Morphological and molecular data on Pseudozoogonoides ugui Shimazu, 1974 (Digenea: Microphalloidea: Zoogonidae) ex Pseudaspius hakonensis (Günther, 1877) and taxonomic problems in Zoogoninae genera.

New morphological and molecular data were generated for trematodes recovered from the intestines of the fish Pseudaspius hakonensis from two locations in the south of the Russian Far East. Morphologically, these trematodes are identical to Pseudozoogonoides ugui (Microphalloidea: Zoogonidae) from Japan. According to results of phylogenetic analysis based on 28S rDNA sequence data, P. ugui was closely related to Zoogonoides viviparus, and P. subaequiporus appears as a sister taxon to these two species. Genetic distance values, calculated based on both 28S rDNA and ITS2 rDNA, between P. ugui and Z. viviparus represents an interspecific differentiation level. Our results have an ambiguous explanation, indicating that the implication of the presence of one or two compact vitellarial aggregations for the differentiation of Zoogonoides and Pseudozoogonoides should be reconsidered or that our results open up the question of the taxonomical status of trematodes previously denoted as Z. viviparus and P. subaequiporus.

A report on the complete mitochondrial genome of the trematode Azygia robusta Odhner, 1911, its new definitive host from the Russian Far East, and unexpected phylogeny of Azygiidae within Digenea, as inferred from mitogenome sequences.

New data on the complete mitochondrial genome of Azygia robusta (Azygiidae) were obtained by the next-generation sequencing (NGS) approach. The mitochondrial DNA (mtDNA) of A. robusta had a length of 13 857 bp and included 12 protein-coding genes, two ribosomal genes, 22 transfer RNA genes, and two non-coding regions. The nucleotide sequences of the complete mitochondrial genomes of two A. robusta specimens differed from each other by 0.12 ± 0.03%. Six of 12 protein-coding genes demonstrated intraspecific variation. The difference between the nucleotide sequences of the complete mitochondrial genomes of A. robusta and Azygia hwangtsiyui was 26.95 ± 0.35%; the interspecific variation of protein-coding genes between A. robusta and A. hwangtsiyui ranged from 20.5 ± 0.9% (cox1) to 30.7 ± 1.2% (nad5). The observed gene arrangement in the mtDNA sequence of A. robusta was identical to that of A. hwangtsiyui. Codon usage and amino acid frequencies were highly similar between A. robusta and A. hwangtsiyui. The results of phylogenetic analyses based on mtDNA protein-coding regions showed that A. robusta is closely related to A. hwangtsiyui (belonging to the same suborder, Azygiida) that formed a distinct early-diverging branch relative to all other Digenea. A preliminary morphological analysis of paratypes of the two azygiid specimens studied showed visible morphological differences between them. The specimen extracted from Sakhalin taimen (Parahucho perryi) was most similar to A. robusta. Thus, we here provide the first record of a new definitive host, P. perryi, for A. robusta and also molecular characteristics of the trematode specimens.

The first modern morphological description of Cercaria pennata and molecular evidence of its synonymy with Pronoprymna ventricosa in the Black Sea.

During the parasitological examination of molluscs Abra segmentum obtained from the Black Sea basin, parthenitae belonging to the family Faustulidae were found. The cercariae were obtained by natural emergence and were studied using differential interference contrast microscopy and scanning electron microscopy. Specimens resemble Cercaria pennata ex Tapes rugatus which was described from the Sevastopol area, in the shape and length of the body, tail length, location and shape of internal organs, suckers, pharynx, testicular rudiments, and the number and position of longitudinal lamellae on the tail finlets. To date, there are only limited descriptions of the parthenitae of C. pennata without detailed measurements, thus the taxonomic position of the individuals studied needs thorough revision and molecular verification. According to the molecular analyses, C. pennata was identical to that of published sequences of Pronoprymna ventricosa.

Molecular-genetic approaches to species identif ication of platyhelminthes of the genus Ligophorus (Monogenea) parasitising f lathead mullet.

Mugil cephalus L., 1758 (flathead mullet) is a valuable commercial fish and a promising object of artificial breeding in the Black Sea and the Sea of Azov, and the study of its parasite fauna is important for fishery and mariculture. Monogeneans of the genus Ligophorus are common ectoparasites dwelling on the gills of mullets. Two representatives of this genus parasitise flathead mullet in the Azov-Black Sea region, namely Ligophorus mediterraneus Sarabeev, Balbuena et Euzet, 2005 and Ligophorus cephali Rubtsova, Balbuena, Sarabeev, Blasco- Costa et Euzet, 2006. Morphological identification of these species requires spending much time and a high level of experience in monogenean taxonomy. For quick and correct species identification of these parasites, we have developed a genotyping approach based on the polymerase chain reaction of allele-specific gene sites for various Monogenea species. A fragment of the 28S ribosomal gene, which includes conserved and variable sites, was chosen as a genetic marker. Three approaches were used as follows: amplified fragment length analysis, allelespecific PCR with endpoint detection and allele-specific real-time PCR using SYBR Green intercalating dye. The first approach was by obtaining PCR products of different lengths that were specific either to L. mediterraneus or to L. cephali. This approach was implemented due to the presence of several variable sites located at a distance from each other. The PCR mixture contained three primers: one forward and two reverse. The forward primer was complementary to the conserved site, which did not differ between species. Reverse primers were speciesspecific and, for each species, they were complementary to different DNA regions located 100 bp apart. As a result, L. mediterraneus was characterized by shorter amplicons than L. cephali. For the second and third approaches, a pair of primers was designed according to the following principle: the forward primer was complementary to both species, since it was selected for the conserved gene region. Reverse primers were species-specific and were designed for the 28S variable region. The two parasite species were distinguished by three-point mutations. Thus, one pair of primers was complementary to L. mediterraneus, the other, to L. cephali. The amplified fragment length analysis and the allele-specific real-time PCR demonstrated 100 % coincidence of genotyping results compared with Sanger sequencing. The developed genotyping protocols can be used not only to distinguish two species of Ligophorus from flathead mullet in ecological studies and veterinary practice but also for further development of similar approaches for other monogeneans, among which there are many pathogenic species.

Characterization of complete mitochondrial genome and ribosomal operon for Carassotrema koreanum Park, 1938 (Digenea: Haploporidae) by means of next-generation sequencing data.

We obtained new data on the complete mitochondrial DNA (mtDNA) and the ribosomal operon of the trematode Carassotrema koreanum (Digenea: Haploporata: Haploporidae), an intestinal parasite of Carassius auratus, using next-generation sequencing. The mtDNA of C. koreanum contained 13,965 bp, including 12 protein-coding genes, two ribosomal genes, 22 transport RNA (tRNA) genes and a non-coding region. The ribosomal operon of C. koreanum was 10,644 bp in length, including ETS1 (1449 bp), 18S ribosomal RNA (rRNA) gene (1988 bp), ITS1 ribosomal DNA (rDNA) (558 bp), 5.8S rRNA gene (157 bp), ITS2 rDNA (274 bp), 28S rRNA gene (4152 bp) and ETS2 (2066 bp). Phylogenetic analysis based on mtDNA protein-coding regions showed that C. koreanum was closely related to Parasaccocoelium mugili, a species from the same suborder Haploporata. Bayesian phylogenetic tree topology was the most reliable and confirmed the validity of the Haploporata. The results of sequence cluster analysis based on codon usage bias demonstrated some agreement with the results of the phylogenetic analysis. In particular, Schistosoma spp. were differentiated from the other members of Digenea and the members of Pronocephalata were localized within the same cluster. Carassotrema koreanum and P. mugili fell within different clusters. The grouping of C. koreanum and P. mugili within the same cluster was obtained on the basis of frequencies of 13 specified codons, of which three codon pairs were degenerate. A similarity was found between two haploporid species and two Dicrocoelium spp. in the presence of TTG start codon of the mitochondrial nad5 gene. Our results confirmed the taxonomical status of the Haploporata identified in the previous studies and revealed some characteristic features of the codon usage in its representatives.

Chelatrematidae n. fam., a new family of digenetic trematodes from the South Western Ghats, India, erected on the basis of morphological and molecular studies.

On the basis of the morphological characterization of Chelatrema neilgherriensis Manjula & Janardanan, 2006 recovered from the freshwater fish Barilius gatensis (Valenciennes, 1844) in the Wayanad region of the Western Ghats, the diagnostic features of the genus Chelatrema Gupta & Kumari, 1973 have been modified. Based on the phylogenetic analysis of C. neilgherriensis and comparative morphology studies relative to members of other families of Gorgoderoidea Looss, 1901, this genus is placed in a new family Chelatrematidae n. fam. The studies revealed the molecular and morphological closeness of Chelatrema with Paracreptatrematina limi Amin & Myer, 1982, and the latter is transferred to this new family. Hence the new family Chelatrematidae n. fam. comprises the genera Chelatrema and Paracreptatrematina.

New trematode species Lecithostaphylus halongi n. sp. (Zoogonidae, Microphalloidea) and Gymnotergestia strongyluri n. sp. (Fellodistomidae, Gymnophalloidea) from beloniform fishes in Vietnam.

In this study we described two new trematode species, Lecithostaphylus halongi n. sp. (Zoogonidae, Lecithostaphylinae) and Gymnotergestia strongyluri n. sp. (Fellodistomidae, Tergestiinae), on the basis of morphological and molecular data. Adult worms of these two species were collected from, respectively, Hemiramphus spp. (Hemiramphidae) and Strongylura strongylura (Belonidae) caught in the coastal waters of Vietnam. Adult worms of L. halongi n. sp. are morphologically close to Lecithostaphylus gibsoni Cribb, Bray & Barker, 1992 ex Abudefduf whitleyi from Heron Island and Lecithostaphylus depauperati Yamaguti, 1970 ex Hemiramphus depauperatus from Hawaii, but differ from these species in having a larger cirrus sac and a different arrangement of vitelline fields. They also differ from Lecithostaphylus brayi Cabañas-Granillo, Solórzano-García, Mendoza-Garfias & Pérez-Ponce de León, 2020 in the 28S ribosomal DNA (rDNA) sequence data at the interspecific level. Adult worms of G. strongyluri n. sp. ex S. strongylura are morphologically similar to Gymnotergestia chaetodipteri, the only previously known species of this genus, described from Chaetodipterus faber in Jamaica. The new species differs from G. chaetodipteri in body shape, testicular arrangement and the size of the pharynx and eggs. The 28S rDNA-based phylogenetic analysis indicates that G. strongyluri n. sp. is closely related to Tergestia spp., rendering Tergestia paraphyletic. Genetic divergence values between G. strongyluri n. sp. and Tergestia spp. are similar to those among species in the genera Tergestia, Steringophorus and Proctoeces. Our molecular results indicate that G. strongyluri n. sp. and Tergestia spp. may belong the same genus, but additional molecular data are needed for the final conclusion.

First rDNA sequence data for Haplosplanchnus pachysomus (Digenea: Haplosplanchnidae) ex Mugil cephalus from the Black Sea, and molecular evidence for cryptic species within Haplosplanchnus pachysomus (Digenea: Haplosplanchnidae) in Palaearctic and Indo-West Pacific regions.

Adult trematodes, morphologically similar to Haplosplanchnus pachysomus (Eysenhardt, 1829), were extracted from the intestine of Mugil cephalus Linnaeus, 1758, collected in the Black Sea basin. Morphological, morphometric and 28S ribosomal DNA (rDNA) partial sequence data were obtained for these trematodes following comparative analysis with previous data on this species. Worms from this study were morphologically identical to all previously reported H. pachysomus specimens from different locations. The results of the morphometric analysis indicated general similarity between H. pachysomus from the Black Sea and trematodes from Vietnam and Australia. Trematodes from the Black Sea and specimens from Spain were identical based on 28S rDNA partial sequences; however, these sequences differed from that of H. pachysomus from Vietnam and Australia by eight fixed substitutions. Overall, our results indicate that H. pachysomus from Spain and the Black Sea and from Vietnam and Australia can presently be considered as two cryptic species, one in Palaearctic and one in Indo-West Pacific regions. Our results provide a molecular base for including Haplosplanchnus purii in the genus Provitellotrema or to consider H. purii, P. crenimugilis and H. pachysomus within the same genus.

New species of Parasaccocoelium (Haploporidae) and new genus Pseudohaplosplanchnus (Haplosplanchnidae) from mullet fish in the Far East of Russia and Vietnam: morphological and molecular data.

A description and the molecular characterization of two new species in the Haploporidae and Haplosplanchnidae families are provided herein. Parasaccocoelium armatum n. sp. was collected from the intestine of a Mugil cephalus Linnaeus, 1758 from the Primorsky region, Russia, and Pseudohaplosplanchnus catbaensis n. g. n. sp. was collected from Moolgarda seheli (Forsskål, 1775) in the coastal waters of Cat Ba Island, Vietnam. The morphological features of P. armatum n. sp. closely resemble those of Parasaccocoelium polyovum, but these species differ from one another by hermaphroditic sac and vitellaria area length and by maximal egg size. The main difference between P. armatum n. sp. and P. polyovum is the presence of an armed hermaphroditic duct in the new species. Molecular data support the case for inclusion of the studied trematodes in P. armatum n. sp. Worms P. catbaensis n. g. n. sp. from the mullet from Vietnam are morphologically close to Haplosplanchnus (Haplosplosplanchninae). The only difference between P. catbaensis n. g. n. sp. and species of Haplosplanchnus is the presence of few (1-7) large eggs, measuring 135-142 × 92-104 µm, versus numerous small eggs with a maximal size of 75 × 50 µm. Phylogenetic analysis showed that there is a contradiction between the morphological similarity of the worms and their position in the Haplosplanchnidae system, based on the genetic data. Results of this study indicate that P. catbaensis n. g. n. sp. is genetically distant from other representatives of Haplosplanchnus, despite their morphological similarity. According to the molecular data, P. catbaensis n. g. n. sp. is close to Hymenocotta mulli Manter, 1961 (Hymenocottinae). However, these species are considerably different to each other morphologically. Molecular data argue for the possibility of establishing a new subfamily for P. catbaensis n. g. n. sp. However, considering earlier studies of Haplosplanchnidae, we support the view that creating new subfamilies within this family is unreasonable because of the lack of molecular data for most haplosplanchnid species, which are necessary to resolve the problematic systematics and phylogeny of this family.

New species and new genus of Pseudohaploporinae (Digenea): Pseudohaploporus pusitestis sp. n. and Parahaploporus elegantus n. g., sp. n. (Digenea: Pseudohaploporinae) from Vietnamese mullet fish.

Two new species of Pseudohaploporinae, Pseudohaploporus pusitestis sp. n. and Parahaploporus elegantus n. g., sp. n., are described from intestines of the Vietnamese mullet fish Moolgarda seheli and Osteomugil cunnesius, respectively. Pseudohaploporus pusitestis sp. n. differs from two known Pseudohaploporus species, P. vietnamensis and P. planiliza, by the absence of a diverticulate hermaphroditic duct and muscular sphincters at the proximal end of the hermaphroditic sac. Metrically, P. pusitestis sp. n. is close to P. vietnamensis and differs from this species and from P. planilizum by lower maximum sizes of most parameters. Parahaploporus elegantus n. g., sp. differs from representatives of Pseudohaploporus by the presence of a single testis and the armament of hermaphroditic duct and is morphologically close to trematodes of the genus Haploporus. However, P. elegantus n. g differs from all known Haploporus species from mugilids of the Indo-West Pacific by the structure of the armament of the hermaphroditic duct and also by size of body, organs and eggs. The validity of designating two new species and a new genus of trematodes is supported by ITS and 28S rDNA sequence data. Phylogenetic reconstructions showed that the new trematodes belong to the Pseudohaploporinae, which formed a well-supported cluster within the monophyletic Haploporidae.

Redescription and supplementary molecular characteristics of Aspidogaster ijimai Kawamura, 1915 (Trematoda, Aspidogastrea, Aspidogastridae), a parasite of Cyprinus carpio Linnaeus, 1758 s. lato (Actinopterygii) and freshwater bivalves in East Asia.

A detailed morphological description and ribosomal DNA (rDNA) sequence molecular data for Aspidogaster ijimai from Japan are provided. Morphological analysis, including a description of the cirrus-sac, indicate the conspecificity of Japanese and continental East Asian A. ijimai specimens. Analysis of ITS1-5.8S-ITS2 rDNA sequences of Japanese, Chinese and Russian specimens confirmed the morphological results. Phylogenetic analysis using ITS rDNA sequences confirmed that A. ijimai is a sister species for Aspidogaster chongqingensis. Median-joining network analysis showed an initial molecular differentiation step of Russian specimens from group of Japanese-Chinese samples. Our 28S rDNA results based on maximum likelihood (ML) and Bayesian (BI) phylogenetic analyses indicated well-supported monophyly of the Aspidogaster conchicola + A. ijimai group, a finding that indicates that these species are congeneric. At the same time, our data demonstrated that the genus Lobatostoma is paraphyletic and the family Aspidogastridae is polyphyletic, results that confirm previous studies.

A new subfamily, Pseudohaploporinae subfam. n. (Digenea: Haploporidae), with morphometric and molecular analyses of two new species: Pseudohaploporus vietnamensis n. g., sp. n. and Pseudohaploporus planilizum n. g., sp. n. from Vietnamese mullet.

Adult specimens representing Pseudohaploporus vietnamensis n. g., sp. n., Pseudohaploporus planilizum n. g., sp. n., and Pseudohaploporus sp. were collected from the intestines of Osteomugil engeli (Bleeker, 1858), Moolgarda seheli (Forsskål, 1775), and Planiliza subviridis (Valenciennes, 1836) from Vietnam. The new genus Pseudohaploporus n. g. is similar to the genus Haploporus Looss, 1902 in morphology and host-specificity. Species of Pseudohaploporus and Haploporus both possess an elongated body, a long oesophagus, an intestinal bifurcation behind the ventral sucker, a large hermaphroditic bursa, vitellarium consisting of two compact accumulations, concretions in the excretory vesicle, and numerous small eggs that contain miracidia with a pigment eye, and both infect mullet. In contrast to Haploporus, both species of the genus Pseudohaploporus possess two testes. These species differ from each other morphometrically. Pseudohaploporus planilizum sp. n., in comparison with P. vietnamensis sp. n., possess a larger maximum and mean size for the body length, oral and ventral suckers, pharynx, oesophagus length, hermaphrodite bursa, and egg width. Molecular data (ITS and 28S rDNA sequences) support the validity of the new genus and the two new species and indicate the presence of another representative of this genus, Pseudohaploporus sp. Phylogenetic reconstructions show that Pseudohaploporus formed a well-supported clade within a large clade that represents the Haploporidae Nicoll, 1914. Based on morphological and molecular data for P. vietnamensis sp. n. and P. planilizum sp. n., we erected a new subfamily, Pseudohaploporinae subfam. n., which appears to be a sister taxon to Haploporinae Nicoll, 1914. Molecular data show that Pseudohaploporus differ from other haploporids at a level similar to that between other subfamilies.

Morphological and molecular data for species of Lecithaster Lühe, 1901 and Hysterolecithoides Yamaguti, 1934 (Digenea: Lecithasteridae) from fish of East Asia and phylogenetic relationships within the Hemiuroidea Looss, 1899.

Four representatives of the genus Lecithaster and one representative of the genus Hysterolecithoides were found during investigation of the trematode fauna of fish species in Vietnamese, Japanese and eastern coastal waters of the Russian Far East. Based on morphometric data, adult trematodes from Vietnamese Strongylura strongylura and Russian Acanthogobius flavimanus were identified as Lecithaster confusus, trematodes from Vietnamese Hemirhamphus marginatus as L. sayori and from osmerid fishes as L. salmonis. Further, a single specimen of Lecithaster sp. and representatives of Hysterolecithoides epinepheli were found in Vietnamese Siganus fuscescens. Morphological and molecular data, including 18S ribosomal DNA (rDNA) V4 fragment, 28S rDNA D1-D3 fragment, internal transcribed spacers (ITS) and a mitochondrial COI gene fragment were analysed for Lecithaster spp. The results revealed that L. sayori and L. salmonis are not synonyms of L. stellatus and L. gibbosus, respectively, but that Hysterolecithoides frontilatus and H. guangdongensis are junior synonyms of H. epinepheli. The 28S-rDNA-based phylogenetic tree of Hemiuroidea showed a distinct position for the genus Lecithaster with internal differentiation into three subclades, including L. confusus, L. sayori and Lecithaster sp. within the first subclade, L. mugilis and L. sudzuhensis within the second subclade and L. salmonis and L. gibbosus within the third subclade. Bayesian phylogenetic reconstructions of Hemiuroidea showed four clades for members of Hemiuridae and Lecithasteridae. The first clade consisted of Hemiuridae representatives and the second clade represented the genus Lecithaster. The third clade included genera Aponurus and Lecithophyllum (Lecithasteridae) and the fourth clade combined members of lecithasterid Quadrifoliovariinae and Hysterolecithinae and hemiurid Opisthadeninae and Bunocotylidae with high statistical support.

Diversity of the genus Bunodera Railliet, 1896 (Trematoda: Allocreadiidae) in the northern part of Eastern Europe and North-eastern Asia, estimated from 28S rDNA sequences, with a description of Bunodera vytautasi sp. nov.

Phylogenetic relationship reconstruction and taxonomical analysis of trematodes of the genus Bunodera was carried out using 28S rDNA partial sequences along with a description and molecular characterisation of a new species, B. vytautasi sp. nov. A new species is reported from the intestine of Pungitius pungitius (Linnaeus, 1758), collected from Magadan Region, Russia. The diagnostic characters of B. vytautasi sp. nov. are the extension of vitelline fields from the oral sucker or posterior edge of the pharynx to the posterior extremity of the body, the confluence of vitelline fields within the forebody, the extension of the cirrus-sac to the posterior third of ventral sucker or further backwards with an outlet beyond the posterior edge of the sucker, and the presence of a unipartite internal seminal vesicle. Both Bayesian and Maximum Likelihood trees shared the same tree topology, in which the genus Bunodera was shown to be monophyletic. Representatives of the genus Bunodera were distributed into three well-supported clades: percid-infecting species (Eurasian species B. luciopercae and B. acerinae and North American B. luciopercae s.l.), gasterosteid-infecting species (amphi-Pacific B. mediovitellata and North American B. inconstans and B. eucaliae) and percid/gasterosteid-infecting species (Asiatic B. vytautasi sp. nov. and North American B. sacculata). Eurasian B. luciopercae and B. acerinae were more closely related to each other than to North American B. luciopercae s.l.

Phylogenetic relationships among European and Asian representatives of the genus Aspidogaster Baer, 1827 (Trematoda: Aspidogastrea) inferred from molecular data.

In the present study, phylogenetic relationships of European and Far Eastern representatives of the genus Aspidogaster Baer, 1827 were analysed: A. conchicola Baer, 1827, A. limacoides Diesing, 1834, A. ijimai Kawamura, 1915 and A. chongqingensis Wei, Huang & Dai, 2001. Based on ITS1-5.8S-ITS2 rDNA sequence data, an obvious differentiation was seen between specimens of A. limacoides sensu stricto from the European part of Russia and A. limacoides sensu Chen et al., 2010 from China (13.7%); the latter parasites were recognized as A. chongqingensis. Aspidogaster chongqingensis was more closely related to A. ijimai than to A. limacoides s. str. Specimens of A. ijimai from the Amur River, Khanka Lake (Russian Far East) and China were grouped into a single clade with low intra specific molecular differentiation (d = 0-0.3%). Specimens of A. conchicola from the European part of Russia, the Russian Far East and China also formed a single distinct clade. Genetic differentiation between European and Chinese samples of this species was two times lower (d = 0.45%) than between Russian Far East and European or Chinese samples (d = 0.96%), suggesting a long-term separate existence of A. conchicola in the Russian Far East.

Morphometric and molecular analyses of two digenean species from the mullet: Skrjabinolecithum spinosum n. sp. from the Russian southern Far East and Unisaccus tonkini n. sp. from Vietnam.

Adults of Skrjabinolecithum spinosum n. sp. were discovered in Mugil cephalus from the Gulf of Peter the Great in southern Far-East Russia. Additionally, adults of Unisaccus tonkini n. sp. were found in the intestine of Moolgarda cunnesius and Moolgarda seheli from the coastal waters of Cat Ba Island, Tonkin Bay, northern Vietnam. Skrjabinolecithum spinosum n. sp. possesses a larger body, and ventral and oral sucker size in comparison with Skrjabinolecithum vitellosum, a smaller pharynx size and body length/width rate ratio in comparison to Skrjabinolecithum pyriforme, a smaller body length and prepharynx size in comparison to Skrjabinolecithum lobolecitum and a smaller pharynx length and egg size in comparison to Skrjabinolecithum indicum and S. lobolecitum. The new species also differs from S. indicum, S. lobolecitum and S. vitellosum by the form of the testis, and from the last two species by the presence of a two-branched intestine. The morphometric parameters of S. spinosum n. sp. are similar to those of Skrjabinolecithum spasskii. However, S. spinosum n. sp., unlike S. spasskii, has an armed hermaphroditic duct. Unisaccus tonkini n. sp. is similar to Unisaccus spinosus (Martin, 1973), Unisaccus brisbanensis (Martin, 1973) and Unisaccus overstreeti (Ahmad, 1987) in body size but differs in oral sucker, pharynx and hermaphroditic sac size from U. spinosus, and in ventral sucker and ovary size from U. brisbanensis and U. overstreeti. Bayesian phylogenetic analysis, based on combined data of internal transcribed spacer 2 (ITS2) and partial 28S rRNA gene sequences, confirmed the validity of S. spinosum n. sp. and U. tonkini n. sp. Analysis of interrelationships of the family Haploporidae, including molecular data on new species, showed that the Waretrematinae subfamily is more heterogeneous in comparison with Haploporinae and Forticulcitinae, and includes U. tonkini n. sp.

Phylogenetic relationships of Hemiuridae (Digenea: Hemiuroidea) with new morphometric and molecular data of Aphanurus mugilis Tang, 1981 (Aphanurinae) from mullet fish of Vietnam.

Adult Aphanurus mugilis Tang, 1981 worms were detected in the intestine of Moolgarda engeli in the shallow waters off Cat Ba Island, Vietnam. Tang (1981) first described this species in Mugil cephalus off China. The worms in Vietnamese mullet were identical to Chinese specimens in a number of morphometric characteristics, with the exception of body and ovary size. In the present study, morphological characteristics, and the first molecular data for A. mugilis are provided. Additionally, molecular phylogenetic analysis of the family Hemiuridae was performed. The results of our molecular phylogenetic study indicate that the presence or absence of an ecsoma was not associated with molecular data for hemiurid subfamilies differentiation. The basal position of Bunocotylinae on the molecular-based phylogenetic tree indicated a primordial nature of ecsoma of hemiurid trematodes. Considerable molecular differentiation of Bunocotylinae from other hemiurids indicated the possibility of the recognition of the family Bunocotylidae Dollfus, 1950. Assuming that Machidatrema chilostoma is considered within the Bunocotylinae, the paraphyly of the Lecithasterinae was supported.

[Divergence of paralogous growth-hormone-encoding genes and their promoters in Salmonidae].

In many fish species, including salmonids, the growth-hormone is encoded by two duplicated paralogous genes, gh1 and gh2. Both genes were already in place at the time of divergence of species in this group. A comparison of the entire sequence of these genes of salmonids has shown that their conserved regions are associated with exons, while their most variable regions correspond to introns. Introns C and D include putative regulatory elements (sites Pit-1, CRE, and ERE), that are also conserved. In chars, the degree of polymorphism of gh2 gene is 2-3 times as large as that in gh1 gene. However, a comparison across all Salmonidae species would not extent this observation to other species. In both these chars' genes, the promoters are conserved mainly because they correspond to putative regulatory sequences (TATA box, binding sites for the pituitary transcription factor Pit-1 (F1-F4), CRE, GRE and RAR/RXR elements). The promoter of gh2 gene has a greater degree of polymorphism compared with gh1 gene promoter in all investigated species of salmonids. The observed differences in the rates of accumulation of changes in growth hormone encoding paralogs could be explained by differences in the intensity of selection.

Morphometric and molecular data of the two digenean species Lasiotocus lizae Liu, 2002 (Monorchiidae) and Paucivitellosus vietnamensis sp. n. (Bivesiculidae) from mullet fish in Tonkin Bay, Vietnam.

Adults of Lasiotocus lizae Liu, 2002 (Monorchiidae) were found in the mullet Liza longimanus (Günther) from Tonkin Bay, near Cat Ba Island, Vietnam. In this region, flukes belonging to the genus Paucivitellosus (Bivesiculudae) were found in Liza subviridis (Valenciennes), Liza engeli (Bleeker) and Valamugil seheli (Forskåi). Results of investigations showed that morphological features of L. lizae (like L. glebulentus Overstreet, 1971) do not meet the criteria of the genus Lasiotocus. In addition, L. lizae is highly differentiated from other species of Lasiotocus from which molecular data were obtained, including L. arrhichostoma Searle, Cutmore et Cribb, 2014 and L. typicum (Nicoll, 1912). Phylogenetic analyses revealed that L. lizae differs considerably from other species of the genus Lasiotocus presented in the GenBank database. We have identified a new species of the genus Paucivitellosus - P. vietnamensis sp. n. - from L. subviridis, which differs from P. fragilis Coil, Reid et Kuntz, 1965 by metrical and molecular (18S rRNA) data, and from P. hanumanthai Mani, 1989 by metric features. Our results also show considerable molecular differentiation between P. vietnamensis sp. n. and Paucivitellosus spp. recovered from L. engeli and V. seheli in Vietnam.

Morphometric and molecular analyses of Skrjabinolecithum pyriforme n. sp. (Digenea: Haploporidae) in mullet fish from the Primorsky Region, Russia.

A new species of Skrjabinolecithum (Digenea: Waretrematinae), Skrjabinolecithum pyriforme n. sp., has been found in the intestines of Liza haematocheila and Mugil cephalus from the Primorsky Region, Russia. These worms differ from S. vitellosum and S. lobolecithum by the presence of two caeca, as do S. spasskii, S. indicum and S. bengalensis. These species differ morphologically from S. pyriforme n. sp. by a number of features, including body width, oral sucker, pharynx, eggs and ratio of length and width of the body. The most similar species to S. pyriforme n. sp. is Platydidymus flecterotestis (Zhukov, 1971) with some differences in maximal body length, testis and egg sizes. The results of molecular analysis confirmed that this new species belongs to Skrjabinolecithum on the basis of close relationships with S. spasskii-type species.