Saccocoelioides is a genus within the family Haploporidae and is distributed throughout the Americas. The recent application of molecular techniques has facilitated the reorganization of this genus and the description of new species, resulting in a current total of 28 species. In Argentina, 11 species have been identified; however, the validity of Saccocoelioides bacilliformis and Saccocoelioides octavus remains controversial. Recently, specimens of digeneans were collected from two distinct geographical locations: the Paraná River basin (Misiones province) and the Río de la Plata basin (Buenos Aires province). These specimens were found parasitizing two different fish species: Bryconamericus sylvicola and Bryconamericus iheringi. Sequences of the 28S and COI genes were obtained for both digeneans from the two collection sites and hosts. The morphological study, combined with genetic sequences, confirmed their assignment to the genus Saccocoelioides. The COI and 28S gene sequences obtained from specimens at both collection sites exhibited genetic distances of 0.13% and 3.41%, respectively, indicating that these Saccocoelioides specimens represent a single species. The comprehensive taxonomic study of the newly collected specimens, along with an examination of the type material of S. bacilliformis and S. octavus, allows us to confirm the validity of both species. Argentina is home to more than 550 species of freshwater fauna, and it is likely that new species of Saccocoelioides remain to be discovered in underexplored regions of the basin, such as the Uruguay, Pilcomayo and Bermejo rivers.

Here we describe a new species of the genus Saccocoelioides found parasitizing Astyanax dissimilis Garavello & Sampaio, Psalidodon bifasciatus (Garavello and Sampaio) and Bryconamericus ikaa Casciotta, Almirón & Azpelicueta from the Iguazu National Park, Misiones province, Argentina. Saccocoelioides miguelmontesi n. sp. was studied based on morphological and molecular (28S rDNA and COI mtDNA sequences) data. The COI mtDNA tree indicated that the specimens collected from the three fish hosts are conspecific, with an intragroup p-distance of 0%. The new species shows an intermediate morphological configuration between the diminutive and robust forms described for Saccocoelioides by Curran (2018). Although, in the 28S rDNA tree, it is placed in a well-supported clade with the two robust species analysed (S. elongatus and S. magnus; p-distance of 1 and 2%, respectively), it differs from the robust group by the range of body size, mature egg size, oral and ventral sucker size, sucker ratio, oral sucker to pharynx ratio, and post-cecal or post-testis/body length percentage. Our results led us to redefine the robust group as having eggs shorter or equal in length to the pharynx. Saccocoelioides miguelmontesi n. sp. the 10th species reported from Argentina and the 7th species within the robust group.

Hemiurid digeneans conspecific with Stomachicola muraenesocis Yamaguti, 1934 (the type species of the genus Stomachicola Yamaguti, 1934) were collected from the stomach of the daggertooth pike conger Muraenesox cinereus (Forsskål) off the Persian Gulf of Iran. This study aimed to provide a detailed characterization of Stom. muraenesocis, including measurements, illustrations and scanning electron microscopy (s.e.m.) representations. Comparisons with the original and previous descriptions revealed morphological and metrical variations in several features (i.e. body size and shape, arrangement of reproductive organs, soma to ecsoma length ratio, position of genital opening, number of vitelline tubules and extension of uterine coils) between Stom. muraenesocis from different hosts and localities. This study presents the first molecular sequence data associated with the small (18S) and large (28S) subunit nuclear ribosomal RNA genes (rDNA) for Stom. muraenesocis. Phylogenetic analyses of the 18S dataset placed Stom. muraenesocis as sister lineage to a clade formed of a group of species of Lecithaster Lühe, 1901 (Lecithasteridae Odhner, 1905). In contrast, phylogenetic analyses based on the 28S consistently recovered a sister relationship between Stom. muraenesocis and representatives of the Hemiuridae Looss, 1899. Further comprehensive phylogenetically based classification in light of morphology and taxonomic history of the Hemiuridae and Lecithasteridae is required to infer phylogenetic affinities and historical biogeography of Stomachicola. A comprehensive list of previously reported species of Stomachicola together with their associated hosts, localities and morphometric data is provided.

Phyllodistomum pepirense n. sp. is described from the urinary bladder of Hoplias malabaricus (Bloch, 1794), sampled in the Jacaré-Pepira River in São Paulo state, Brazil. The isolates of the new species were recovered as a monophyletic group in the phylogenetic analysis of the 28S rRNA gene, which showed the new species as the sister taxa of Phyllodistomum virmantasi Pinacho-Pinacho, Sereno-Uribe, Hernández-Orts, García-Varela & Pérez-Ponce de León, 2021, a species sampled from an eleotrid fish in Southeastern Mexico. The new species differs morphologically from P. virmantasi by having a larger body size, slightly lobed testes and ovary, a mostly intercaecal uterus, slightly diverticulated caeca, and vitelline masses irregularly shaped. The new species is also readily distinguished from other species of Phyllodistomum Braun, 1899 reported from freshwater fishes in Brazil – namely, P. rhamdiae Amato & Amato, 1993 and P. spatula Odhner, 1902. The new species is herein described based on morphological characteristics, molecular data from D1–D3 domains of the 28S rRNA gene, host association, and geographical distribution.

Furcocercariae of the genus Neodiplostomum Railliet, 1919 (Diplostomidae Poirier, 1886) were found in freshwater snails Helicorbis sujfunensis Starobogatov, 1957 (Planorbidae Rafinesque, 1815) collected from three localities in the Russian southern Far East. For the trematodes from each locality, frogs played the role of the second intermediate host, and rats were the definitive host. Chickens were insusceptible to infection. The morphological and molecular data obtained for these trematodes indicated they were representatives of the same species. The experimentally-derived adult individuals were morphometrically similar to the East Asian Neodiplostomum seoulense (Seo, Rim, Lee, 1964), Neodiplostomum oriolinum Oschmarin, 1963, Neodiplostomum leei Chai and Shin, 2002, and Neodiplostomum boryongense Shin et al., 2008. Analysis of available data on the life cycle, developmental stage morphology, and molecular genetic characteristics of East Asian Neodiplostomum revealed a lack of information for objective assessment of the species status of neodiplostomula found in the East Asia region. Based on the considerations above and the data for the cox1 marker, we named the trematode Neodiplostomum cf. seoulense (Seo, Rim, Lee, 1964) sensu Pyo et al., 2014. In a phylogenetic reconstruction based on nuclear and mitochondrial markers, neodiplostomulas clustered into geographically related groups: South American, North American, European, and East Asian, with the former occupying an external position in the tree, which may indicate South America as a center of Neodiplostomum speciation.

Morphological discrimination of species is problematic in many digenean taxa. Parasites of marine fish from the genus Lecithaster Lühe, 1901 are a good example of this. Our goal was to understand which species of Lecithaster infect fish in the White Sea, and reveal their life cycles. We collected specimens of maritae from nine fish species, analysed their morphology and sequenced 28S ribosomal DNA and internal transcribed spacer 2 (ITS2). Contrary to previous accounts, all of them belong to a single species, Lecithaster salmonis Yamaguti, 1934, which was previously only recorded from the Pacific. Morphologically, our maritae specimens were highly variable, sharing characters of L. salmonis, Lecithaster confusus Odhner, 1905 and Lecithaster gibbosus (Rudolphi, 1802) Lühe, 1901. This variability did not correlate with the moderate differences in ITS2 among the specimens, and neither did the fish host species. Members of the subfamily Salmoninae appear to be the best suited definitive hosts, judging from the intensity rates. The intermediate hosts were also discovered: the first is Cryptonatica affinis (Gmelin, 1791) and the second are planktonic copepods. These lifecycle data from the White Sea are consistent with L. salmonis species identification and with the distribution of this species in the North Pacific. The geographical range of L. salmonis seems to be interrupted, and we discuss possible ways of L. salmonis expansion.

Trematode life cycles involve molluscs as first intermediate hosts. The identification of trematodes based on the morphology of cercariae released from molluscs is challenging because the identification relies on adult forms obtained from their definitive hosts. Several studies have recently genetically characterized these larval forms establishing a link with adults sampled from their vertebrate hosts, allowing their identification at species level. In Los Tuxtlas tropical rainforest, in south-eastern Mexico, 57 species of trematodes have been reported from wildlife vertebrates; however, studies evaluating the diversity of trematode cercariae in molluscs are lacking. Here, we studied 11 species of molluscs to assess the diversity of cercariae in two lakes of Los Tuxtlas, Veracruz, Mexico; six of the 11 species were infected. Twelve cercarial morphotypes were collected and characterized morphologically. Sequences of the 28S ribosomal RNA gene were generated to allocate each morphotype into a family using the most recent phylogenetic classification of the Digenea as a framework; molecular work revealed 16 genetic lineages; some cercariae were identified up to genus (Apharyngostrigea, Ascocotyle, Centrocestus, Echinochasmus, Lecithodendrium and Posthodiplostomum), and some to species levels (Gorgoderina rosamondae, Langeronia macrocirra, Oligogonotylus manteri and Phyllodistomum inecoli) based on their phylogenetic position within the tree, and the genetic distance with respect to other sequenced congeners. Therefore, the cercarial morphotypes in the present study represent at least 16 putative species. Our study contributes to a better understanding of the trematode diversity in an area of high vertebrate species diversity, and to the knowledge of trematode life cycles.

Digeneans (Trematoda: Digenea) are endoparasites that present a complex life cycle, generally involving an intermediate invertebrate host and a vertebrate host. There is limited information about which species of molluscs may act as intermediate hosts in the upper Paraná River floodplain (UPRF), where Aylacostoma chloroticum can be considered a potential candidate. The study of digeneans in this region is important because some of these parasites are potentially zoonotic, and, therefore, are relevant to public health. However, the correct identification of these organisms during the larval stages is difficult because of the lack of morphologically diagnostic characteristics. The objective of this study was to identify and molecularly characterize the larval stages of digeneans found in A. chloroticum in the UPRF, using the mitochondrial marker of subunit I of cytochrome c oxidase and the 28S nuclear marker. The molluscs were examined in the laboratory and three morphotypes of cercariae were found. DNA was extracted from the specimens obtained and was then amplified and sequenced. The morphotypes exhibited high genetic similarities with Pseudosellacotyla, Paralecithodendrium and Philophthalmus, indicating that these organisms belong to these genera. This is the first record of larval stages of these genera in molluscs collected in the UPRF.

Neoechinorhynchus is one of the most speciose genera of acanthocephalans, with approximately 116 described species. A recent study, aimed at establishing the genetic diversity of Neoechinorhynchus in Middle American freshwater fishes, validated nine species molecularly and morphologically and revealed the existence of 10 putative candidate species. Neoechinorhynchus golvani, a parasite commonly found in cichlids throughout Middle America with an allegedly large intraspecific morphological variability, was found to represent a species complex; species delimitation methods uncovered three additional genetic lineages. Here, we re-analyse the morphological and molecular data for N. golvani species complex infecting cichlids in that geographical area. A multivariate analysis of variance (MANOVA) was conducted particularly for the length of apical, middle and posterior hooks of the species/lineages of Neoechinorhynchus in cichlids, revealing morphological variation in the length of apical hooks for Lineage 8, although no morphological distinction was observed for Lineages 9 and 10. A new concatenated phylogenetic analysis of one mitochondrial and two ribosomal DNA genes was used to further corroborate the species delimitation among lineages; Neoechinorhynchus Lineage 8 was found to be morphologically and genetically distinct from its sister taxa, N. golvani and other two undescribed genetic lineages, and is formally described as a new species. Neoechinorhynchus costarricense n. sp. is described from the intestines of eight species of cichlids in Costa Rica. The new species is distinguished from the other species/lineages of Neoechinorhynchus in cichlids mainly by the size of the apical hooks of the proboscis.

Five of the 13 known species of Mammomonogamus have been described in members of the family Felidae, including domestic cats, making felids the most frequent hosts of Mammomonogamus. The occurrence of Mammomonogamus in felids is geographically scattered and information on the life cycle and other aspects of infections is lacking. The paucity of data opens the questions on possible conspecificity of some of the described species of Mammomonogamus and on the existence of possible reservoirs for infections in domestic cats in geographically isolated endemic foci of infection. To test such hypotheses, we compared sequences of mitochondrial and nuclear markers obtained from Mammomonogamus adults or eggs collected from domestic cats in three geographically distant localities. Based on morphology, geographic origin and site of infection, the worms examined can be referred to as Mammomonogamus ierei and Mammomonogamus auris. Phylogenetic analyses of both mitochondrial and ribosomal DNA markers showed monophyly of the genus Mammomonogamus and suggested the existence of at least two species in cats. Review of the literature, the existence of several species and the discontinuous geographic distribution of Mammomonogamus infections in domestic cats suggest an historical spillover of infection from wild reservoirs, presumably wild felids.

The soft scales (Hemiptera: Coccoidea: Coccidae) are a group of sap-sucking plant parasites, many of which are notorious agricultural pests. The quarantine and economic importance of soft scales necessitates rapid and reliable identification of these taxa. Nucleotide sequences of the mitochondrial cytochrome c oxidase subunit I (COI) gene (barcoding region) and 28S rDNA were generated from 340 individuals of 36 common soft scales in China. Distance-based [(best match, Automated Barcode Gap Discovery (ABGD)], tree-based (neighbor-joining, Bayesian inference), Klee diagrams, and general mixed Yule coalescent (GMYC) models were used to evaluate barcoding success rates in the data set. Best match showed that COI and 28S sequences could provide 100 and 95.52% correct identification, respectively. The average interspecific divergences were 19.81% for COI data and 20.38% for 28S data, and mean intraspecific divergences were 0.56 and 0.07%, respectively. For COI data, multiple methods (ABGD, Klee, and tree-based methods) resulted in general congruence with morphological identifications. However, GMYC analysis tended to provide more molecular operational taxonomic units (MOTUs). Twelve MOTUs derived from five morphospecies (Rhodococcus sariuoni, Pulvinaria vitis, Pulvinaria aurantii, Parasaissetia nigra, and Ceroplastes rubens) were observed using the GMYC approach. In addition, tree-based methods showed that 28S sequences could be used for species-level identification (except for Ceroplastes ceriferus – Ceroplastes pseudoceriferus), even with low genetic variation (<1%). This report demonstrates the robustness of DNA barcoding for species discrimination of soft scales with two molecular markers (COI and 28S) and provides a reliable barcode library and rapid diagnostic tool for common soft scales in China.

The cyanobacteriosponge Terpios hoshinota is expanding its range across the Indo-Pacific. This species can have massive outbreaks on coral reefs, actively kill corals it overgrows and affect the entire benthic community. Although it has received much attention on the post-outbreak follow-up, little is known about its ecology, habitat preferences, and the possible environmental triggers that cause its outbreaks. We present a baseline study in the Spermonde Archipelago, Indonesia, where T. hoshinota was first observed in 2012. We surveyed 27 reefs and recorded patches between 14 and 217 cm2, at four reef sites (~15% cover). The sponge was found on both mid-shelf and outer shelf reefs but not close to the coast and the city of Makassar. Differences in benthic community structure, as well as spatial variables relating to the on-to-offshore gradient in the Spermonde archipelago, neither constrained nor promoted its expansion. Patches of the sponge were mostly overgrowing branching corals, belonging to Acroporidae species. Genetic variation within T. hoshinota was studied by sequencing partitions of the mitochondrial CO1 and nuclear ribosomal 28S gene. Two haplotypes were found within the Spermonde archipelago, which differed from the CO1 sequence in GenBank. The present study provides an indication of habitat preferences of T. hoshinota in non-outbreak conditions, although it is still unclear which environmental conditions may lead to the onset of its outbreaks.

Pseudococcidae (mealybugs) is a large taxonomic group, including a number of agronomic pests. Taxonomic identification of mealybug species is a recurrent problem and represents a major barrier to the establishment of adequate pest management strategies. We combined molecular analysis of three DNA markers (28S-D2, cytochrome oxidase I and internal transcribed spacer 2) with morphological examination, for the identification of 176 specimens collected from 40 mealybug populations infesting various crops and ornamental plants in Egypt and France. This combination of DNA and morphological analyses led to the identification of 17 species: seven in Egypt (Planococcus citri (Risso), Planococcus ficus (Signoret), Maconellicoccus hirsutus (Green), Ferrisia virgata (Cockerell), Phenacoccus solenopsis Tinsley, Phenacoccus parvus Morrison and Saccharicoccus sacchari (Cockerell)) and 11 in France (Planococcus citri, Pseudococcus viburni Signoret, Pseudococcus longispinus (Targioni-Tozzetti), Pseudococcus comstocki (Kuwana), Rhizoecus amorphophalli Betrem, Trionymus bambusae (Green), Balanococcus diminutus (Leonardi), Phenacoccus madeirensis Green, Planococcus vovae (Nasonov), Dysmicoccus brevipes (Cockerell) and Phenacoccus aceris Signoret), Pl. citri being found in both countries. We also found genetic variation between populations considered to belong to the same species, justifying further investigation of the possible occurrence of complexes of cryptic taxa.

Dryocosmus kuriphilus Yasumatsu (Hymenoptera: Cynipidae) develops in chestnut buds that remain asymptomatic from oviposition (June–July) until budburst; it is, thus, easily spread by plant material used in propagation. Therefore, it is particularly interesting to identify infested plant batches before their movement. Unfortunately, a non-destructive method for checking buds has not yet been developed, and the only technique available is the screening of a bud sample. The visual investigation is long and requires highly skilled and trained staff. The purpose of this work was to set up an effective and fast method able to identify the presence of first instar larvae of D. kuriphilus in a large number of chestnut buds by PCR. Four primer pairs were designed on nuclear and mitochondrial sequences of a set of seven gall wasp taxa and tested on five different cynipid's DNA. Nested diagnostic PCR was carried out on DNA extracted from samples of 2 g buds simulating four levels of infestation (larvae were added to uninfested buds); 320 bp amplicon of 28S sequence was chosen as a marker to detect one larva out of 2 g buds. The method showed a potential efficiency of 5000 to 15,000 buds per week, depending on bud size.

Kudoid parasites are significant pathogens of marine fish. In the past, specific identification has been difficult due to a paucity of detailed morphological and biological information provided in the original description of some species. However, the introduction of DNA analysis has dramatically improved diagnosis. For morphological characterization, this paper proposes a new set of schematics including additional measurements to give a more thorough description of spore morphology, and provides evidence for uniqueness of 4 new species: Kudoa gunterae n. sp. (from 10 pomacentrid species and 1 apogonid species), K. kenti n. sp. (from 4 pomacentrid species), K. paraquadricornis n. sp. (from 4 carangid species), and K. whippsi n. sp. (from 8 pomacentrid species and 1 apogonid species). Subtle morphological differences found between closely related species were reflected in their genetics, with increased resolution provided by the large subunit, compared with that of the small subunit, of the ribosomal DNA gene region. This article proposes comprehensive requirements for species descriptions within the Kudoidae that incorporate biology, morphology, and genetic sequence.