DESCRIPTIONS AND PHYLOGENETIC AFFINITIES OF TWO NEW SPECIES OF RHABDIAS STILES AND HASSALL, 1905 (NEMATODA: RHABDIASIDAE) FROM NORTH AMERICAN FROGS: ARE WE STILL ONLY SCRATCHING THE SURFACE?

Two new species of lung-dwelling nematodes are described from North American frogs: Rhabdias aurorae n. sp. from Rana aurora and Rhabdias conni n. sp. from Rana clamitans and Rana catesbeiana from Arkansas; the latter species was also found in Oklahoma and Georgia. Rhabdias aurorae n. sp. differs from other Nearctic congeners in the combination of the following characteristics: buccal capsule 22-25 µm wide, elongated tail covered with inflated cuticle, esophagus with prominent dilatation in anterior part and 6 small circumoral lips. Rhabdias conni n. sp. is morphologically closest to Rhabdias ranae Walton, 1929 and Rhabdias joaquinensisIngles, 1936; it differs from them in the shape of lateral pseudolabia, the dimensions of the body, and the egg size. Both new species were found to be significantly different from the Nearctic congeners in the nucleotide sequences of nuclear ribosomal DNA (18S-ITS-28S region), 12S, and CO1 mitochondrial genes. The 2 new species differ from other currently sequenced Nearctic congeners by 1.1-2.7% of nucleotide positions in the nuclear rDNA region, 1.3-3.4% in the 12S gene, and 3.4-9.4% in CO1 gene. Molecular phylogenetic analysis based on nuclear ribosomal DNA sequences placed both new species into the clade consisting of Nearctic and Neotropical Rhabdias spp. The position of Rh. aurorae n. sp. within the clade is uncertain because of a polytomy, but Rh. conni n. sp. is nested within the "Rh. joaquinensis complex" related to Rh. ranae and Rhabdias tarichaeKuzmin, Tkach, and Snyder, 2003. The phylogenetic analysis based on nuclear ribosomal DNA sequences has revealed 3 evolutionary host-switching events from anuran to caudatan hosts among Rhabdias spp. that occurred in the Nearctic and Palearctic. The molecular phylogeny also suggests that Rhabdias may have originally evolved in what is now Africa.

Description of Ornithodoros (Pavlovskyella) tartakovskyi using scanning electron microscopy, with notes on the morphology of Pavlovskyella sensu stricto and Theriodoros subgenera.

Accumulation of DNA sequence data and its use in systematics of the family Argasidae reveals new incongruencies between genera and subgenera, since several groups defined by classical taxonomy appear to be paraphyletic, which is the case of the subgenus Pavlovskyella. In order to identify morphological characters unique to one of the monophyletic groupings within Pavlovskyella and improve its system, we describe all active stages of Ornithodoros (Pavlovskyella) tartakovskyi, a species with an incomplete original description. Larvae, nymphs, males and females from Iran were examined with scanning electron microscopy (SEM). The larva of O. (P.) tartakovskyi lacks dorsal plate, posteromedian seta, postcoxal setae and a spinose area in palpal article I; the dorsal surface has 13 pairs of setae and the hypostome has two short rows of denticles in the apex only. The first nymphal instar (N1) has a micromammillated body with faint dorsal and ventral disks. The second nymphal instar (N2) shows mammillated body with an incipient hood, and dorsal disks outlined as in subsequent instars. Both N1 and N2 have a small patch of glabrous surface in the anterior margin of preanal groove, absent in posterior stages. Third nymphal instar (N3) has a small hood and lack cheeks, which are incipient in the fourth nymphal instar (N4). The four nymphal instars (N1-N4) have blunt hypostomes, striated capitula, non-mammillated legs, and 3 humps on tarsi I. Females and males of O. (P.) tartakovskyi have mammillated bodies, small cheeks not covering the capitulum, the anteromedian disk is placed anteriorly to anterior central disks, and a posterior median file of disks is merged with the median disk. We describe all postembryonic stages of O. (P.) tartakovskyi providing morphological characters that define Pavlovskyella sensu stricto and Theriodoros, a sister subgenus that includes similarly shaped species.

Proterodiplostomid no longer: Molecular phylogeny reveals the true position of Proalarioides (Digenea: Diplostomoidea).

Proalarioides Yamaguti, 1933 (Digenea Carus, 1863: Diplostomoidea Poirier, 1886) is a small genus of proterodiplostomids parasitic in the intestines of snakes in Asia. Only two species are considered valid: Proalarioides serpentis Yamaguti, 1933 and Proalarioides tropidonotis Vidyarthi, 1937. Unlike other proterodiplostomids, Proalarioides spp. possess pseudosuckers and lack the paraprostate, otherwise extremely characteristic of the Proterodiplostomidae Dubois, 1936. In the present study, we describe the morphology of progenetic metacercariae of a Proalarioides sp. from bicolored frog, Clinotarsus curtipes (Jerdon), collected in India and provide the first DNA sequences from any member of the genus. These specimens differ from previously described metacercariae and adults of P. serpentis and P. tropidonotis in several ways, including body and organ sizes, sucker ratios, and distribution of vitellarium. The newly generated partial large ribosomal subunit (28S) rRNA gene sequence was used to test the phylogenetic position of the genus among other major lineages of diplostomoideans. Our 28S phylogeny clearly demonstrated Proalarioides sp. to be well-separated from other members of the Proterodiplostomidae. Based on morphological and molecular evidence, we transfer Proalarioides out of the Proterodiplostomidae into the Diplostomidae Poirier, 1886.

Uncovering further diversity of Ochoterenatrema Caballero, 1943 (Digenea: Lecithodendriidae) in South American bats.

Ochoterenatrema Caballero, 1943 is a genus of lecithodendriid digeneans that prior to this study included 8 species parasitic in bats in the Western Hemisphere. Species of Ochoterenatrema possess a unique morphological feature in form of the pseudogonotyl on the sinistral side of the ventral sucker. In this study, we describe 2 new species of Ochoterenatrema from bats in Ecuador. The new species are readily differentiated from their congeners by a combination of morphological characters, including the distribution of vitelline follicles, length of oesophagus, sucker ratio and the body shape, among other features. We have generated partial nuclear 28S rDNA and mitochondrial cox1 gene DNA sequences from both new species. The newly obtained sequences were used to differentiate among species and study the phylogenetic interrelationships among Ochoterenatrema spp. The internal topology of the clade was weakly supported, although the cox1 tree was much better resolved than the 28S tree. Comparison of sequences revealed 0-1.2% interspecific divergence in 28S and 3.3-20.5% interspecific divergence in cox1 among Ochoterenatrema spp. The new findings demonstrate that bats in South America likely harbor multiple additional undescribed species of Ochoterenatrema. More extensive sampling from broader geographic and host ranges, especially in North America, should allow for a better understanding of the evolution of host associations and morphological traits of this lineage of lecithodendriid digeneans.

Checklist of helminth parasites of Chiroptera of North America north of Mexico.

A parasite-host, host-parasite and distribution-based checklist of helminths found in bats (Chiroptera) of North America north of Mexico is presented. The parasite-host checklist includes a total of 93 species (including records without a species identification) of helminth parasites reported in the literature from 30 species of bats. These include 54 trematodes, 11 cestodes, and 28 nematodes. Each helminth species is listed under its most current accepted name, with all known synonyms, distribution by state/province, and references for each geographic location. Lists of helminths reported from individual species of bats as well as states of the United States and provinces/territories of Canada are also provided. The following new combinations are proposed: Paralecithodendrium alaskensis (Neiland, 1962) n. comb. for Prosthodendrium alaskensis Neiland, 1962; Paralecthodendrium longiforme (Bhalerao, 1926) n. comb. for Lecithodendrium longiforme Bhalerao, 1926; and Paralecithodendrium singularium (Byrd & Macy, 1942) n. comb. for Prosthodendrium singularium Byrd & Macy, 1942. The state of knowledge of helminths of bats in North America is briefly discussed.

TAXONOMIC REASSESSMENT AND MORPHOLOGICAL REDESCRIPTION OF NEMATOSTRIGEA SERPENS ANNULATA (DIGENEA: STRIGEIDAE) FROM OSPREY IN NORTH AMERICA.

Digenean trematodes in the strigeid genus Nematostrigea are parasites of various birds, most often ospreys (Pandion haliaetus). Nematostrigea currently contains 2 species, Nematostrigea serpens and Nematostigea hepatica. Nematostrigea serpens is divided into 2 subspecies: N. serpens serpens from the Palearctic and Indomalayan realms and N. serpens annulata from the Nearctic realm. In the present work, we studied the type material of N. serpens annulata and collected new specimens from an osprey in Wisconsin close to the type locality. The original description and illustration of N. serpens annulata were incomplete. Herein we provide a detailed redescription and new illustrations of N. serpens annulata on the basis of the type material as well as newly collected and sequenced specimens. Nematostrigea serpens annulata has a distinct tegumental thickening near the mid-length of the genital cone, which is absent in its congeners. Partial sequences of the large ribosomal subunit (28S) DNA and cytochrome c oxidase subunit 1 (COI) mitochondrial DNA gene sequences were generated from our specimens of N. serpens annulata. Comparisons with previously published sequences of N. serpens serpens from the Palearctic demonstrated 2.2% difference in 28S and 3.7% in COI. On the basis of morphological and molecular comparisons, we elevate N. serpens annulata to species status and recognize it as Nematostrigea annulata Dubois and Rausch, 1948.

Jumping continents and major host lineages: phylogeny and diversity of the enigmatic Cryptotropidae (Platyhelminthes: Digenea).

Members of several genera in the digenean superfamily Microphalloidea, namely Renschetrema, Rohdetrema, Cryptotropa, Cephalouterina and Pseudocryptotropa, are characterized by an unusual dorsal position of the genital atrium. In the absence of phylogenetic data, their systematic position has been unstable. In the most recent taxonomic revision of the Microphalloidea, they were housed in three different families. We studied the morphology and obtained DNA sequences of several microphallolideans with a dorsal genital pore, collected from vertebrates in Southeast Asia and South America. We used sequences of the nuclear 28S gene to infer a superfamily-level phylogeny and sequences of the mitochondrial cox1 gene for family-level phylogeny and species-level comparisons. Based on the combination of molecular phylogenetic data and morphological features, we restore the Cryptotropidae and synonymize the Renschetrematidae with the Cryptotropidae. We erect new genera Paracryptotropagen. nov. and Armadoatriumgen. nov., provide amended diagnoses of the Cryptotropidae and Renschetrema and provide keys to the identification of genera within the family. We resurrect the genus Novetrema, previously synonymized with Pseudocryptotropa, and remove Renschetrema indicum from Renschetrema. We describe four new species from lizards in Vietnam and birds in the Philippines and Peru. The unique morphological features and unusually broad host associations and geographical distributions of cryptotropids are discussed.

TAPEWORMS (CESTODA: OPHIOTAENIA) FROM THE NORTHERN COTTONMOUTH (AGKISTRODON PISCIVORUS).

The northern cottonmouth, Agkistrodon piscivorus (Lacépède, 1789) (Viperidae: Crotalinae), occurs in the southeastern United States and is one of the few semiaquatic vipers in the world. Three proteocephalid tapeworms of the genus OphiotaeniaLa Rue, 1911 (Cestoda), have been described from this venomous snake. A critical evaluation of type specimens and tapeworms recently collected from A. piscivorus has revealed that only 2 species, Ophiotaenia marenzelleri (Barrois, 1898) and Ophiotaenia grandisLa Rue, 1911 (new synonym Ophiotaenia agkistrodontis [Harwood, 1933]), are specific parasites of this crotaline viper; both species are redescribed here. Ophiotaenia grandis was originally described from a mixture of 2 species: 'true' O. grandis, which is indistinguishable from O. agkistrodontis described 22 yr later and thus considered to be a junior synonym of O. grandis, and O. marenzelleri. Ophiotaenia marenzelleri, also reported from the pygmy rattlesnake, Sistrurus miliarius (Linnaeus, 1766), is a larger tapeworm with a massive scolex, a large cirrus sac, a very large, annular vaginal sphincter near the equatorial gonopore, and an oval, 3-layered embryophore surrounded by a nearly square hyaline outer membrane. Ophiotaenia grandis is much smaller and more slender and has a narrower scolex, a smaller cirrus sac and vaginal sphincter, a 2-layered embryophore, and a distinctly pre-equatorial gonopore. In addition to O. marenzelleri and O. grandis, other Ophiotaenia species typical of semiaquatic snakes (Colubridae: Natricinae) may be present in the northern cottonmouth, which serves only as a postcyclic or occasional host. There is also a tabular summary of 18 species of Ophiotaenia from semiaquatic snakes worldwide, with information on their hosts, distribution, and taxonomically important characters, including key measurements.

A global survey of tapeworms (Cestoda: Proteocephalidae) of 'true' frogs (Amphibia: Ranidae), including a tabulated list of all proteocephalids parasitising amphibians.

Proteocephalid tapeworms of frogs of the family Ranidae ('true' frogs) are reviewed with emphasis on their species diversity, host specificity and geographical distribution. New molecular data (nuclear lsrDNA and mitochondrial COI sequences) are presented for tapeworms of four species of ranid frogs in North America, including the poorly known Ophiotaenia saphena Osler, 1931 of Rana clamitans Latreille and R. catesbeiana (Shaw), which is redescribed using new material from Arkansas, USA. Tapeworms of R. sphenocephala (Cope) and R. pipiens Schreber, the latter previously identified as O. saphena, represent another, putative new species, but are not formally described due to insufficient available material. Proteocephalus papuensis Bursey, Goldberg et Kraus, 2008 from Sylvirana supragrisea (Menzies) is transferred to Ophiotaenia La Rue, 1911 as a new combination. After a critical review of the literature, only nine nominal species of Ophiotaenia are recognised as valid, which is in contrast to the large number of ranid frogs (> 440 spp.). The reasons for this striking disparity are briefly discussed, and a key based on morphology is presented for the identification of all species of Ophiotaenia from the Ranidae. Molecular data are available for only two taxa from North America that form a monophyletic group. The relationships among tapeworms of ranid frogs occurring in other zoogeographical regions are not yet known. The taxonomic status of Batrachotaenia Rudin, 1917, which was erected to accommodate proteocephalids from amphibians, is also discussed. To facilitate future studies, a tabulated summary of all 32 species of proteocephalids belonging to three genera reported from amphibians (frogs and salamanders) is presented, with information on their hosts, distribution, and taxonomically important characters, including key measurements.

Non-Invasive Wildlife Disease Surveillance Using Real Time PCR Assays: The Case of the Endangered Galemys pyrenaicus Populations from the Central System Mountains (Extremadura, Spain).

The Iberian desman (Galemys pyrenaicus) is a small semi-aquatic mammal that inhabits mountainous areas from the centre to the north of the Iberian Peninsula and the Pyrenees and is listed as endangered because it has suffered a serious decline. Since 1960, only three species of digeneans (Omphalometra flexuosa, Maritrema pyrenaica and Mathovius galemydis) and two nematodes (Aonchotheca galemydis and Paracuaria hispanica) have been reported from the desman, but no further information on health status and no data from Extremadura has been available. The aim of our study was to characterise the diversity and distribution of parasites and microbiomes of desmans in different areas of the Central System of Extremadura. Between 2019 and 2021 we collected 238 fecal samples and one tissue (intestine) sample that was obtained from a dead desman. DNA templates were processed by commercial or customised real-time PCR using TaqMan probes. Representative data were obtained for Cryptosporidium spp., Omphalometra spp., Eimeria spp., Salmonella spp., Staphylococcus spp. and Leptospira spp. Omphalometra spp. was studied using a newly developed PCR test. The screening of the dead desman allowed us to obtain, for the first time, a partial sequence of the 18SrDNA. This study is the most complete study of the desman, allowing us to identify parasites and the microbiome in populations of G. pyrenaicus using non-invasive sampling.

Elucidating the life cycle of opossum parasites: DNA sequences reveal the involvement of planorbid snails as intermediate hosts of Rhopalias spp. (Trematoda: Echinostomatidae) in Brazil.

Echinostomatid digeneans belonging to the genus Rhopalias are intestinal trematodes found mainly in opossums in the New World. The genus comprises seven species, but their life cycles and intermediate hosts have been unknown until now. During our long-term study carried out in freshwater habitats within the state of Minas Gerais, Southeast Brazil, echinostomatid cercariae lacking collar spines were found in planorbid snails Biomphalaria glabrata, Biomphalaria straminea, Drepanotrema lucidum and Gundlachia ticaga in six different batches of snail samples collected between 2010 and 2019. Morphologically, the larvae reported herein are morphologically consistent with each other and characterized by the presence of 2-3 large ovoid or spherical corpuscles in each main duct of the excretory system, resembling to Cercaria macrogranulosa previously described from the same region of Brazil. Partial sequences of the ITS (ITS1-5.8S-ITS2) region and 28S gene of the nuclear ribosomal RNA operon, and partial sequences of mitochondrial nad1 and cox1 genes were obtained and compared with the data available for members of the family Echinostomatidae. Nuclear markers indicate that all samples of cercariae evaluated in the present study can be assigned to Rhopalias, but distinct from North American isolates of Rhopalias macracanthus, Rhopalias coronatus and Rhopalias oochi (divergence 0.2-1.2% in 28S and 0.8-4.7% in ITS). The lack of differences verified in both 28S and ITS in 5 out 6 studied samples suggested that they belong to the same species. However, nad1 sequences revealed that our cercariae correspond to three distinct species of Rhopalias (interspecific divergence: 7.7-9.9%), named here as Rhopalias sp. 1, found in B. straminea and G. ticaga, Rhopalias sp. 2 found in B. glabrata and D. lucidum, and Rhopalias sp. 3 also found in D. lucidum. They also differ by 10.8-17.2% from a North American isolate of R. macracanthus sequenced in this study. The cox1 sequences obtained for Rhopalias sp. 1 and Rhopalias sp. 2 (but not Rhopalias sp. 3) reveal that they are distinct from North American isolates of R. macracanthus (genetic divergence 16.3-16.5% and 15.6-15.7%, respectively), R. coronatus (9.2-9.3% and 9.3-9.5%) and Rhopalias oochi (9.0% and 9.5-10.1%). Encysted metacercariae with general morphology similar to that of the body of cercariae were found in tadpoles of Rhinella sp. from the same stream where snails harbored Rhopalias sp. 2, suggesting that the amphibians could act as second intermediate hosts of species of Rhopalias. Data obtained provide the first insights into the life cycle of this unusual echinostomatid genus.

Description and Phylogenetic Relationships of Anhingatrema n. gen. (Digenea: Diplostomidae) with Two New Species from New World Anhingas (Aves: Anhingidae).

PURPOSE: The Diplostomidae is a globally distributed family of digeneans that parasitize a wide variety of tetrapod definitive hosts. Recent molecular phylogenetic studies have revealed unknown diplostomid diversity in avian hosts throughout the New World. Herein, we provide descriptions of a novel genus of diplostomids with two new species. METHODS: Two species of diplostomids belonging to the new genus were collected from anhinga birds in Mississippi (USA) and Brazil. Partial nuclear 28S ribosomal and mitochondrial cox1 genes were sequenced. Ribosomal data were used for phylogenetic inference. RESULTS: Both species of Anhingatrema n. gen. were positioned in a 100% supported, monophyletic clade in the phylogenetic tree. The molecular phylogenetic position and a combination of morphological features (e.g., presence of pseudosuckers, testes shape and orientation) supported erection of the new genus. Anhingatrema overstreeti n. sp. and Anhingatrema cararai n. sp. are morphologically similar, but differ in size of and ratios associated with pseudosuckers. The two species differ by 2% of 28S sequences and 13.8% of cox1 sequences. Comparison of DNA sequences revealed that Diplostomidae gen. sp. in GenBank (MZ314151) is conspecific with An. overstreeti n. sp. CONCLUSION: Anhingatrema n. gen. is the sixth genus of diplostomids known from anhingas worldwide. Anhingatrema cararai n. sp. is the first diplostomid to be reported from anhingas in South America. Combined with previous studies, the molecular phylogenies revealed at least two host switches to anhingas from other birds during the evolutionary history of the Diplostomidae.

A NEW GENUS OF DIPLOSTOMIDS (DIGENEA: DIPLOSTOMOIDEA) FROM THE NILE CROCODILE IN SOUTH AFRICA WITH A KEY TO DIPLOSTOMID GENERA.

The Diplostomidae Poirier, 1886 is a large family of digeneans within the superfamily Diplostomoidea Poirier, 1886. Members of the family are distributed worldwide and parasitize a diversity of tetrapod definitive hosts. Notably, only 2 mature diplostomids are known from crocodilians and both are suggested to be accidental infections. In this study, we use morphological and molecular data to describe Neofibricola n. gen. from a Nile crocodile Crocodylus niloticus collected in South Africa. We provide a description of adults and metacercariae of the type species, Neofibricola smiti n. sp., and metacercariae of a likely congeneric species. We generated partial 28S and internal transcribed spacer region ribosomal deoxyribonucleic acid (DNA) and cytochrome c oxidase 1 subunit mitochondrial DNA for both species and utilized the newly generated 28S sequences to examine phylogenetic affinities of these new taxa. In addition, we provide a new key to diplostomid genera, considering the substantial systematic changes and newly erected genera since the previously published key to diplostomid genera.

Molecular phylogeny supports invalidation of Didelphodiplostomum and Pharyngostomoides (Digenea: Diplostomidae) and reveals a Tylodelphys from mammals.

Alaria, Didelphodiplostomum and Pharyngostomoides are among genera of diplostomid digeneans known to parasitize mammalian definitive hosts. Despite numerous recent molecular phylogenetic studies of diplostomids, limited DNA sequence data is available from diplostomids parasitic in mammals. Herein, we provide the first 28S rDNA and cox1 mtDNA sequences from morphologically identified, adult specimens of Didelphodiplostomum and Pharyngostomoides. Newly generated 28S sequences were used to infer the phylogenetic interrelationships of these two genera among other major lineages of diplostomoideans. The phylogeny based on 28S and a review of morphology clearly suggests that Pharyngostomoides should be considered a junior synonym of Alaria, while Didelphodiplostomum should be considered a junior synonym of Tylodelphys. Pharyngostomoides procyonis (type species), Pharyngostomoides adenocephala and Pharyngostomoides dasyuri were transferred into Alaria as Alaria procyonis comb. nov., Alaria adenocephala comb. nov. and Alaria dasyuri comb. nov.; Didelphodiplostomum variabile (type species) and Didelphodiplostomum nunezae were transferred into Tylodelphys as Tylodelphys variabilis comb. nov. and Tylodelphys nunezae comb. nov. In addition, Alaria ovalis comb. nov. (formerly included in Pharyngostomoides) was restored and transferred into Alaria based on a morphological study of well-fixed, adult specimens and the comparison of cox1 DNA sequences among Alaria spp. The diplostomid genus Parallelorchis was restored based on review of morphology.

Phylogenomics and Diversification of the Schistosomatidae Based on Targeted Sequence Capture of Ultra-Conserved Elements.

Schistosomatidae Stiles and Hassall 1898 is a medically significant family of digenetic trematodes (Trematoda: Digenea), members of which infect mammals or birds as definitive hosts and aquatic or amphibious gastropods as intermediate hosts. Currently, there are 17 named genera, for many of which evolutionary interrelationships remain unresolved. The lack of a resolved phylogeny has encumbered our understanding of schistosomatid evolution, specifically patterns of host-use and the role of host-switching in diversification. Here, we used targeted sequence capture of ultra-conserved elements (UCEs) from representatives of 13 of the 17 named genera and 11 undescribed lineages that are presumed to represent either novel genera or species to generate a phylogenomic dataset for the estimation of schistosomatid interrelationships. This study represents the largest phylogenetic effort within the Schistosomatidae in both the number of loci and breadth of taxon sampling. We present a near-comprehensive family-level phylogeny providing resolution to several clades of long-standing uncertainty within Schistosomatidae, including resolution for the placement of the North American mammalian schistosomes, implying a second separate capture of mammalian hosts. Additionally, we present evidence for the placement of Macrobilharzia at the base of the Schistosoma + Bivitellobilharzia radiation. Patterns of definitive and intermediate host use and a strong role for intermediate host-switching are discussed relative to schistosomatid diversification.

Phylogenetic relationships of the Renschetrematidae Yamaguti, 1971 with description of a new species of Renschetrema Rohde, 1964 from the Philippines.

Renschetrematidae is a very small family of digeneans parasitic in bats in southern and Southeast Asia. According to the original descriptions and the latest revision, its representatives are characterized by the presence of several unusual characters. Among them are the dorsal position of the genital pores, separate male and female genital pores, and the presence of an accessory sac (stylet pouch) associated with terminal genitalia and containing a stylet-like structure. Prior to our study, the phylogenetic relationships of the Renschetrematidae were unknown and DNA sequence data were absent from any of its representatives. In this work, we present the description of a new species of Renschetrema from bats in the Philippines. We also evaluate the phylogenetic affinities of the Renschetrematidae using newly obtained partial sequences of the 28S rRNA gene from Renschetrema specimens collected in the Philippines and Southeast China. The new species differs from previously described species by the position and orientations of the stylet pouch, presence of accessory spines around genital atrium, position of gonads, body shape and relative size of pharynx and oral sucker. Our phylogenetic analysis supports the status of the Renschetrematidae as an independent family within the superfamily Microphalloidea. In the phylogenetic tree, the Renschetrematidae appeared as an independent family-level lineage, basal to the remaining taxa within the Microphalloidea. Detailed examination of our specimens revealed significant errors in the current diagnoses of the genus Renschetrema and the Renschetrematidae. Amended diagnoses of the genus and the family are provided.

Phylogenetic relationships of Ochoterenatrema Caballero, 1943 (Digenea: Lecithodendriidae) with descriptions of two new species.

Ochoterenatrema is a small genus of lecithodendriid digeneans that includes six species parasitic in Neotropical bats in North and South America. Members of this genus are characterized by the presence of a pseudogonotyl formed by thickened tegument on the left side of the ventral sucker. We examined morphology of specimens belonging to five species of Ochoterenatrema from Brazil, Ecuador and the USA and describe two new Ochoterenatrema spp. (Ochoterenatrema sphaerula n. sp. and Ochoterenatrema gracilis n. sp.). These new species are readily differentiated from their congeners by a combination of morphological characters, including the distribution of vitelline follicles and the body shape. We have provided the first DNA sequences for Ochoterenatrema. Newly generated partial nuclear 28S rDNA and mitochondrial cox1 gene sequences were used to differentiate among species and study the phylogenetic affinities of Ochoterenatrema spp., including one of the new species, O. gracilis n. sp., as well as Ochoterenatrema diminutum, Ochoterenatrema fraternum and Ochoterenatrema cf. labda. The phylogeny of the Microphalloidea based on 28S had well-supported topology, particularly at the family level. The Ochoterenatrema clade was strongly supported; however, the internal topology of the clade was weakly supported. Comparison of sequences revealed 0.4-1.3% interspecific divergence in 28S and 9.1-19.7% interspecific divergence in cox1 among Ochoterenatrema spp. We hypothesize that extremely diverse fauna of bats in South and Central America harbors multiple undescribed species of Ochoterenatrema. Several lecithodendriid genera from bats and other hosts are yet to be included in future molecular phylogenetic analyses to test the monophyly of this extremely diverse digenean family and analyze evolutionary affinities of its constituent taxa.

The life cycle of Philophthalmus aylacostoma n. sp. (Trematoda: Philophthalmidae), a new eye fluke species transmitted by Aylacostoma spp. (Gastropoda: Thiaridae) in Brazil.

Philophthalmus is a cosmopolitan genus of digeneans that includes ocular parasites of birds and mammals. Despite broad distribution and veterinary importance of these digeneans, there are still gaps in knowledge about their diversity and biology, especially in South America. Herein, we conducted morphological, life cycle, and molecular studies of megalurous cercariae found in aquatic gastropod molluscs Aylacostoma chloroticum and A. tuberculatum collected in the São Francisco River, Brazil. Adult parasites reared experimentally in the eyes of chicks are described here as Philophthalmus aylacostoma n. sp. The new species differs from its congeners known in the Americas by a combination of traits, including the sucker width ratio, the oral sucker to pharynx width ratio, egg size, and the type of vitellarium in adult forms. The new species is morphologically closest to Philophthalmus megalurus, from which it differs by the smaller body and larger eggs, as well as by the measurements of cercariae and the family of snails that act as the intermediate host. Molecular phylogenetic analysis based on 28S rDNA and comparison of cox1 sequences confirm that P. aylacostoma n. sp. is distinct from four previously sequenced named species of the genus. Moreover, cox1 sequences revealed conspecificity of our specimens with an isolate of Philophthalmus sp. previously reported, also in thiarid snails, in Paraná River, Brazil. The interspecific divergence in cox1 between the new species and other species with sequences available for comparison varied between 12 and 15%.

Host foraging behavior and nest type influence prevalence of avian haemosporidian parasites in the Pantanal.

Avian haemosporidians from the genera Plasmodium and Haemoproteus are vector transmitted parasites. A growing body of evidence suggests that variation in their prevalence within avian communities is correlated with a variety of avian ecological traits. Here, we examine the relationship between infection probability and diversity of haemosporidian lineages and avian host ecological traits (average body mass, foraging stratum, migratory behavior, and nest type). We used molecular methods to detect haemosporidian parasites in blood samples from 642 individual birds of 149 species surveyed at four localities in the Brazilian Pantanal. Based on cytochrome b sequences, we recovered 28 lineages of Plasmodium and 17 of Haemoproteus from 31 infected avian species. Variation in lineage diversity among bird species was not explained by avian ecological traits. Prevalence was heterogenous across avian hosts. Bird species that forage near the ground were less likely to be infected by Haemoproteus, whereas birds that build open cup nests were more likely infected by Haemoproteus. Furthermore, birds foraging in multiple strata were more likely to be infected by Plasmodium. Two other ecological traits, often related to host resistance (body mass and migratory behavior), did not predict infection probability among birds sampled in the Pantanal. Our results suggest that avian host traits are less important determinants of haemosporidian diversity in Pantanal than in other regions, but reinforces that host attributes, related to vector exposure, are to some extent important in modulating infection probability within an avian host assemblage.