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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.

Trematodes can adversely impact the health and survival of wild animals. The trematode family Cyclocoelidae, which includes large digenean bird parasites, lacks molecular analysis, and reclassifications have not been supported. This study produced the first fully assembled and annotated mitochondrial genome sequence for the trematode Morishitium polonicum . The whole length of the M. polonicum (GenBank accession number: OP930879) mitogenome is 14083 bp, containing 22 transfer ribonucleic acids (tRNAs), 2 ribosomal RNAs (rRNAs, rrnL and rrnS ), and a noncoding control section (D-loop) 13777 to 13854 bp in length. The 12 PCG areas have 3269 codons and a total length of 10053 bp, which makes up 71.38% of the mitochondrial genome ’ s overall sequence. Most (10/12) of the PCGs that code for proteins begin with ATG, while the nad4L and nad1 genes have a GTG start codon. Phylogenetic analysis using the concatenated nucleotide sequences of 12 PCGs, and the ML tree analysis results showed that M. polonicum is more closely related to with Echinostomatidae and Fasciolidae, which indicates that the family Cyclocoelidae is more closely associated with Echinochasmidae. This study provides mtDNA information, and analysis of mitogenomic structure and evolution. Moreover, we aimed to understand the phylogenetic relationships of this fluke.

In light of the morphological and molecular data for cyclocoelids observed from the air sacs of Mareca strepera (Linnaeus) (Anatidae, Anseriformes) caught in the southern region of the Russian Far East, we suggest new insights into the systematics of the family Cyclocoelidae. A comparative morphological and phylogenetic analyzes revealed that new cyclocoelids represented the new genus and species Paracyclocoelum lobatum. Based on the 28S rRNA gene fragment we showed the significant genetic divergence of P. lobatum from the type species of the type genus for the family, Cyclocoelum mutabile (Zeder, 1800) Brandes, 1892 and along with the confusing morphological similarity by the prepharyngeal position of the genital pore it most likely indicate homologous development of the reproductive system of Paracyclocoelum and Cyclocoelum. Here, we provide a new dichotomous key for five cyclocoelid genera from the subfamily Cyclocoelinae including Paracyclocoelumn. g. The new genus Paracyclocoelum had sister relationship to the cyclocoelin genus Circumvitellatrema. Based on the polyphyletic interrelationships of Cyclocoelum and Circumvitellatrema the Cyclocoelinae were assigned with the status sensu lato.

The complete mitochondrial sequence of 17,030 bp was obtained from Echinostoma revolutum and characterized with those of previously reported members of the superfamily Echinostomatoidea, i.e. six echinostomatids, one echinochasmid, five fasciolids, one himasthlid, and two cyclocoelids. Relationship within suborders and between superfamilies, such as Echinostomata, Pronocephalata, Troglotremata, Opisthorchiata, and Xiphiditata, are also considered. It contained 12 protein-coding, two ribosomal RNA, 22 transfer RNA genes and a tandem repetitive consisting non-coding region (NCR). The gene order, one way-positive transcription, the absence of atp 8 and the overlapped region by 40 bp between nad 4L and nad 4 genes were similar as in common trematodes. The NCR located between tRNA Glu ( trn E) and cox 3 contained 11 long (LRUs) and short repeat units (SRUs) (seven LRUs of 317 bp, four SRUs of 207 bp each), and an internal spacer sequence between LRU7 and SRU4 specifying high-level polymorphism. Special DHU-arm missing tRNAs for Serine were found for both tRNA S1(AGN) and tRNA S2(UCN) . Echinostoma revolutum indicated the lowest divergence rate to E. miyagawai and the highest to Tracheophilus cymbius and Echinochasmus japonicus . The usage of ATG/GTG start and TAG/TAA stop codons, the AT composition bias, the negative AT-skewness, and the most for Phe/Leu/Val and the least for Arg/Asn/Asp codons were noted. Topology indicated the monophyletic position of E. revolutum to E. miyagawai . Monophyly of Echinostomatidae and Fasciolidae was clearly solved with respect to Echinochasmidae, Himasthlidae, and Cyclocoelidae which were rendered paraphyletic in the suborder Echinostomata.

Typhlocoelum cucumerinum is a tracheal parasite of birds widely distributed across the globe. Nevertheless, aspects of the biology of this cyclocoelid are still poorly understood. Herein, we report the finding of T. cucumerinum in definitive and intermediate hosts from an urban waterbody of Brazil. The parasite was initially detected during the necropsy of domestic Muscovy ducks (Cairina moschata) found dead in the locality. Coproparasitological tests in live animals revealed that 12/47 (25.53%) Muscovy ducks and 2/8 (25%) mallards (Anas platyrhynchos platyrhynchos) were infected with T. cucumerinum. Moreover, rediae and metacercariae morphologically similar to T. cucumerinum were found in 3/248 (1.33%) Biomphalaria straminea collected in the same waterbody frequented by the birds. The conspecificity between the adult and the larval stages was confirmed molecularly (100% similarity in Cox-1). Moreover, the phylogenetic position of T. cucumerinum was determined for the first time based on partial fragments of the 28S, Cox-1 and Nad-1 genes. The species grouped with other members of the subfamily Typhlocoelinae with sequences available, but the data obtained do not support the distinctiveness of the genera Typhlocoelum and Tracheophilus. Further studies involving a broader range of species can result in taxonomic rearrangements in Typhlocoelinae.

The taxonomic status of many species of the family Cyclocoelidae is still unclear. Two species of cyclocoelids, Harrahium obscurum and Morishitium dollfusi, were collected from the air sacs of birds (Turdus merula and Tringa ochropus) inhabiting the Middle Volga region (European Russia). Here, we provide the first detailed morphological description of these cyclocoelids and combine it with the first molecular phylogenetic analysis of Cyclocoelidae from birds in Russia based on partial sequences of their 28s rDNA and coI mtDNA genes. Specimens of both flatworm species from different host individuals differ slightly in body shape and size, which probably reflects host-induced intraspecific variability. For the first time, we have shown that a stable morphological character such as the length of the vitelline fields in the studied digeneans is variable at the species level and cannot be used in their morphological diagnosis.

Tracheophilus cymbius (Trematoda: Cyclocoelidae) is a common tracheal fluke of waterfowl, causing serious loss in the poultry industry. However, taxonomic identification of T. cymbius remains controversial and confused. Mitochondrial (mt) genomes can provide genetic markers for the identification of closely related species. We determined the mt genome of T. cymbius and reconstructed phylogenies with other trematodes. The T. cymbius mt genome is 13,760 bp in size, and contains 12 protein-coding genes ( cox 1 – 3, nad 1 – 6, nad 4L, cyt b and atp 6), 22 transfer RNA (tRNA) genes, two ribosomal RNA genes and one non-coding region. All are transcribed in the same direction. The A + T content is 62.82%. ATG and TAG are the most common initiation and termination codons, respectively. Phylogenetic analyses of concatenated nucleotide sequences show T. cymbius grouping in suborder Echinostomata, and clustering together, with high statistical support, as a sister taxon with Echinochasmus japonicus (Echinochasmidae), the two forming a distinct branch rooted to the ancestor of all Echinostomatidae and Fasciolidae species. This is the first report of the T. cymbius mt genome, and the first reported mt genome within the family Cyclocoelidae. These data will provide a significant resource of molecular markers for studying the taxonomy, population genetics and systematics of trematodes.

Trematodes of the family Cyclocoelidae are parasites mainly of the respiratory system of birds and present a cosmopolitan distribution. Although infection with these flukes can result in pathological changes and even bird death, information on their life cycles is scarce and almost entirely based on experimental infection data. Thus, the generation of knowledge on the mollusks that act as natural intermediate hosts of cyclocoelids is necessary and can aid control measures against these air sac trematodes. In the present study, gastropod mollusks collected in an urban stream from Belo Horizonte, Minas Gerais, Brazil, were subjected to the compression technique for the detection of non-emerging larval trematodes. Tailless cercariae with confluent ceca were found in 8/30 (26.7%) specimens of Biomphalaria glabrata and 3/33 (9.1%) specimen of Physella acuta. Samples of the cercariae were subjected to morphological characterization and genetic study (28S, Cox-1, and Nad-1). For comparative purposes, adult trematodes previously collected in the air sac of a common gallinule (Gallinula galeata) found dead in another waterbody from the same region were also characterized. The molecular sequences obtained revealed a high degree of similarity (100% in 28S, 99.2% in Cox-1, and 99.5% in Nad-1) between larval stages found in mollusks and adult parasites found in G. galeata and morphologically identified as Cyclocoelum mutabile. The conspecificity with this widely distributed cyclocoelid was also corroborated by phylogenetic analysis and comparison with isolates of this species previously characterized in Peru and the Czech Republic (99.4–100% and 96.7–97.0% of similarity in Nad-1, respectively). Thus, the integrative analysis carried out in the present work enabled us to identify C. mutabile in mollusks in South America for the first time. The finding of B. glabrata and P. acuta as new intermediate hosts corroborates the importance of freshwater gastropods in the transmission of C. mutabile, as well as the low specificity to the mollusk group, as previously characterized through experimental studies.

Cyclocoelidae Stossich, 1902 are medium-sized to large digenean bird parasites. Although these parasites bear few visible autapomorphic signs, and their diagnostic characters are unstable in response to the pressure applied during preparation, the numerous hitherto suggested re-classifications within the family have not been supported by any molecular analysis. We analyse here cyclocoelids found during the extensive examination of central European birds performed from 1962 to 2016, provide comparative measurements, host spectra, prevalence and intensity, and provide and analyse sequences of four DNA loci of five of the cyclocoelid species. Cyclocoleum Brandes, 1892 appears paraphyletic; thus we suggest the re-classification of Cyclocoleum obscurum (Leidy, 1887) as Harrahium obscurum (Leidy, 1887) Sitko and Heneberg comb. n. Molecular phylogenetics questioned also the validity of Cyclocoelinae Stossich, 1902 and Hyptiasminae Dollfus, 1948, which formed a single clade, whereas Allopyge Johnston, 1913, Prohyptiasmus Witenberg, 1923 and Morishitium Witenberg, 1928 formed another clade. Haematotrephinae Dollfus, 1948 are newly characterized as having a pretesticular or intertesticular ovary that forms a triangle with the testes. Analyses of non-European genera of the Cyclocoelidae and an examination of the position of families within Echinostomata La Rue, 1926 are needed.

Based on a morphological approach, five species of cyclocoelids (Digenea: Cyclocoelidae) are revised based on material from birds in Mexico. The species studied are: Cyclocoelum cf. leidyi Harrah, 1922 from the black-necked stilt Himantopus mexicanus Müller (Recurvirostridae); Cyclocoelum mutabile (Zeder, 1800) from the northern jacana Jacana spinosa L. (Jacanidae) and the willet Tringa semipalmata Gmelin (Scolopacidae); Cyclocoelum pseudomicrostomum Harrah, 1922 from the American coot Fulica americana Gmelin (Rallidae); Selfcoelum lamothei Blend & Dronen, 2008, from the long-billed curlew Numenius americanus Bechstein (Scolopacidae); and Neohaematotrephus arayae Zamparo, Brooks, Causey & Rodriguez, 2003 from J. spinosa. New morphological data, illustrations and measurements are presented for these trematodes. Three species, insufficiently described, i.e. C. cf. leidyi, C. mutabile and C. pseudomicrostomum, are redescribed in detail. Our results indicate morphological variation in some important features used in the diagnosis of Cyclocoelum Brandes, 1892 (i.e. the presence or absence of oral sucker, the position of the genital pore relative to the pharynx, and the posterior extension of the uterus) and Selfcoelum Dronen, Gardner & Jiménez, 2006 (i.e. anterior extension of vitelline follicles). Finally, this study provides the first molecular data for the large subunit of the ribosomal RNA (28S rRNA) gene for N. arayae.