Helminths found in common species of the herpetofauna in Ukraine.

Background: Only a few comprehensive studies have been carried out on parasites in amphibians and reptiles in Ukraine. This has resulted in identifying over 100 helminth species across these vertebrate groups. However, most of the studies were performed in the 20th century and the taxonomy of many parasites and their hosts has changed ever since, in addition to the discovery of new species and registrations of species that had not been previously known for Ukraine. In recent decades, there have been very few publications on helminths from amphibian or reptile hosts in this region. Notably, just one of these recent studies is a faunistic study, providing a list of helminths found in two species of green frogs - Pelophylaxridibundus (Pallas, 1771) and Pelophylaxesculentus (Linnaeus, 1758). Therefore, it is clear that publishing datasets of modern records of helminths in these vertebrate groups, based on modern taxonomy, is an essential step in further studies of their parasitic diversity. Additionally, such study is important in terms of global climate change, the growing number of possibilities of invasion of alien species (both hosts and parasites) that might potentially become a threat to native biota and growing anthropogenic pressure on local populations of hosts that affect the parasites as well. In future, this study is planned to be used for the creation of a checklist of helminths of the herpetofauna of Ukraine. The present dataset is an inventory of various species of helminths parasitising common species of the herpetofauna in central, northern, western and southern Ukraine recorded during field studies in the 2021-2023 period. New information: The dataset is the first one to represent the up-to-date and unified data on helminths of reptiles and amphibians of Ukraine. Previously, records of this group of organisms with reference to their hosts were presented as several separate records within the country. Currently, this is the largest dataset presenting geocoded records of non-human-related helminths in the fauna of Ukraine. It reports helminth species from 15 hosts (205 individuals), including eight amphibians and seven reptilian species found in various Ukrainian regions. A total of 47 helminth species have been documented in the research and during 2021-2023 period on the territory of northern (Kyiv and Zhytomyr), western (Lviv, Zakarpattia Ivano-Frankivsk), central (Vinnytsia, Dnipropetrovsk, Cherkasy, Zaporizhzhia and Poltava) and southern (Odesa) regions of Ukraine. The identified helminth species belong to the following phyla: Acanthocephala (Centrorhynchidae (2), Echinorhynchidae (2)); Nematoda (Acuariidae, Anisakidae, Cosmocercidae (3), Dioctophymatidae, Gnathostomatidae (1), Kathlanidae (1), Molineidae (7), Onchocercidae (1), Pharyngodonidae (1), Rhabdiasidae (6), Strongyloididae); Platyhelminthes (Diplodiscidae (1), Diplostomidae (2), Encyclometridae (1), Haematoloechidae (1), Leptophallidae (2), Macroderidae (1), Mesocestoididae, Opisthorchiidae (2), Plagiorchiidae (3), Pleurogenidae (2), Polystomatidae (3), Proteocephalidae (1), Strigeidae (1) and Telorchiidae (3)). Only some helminths in the dataset were not identified to species level. Material is stored in the collection of the department of Parasitology of the I. I. Schmalhausen Institute of Zoology NAS of Ukraine.

Cosmocerca goroensis n. sp. (Nematoda: Cosmocercidae) from South Africa and its phylogenetic relationships with other cosmocercids based on partial 28S sequences.

Nematodes of the genus Cosmocerca are commonly found in various amphibians in South Africa and in most cases are identified as C. ornata. However, after detailed morphological studies and molecular approaches, three new species of the genus were recently described from three different frogs in South Africa. In present study, we describe another new species - Cosmocerca goroensis parasitising the Northern Pygmy Toad Poyntonophrynus fenoulheti in Soutpansberg mountains, Limpopo province, South Africa. The new species is characterised by prominent sex dimorphism, wide lateral alae, numerous somatic papillae in both sexes, and wide triangularly shaped gubernaculum and simple prominent spicules in males. Cosmocerca goroensis n. sp. distinguished from congeners, previously reported in Southern Africa by the shape of the gubernaculum and arrangement of somatic papillae in males. Morphological differences were confirmed by molecular analysis based on fragments of the 28S gene. Phylogenetic analysis based on the 28S gene fragments, including C. goroensis n. sp. and newly obtained sequence of C. ornata from Pelophylax lessonae from Ukraine, supported previously known data of closer relationships between species of Cosmocerca and Aplectana and more distant with Cosmocercoides spp.

Three new species of Rhabdias Stiles et Hassall, 1905 (Nematoda: Rhabdiasidae) parasitic in Ptychadena spp. (Amphibia: Anura: Ptychadenidae) and an identification key to Rhabdias spp. from Afrotropical anurans.

Three new species of lung-dwelling nematodes are described from the frogs Ptychadena anchietae (Bocage), P. oxyrhynchus (Smith), and P. uzungwensis (Loveridge) in southern Africa. All three species are medium-sized species of Rhabdias Stiles et Hassall, 1905, with the thick-walled buccal capsules measuring 11-13 µm × 6-11 µm, consisting of longer anterior and shorter posterior parts. Rhabdias athos n. sp. and R. porthos n. sp. are characterised by the rounded anterior end of the body and the presence of short dilatation of the oesophagus at its mid-length. Rhabdias porthos n. sp. has distinct excretory glands which are absent in two other species. Rhabdias aramis n. sp. is characterised by the truncated anterior end and the slight constriction of the oesophagus at the level of its mid-length. Phylogenetic analysis based on ITS-28S rDNA sequences placed R. aramis n. sp. in the clade consisting of R. engelbrechti Kuzmin et al., 2017 from South Africa and Eurasian Rhabdias spp., while R. athos n. sp. and R. porthos n. sp. formed a sister group to that clade. Identification key to 14 Rhabdias spp. parasitic in anuran amphibians from the Afrotropical Realm is provided.

A new species of Serpentirhabdias Tkach, Kuzmin et Snyder, 2014 (Nematoda: Rhabdiasidae) parasitic in the herald snake, Crotaphopeltis hotamboeia (Laurenti) (Reptilia: Serpentes: Colubridae) in South Africa.

Serpentirhabdias mamlambo n. sp. is described from the lung of the herald snake, Crotaphopeltis hotamboeia (Laurenti) in KwaZulu-Natal Province, South Africa. The new species is characterised by the small lips, the absence of the buccal capsule, the large excretory glands (1.5-2.3 times longer than the oesophagus), and the numerous eggs in uteri. Serpentirhabdias mamlambo n. sp. differs from other Serpentirhabdias spp. in the presence of M-shaped cuticular folds on dorsal and ventral sides of apical surface, the eye-shaped internal labial papillae, and the distinct dilatation of the oesophagus at its mid-length. Morphological differentiation of S. mamlambo n. sp. from its congeners is supported by molecular evidence based on cox1 mitochondrial gene, and ITS and partial 28S region of nuclear rDNA. Serpentirhabdias mamlambo n. sp. is the first species of the genus Serpentirhabdias described from the Afrotropical Realm.

Two new species of Neofoleyellides (Nematoda: Onchocercidae) parasitising anuran amphibians in South Africa.

The genus Neofoleyellides was recently erected for a single species, Neofoleyellides boerewors from bufonid hosts in South Africa. In present study, we discovered two undescribed species of Neofoleyellides, namely N. steyni n. sp. and N. martini n. sp. parasitising frogs Amietia delalandii and Leptopelis natalensis, respectively. Both species differ from N. boerewors and between each other in shape and relative length of oesophagus, size of spicules, arrangement of genital papillae and morphology of caudal alae in males. Phylogenetic analysis based on concatenated fragments of the 18S ribosomal ribonucleic acid (18S rRNA) and the Cytochrome oxidase c subunit I (COI) genes confirmed both species as Neofoleyellides sister to Icosiellinae and Oswaldofilariinae.

Pseudocapillaria (Ichthyocapillaria) bumpi n. sp. (Nematoda: Capillariidae) Parasitising West African Lungfish Protopterus annectens (Owen, 1839) (Lepidosireniformes: Protopteridae) in Mozambique and Its Phylogenetic Position Within Capillariid Nematodes.

INTRODUCTION: Pseudocapillaria (Ichthyocapillaria) bumpi n. sp. (Nematoda: Capillariidae) is described from specimens infecting the digestive tract of the West African lungfish, Protopterus annectens (Owen) (Lepidosireniformes: Protopteridae). METHODS: Nematodes were collected in Karingani Game Reserve, Southern Mozambique and studied using light microscopy; obtained sequences of the 18S ribosomal (18S rRNA) gene used for phylogenetic studies. RESULTS: The new species is assigned to Pseudocapillaria Freitas, 1959 by having a stichosome consisting of a single row of stichocytes and ventrolateral lobes on the male tail as well as by lacking spines on the specular sheath, caudal alae, a membranous caudal bursa, and a lateral expansion of the caudal end. It is also assigned to the sub-genus Ichthyocapillaria Moravec, 1982 by having a membrane between the ventrolateral lobes and by lacking a vulvar appendage. Pseudocapillaria (I.) bumpi n. sp. differs from its nominal congeners by having a comparatively large body size with relatively long spicule, bearing the thin membrane non-extending further than ventrolateral lobes in males and relatively smaller eggs without protruding polar plugs. It is the 20th species of the genus, the first from the Afrotropical Realm, and the first from any lungfish species. Phylogenetic analyses suggested that P. (I.) bumpi is deeply nested in a clade that associates species of the genera Pearsonema, Aonchotheca and Baruscapillaria.

Spiroxys ankarafantsika Roca et Garcia, 2008 (Nematoda: Gnathostomatidae) and Other Nematodes Parasitising Pelusios spp. (Testudines: Pelomedusidae) from South Africa and Mozambique.

INTRODUCTION: The nematode Spiroxys ankarafantsika was described parasitising freshwater turtles Pelusios castanoides and Pelomedusa subrufra from Madagascar. During parasitological surveys at different localities in Mozambique and South Africa the species was recovered from the digestive tract of Pelusios sinuatus, P. subniger and P. castanoides. METHODS: The species were identified based on the morphology of the anterior extremity (absence of additional teeth on pseudolabia) and the male caudal end (number and arrangement of caudal papillae, size and shape of spicules). Molecular data included a 720 bp sequence fragment of 18S rDNA, the entire internal transcribed spacer 1 (ITS1), 5.8S rRNA, and internal transcribed spacer 2 (ITS), flanked by a 288 bp 18S rDNA sequence and about 1000 bp 28S rDNA sequence, and the mitochondrial cytochrome c oxidase subunit I (cox1) gene marker obtained in the present study. RESULTS: All specimen recovered were identical, based on morphological and molecular data, with the exception of two specimens showing an intraspecific divergence of 9% based on a 694 bp sequence fragment of the cox1 gene. Additionally, a Contracaecum sp. third stage larva and an unidentified larva with mushroom-shaped anterior extremity were found. Descriptions of S. ankarafantsika and the two above-mentioned larval stages, supported by photomicrographs and molecular data 18S rDNA, ITS1-5.8S-ITS2-28S and cox1 gene fragments are presented herein.

New records of Amphibiophilus spp. (Nematoda: Amphibiophilidae) parasitic in Strongylopus grayii (Smith) and Amietia delalandii (Duméril & Bibron) (Amphibia: Anura: Pyxicephalidae) in South Africa, with a description of Amphibiophilus bialatus n. sp.

Nematodes of the genus Amphibiophilus Skrjabin, 1916 are a small group of parasites restricted to pyxicephalid frogs in southern Africa. In the present study, the new species A. bialatus parasitising the clicking stream frog Strongylopus grayii (Smith) as well as two forms parasitising the common river frog Amietia delalandii (Duméril & Bibron) from two distant localities are described. Amphibiophilus bialatus n. sp. clearly differs from the remaining species of the genus by having wide cervical alae, the dorsal oesophageal tooth not reaching the oral opening, and the presence of extra processes on the spicules. Specimens parasitising Am. delalandii in Mpumalanga Province and Limpopo Province, South Africa, differed from other species and from each other in the shape of the gubernaculum, though were almost identical in other characters. Based on morphological and molecular data, specimens from two localities were assigned to Amphibiophilus sp. 1 and Amphibiophilus sp. 2. Pairwise analyses of ITS-28S and cox1 gene fragments are presented for four Amphibiophilus spp.

Description and molecular characterisation of a new nematode species parasitic in the lungs of Strongylopus grayii (Smith) (Anura: Pyxicephalidae) in South Africa.

Rhabdias delangei n. sp. (Nematoda: Rhabdiasidae) is described from the lungs of the clicking stream frog Strongylopus grayii (Smith) in the Western Cape Province of South Africa. The new species is characterised by the nerve-ring being located just posterior to the mid-length of the oesophagus, a pre-equatorial vulva, as well as a thin-walled and apparently underdeveloped rectum. Pairwise analyses based on ITS-28S and partial cox1 gene fragments showed significant differences between R. delangei n. sp. and its congeners from the Afrotropical Realm. Rhabdias delangei n. sp. is the 12th species of Rhabdias Stiles & Hassall, 1905 recorded in Afrotropical amphibians and the second species of the genus found in pyxicephalid hosts. Our phylogenetic analysis based on the ITS-28S region showed that the new species is closely related to the European species R. rubrovenosa (Schneider, 1866). Furthermore, both species are similar with respect to the underdeveloped rectum and pre-equatorial position of vulva.

Neofoleyellides boerewors n. gen. n. sp. (Nematoda: Onchocercidae) parasitising common toads and mosquito vectors: morphology, life history, experimental transmission and host-vector interaction in situ.

Anuran filarial nematodes are restricted to two comparatively small subfamilies (Icosiellinae and Waltonellinae) of the filariae that currently comprise six genera and 41 recognised species. However, the life histories of only five anuran filarial nematodes, proposed as an ancestral group based on molecular phylogenetic studies, have been elucidated. Furthermore, data on the natural vectors (in situ) and parasite transmission is limited. In the current study we elucidate the life history of Neofoleyellides boerewors n. gen. n. sp. parasitising the guttural toad, Sclerophrys gutturalis and the mosquito vectors Uranotaenia (Pseudoficalbia) mashonaensis and Uranotaenia (Pseudoficalbia) montana. Additionally, we report on the unique host-seeking behaviour of the mosquito vectors which locate their toad hosts using their calls. The complex host-vector relationship and specialised host-seeking behaviour by these mosquitoes indicate biases towards host species and male toad infections.

Novel information on the morphology, phylogeny and distribution of camallanid nematodes from marine and freshwater hosts in South Africa, including the description of Camallanus sodwanaensis n. sp.

Four species of previously known nematodes from the family Camallanidae were found from different hosts in South Africa: Batrachocamallanus xenopodis from the frog Xenopus muelleri, Paracamallanus cyathopharynx and Procamallanus pseudolaeviconchus from the catfish Clarias gariepinus and Spirocamallanus daleneae from the catfish Synodontis zambezensis. In the material collected from various marine fishes, several specimens of nematodes from the genus Camallanus clearly differed from all previously known species. Based on morphological differences these specimens are assigned to a new species, C. sodwanaensis. Molecular data of 18S and 28S rDNA and COI sequences are provided for the collected species and a phylogenetic analyses based on 28S gene fragmets are presented.

Serpinema cayennensis n. sp. (Nematoda: Camallanidae), a parasite of the freshwater turtle Rhinoclemmys punctularia Daudin (Reptilia: Testudines: Geoemydidae) from French Guiana: morphology and phylogenetic relationships with other turtle-parasitising camallanids.

The genus Serpinema was erected from the genus Camallanus to include species parasitising freshwater turtles. Following this, the taxonomic status of Serpinema was challenged by different authors considering it as a subgenus or junior synonym of Camallanus. Several specimens of these nematodes were retrieved from the spot-legged wood turtle Rhinoclemmys punctularia from Cayenne, French Guiana. These specimens clearly differed from previously reported Serpinema and Camallanus species parasitising turtles by the number of caudal papillae, shape of spicules and development of vulvar lips. Based on morphological characters the nematodes were assigned to the genus Serpinema and described as a new species S. cayennensis n. sp. The description followed by the molecular data analyses based on 28S, 18S rDNA and cytochrome oxidase subunit 1 (COI) gene sequences.

New information on morphology and molecular data of camallanid nematodes parasitising Xenopus laevis (Anura: Pipidae) in South Africa.

Three species of nematodes from the Camallanidae that are known to infect Xenopus laevis Daudin (Anura: Pipidae) were collected from several localities across South Africa. New data on morphology, partial 28S and cox1 genes, infection levels and distribution are presented herein. The most common species, Batrachocamallanus slomei Southwell et Kirshner, 1937, from the stomach and less often oesophagus, was found in eight localities. Camallanus kaapstaadi Southwell et Kirshner, 1937, also from the oesophagus, was found in two localities and C. xenopodis Jackson et Tinsley, 1995, from the intestine, at a single locality. New localities for both C. kaapstaadi and C. xenopodis provide a geographical range extension. Males of C. xenopodis are described for the first time herein. The existence of a left spicule in the males of both the species of Camallanus Railliet and Henry, 1915 is confirmed and measurements are provided. Although C. xenopodis is distinguished from C. mazabukae Kung, 1948 in the present study, we suggest greater sampling effort in other African amphibians to confirm the species status of the latter taxon. Finally, the new molecular data showed distant relationships between collected species of Camallanus and species parasitising fish and freshwater turtles.

Molecular and morphological characterisation of Rhabdias picardiae Junker, Lhermitte-Vallarino et Bain, 2010 (Nematoda: Rhabdiasidae) from Delaland's River Frog, Amietia delalandii (Duméril et Bibron, 1841) (Amphibia: Pyxicephalidae) in South Africa.

Rhabdias picardiae previously known from the Guttural Toad, Sclerophrys gutturalis (Bufonidae) is described from a new host, Delaland's River Frog, Amietia delalandii (Pyxicephalidae). The species identification is confirmed by analysis of the COI gene sequence and morphological comparison with the original description. Morphological details of apical structures in adult parasitic R. picardiae, namely, the shape and position of lips, the number and position of external labial papillae, and the buccal capsule morphology are described using light and scanning electron microscopy. The buccal capsule width and length, the distance from anterior end to nerve ring and excretory pore, and ratios of distance to vulva to body length and body length to width were identified as less variable metrical characters in the studied sample of 30 specimens. Adult specimens demonstrated differences in the position of lips in relation to the oral opening and position of the buccal capsule in relation to the oesophagus depending on the size (age) of worms. The infective larvae of R. picardiae are described for the first time and characterised by two lateral alae consisting of two ridges, triangular pseudolabia with rounded tops, and presence of ornamentation on the rounded tail tip.

Amphibiophilus mooiensis n. sp. (Nematoda: Amphibiophilidae), a parasite of Amietia delalandii (Duméril & Bibron) (Amphibia: Pyxicephalidae) from South Africa.

Four species of the genus Amphibiophilus Skrjabin, 1916 from pyxycephalid frogs in southern and central Africa are currently recognised as valid. Several specimens of Amphibiophilus were found in material from the common river frog, Amietia delalandii (Duméril & Bibron) (Amphibia: Pyxicephalidae), collected in Potchefstroom (North-West Province, South Africa). These specimens clearly differ from all previously known species by the shape of the distal end of the spicule, the shape of the gubernaculum and the structure of the synlophe. They are, thus, considered as a new species, Amphibiophilus mooiensis. As all other species in the family Amphibiophilidae Durette-Desset & Chabaud, 1981, A. mooiensis n. sp. possesses a number of archaic characters, such as a buccal capsule with a well-developed dorsal oesophageal tooth, six inner labial papillae, six outer labial papillae and four cephalic papillae. Molecular data (cox1 and ITS-28S rDNA sequences) are provided and host and geographical specificity are discussed.

Two new species of Oswaldocruzia (Nematoda, Molineidae) parasitising lizards in Ukraine.

Two new species, Oswaldocruzia lisnykiensis sp. n. and Oswaldocruzia lacertica sp. n., are described from Anguis fragilis L. (Reptilia: Anquidae) and Lacerta agilis L. (Reptilia: Lacertidae) respectively. Both species belong to the Palaearctic group of species which have spicules divided in three main branches (blade, fork and shoe) with fork division above its distal third. The new species differs from all previously known Palaearctic species by several morphological characters: type of caudal bursa (type III in O. lisnykiensis and type I in O. lacertica), shape of spicules (blade distally divided into four tips and specific shape of shoe) and synlophe structure (narrow cervical alae consist of three crests in both species). Illustrated descriptions of O. lisnykiensis and O. lacertica based on 63 and 38 specimens respectively are presented. Host specificity and geographical distribution of the species are discussed. O. dispar is considered as a species inquirenda.