NEW GENUS AND SPECIES OF CYCLOCOELIDAE STOSSICH, 1902 (PLATYHELMINTHES: DIGENEA) INFECTING THE NASOPHARYNGEAL CAVITY OF CANADA GOOSE, BRANTA CANADENSIS (ANSERIFORMES: ANATIDAE) FROM WESTERN ALABAMA.

While surveying the parasites of birds associated with western Alabama aquaculture ponds, we collected several specimens of Anativermis normdroneni n. gen., n. sp. (Digenea: Cyclocoelidae) from the nasopharyngeal cavity of a Canada goose, Branta canadensis (Linnaeus, 1758) (Anseriformes: Anatidae). These flukes were heat killed and fixed in neutral buffered formalin for morphology or preserved in 95% ethanol for DNA extraction. Anativermis resembles Morishitium (Witenberg, 1928) by having testes that are spheroid with smooth margins and located in the posterior quarter of the body, an anterior testis that is lateral to the midline and abuts the respective cecum, a posterior testis that is medial (testes diagonal) and abuts the cyclocoel, a genital pore that is immediately postpharyngeal, and a vitellarium that is discontinuous posteriorly. The new genus differs from Morishitium and is unique among all other cyclocoelid genera by having the combination of a body that is broadest in the anterior body half, a posterior body end that is more sharply tapered than the anterior body end, an ovary that nearly abuts the posterior testis, a vitellarium that is asymmetrical and distributes from the area immediately posterior to the cecal bifurcation posteriad to approximately the level of the ovary, and uterine loops extending dorsolateral to the ceca and filling the space between the ceca and the respective body margin for nearly the entire body length. The new genus was recovered as a distinct lineage in separate 28S, 18S, and ITS2 phylogenetic analyses. This is the first report of a cyclocoelid infecting the Canada goose and of a cyclocoelid from Alabama.

Characterization of the complete mitochondrial genome of Plagiorchis multiglandularis (Digenea, Plagiorchiidae): Comparison with the members of Xiphidiatan species and phylogenetic implications.

Plagiorchis multiglandularis Semenov, 1927 is a common fluke of birds and mammals, with significant impacts on animals and also human health. However, the systematics of Plagiorchiidae remain ambiguous. In the present study, the complete mitochondrial (mt) genome of P. multiglandularis cercariae was sequenced and compared with other digeneans in the order Xiphidiata. The complete circular mt genome of P. multiglandularis was 14,228 bp in length. The mitogenome contains 12 protein-coding genes and 22 transfer RNA genes. The 3' end of nad4L overlaps the 5' end of nad4 by 40 bp, while the atp8 gene is absent. Twenty-one transfer RNA genes transcribe products with conventional cloverleaf structures, while one transfer RNA gene has unpaired D-arms. Comparative analysis with related digenean trematodes revealed that A + T content of mt genome of P. multiglandularis was significantly higher among all the xiphidiatan trematodes. Phylogenetic analyses demonstrated that Plagiorchiidae formed a monophyletic branch, in which Plagiorchiidae are more closely related to Paragonimidae than Prosthogonimidae. Our data enhanced the Plagiorchis mt genome database and provides molecular resources for further studies of Plagiorchiidae taxonomy, population genetics and systematics.

First molecular assessment of two digenean parasites of the lancehead snake Bothrops moojeni Hoge, 1966 (Serpentes, Viperidae) in Brazil.

Two digenean species, Infidum infidum Faria, 1910 (Dicrocoeliidae) and Travtrema stenocotyle Cohn, 1902 (Plagiorchiidae), were collected in the large pit viper Bothrops moojeni Hoge, 1966 from Reserva Particular do Patrimônio Natural Cisalpina, municipality of Brasilândia, Mato Grosso do Sul State, Brazil. In this study, we provide the first molecular characterisation using the 28S rDNA and phylogenetic position data of these two common digeneans from B. moojeni. The molecular framework revealed topologies with strongly supported clades using maximum likelihood and Bayesian inference methods, positioned I. infidum among Plagiorchiidae and not among Dicrocoeliidae as expected and T. stenocotyle (Plagiorchiidae) surprisingly grouped as a sister group to Allassogonoporidae, Microphallidae, Pleurogenidae, and Prosthogonimidae, not related to plagiorchids. Our molecular phylogenetic data showed that these species may not correspond to their assigned families and encourage future studies on the systematic of these understudied groups.

Characterization of the complete mitochondrial genome of Plagiorchis maculosus (Digenea, Plagiorchiidae), Representative of a taxonomically complex digenean family.

Despite the highly divergent morphology, pathogenicity and worldwide distribution of digenean parasites belonging to one of the largest families, the Plagiorchiidae, there are no complete mitochondrial (mt) genomes published to date for plagiorchiids. In this study, we obtained nuclear ribosomal DNA (ITS region and 28S rDNA) sequences and the complete mt genome sequences of Plagiorchis maculosus (Rudolphi 1802) Braun, 1902, and assessed its phylogenetic relationship with other xiphidiates, based on the mtDNA sequences. The obtained ITS and 28S rDNA sequences were identical to the corresponding sequences of P. maculosus available in GenBank. The complete mitochondrial genome of P. maculosus (14,124 bp) contained 36 genes (atp8 is absent) and a long non-coding region (NCR) with two sets of repeated sequences of 283 nucleotides each. The phylogenetic tree resulting from Bayesian inference (BI) analyses based on concatenated nucleotide sequences of all 36 genes of P. maculosus and other xiphidiates mitochondrial genomes, indicated that P. maculosus (and the Plagiorchiidae) is phylogenetically closest to the Brachycladiidae and Paragonimidae. The present study describes the first mitochondrial genome from the type genus of the family Plagiorchiidae. The overall gene arrangement, nucleotide composition, A + T contents, AT and GC skew and codon usage with relative synonymous codon usage (RSCU) for 12 PCGs are described. Characterization of mitochondrial genomes from additional plagiorchiid taxa is necessary to make further progress in phylogenetic and epidemiological studies of these digeneans as well as accurate diagnostics of these parasites including those parasitic in humans.

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.

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

The ultrastructure of the mature spermatozoon of the type genus of the Plagiorchiidae Plagiorchis elegans (Rudolphi, 1802), a parasite of the Golden hamster, Mesocricetus auratus is described. This study is the first ultrastructural study of the spermatozoon of a Plagiorchis, the second of a plagiorchiid species and only the third in the Plagiorchioidea. Previously data on spermatozoon ultrastructure existed only for the plagiorchiid Enodiotrema reductum and the omphalometrid Rubenstrema exasperatum. The mature spermatozoon of P. elegans exhibited the general pattern described in most digenean species, namely two axonemes of the 9 + "1" Trepaxonemata pattern, nucleus, mitochondria, external ornamentation of the plasma membrane, spine-like bodies, and glycogen granules. However, the rather typical expansion of the plasma membrane is not found in P. elegans. Another peculiarity of the spermatozoon of P. elegans is the presence of a structure called thin cytoplasm termination. Spermatozoon ultrastructure of P. elegans is compared with that of E. reductum and R. exasperatum. Spermatozoon of P. elegans conforms to the general pattern described in E. reductum. Thus, this study further expands our knowledge on the spermatozoon ultrastructure among the members of the Plagiorchioidea, one of the most phylogenetically derived groups of the digenea.

New record of Haplometroides intercaecalis (Digenea, Plagiorchiidae) infecting a Brazilian snake

Phalotris matogrossensis (Serpentes, Colubridae) was reported as a new host for Haplometroides intercaecalis (Digenea, Plagiorchiidae). The host snake was obtained from the municipality of Anastácio, state of Mato Grosso do Sul, Brazil. One specimen of H. intercaecalis was recovered from the esophagus of the host and identified by the intercecal position of the vitellaria in the pre-acetabular region. This paper describes the second report of the occurrence of this trematode in fossorial snakes of the genus Phalotris in the state of Mato Grosso do Sul, Brazil.

New reports on parasitism by Haplometroides buccicola (Digenea, Plagiorchiidae) in Brazilian snakes

The occurrence of Haplometroides buccicola (Digenea, Plagiorchiidae) in the esophagus of two Brazilian snakes is reported in the present study. The trematodes were collected from one Micrurus corallinus (Elapidae) and one Phalotris lativittatus (Colubridae); both snakes were found in Botucatu city, São Paulo State, Brazil. Morphological and morphometric analyses of the trematodes are presented. For the first time Micrurus corallinus has been recorded as a host for H. buccicola and this is the second time that P. lativittatus has been reported as a host for this trematode species.