Immunosenescence patterns differ between populations but not between sexes in a long-lived mammal.

In animals, physiological mechanisms underlying reproductive and actuarial senescence remain poorly understood. Immunosenescence, the decline in the ability to display an efficient immune response with increasing age, is likely to influence both reproductive and actuarial senescence through increased risk of disease. Evidence for such a link has been reported from laboratory animal models but has been poorly investigated in the wild, where variation in resource acquisitions usually drives life-history trade-offs. We investigated immunosenescence patterns over 7 years in both sexes of two contrasting roe deer populations (Capreolus capreolus). We first measured twelve immune markers to obtain a thorough identification of innate and adaptive components of immunity and assessed, from the same individuals, the age-dependent variation observed in parasitic infections. Although the level of innate traits was maintained at old age, the functional innate immune traits declined with increasing age in one of two populations. In both populations, the production of inflammatory markers increased with advancing age. Finally, the adaptive response declined in late adulthood. The increasing parasite burden with age we reported suggests the effective existence of immunosenescence. Age-specific patterns differed between populations but not between sexes, which indicate that habitat quality could shape age-dependent immune phenotype in the wild.

Trichobilharzia anseri n. sp. (Schistosomatidae: Digenea), a new visceral species of avian schistosomes isolated from greylag goose (Anser anser L.) in Iceland and France.

Parasitological investigations carried out on birds in Iceland and France highlight the presence of four species of avian schistosomes from greylag geese (Anser anser L.): the european nasal species Trichobilharzia regenti and three visceral species, among which an unknown species isolated from blood vessels of the large intestine and liver. Morphological and molecular analyzes of different parasite stages (eggs, adults) revealed new species of Trichobilharzia genus ­ Trichobilharzia anseri sp. nov. Studies on host-parasite relationship under natural conditions, showed that the life-cycle includes the snail Radix balthica (syn. R. peregra) as intermediate host. The cercariae, already isolated in Iceland from two ponds of the Reykjavik capital area ­ the Family park and Tjörnin Lake ­ are the same as those isolated in 1999 by Kolárová et al. during the first study on Icelandic parasitic agents of cercarial dermatitis.

Fatal Parastrongylus dujardini infection in captive callitrichids.

Intravascular nematodes were considered the cause of death of 14 captive callitrichids. All animals were captive born at zoos in France and died with little or no premonitory signs of disease. No consistent gross lesions were observed at necropsy, although in certain cases intracardiac adult parasites were noted. The most significant histologic findings were verminous pneumonia and pulmonary endarteritis. In all cases except one, intravascular adult nematodes were observed with eggs and larvae in the lungs. Adult nematodes were obtained from 8 animals and in all cases were identified as Parastrongylus dujardini. To the authors' knowledge, this is the first report of intravascular angiostrongylosis with primary cardiopulmonary location in callitrichids in France.

A comparison of the physiological status in parasitized roe deer (Capreolus capreolus) from two different populations.

Studies of the impact of parasites on host performance have mainly focused on body mass, a phenotypic trait that responds relatively slowly to the presence of parasites, and the expectedly faster response of physiological parameters has been mostly overlooked. We filled the gap by measuring the impact of endoparasites on four hematological/biochemical parameters (hematocrit, albumin, creatinine and fructosamine) in two contrasting free-living populations of roe deer. We generally found negative relationships between parasites and physiological parameters. Our findings also indicate little role of host sex on parasite impact and strongest parasite effects on young and senescent hosts.

New data in France on the trematode Alaria alata (Goeze, 1792) obtained during Trichinella inspections.

The trematode Alaria alata is a cosmopolite parasite found in red foxes (Vulpes vulpes), the main definitive host in Europe. In contrast only few data are reported in wild boars (Sus scrofa), a paratenic host. The aim of this paper is to describe the importance and distribution of Alaria alata mesocercariae in wild boars, information is given by findings of these larvae during Trichinella mandatory meat inspection on wild boars' carcasses aimed for human consumption. More than a hundred cases of mesocercariae positive animals are found every year in the East of France. First investigations on the parasite's resistance to deep-freezing in meat are presented in this work.

Molecular diversity of Trichobilharzia franki in two intermediate hosts (Radix auricularia and Radix peregra): a complex of species.

Recently, the systematic use of the molecular approach as a complement to the other approaches (morphology, biology, life cycle) has brought help for the identification of species considered as different in the past to be regrouped and synonymised, and distinctions to be drawn between species similar at the morphological level. Among these species, we tried to clarify the situation of Trichobilharzia frankiMüller and Kimmig, 1994, species that today include more than 50 haplotypes notably coming from larval stages isolated from intermediate hosts belonging to gastropods of the Radix genus. Cercariae were isolated in France and Iceland from various molluscs, before being analyzed, with their hosts, by molecular analysis of various fields such as the D2 and ITS of the ribosomal DNA and the COX1 of mitochondrial DNA. We thus show the presence of two clades depending on the specificity of their intermediate host in which they were isolated (Radix auricularia or Radix peregra), thus allowing separation of the species T. franki that had been described in the past as a probable new species.

Genetic characterization of Toxoplasma gondii from wild boar (Sus scrofa) in France.

Toxoplasma gondii strains isolated from domestic animals and humans have been classified into three clonal lineages types I-III, with differences in terms of pathogenicity to mice. Much less is known on T. gondii genotypes in wild animals. In this report, genotypes of T. gondii isolated from wild boar (Sus scrofa) in France are described. During the hunting seasons 2002-2008, sera and tissues of individuals from two French regions, one continental and one insular, were tested for Toxoplasma infection. Antibodies to T. gondii were found in 26 (17.6%) of 148 wild boars using the modified agglutination test (MAT, positivity threshold: 1:24). Seroprevalence was 45.9% when considering a threshold of 1:6. Hearts of individuals with a positive agglutination (starting dilution 1:6) (n=60) were bioassayed in mice for isolation of viable T. gondii. In total, 21 isolates of T. gondii were obtained. Genotyping of the isolates using 3 PCR-restriction fragment length polymorphism markers (SAG1, SAG2 and GRA7) and 6 microsatellite loci analysis (TUB2, TgM-A, W35, B17, B18 and M33) revealed that all belonged to type II lineage. These results underline that wild boar may serve as an important reservoir for transmission of T. gondii, and that strains present in wildlife may not be different from strains from the domestic environment.

Avian schistosomes in French aquatic birds: a molecular approach.

The prevalence of human cercarial dermatitis (HCD) caused by bird schistosomes appears to be increasing in France, in light of the impact of tourism combined with high densities of wild aquatic hosts in freshwater areas. The present work expands our knowledge of schistosome systematics by including samples of bird schistosomes collected from their natural hosts in France. Heads (318) and viscera (81) of aquatic birds belonging to 16 species from five orders, collecting during the hunting seasons or found dead, were autopsied for nasal and visceral schistosomes. Eggs and/or adults were analysed by molecular methods using the D2 domain and the second internal transcribed spacer (ITS-2) region of rDNA to determine species. Even if nasal eggs were polymorphic according to the host, all haplotypes were similar to that of Trichobilharzia regenti. Marked diversity of visceral species was observed. Final hosts under natural conditions were reported. For the first time, Trichobilharzia franki is reported in its natural bird hosts, Anas platyrhynchos, Anas crecca, Aythya fuligula and Cygnus olor. We also identified T. szidati in A. crecca and Anas clypeata. Bilharziella polonica was found in six species of aquatic birds, including Grus grus. This finding is the first record of bird schistosomes in this aquatic bird. Three new taxa of visceral schistosomes in Anser anser are strongly suspected according to their haplotypes. Futhermore, a new haplotype of visceral schistosomes isolated in Cygnus olor and similar to Allobilharzia visceralis was identified.

[Molecular and morphological characterisation of two closely related species belonging to the subgenus Phlebotomus chabaudi Croset, Abonnenc & Rioux, 1970 et P. riouxi Depaquit, Killick-Kendrick & Léger, 1998 (Diptera: Psychodidae)]. / Caractérisation moléculaire et morphologique de deux espèces affines de Paraphlebotomus: Phlebotomus chabaudi Croset, Abonnenc & Rioux, 1970 et P. riouxi Depaquit, Killick-Kendrick & Léger, 1998 (Diptera: Psychodidae).

In 1998, the description of Phlebotomus riouxi emphasised the difficulty to differentiate the female from the closely related P. chabaudi, a suspected vector of Leishmania killicki in several foci in Tunisia. In order to be able to distinguish the females of these two species, we started a molecular study based on 37 Algerian and Tunisian specimens. The alignment of the sequences of the cytochrome b of the mitochondrial DNA and their analysis using Neighbor-Joining, maximum likelihood and maximum parsimony shows the individualisation of two species including an intraspecific variability. Following a morphological approach, it is not possible to distinguish the females on the basis of their spermathecae. A new character is proposed: the presence of anterolateral teeth of the pharyngeal armature for P. chabaudi, never observed in P. riouxi. However, a molecular typing is necessary at the present time for a sure identification of the females.

Geographical variation in populations of Phlebotomus (Paraphlebotomus) caucasicus (Diptera: Psychodidae) in Iran.

A comparative morphological and molecular study was carried out on 11 different populations of Phlebotomus (Paraphlebotomus) caucasicus Marzinovsky 1917 caught in 7 provinces in Iran (2004-2005). Differences in the implantation level of the two distal spines of the style, the number of setae of the basal lobe of coxite, and the length of the third antennal segment, revealed the existence of two morphotypes within P. (Pa) caucasicus, a species having a confused history if we take into account an unclear synonymisation with Phlebotomus (Paraphlebotomus) grimmi Porchinsky 1874. Sequencing of mtDNA (a fragment of cytchromeB gene, tRNA for serine gene and a fragment of NADH1 gene) and Neighbour-Joining analysis showed a partial correlation between morphotypes and haplotypes. We also found a correlation between the latter and the geographical origin of the specimens. These results need further studies in order to appreciate the role of each morphotype/haplotype in the transmission of Leishmania major.

Seasonal morphological variations in bird schistosomes.

The present work is a contribution to the systematics of Bilhorziello and Dendritobilharzio. Wildfowl was killed in hunting seasons or found dead in Champagne-Ardenne region, France, and autopsied with focus on schistosomes. Seven Anas plotyrhynchos (mallards), one Ardeo cinerea (grey heron) and two Cygnus olor (mute swans) were parasitized by Bilharziella. One C. olor was parasitized by Dendritobilharzia. Depending on season and hosts, various morphological forms of Bilharziello suggesting several species were observed. The differences in male and female worms concerned the morphology of genital apparatus, the spination on suckers, the body size and proportions. However, the comparison of DNA sequences led to a conclusion that these forms belonged to one species, Bilharziella polonica (Kowalewski, 1895). The morphological features and the body sizes of our samples of Dendritobilharzia seemed to differ from the type species of D. pulverulenta (Braun, 1901). Nevertheless, molecular analysis confirmed identity. We hypothesize that the differences in Bilharziella and Dendritobilharzia might be linked to internal host factors (e.g. hormonal levels), and influenced by season, host, and worm age. The definition of the genera Bilharziello and Dendritobilharzio was amended.

Phlebotomine sand flies from Ouagadougou, Burkina Faso: first record of Phlebotomus (Larroussius) longicuspis south of the Sahara.

During a brief entomological survey carried out in Ouagadougou, Burkina Faso, in March 2002, 178 phlebotomine sand flies (Diptera, Psychodidae) were caught using CDC miniature light traps. They were identified as Phlebotomus duboscqi Neveu-Lemaire, P. longicuspis Nitzulescu, Sergentomyia adleri (Theodor), S. affinis vorax (Parrot), S. antennata (Newstead), S. bedfordi (Newstead), S. christophersi (Sinton), S. fallax (Parrot), S. magna (Sinton) and S. schwetzi (Adler, Theodor & Parrot). This is the first record south of the Sahara of P. longicuspis sensu lato. This proven vector of Leishmania infantum Nicolle is a species closely related to P. perniciosus. It was identified by morphology and by the sequencing of a fragment of mitochondrial cytochrome b. There was 100% sequence homology with typical Tunisian specimens, and all characters identified the specimen as P. longicuspis sensu stricto. Epidemiological and biogeographical consequences are discussed.

[Phlebotomus from Madagascar (Diptera: Psychodidae). II--Description of the female Phlebotomus (Anaphlebotomus) fertei Depaquit, Léger & Robert, 2002; description of the male and new data for the female Phlebotomus (Anaphlebotomus) berentiensis (Léger & Rodhain, 1978) comb. nov]. / Les phlébotomes de Madagascar (Diptera: Psychodidae). II--Description de la femelle de Phlebotomus (Anaphlebotomus) fertei Depaquit, Léger & Robert, 2002; description du mâle et redescription de la femelle de Phlebotomus (Anaphlebotomus) berentiensis (Léger & Rodhain, 1978) comb. nov.

The female of Phlebotomus (Anaphlebotomus) fertei Depaquit, Léger & Robert, 2002 is described from Madagascar. The spermathecae have separated ducts, a body with an enlarged-sclerified dilatation and a deriviated large bladder with thin walls and long head. P. fertei female has the following characters: long antennal segments, pharyngeal armature with small teeth and cibarium with 15 to 30 denticles, and very long legs. Its remarkable spermatheca is closely related to that of P. berentiensis. The status of P. (Anaphlebotomus) berentiensis (Léger & Rodhain, 1978) is revised into this paper. The female is redescribed and the male is described for the first time. The latter has an unusual antennal formula (2/III-XII), a cibarium with four teeth and about 30 denticles, no tuft of setae on the coxite, four spines on the style, including a very long proximal one. Males and females were linked to each other based on morphological and molecular characteristics (second internal transcribed spacer of the ribosomal DNA and ND4 gene of the mitochondrial DNA). The inclusion of these taxa into the subgenus Anaphlebotomus Theodor, 1948 is exclusively based on male characters as listed by Theodor (style with four spines, no basal lobe on the coxite, paramere with two or three branches). The very heterogeneous characters of the females do not allow their use for the definition of this subgenus, which is probably not homogeneous.

Phylogeny of the Trichostrongylina (Nematoda) inferred from 28S rDNA sequences.

We produced a molecular phylogeny of species within the order Strongylida (bursate nematodes) using the D1 and D2 domains of 28S rDNA, with 23 new sequences for each domain. A first analysis using Caenorhabditis elegans as an outgroup produced a tree with low resolution in which three taxa (Dictyocaulus filaria, Dictyocaulus noerneri, and Metastrongylus pudendotectus) showed highly divergent sequences. In a second analysis, these three species and C. elegans were removed and an Ancylostomatina, Bunostomum trigonocephalum, was chosen (on the basis of previous morphological analyses) as the outgroup for an analysis of the phylogenetic relationships between and within the Strongylina (strongyles) and Trichostrongylina (trichostrongyles). A very robust tree was obtained. The Trichostrongylina were monophyletic, but the Strongylina were paraphyletic, though this requires confirmation. Within the Trichostrongylina, the three superfamilies defined from morphological characters are confirmed, with the Trichostrongyloidea sister group to a clade including the Molineoidea and Heligmosomoidea. Within the Trichostrongyloidea, the Cooperiidae, Trichostrongylidae, and Haemonchidae were polytomous, the Haemonchinae were monophyletic, but the Ostertagiinae were paraphyletic. The sister-group relationships between Molineoidea and Heligmosomoidea were unsuspected from previous morphological analysis. No unequivocal morphological synapomorphy could be found for the grouping Molineoidea + Heligmosomoidea, but none was found which contradicted it.

[Phlebotomines of the Isle of Cyprus. III. Species inventory ]. / Les phlébotomes de l'Ile de Chypre. III--Inventaire faunistique.

Three surveys on canine leishmaniasis were carried out in Cyprus (1993, 1998 and 1999) emphasise the presence of eleven species of phlebotomine sandflies: Phlebotomus (Phlebotomus) papatasi, P. (Paraphlebotomus) alexandri, P. (Pa.) jacusieli (first mention in Cyprus), P. (Pa.) sergenti, P. (Larroussius) galilaeus, P. (L.) tobbi, P. (Transphlebotomus) economidesi, P. (T.) mascittii, Sergentomyia (Sergentomyia) azizi (its specific statute is validated by the authors), S. (S.) fallax et S. (S.) minuta. P. (Adlerius) kyreniae was not caught during the surveys. The authors propose hypothesis of settlement of the island by phlebotomine sandflies according this species inventory. A first migration period took probably place during the Miocene time and a second one during the Pleistocene time.

[The subgenus Paraphlebotomus (Phlebotomus--Phlebotominae-- Psychodidae--Diptera): a review. Morphological and molecular studies]. / Révision du sous-genre Paraphlebotomus (Phlebotomus - Phlebotominae - Psychodidae - Diptera). Approches morphologique et moléculaire.

A systematic review of the subgenus Paraphlebotomus (a group of phlebotomine sandflies which contains several vectors of leishmaniasis) is proposed. In the morphological approach, we study the available types and a lot of sandflies coming from different geographic areas. The taxonomic status of each species is revised. We consider P. marismortui as a synonym of P. alexandri. We suppose that P. mofidii is a valid species. P. similis is clearly differentiated from P. sergenti. A new species is described from North Africa: P. riouxi. We discuss the validity of P. saevus. Two molecular phylogenies based on ribosomal DNA sequences are proposed. The first one used the D2 domain of the 28 S sub-unit. It emphasises a very close relation between the subgenera Paraphlebotomus and Phlebotomus, a likely paraphyly of the genus Phlebotomus and the position of the genera Sergentomyia and Lutzomyia as two sister groups. The second one used the ITS 2. It emphasises the informative capacity of this gene for the resolution of the phylogenetic relations within the subgenera Paraphlebotomus.

Status and origin of Haemonchinae (Nematoda: Trichostrongylidae) in deer: a survey conducted in France from 1985 to 1998.

During our investigations on helminthofauna in roe deer (Capreolus capreolus) and red deer (Cervus elaphus) in France (1985-1998) we isolated nematodes not only of the genus Haemonchus Cobb, 1898 but also of the genus Ashworthius Le Roux 1930, both of which belong to the same subfamily of Haemonchinae. The prevalence of Ashworthius was 22% (65/294) in roe deer and 40% (10/25) in red deer. H. contortus was not found in red deer, whereas its prevalence was only 3% (9/294) in roe deer. These data contrast with those observed in other European countries. The presence of A. sidemi in red and roe deers in France suggests a case of parasitism imported via sika deer. A hypothesis of their spread is proposed.

Molecular systematics of the phlebotomine sandflies of the subgenus Paraphlebotomus (diptera, psychodidae, phlebotomus) based on ITS2 rDNA sequences. Hypotheses Of dispersion and speciation.

Phylogenetic Paraphlebotomus relationships are inferred by a study based on the sequences of ITS2, which has been sequenced in nine Paraphlebotomus species: P. alexandri, P. andrejevi, P. jacusieli, P. kazeruni, P. mireillae, P. mongolensis, P. saevus, P. sergenti and P. similis and in two out-groups species of the subgenus Phlebotomus: P. papatasi and P. duboscqi. Paraphlebotomus alexandri appears as the sister group of all other Paraphlebotomus sandflies. Among the other species, three groupings are clearly highlighted: andrejevi and mongolensis; mireillae and saevus; jacusieli, kazeruni, sergenti and similis. These groupings are related to speculations about the migration of Paraphlebotomus from a centre of dispersion located in the Middle East sometime from the early Eocene to the late Miocene.

[Phlebotomine sandflies (Diptera-Psychodidae) of the isle of Cyprus. I--Description of Phlebotomus (Transphlebotomus economidesi n. sp]. / Les phlébotomes (Diptera-Psychodidae) de l'île de Chypre. I--Description de Phlebotomus (Transphlebotomus) economidesi n. sp.

Description of a new palearctic species from Cyprus: Phlebotomus (Transphlebotomus) economidesi. The deposited holotype is a female because of the particularity of the streaked spermathecae and its non dilated ducts. Four to six spines are implanted on the style of the male, but its differential diagnosis with the closely related species P. mascittii and P. canaaniticus is difficult. The distributions of these Transphlebotomus species are mentioned.