, whereas both copepod species and the Isopoda revealed highest rates of infestation corresponding with an increase of water temperature. Four species have a significant indicator value for site location (D. aequans, L. kroyeri, C. minimus, and C. oestroides). The fact that the farm 6 was isolated on the east coast of Corsica may not have allowed the parasite to infect other farms. The presence of copepods on a single farm can also be explained according to salinity variations. Data for species composition and infection levels should help to improve the monitoring and management of parasitism in cultured sea bass populations.

Influence of site, season, silvering stage, and length on the parasites of the European eel Anguilla anguilla in two Mediterranean coastal lagoons of the island of Corsica, France using indicator species method.

The parasites of 425 European eels, Anguilla anguilla, were studied between 2009 and 2012 in two Mediterranean coastal lagoons of the island of Corsica, France. An indicator value (IndVal) method was used for analysis, which combines measures of fidelity and specificity. Because of its resilience to detect changes in abundance, IndVal is an effective ecological bioindicator. The IndVal method demonstrated that site, season, silvering stage, and length could influence the occurrence of parasite species in European eel. A randomization test identified ten parasite species as having a significant indicator value for site (lagoons differed principally in salinity: oligohaline to polyhaline for the Biguglia lagoon and polyhaline to euhaline for the Urbino lagoon; the digeneans Bucephalus anguillae and Lecithochirium musculus, the cestodes Bothriocephalus claviceps, Proteocephalus macrocephalus, and larvae of Myzophyllobothrium sp., the nematodes Anguillicoloides crassus, and encysted larvae of Contracaecum sp., the acanthocephalan Acanthocephaloides incrassatus, the monogenean Pseudodactyogyrus anguillae, and the copepod Ergasilus gibbus); one parasite species for the spring season (the acanthocephalan A. incrassatus); six parasite species for silvering stage (yellow, pre-silver, silver; the trematodes B. anguillae and Deropristis inflata, encysted larvae of the nematode Contracaecum sp., the acanthocephalan A. incrassatus, the monogenean P. anguillae, and the copepod E. gibbus); and three parasite species for some of the five length classes (the cestode P. macrocephalus, encysted larvae of the nematode Contracaecum sp., and the monogenean P. anguillae). Data for species composition and infection levels should help to improve the management of parasitism in the populations of European eels.

Ultrastructural study of spermiogenesis and the spermatozoon of Microcotyle pancerii (Monogenea: Polyopisthocotylea: Microcotylidae), parasite of meagre Argyrosomus regius (Pisces: Teleostei).

The present work deals with the ultrastructure of spermiogenesis and the spermatozoon of Microcotyle pancerii, a gill parasite of meagre Argyrosomus regius collected in Corsican fish farms. Spermiogenesis was rather similar to that observed in other polyopisthocotylean Monogenea. The intercentriolar body was different from that described in digeneans. The nuclear condensation occurred in 2 successive stages. First, during the nuclear migration in the median cytoplasmic process, the nucleus developed a honeycomb-like appearance. Then, after the flagellar fusion, a discontinuous twisting of the chromatin appeared along the nucleus, with this process ending in total nuclear condensation. The structure of the spermatozoon is characterized by 2 axonemes (9 + "1" pattern), a single and continuous field of cortical microtubules, a mitochondrion, and a nucleus. Our findings were compared with various ultrastructural features in order to highlight variability within the group.

Ultrastructural study of spermiogenesis and the spermatozoon of Crepidostomum metoecus (Digenea: Allocreadiidae), a parasite of Salmo trutta (Pisces: Teleostei).

Spermiogenesis and the spermatozoon of Crepidostomum metoecus, an intestinal parasite of brown trout Salmo trutta, were studied by transmission electron microscopy. Spermiogenesis begins with the formation of a differentiation zone in front of 2 centrioles associated by an intercentriolar body. Each centriole is linked to a striated rootlet, and gives rise to a flagellum. The rotation of flagella is greater than 90 degrees; their fusion with the median cytoplasmic extension is proximodistal and asynchronous. The spermatozoon is formed after constriction of arched membranes. The spermatozoon possesses 2 axonemes of the 9 + "1" pattern, a nucleus, mitochondria, and glycogen. A major feature is the presence, in the anterior part, of external ornamentation and a lateral expansion associated with spinelike bodies. Another attribute is the presence of 2 mitochondria rather than just 1, as in most of the digenean spermatozoa. To our knowledge, this study is the first undertaken with a species of the Allocreadiidae.

Spermiogenesis and spermatozoon ultrastructure of Nicolla wisniewskii (Digenea: Opecoelidae), an intestinal parasite of brown trout Salmo trutta (Pisces: Teleostei).

Spermiogenesis and ultrastructure of spermatozoon of Nicolla wisniewskii (Digenea, Opecoelidae), an intestinal parasite of Salmo trutta, were studied by electron microscopy. Spermiogenesis follows the general pattern found in the Digenea. It begins with the formation of a differentiation zone, including striated rootlets associated with 2 centrioles and an intercentriolar body. The flagella undergo a rotation of greater than 90 degrees. Then, their fusion with the median cytoplasmic process is proximodistal and asynchronous. A peculiarity was observed before the fusion of flagella, i.e., the attachment zones joined as 2 pairs by an electron-dense bridge. The mature spermatozoon is characterized by 2 axonemes, cortical microtubules, a nucleus, 2 mitochondria, external ornamentation, and spinelike bodies. At the posterior end of flagella, the spermatozoon is also characterized by the presence of a central element of the axoneme and without the 9 microtubule doublets. These results were compared with those of the other digeneans and, in particular, with other species of Opecoelidae. It appears that the number of cortical microtubules and their localization in the spermatozoon may be an interesting feature of their phylogeny.

Outbreak of urogenital schistosomiasis in Corsica (France): an epidemiological case study.

BACKGROUND: Schistosomiasis is a snail-borne parasitic disease endemic in several tropical and subtropical countries. However, in the summer of 2013, an unexpected outbreak of urogenital schistosomiasis occurred in Corsica, with more than 120 local people or tourists infected. We used a multidisciplinary approach to investigate the epidemiology of urogenital schistosomiasis in Corsica, aiming to elucidate the origin of the outbreak. METHODS: We did parasitological and malacological surveys at nine potential sites of infection. With the snails found, we carried out snail-parasite compatibility experiments by exposing snails to schistosome larvae recovered from the urine of a locally infected Corsican patient. Genetic analysis of both mitochondrial (cox1) and nuclear (internal transcribed spacer) DNA data from the Schistosoma eggs or miracidia recovered from the infected patients was conducted to elucidate the epidemiology of this outbreak. FINDINGS: We identified two main infection foci along the Cavu River, with many Bulinus truncatus snails found in both locations. Of the 3544 snails recovered across all sites, none were naturally infected, but laboratory-based experimental infections confirmed their compatibility with the schistosomes isolated from patients. Molecular characterisation of 73 eggs or miracidia isolated from 12 patients showed infection with Schistosoma haematobium, S haematobium-Schistosoma bovis hybrids, and S bovis. Further sequence data analysis also showed that the Corsican schistosomes were closely related to those from Senegal in west Africa. INTERPRETATION: The freshwater swimming pools of the Cavu River harbour many B truncatus snails, which are capable of transmitting S haematobium-group schistosomes. Our molecular data suggest that the parasites were imported into Corsica by individuals infected in west Africa, specifically Senegal. Hybridisation between S haematobium and the cattle schistosome S bovis had a putative role in this outbreak, showing how easily and rapidly urogenital schistosomiasis can be introduced and spread into novel areas where Bulinus snails are endemic, and how hybridisation could increase the colonisation potential of schistosomes. Furthermore our results show the potential risk of schistosomiasis outbreaks in other European areas, warranting close monitoring and surveillance of all potential transmission foci. FUNDING: WHO, ANSES, RICET, and the Ministry of Health and Consumption.

Ultrastructure of spermiogenesis and the spermatozoon of Macracanthorhynchus hirudinaceus (Pallas, 1781) (Acanthocephala: Archiacanthocephala), a parasite of the wild boar Sus scrofa.

The present paper describes the ultrastructure of spermiogenesis and the spermatozoon of Macracanthorhynchus hirudinaceus, an acanthocephalan parasite of the wild boar Sus scrofa. At the beginning of spermatogenesis, spermatocytes exhibit synaptonemal complexes and 2 centrioles. In the spermatid, only 1 centriole remains, generating a flagellum with a 9+2 pattern. Another ultrastructural feature observed during the spermiogenesis of M. hirudinaceus is the condensation of the chromatin, forming a "honeycomb" structure in the old spermatid and a homogeneous, electron-dense structure in the spermatozoon. The mature spermatozoon of M. hirudinaceus presents a reversed anatomy, as has been described previously in other species of the Acanthocephala. The spermatozoon is divided into 2 parts: an axoneme, and a nucleocytoplasmic derivative. The spermatozoon flagellum exhibits a 9+2 or 9+0 pattern. The process of spermiogenesis and the ultrastructural organization of the spermatozoon of M. hirudinaceus are compared with available data regarding other acanthocephalan species.

Topography and ultrastructure of the tegument of Aphallus tubarium (Rodolphi, 1819) Poche, 1926 (Digenea: Cryptogonimidae), intestinal parasite of the common Dentex dentex (Linnaeus 1758) from Valinco Gulf.

The tegument ultrastructure of the intestinal fluke Aphallus tubarium was studied for the first time with the use of scanning and transmission electron microscopy. New details on morphology were recorded. The ultrastructural study revealed that the tegument of A. tubarium had a syncytial organization with a distal cytoplasm lying over a basal matrix and cytons. The surface of the tegument is covered with pectinate spines arranged quincuncially. As anterior-posterior differences were observed, particular attention was given to spines. Spines decrease in size and density from the anterior part of body to posterior part. Two types of sensory structures were identified, uniciliated and dome-shaped. Type 1 sensory receptors were outgrowths bearing groups of papillae with shorter and rigid apical seta visible on the anterior part of body surface, encircling the worm. Type 2 sensory receptors was dome-shaped papillae devoid of cilia, found mainly around the oral sucker. Diagrams of spines and sensory receptors were made to help in understanding the nature of these structures. Surface morphology may prove to be useful in distinguishing Aphallus spp with other Cryptogonimidae.

Vitellogenesis of the digenean Plagiorchis elegans (Rudolphi, 1802) (Plagiorchioidea, Plagiorchiidae).

The ultrastructural organization of vitellogenesis of Plagiorchis elegans (Rudolphi, 1802), experimentally obtained from the golden hamster Mesocricetus auratus (Linnaeus, 1758), is described using transmission electron microscopy. This study is the first ultrastructural study of vitellogenesis in a member of the superfamily Plagiorchioidea. The four stages usually observed during vitellogenesis are described: stage I, cytoplasm of the vitellocytes mainly filled with ribosomes and few mitochondria; stage II, beginning of the synthetic activity; stage III, active synthesis of the shell globule clusters; stage IV, vitellocytes are filled with shell globule clusters and contain several lipid droplets, and glycogen granules are grouped around clusters and droplets. Vitellogenesis in P. elegans is compared with that of other Digenea. The differences among P. elegans and previously studied digeneans include, but are not limited to the occurrence of dense coiled endoplasmic reticulum saccules and the concentration of glycogen in the mesenchyme, which may be considered as a fifth stage of maturation of the vitelline glands. This peculiarity was not observed in all trematodes, which clearly indicates differences in the vitellogenesis in various digenean lineages at different stages of maturation of their vitelline cells.

Ultrastructural study of vitellogenesis and oogenesis of Metadena depressa (Stossich, 1883) Linton, 1910 (Digenea, Cryptogonimidae), intestinal parasite of Dentex dentex (Pisces, Teleostei).

The ultrastructural organization of the female reproductive system of Metadena depressa, digenean intestinal parasite of Sparidae (Dentex dentex), was investigated by electron microscopy. The vitellogenesis is divided into four stages: stage I, vitellocytes have a cytoplasm mainly filled with ribosomes and few mitochondria; stage II, beginning of the synthetic activity; stage III, active shell globule clusters synthesis; stage IV, mature vitellocytes are filled with shell globule clusters and generally contain several large lipid droplets. Glycogen granules are grouped at the periphery of the cell. The three stages of the oogenesis process take place in the ovary: stage I, oogonia are undifferentiated small cells located at the periphery of the organ; stage II, primary oocytes possess a higher nucleo-cytoplasmic ratio and a nucleus with a nucleolus and synaptonemal complexes indicating the zygotene-pachytene stage of the first meiotic division; stage III, mature oocytes are located in the proximal region of the organ and possess a cytoplasmic chromatoid body and cortical granules in a monolayer close to the periphery of the cell.

Topography and ultrastructure of the tegument of Lecithochirium musculus (Digenea: Hemiuridae), a parasite of the European eel Anguilla anguilla (Osteichthyes: Anguillidae).

The tegumental ultrastructure of the stomach fluke Lecithochirium musculus was studied using scanning and transmission electron microscopy. The surface of the tegument was smooth and covered by transverse cytoplasmic ridges. Cobblestone-like units of the tegument were observed on the ventral surface. Invagination and evagination of the ecsoma induced variations in the tegumental surface. The ultrastructural study revealed that the tegument of L. musculus had a typical syncytial organization with a distal cytoplasm lying over a basal matrix and cytons. Two types of intra-tegumental sensory structures were observed. Type 1 sensory receptor was a domed-like fusiform structure consisting of a smooth elevation of the tegument. Four receptors of this type were observed on the anterior dorsal surface of the fluke. Three nerve bulbs filled with electron-lucent material and mitochondria composed this receptor. Hemispherical electron-dense collars were observed at the top of the nerve bulbs. Striated rootlets laid just beneath the hemispherical electron-dense collars. Type 2 sensory receptor presented two morphological variations, i.e., a bulb-like monolobed structure, and a bulb-like bilobed structure observed at two different degrees of evagination. For both variations, the nerve bulb enclosed mitochondria, electron-lucent material, and a conical electron-dense collar from which extended a striated rootlet. Numerous sensory receptors of this type were observed around the ventral sucker.

Ultrastructural study of vitellogenesis of Aphallus tubarium (Rudolphi, 1819) Poche, 1926 (Digenea: Cryptogonimidae), an intestinal parasite of Dentex dentex (Pisces: Teleostei).

Vitellogenesis of Aphallus tubarium, an intestinal parasite of Sparidae (Dentex dentex ), was studied by transmission electron microscopy. The ultrastructural features allowed us to distinguish 4 stages in the vitellogenesis process. In stage 1, vitellocytes have a cytoplasm mainly filled with ribosomes, but few mitochondria. In stage 2, there is an increase in amount of endoplasmic reticulum and few Golgi complexes. There is production of shell globules that coalesce into clusters in stage 3; some glycogen particles are observed via the Thiéry method. Finally, in stage 4, mature vitellocytes are filled with shell globule clusters and generally contain a large lipid droplet. Glycogen particles are grouped at the periphery of the cell.

Topography and ultrastructure of the tegument of Bucephalus anguillae (Digenea: Bucephalidae), a parasite of the European eel Anguilla anguilla (Osteichthyen: Anguillidae).

The tegumental ultrastructure of the intestinal fluke Bucephalus anguillae was studied with the use of scanning and transmission electron microscopy. The surface of the tegument is covered by transverse ridges from which protrude numerous closely packed, digitated, and claw-shaped spines. Cobblestone-like units of the tegument were observed on the crescent-shaped formation of the rhynchus and at the posterior part of the body. Three types of sensory structures were examined, i.e., 2 uniciliated receptors and 1 without cilia. As anterior-posterior differences were observed, particular attention was given to spines and sensory receptors. Spine insertion zones and average cilia length are variable between anterior and posterior tegument areas. Ultrastructural study revealed that the tegument of B. anguillae has a typical syncytial organization with a distal cytoplasm lying over a basal matrix and cytons below. Cytoplasmic bridges allowed transit of secretory vesicles and granules. Diagrams of spines and sensory receptors were made to help in understanding the nature of these structures.

Spermiogenesis and sperm ultrastructure of Deropristis Inflata Molin, 1859 (Digenea, Deropristidae), a parasite of Anguilla anguilla.

To our knowledge, this paper describes the first ultrastructural results on spermiogenesis and the spermatozoon of a Digenea belonging to the family Deropristidae, Deropristis inflata. Spermiogenesis follows the usual pattern found in the digeneans, but a single noteworthy characteristic concerns the centriole, which presents a "prominent pear-shaped electron-dense region." Spermiogenesis in D. inflata begins with the formation of a differentiation zone. The two centrioles give rise to flagella. These two flagella undergo a rotation of 90 degrees and fuse with the median cytoplasmic process. The proximo-distal fusion occurs at the level of attachment zones. The mature spermatozoon of D. inflata possesses five regions and presents all the features found in the digenean gamete. However, several characteristics allow us to distinguish the spermatozoon of D. inflata from other digenetic trematoda, namely the external ornamentations of the plasmic membrane, the anterior and posterior extremities. Furthermore, the distal part of the nucleus in front of those of the mitochondria appears such a distinctive criterion in the spermatozoon of this species. Our ultrastructural investigations produce new data on the reproduction of D. inflata, which may be useful for the understanding of the phylogenetic relationships in the Deropristidae family.

Ultrastructural study of the spermatozoon of Pronoprymna ventricosa (Digenea, Baccigerinae), parasite of the twaite shad Alosa fallax Lacepede (Pisces, Teleostei).

In this paper, we describe the ultrastructural organisation of the spermatozoon of a Digenea Baccigerinae Pronoprymna ventricosa. This spermatozoon possesses the elements found in the others digeneans: two axonemes with 9 + "1" pattern, a mitochondrion, a nucleus, cortical microtubules and external ornamentations. However, this spermatozoon presents two particularities. The first is the presence of a single field of cortical microtubules disposed in the ventral side (mitochondrion side) of the spermatozoon. In the other digeneans, two fields of cortical microtubules are observed between the two axonemes. The second is the presence of external ornamentations not associated with cortical microtubules. These two points have, to our knowledge, never been described in another digenean. Moreover, a separation in two groups of the digenean spermatozoa is proposed in relation to the localisation of the external ornamentations. We propose to distinguish a first group exhibiting these elements in the proximal part of the spermatozoon and a second one with external ornamentations in a second part of the gamete.

Ultrastructural study of the spermatozoon of Nicolla testiobliquum (Digenea, Opecoelidae) parasite of brown trout Salmo trutta (Pisces, Teleostei).

The present paper describes the spermatozoon of Nicolla testiobliquum (Digenea, Opecoelidae), an intestinal parasite of brown trout Salmo trutta, studied by transmission electron microscopy. The mature spermatozoon possess two axonemes of 9 + "1" pattern, two mitochondria, a nucleus, external ornamentation of the plasma membrane, spine-like bodies and cortical microtubules. A comparative study between N. testiobliquum and Nicolla wisniewskii shows mainly two different dispositions of the two mitochondria in these two species of a same genera. Moreover, the comparison of each of these spermatozoon features with others digeneans, in general, and other Opecoelidae in particular, is also presented. It appears that the organisation of the posterior part of the spermatozoon, the disposition of cortical microtubules and external ornamentation associated with spine-like bodies could be interesting elements for phylogeny.

Ultrastructure of spermiogenesis and spermatozoon of Leptorhynchoides plagicephalus (Acanthocephala, Palaeacanthocephala), a parasite of the sturgeon Acipenser naccarii (Osteichthyes, Acipenseriformes).

This paper describes the ultrastructure of spermiogenesis and the spermatozoon of Leptorhynchoides plagicephalus, an acanthocephalan parasite of the sturgeon Acipenser naccarii, a species which is under the threat of extinction. At the beginning, spermiogenesis in L. plagicephalus is characterized by the presence of a single centriole in the early spermatid. This centriole generates a flagellum with a 9+0 pattern. Another ultrastructural feature observed during the spermiogenesis of L. plagicephalus is the condensation of chromatin to form an "intranuclear wall". The mature spermatozoon of L. plagicephalus presents a reversed anatomy, as observed in other species of the Acanthocephala. The spermatozoon is divided into two parts: an axoneme and a nucleocytoplasmic derivative. The pattern of spermiogenesis and the ultrastructural organization of the spermatozoon of L. plagicephalus are compared with information available on other acanthocephalan species. The appearance of an "intranuclear wall" observed during the present study represents the first record within the Acanthocephala and is unknown from other animal taxa.