Nuclear rRNA Research Articles

Molecular taxonomic characterization and infra-specific diversity of entomopathogenic Beauveria bassiana fungi from Argentina

It has been the aim of this study to molecular-taxonomically identify 15 Beauveria isolates collected from different geographical regions and insect hosts in Argentina and to investigate the levels of inter- and intra-specific diversity across this set of isolates. Based on phylogenetic analyses of EF1A-RPB1-RPB2 concatenated genes and BLOC markers, all Beauveria strains were identify as Beauveria bassiana. Within the B. bassiana clades of both phylogenies, isolates from Argentina were not clustered according to geographic origin or host. The 15 fungal isolates were further analyzed by PCR amplification of the intron insertion hot spot region of the nuclear 28S rRNA encoding sequence. By intron sequence and position, seven different group-I intron combinations termed variants A, B1, B2, C, D, E and F were found in the 15 isolates under study. Variants B1/B2 consisting of a single 28Si2 intron were found in ten isolates, whereas variant A occurred twice and variants C through F were unique across the set of isolates under study. The determination of the different introns and intron combinations in the 28S rRNA gene is a powerful tool for achieving infraspecific differentiation of B. bassiana isolates from Argentina.

Isospora and Lankesterella Parasites (Eimeriidae, Apicomplexa) of Passeriform Birds in Europe: Infection Rates, Phylogeny, and Pathogenicity.

Wild birds are common hosts to numerous intracellular parasites such as single-celled eukaryotes of the family Eimeriidae (order Eucoccidiorida, phylum Apicomplexa). We investigated the infection rates, phylogeny, and pathogenicity of Isospora and Lankesterella parasites in wild and captive passerine birds. Blood and tissue samples of 815 wild and 15 deceased captive birds from Europe were tested using polymerase chain reaction and partial sequencing of the mitochondrial cytochrome b and cytochrome c oxidase I and the nuclear 18S rRNA gene. The infection rate for Lankesterella in wild birds was 10.7% compared to 5.8% for Isospora. Chromogenic in situ hybridization with probes targeting the parasites' 18S rRNA was employed to identify the parasites' presence in multiple organs, and hematoxylin-eosin staining was performed to visualize the parasite stages and assess associated lesions. Isospora parasites were mainly identified in the intestine, spleen, and liver. Extraintestinal tissue stages of Isospora were accompanied by predominantly lymphohistiocytic inflammation of varying severity. Lankesterella was most frequently detected in the spleen, lung, and brain; however, infected birds presented only a low parasite burden without associated pathological changes. These findings contribute to our understanding of Isospora and Lankesterella parasites in wild birds.

Vicariance and cryptic diversity revealed by molecular phylogenetic analyses of estuarine Gammarus species (Crustacea: Amphipoda) due to formation of the Labrador Current

The metapopulation of the estuarine species Gammarus tigrinus along the east coast of the United States has been hypothesised to represent two cryptic species divided biogeographically off the coast of North Carolina, USA. This divergence has been attributed to a strong temperature gradient created by the formation of the cold Labrador Current c. 3.0 million years ago. In addition, the northern phylogeographic clade of G. tigrinus has been demonstrated to be invasive in estuarine habitats across a large portion of northern Europe. Recent collections of G. tigrinus from Florida and Maryland, USA, allow for new approaches to test this hypothesis. Using the nuclear 18S and 28S rRNA, and mitochondrial 16S rRNA and cytochrome c oxidase subunit I genes, species delimitation models provide support that the genetic divergence of the northern and southern clades is equivalent to species level. In addition, molecular clock data demonstrate that this phylogeographic divergence coincides with the formation of the Labrador Current. Furthermore, the collections of G. daiberi from Florida, a species with biogeographical and ecological characteristics similar to those of G. tigrinus, provide independent support for the hypothesis. The potential for invasive species to be cryptic highlights the need for accurate identification of taxa to ensure that appropriate biogeographical assessment of potential source populations and mechanisms of dispersal can be made.

Paratrypaea Komai & Tachikawa, 2008 (Decapoda: Axiidea: Callianassidae): more than two or three species

Abstract Paratrypaea Komai & Tachikawa, 2008 has for more than a decade been thought to contain only two species, both widespread throughout the Indo-West Pacific, or briefly three species. Molecular studies using mitochondrial 16S rRNA and 12S rRNA plus nuclear H3 and 18S rRNA genes, repeated here with samples from more individuals, have shown well separated clades within the genus. Morphological examination has found differences, especially in the form of the adult male cheliped, between the clades leading to the conclusion that Paratrypaea comprises at least eight species of which four are new and described herein. Not all closely related clades could be morphologically distinguished. Some species are narrowly and others widely distributed. The species are: P. baliensissp. nov. from Indonesia; P. bouvieri (Nobili, 1904) from the Red Sea and Indian Ocean; P. chichijimaensis (Sakai, 2015) comb. nov. from Japan; P. dworschakisp. nov. from the Hawaiian Islands; P. maldivensis (Borradaile, 1904) from the eastern Indian Ocean, Papua New Guinea, and possibly western Pacific; P. niuginiensissp. nov. from Papua New Guinea; P. rectangularis (Ngoc-Ho, 1991) from French Polynesia to the Red Sea; and P. rempiensissp. nov. from Papua New Guinea. A published partial 16S sequence from Japan has provided evidence of one more species than those proposed here. Paratrypaea cristata (Borradaile, 1910) comb. nov. remains a species inquirenda.

Description of Naviculavanseea sp. nov. (Naviculales, Naviculaceae), a new species of diatom from the highly alkaline Lake Van (Republic of Türkiye) with complete characterisation of its organellar genomes and multigene phylogeny.

The current article describes Naviculavanseeasp. nov., a new species of diatom from Lake Van, a highly alkaline lake in Eastern Anatolia (Türkiye). The description is based on light and scanning electron microscopy performed on two monoclonal cultures. The complete nuclear rRNA clusters and plastid genomes have been sequenced for these two strains and the complete mitogenome for one of them. The plastome of both strains shows the probable loss of a functional ycf35 gene. They also exhibit two IB4 group I introns in their rrl, each encoding for a putative LAGLIDADG homing endonuclease, with the first L1917 IB4 intron reported amongst diatoms. The Maximum Likelihood phylogeny inferred from a concatenated alignment of 18S, rbcL and psbC distinguishes N.vanseea sp. nov. from the morphologically similar species Naviculacincta and Naviculamicrodigitoradiata.

Two new species of Pseudocrangonyx (Amphipoda: Pseudocrangonyctidae) from the hyporheic zones in South Korea

Pseudocrangonyx Akatsuka & Komai, 1922, is the most diverse group of subterranean amphipods in the groundwater communities of Far East Asia and is related to the biogeographic origin and evolution of subterranean amphipod fauna in the region. In this study, we describe two new subterranean species of pseudocrangonyctid amphipods, P. seomjinensis sp. nov. and P. danyangensis sp. nov., from the hyporheic zones in South Korea. These new species can be distinguished from their congeners by the length of the uropod 2 outer ramus, the number of accessory flagella of antenna 1, and telson shape. Additionally, we determined sequences of the nuclear 28S rRNA and the mitochondrial cytochrome c oxidase subunit I gene of the new species for molecular analyses.

A new parasitic barnacle (Crustacea, Cirripedia, Rhizocephala, Mycetomorpha) from the abyssal zone in the northwestern Pacific

I describe the parasitic barnacle Mycetomorpha abyssalissp. nov. from the crangonid shrimp Sclerocrangon zenkevitchi collected from 3893–3890 m depth off the eastern coast of Iwate, Japan, northwestern Pacific. This is the first Mycetomorpha rhizocephalan from the abyssal zone and the third species in Mycetomorpha. Mycetomorpha abyssalissp. nov. differs from its congeners M. vancouverensis and M. albatrossi in (1) triangular shield lacking, (2) stalk one-quarter of length from posterior end of externa, (3) mantle opening clearly anterior to stalk, (4) different host genus, and (5) depth range much deeper. I determined partial sequences for the mitochondrial cytochrome c oxidase subunit I (COI) and 16S rRNA genes and nuclear 18S rRNA and 28S rRNA genes from M. abyssalissp. nov. for future DNA barcoding and phylogeny. Kimura 2-parameter distances between M. abyssalissp. nov. and M. vancouverensis were 21.2% (16S), 0.6% (18S), and 1.5% (28S).

The invasive land flatworm Arthurdendyus triangulatus has repeated sequences in the mitogenome, extra-long cox2 gene and paralogous nuclear rRNA clusters

Using a combination of short- and long-reads sequencing, we were able to sequence the complete mitochondrial genome of the invasive ‘New Zealand flatworm’ Arthurdendyus triangulatus (Geoplanidae, Rhynchodeminae, Caenoplanini) and its two complete paralogous nuclear rRNA gene clusters. The mitogenome has a total length of 20,309 bp and contains repetitions that includes two types of tandem-repeats that could not be solved by short-reads sequencing. We also sequenced for the first time the mitogenomes of four species of Caenoplana (Caenoplanini). A maximum likelihood phylogeny associated A. triangulatus with the other Caenoplanini but Parakontikia ventrolineata and Australopacifica atrata were rejected from the Caenoplanini and associated instead with the Rhynchodemini, with Platydemus manokwari. It was found that the mitogenomes of all species of the subfamily Rhynchodeminae share several unusual structural features, including a very long cox2 gene. This is the first time that the complete paralogous rRNA clusters, which differ in length, sequence and seemingly number of copies, were obtained for a Geoplanidae.

New insights into the molecular phylogeny, biogeographical history, and diversification of Amblyomma ticks (Acari: Ixodidae) based on mitogenomes and nuclear sequences.

Amblyomma is the third most diversified genus of Ixodidae that is distributed across the Indomalayan, Afrotropical, Australasian (IAA), Nearctic and Neotropical biogeographic ecoregions, reaching in the Neotropic its highest diversity. There have been hints in previously published phylogenetic trees from mitochondrial genome, nuclear rRNA, from combinations of both and morphology that the Australasian Amblyomma or the Australasian Amblyomma plus the Amblyomma species from the southern cone of South America, might be sister-group to the Amblyomma of the rest of the world. However, a stable phylogenetic framework of Amblyomma for a better understanding of the biogeographic patterns underpinning its diversification is lacking. We used genomic techniques to sequence complete and nearly complete mitochondrial genomes -ca. 15 kbp- as well as the nuclear ribosomal cluster -ca. 8 kbp- for 17 Amblyomma ticks in order to study the phylogeny and biogeographic pattern of the genus Amblyomma, with particular emphasis on the Neotropical region. The new genomic information generated here together with genomic information available on 43 ticks (22 other Amblyomma species and 21 other hard ticks-as outgroup-) were used to perform probabilistic methods of phylogenetic and biogeographic inferences and time-tree estimation using biogeographic dates. In the present paper, we present the strongest evidence yet that Australasian Amblyomma may indeed be the sister-group to the Amblyomma of the rest of the world (species that occur mainly in the Neotropical and Afrotropical zoogeographic regions). Our results showed that all Amblyomma subgenera (Cernyomma, Anastosiella, Xiphiastor, Adenopleura, Aponomma and Dermiomma) are not monophyletic, except for Walkeriana and Amblyomma. Likewise, our best biogeographic scenario supports the origin of Amblyomma and its posterior diversification in the southern hemisphere at 47.8 and 36.8 Mya, respectively. This diversification could be associated with the end of the connection of Australasia and Neotropical ecoregions by the Antarctic land bridge. Also, the biogeographic analyses let us see the colonization patterns of some neotropical Amblyomma species to the Nearctic. We found strong evidence that the main theater of diversification of Amblyomma was the southern hemisphere, potentially driven by the Antarctic Bridge's intermittent connection in the late Eocene. In addition, the subgeneric classification of Amblyomma lacks evolutionary support. Future studies using denser taxonomic sampling may lead to new findings on the phylogenetic relationships and biogeographic history of Amblyomma genus.

A cyst-forming coccidian with large geographical range infecting forest and commensal rodents: Sarcocystis muricoelognathis sp. nov.

BackgroundThe geographic distribution and host-parasite interaction networks of Sarcocystis spp. in small mammals in eastern Asia remain incompletely known.MethodsExperimental infections, morphological and molecular characterizations were used for discrimination of a new Sarcocystis species isolated from colubrid snakes and small mammals collected in Thailand, Borneo and China.ResultsWe identified a new species, Sarcocystis muricoelognathis sp. nov., that features a relatively wide geographic distribution and infects both commensal and forest-inhabiting intermediate hosts. Sarcocystis sporocysts collected from rat snakes (Coelognathus radiatus, C. flavolineatus) in Thailand induced development of sarcocysts in experimental SD rats showing a type 10a cyst wall ultrastructure that was identical with those found in Rattus norvegicus from China and the forest rat Maxomys whiteheadi in Borneo. Its cystozoites had equal sizes in all intermediate hosts and locations, while sporocysts and cystozoites were distinct from other Sarcocystis species. Partial 28S rRNA sequences of S. muricoelognathis from M. whiteheadi were largely identical to those from R. norvegicus in China but distinct from newly sequenced Sarcocystiszuoi. The phylogeny of the nuclear 18S rRNA gene placed S. muricoelognathis within the so-called S. zuoi complex, including Sarcocystisattenuati, S. kani, S. scandentiborneensis and S. zuoi, while the latter clustered with the new species. However, the phylogeny of the ITS1-region confirmed the distinction between S. muricoelognathis and S. zuoi. Moreover, all three gene trees suggested that an isolate previously addressed as S. zuoi from Thailand (KU341120) is conspecific with S. muricoelognathis. Partial mitochondrial cox1 sequences of S. muricoelognathis were almost identical with those from other members of the group suggesting a shared, recent ancestry. Additionally, we isolated two partial 28S rRNA Sarcocystis sequences from Low’s squirrel Sundasciurus lowii that clustered with those of S. scandentiborneensis from treeshews.ConclusionsOur results provide strong evidence of broad geographic distributions of rodent-associated Sarcocystis and host shifts between commensal and forest small mammal species, even if the known host associations remain likely only snapshots of the true associations.Graphical

Four new species and a ribosomal phylogeny of Rhabdopleura (Hemichordata: Graptolithina) from New Zealand, with a review and key to all described extant taxa.

All eight extant species ofRhabdopleuradescribed between 1869 and 2018 are provisionally accepted as valid based on a review of the literature and new data on two little-known species from the Azores. Additionally, four new species are described from the New Zealand region, increasing global diversity by 50%, and a dichotomous key to all 12 described species is provided based on morphological criteria. The distinction between colony morphologies based on erect-tube inception is regarded as particularly helpful in initial characterization of species. Erect ringed tubes are either produced directly from the surface of creeping-tubes or indirectly, i.e. a short adherent side branch from a creeping tube is interpolated between the creeping tube and an erect tube; such side branches are blind-ending. These two modes of erect-tube origination are here respectively termeddirectandindirect. Species with indirect erect-tube budding are predominant in the North Atlantic whereas species with direct erect-tube budding dominate in New Zealand waters. The only indirect-erect species from New Zealand, Rhabdopleura chathamica n. sp., was discovered on deepwater coral from 10081075 m, constituting the deepest record of the genus to date. Rhabdopleura emancipata n. sp., collected only in a detached state, constitutes a three-dimensional tangled growth that grew freely into the water columna unique morphology hitherto unknown among extant species. Owing to this growth mode, it provided a substratum for epibionts from several phyla. Rhabdopleura francesca n. sp. and Rhabdopleura decipula n. sp. are morphologically very similar but are distinguishable by their distinct placements in a phylogeny based on 16S mitochondrial and 18S nuclear rRNA genes. Phylogenetic reconstructions based on rRNA and mitochondrial genome data contribute to an updated phylogeny of all Rhabdopleura species sequenced thus far, some of which require more molecular sequences and morphological analyses for taxonomic determination.

Terrestrial predatory leeches of the genus Orobdella (Hirudinea: Erpobdelliformes: Orobdellidae) endemic to the Southern Russian Far East: a new species of the genus from Primorsky Krai, Russia.

A quadrannulate species, Orobdella ganini sp. nov., is described from the Lazovsky Nature Reserve in Primorsky Krai, the Southern Russian Far East, Russia. Morphological features of O. ghilarovi Nakano & Prozorova, 2019 from the reserve are also provided leading to an amendment of the species diagnosis. Maximum likelihood and Bayesian inference phylogenetic analyses, which were performed using nuclear 18S rRNA, 28S rRNA, and histone H3, and mitochondrial cytochrome c oxidase subunit I, tRNACys, tRNAMet, 12S rRNA, tRNAVal, 16S rRNA, tRNALeu and NADH dehydrogenase subunit 1 markers, show that O. ganini sp. nov., O. ghilarovi and two species endemic to Hokkaido, Japan form a clade, with the new species sister to a lineage composed of the two Japanese species. A partial cytochrome c oxidase subunit I sequence obtained from a cocoon found in the Lazovsky Nature Reserve reveals that Orobdella leeches deposit cocoons somewhat similar to those deposited by terrestrial blood-sucking leeches of Haemadipsidae.

The chloroplast and mitochondrial genomes of two Gomphonema parvulum (Bacillariophyta) environmental isolates from South Carolina (United States) and Virginia (United States).

Gomphonema parvulum is a cosmopolitan freshwater diatom that is used as an indicator in water quality biomonitoring. In this study, we report the culturing of two geographically separated isolates from southeastern North America, their morphology, and the sequencing and assembly of their mitochondrial and chloroplast genomes. Morphologically, both strains fit G. parvulum sensu lato, but the frustules from a protected habitat in South Carolina were smaller than those cited in the historic data of this species from the same location as well as a second culture from Virginia. Phylogenetic analyses using the rbcL gene placed both within a clade with G. parvulum. Genetic markers, including full chloroplast and mitochondrial genomes and the nuclear small subunit rRNA gene region were assembled from each isolate. The organellar genomes of the two strains varied slightly in size due to small differences in intergenic regions with chloroplast genomes of 121,035 bp and 121,482 bp and mitochondrial genomes of 34,639 bp and 34,654 bp. The intraspecific pairwise identities of the chloroplast and mitochondrial genomes of these two isolates were 97.9% and 95.4%, respectively. Multigene phylogenetic analysis demonstrated a close relationship between G. parvulum, Gomphoneis minuta, and Didymosphenia geminata.

Two new genera and species of the valvatiform hydrobiid snails (Caenogastropoda: Truncatelloidea: Hydrobiidae) from Morocco.

Stygobiont and crenobiont minute gastropods representing the family Hydrobiidae (Caenogastropoda: Truncatelloidea), characterized by the valvatoid low-spired shell, were collected from one spring and four wells in Bouregreg region, NW Morocco. The shells were photographed and measured; shell biometry is illustrated with principal component analysis. Penes were illustrated and described. Mitochondrial cytochrome oxidase subunit 1 (COI) and 16S rRNA, as well as nuclear 18S rRNA sequences were used to infer the phylogeny. The snails represented two genera, both new to science. Their closest relatives were taxa from the Iberian Peninsula, the rough molecular estimate of the time of divergence between these Moroccan and Iberian genera coincided with the Pliocene Flooding, which restored the Strait of Gibraltar to connect the Mediterranean Basin with Atlantic Ocean.

Taxonomic study of the polyphyletic Dudresnaya (Dumontiaceae, Florideophyceae) with descriptions of Dudresnaya ryukyuensis sp. nov. and two new genera, Himehibirhodia and Nudresdaya

SUMMARYThe red algal genus Dudresnaya (Dumontiaceae, Gigartinales) has traditionally been a morphologically well‐defined taxon, but its molecular phylogeny has rarely been studied. To examine the phylogenetic relationships among Dudresnaya species, we generated new partial sequences of mitochondrial cox1, chloroplast rbcL and nuclear 28S rRNA genes from an undescribed Dudresnaya species from Okinawa Island, Japan, alongside five additional described species. Our phylogenetic analyses show that Dudresnaya is genetically diverse and polyphyletic. Based on molecular phylogeny and morphological data, we describe the Okinawan Dudresnaya as a new species, Dudresnaya ryukyuensis, and transferred Dudresnaya minima and Dudresnaya littleri, which were phylogenetically and morphologically distinct from the genuine Dudresnaya, to the new genera Himehibirhodia and Nudresdaya, respectively. Our phylogenetic analyses also showed that the Dumontiaceae is not a monophyletic group including the Gainiaceae and Rhizophyllidaceae (DGR complex). Considering that the DGR complex exhibits female reproductive structures and their post‐fertilization development that are similar to each other, the DGR complex appears to be recognized as the Dumontiaceae sensu lato.

Microeukaryotes have unexpected importance in cold seep food webs through predation and parasitism

Cold seeps are inhabited by a high abundance of chemosynthetic prokaryotes and invertebrates. However, a complete food web between invertebrates and prokaryotes cannot be established without the important link of microeukaryotes. Yet, question such as which microeukaryotes are at work in cold seeps is still mostly open. We, therefore, compared the biological communities in cold seeps and adjacent areas of the South China Sea by the metabarcoding and quantitative PCR (qPCR) of nuclear 18S and 16S rRNA genes, and estimated the potentially ecological roles of microeukaryotes by cross-domain microbial network analysis. The anaerobic methane oxidizing archaea (ANME), sulfate-reducing bacteria (SRB) and Radiolaria accounted for most variations between microbial communities from the cold seeps and adjacent areas. A significant correlation was detected between the copy numbers of 18S rRNA gene and communities of the ANME and SRB in cold seeps, indicating close links between microeukaryotes and chemosynthetic prokaryotes. After adding microeukaryotic nodes microbial network became more complex, and the negative correlation proportion and average path length increased, suggesting increased predation and parasitism. The ANME and SRB groups were mainly negatively correlated with and potentially preyed by Acantharea and Cercozoa, which were then negatively correlated with annelids and mollusks. Parasitic microeukaryotes of Syndiniales widely existed in cold seeps and had close relationships with Polycystinea. Parasitism might cause the death of Polycystinea and the release of Syndiniales spores, which may be an important carbon source for the cold seep biota. Using Mantel analyses, we also demonstrated that trophic relationships are closer in cold seeps than in the adjacently deep-sea sediments, and that microeukaryote-metazoans linkages in cold seeps are closer than those between prokaryotes and metazoans. Overall, our study outlines the main components of food webs in cold seeps and highlights that microeukaryotes have an unexpected importance in the carbon cycle.

The Extraordinary Diversity of Merodon avidus Complex (Diptera: Syrphidae)-Adding New Areas, New Species and a New Molecular Marker.

In this paper, the Merodon avidus (Diptera, Syrphidae) species complex was revised, whereupon we discovered and described four new species for science: Merodon atroavidus Vujić, Radenković et Likov sp. nov., M. magnus Vujić, Kočiš Tubić et Ačanski sp. nov., M. nigroscutum Vujić, Radenković et Likov sp. nov. and M. pseudomoenium Vujić, Kočiš Tubić et Ačanski sp. nov. An integrative taxonomy approach was used to delimit species boundaries. Two molecular markers (the mitochondrial COI gene and nuclear 28S rRNA gene-newly analysed marker for the complex) and geometric morphometry of the wing shape, together with morphological data and distribution, successfully separated all species from the complex. The morphological variability of the analysed species is described and discussed and an illustrated diagnostic key for typical morpho-forms of species from the M. avidus complex is presented. A distribution map of all investigated species from the complex is provided. The level of endemicity of the M. avidus complex was discussed.

Morphological and phylogenetic resolution of Conoideocrella luteorostrata (Hypocreales: Clavicipitaceae), a potential biocontrol fungus for Fiorinia externa in United States Christmas tree production areas

ABSTRACT The entomopathogenic fungus Conoideocrella luteorostrata has recently been implicated in natural epizootics among exotic elongate hemlock scale (EHS) insects in Christmas tree farms in the eastern United States. Since 1913, C. luteorostrata has been reported from various plant feeding Hemiptera in the southeastern United States, but comprehensive morphological and phylogenetic studies of U.S. populations are lacking. The recovery of multiple strains of C. luteorostrata from mycosed EHS in North Carolina provided an opportunity to conduct pathogenicity assays and morphological and phylogenetic studies to investigate genus- and species-level boundaries among the Clavicipitaceae. Pathogenicity assays confirmed that C. luteorostrata causes mortality of EHS crawlers, an essential first step in developing this fungus as a biocontrol. Morphological studies revealed that conidia aligned with previous measurements of the Paecilomyces-like asexual state of C. luteorostrata, with conidiophore morphology consistent with historical observations. Additionally, a Hirsutella-like synanamorph was observed in select C. luteorostrata strains. In both a four-locus, 54-taxon Clavicipitaceae-wide phylogenetic analysis including D1–D2 domains of the nuclear 28S rRNA region (28S), elongation factor 1 alpha (EF1-α), DNA-directed RNA polymerase II subunit 1 (RPB1), and DNA-directed RNA polymerase II subunit 2 (RPB2) and a two-locus, 38-taxon (28S and EF1-α) phylogenetic analysis, all three Conoideocrella species were resolved as strongly supported monophyletic lineages across all loci and both methods (maximum likelihood and Bayesian inference) of phylogenetic inference except for 28S for C. tenuis. Despite the strong support for individual Conoideocrella species, none of the analyses supported the monophyly of Conoideocrella with the inclusion of Dussiella. Due to the paucity of RPB1 and RPB2 sequence data, EF1-α provided superior delimitation of intraspecies groupings for Conoideocrella and should be used in future studies. Further development of C. luteorostrata as a biocontrol against EHS will require additional surveys across diverse Hemiptera and expanded pathogenicity testing to clarify host range and efficacy of this fungus.

Assessing the applicability of nuclear 18S and ITS2-28S rRNA genes for diagnostics of Bactrocera dorsalis (Diptera: Tephritidae) in the Philippines

Fruit flies belonging to the Bactrocera dorsalis species complex pose a significant threat to mangoes and other crops in the Philippines and worldwide. Identifying cryptic species within this complex is challenging, particularly when relying solely on morphological analysis. In this study, we sequenced two fragments of the nuclear 18S and ITS2-28S rRNA genes from specimens of Bactrocera dorsalis Hendel collected in the Philippines to assess their applicability for species diagnostics. Subsequent sequencing and analysis revealed that the 18S and 28S rRNA gene fragments matched B. dorsalis sequences in NCBI but also displayed high similarity with other Bactrocera and insect species. On the other hand, sequences of the ITS2 segment showed hits specific to B. dorsalis. Further analysis of the 18S rRNA gene in fruit flies collected from various sources and host plants in the country suggests conserved sequences among Bactrocera samples, irrespective of collection site and host plant species. In conclusion, our findings suggest that, among the tested nuclear DNA fragments, only the ITS2 demonstrates sufficient species-level nucleotide variation for effective use as a molecular diagnostic marker for B. dorsalis identification.

New insights into a poorly known parasite, Dero lutzi (Oligochaeta: Naididae), associated with tree frogs of the genus Scinax: morphological evaluation and genotypic data.

The oligochaete Dero lutzi follows a life strategy that alternates between free-living periods in aquatic environments and endoparasitic phases. Most occurrences of D. lutzi in anurans are reported in species with arboreal habits, with studies limited to the recording of the oligochaete's presence in the host. Our study recovered specimens of D. lutzi from the tree frogs Scinax fuscovarius and Scinax. nasicus. We performed a morphological assessment of the parasite using light microscopy, for the first time, scanning electron microscopy. Molecular characterization of D. lutzi was carried out using the mitochondrial gene 16S rRNA and the nuclear gene 28S rRNA. Additionally, a phylogenetic tree was constructed to assess the species´position in relation to other group members. In our results, we confirmed the phenotypic morphological characteristics of the endoparasitic phase of D. lutzi. We also presented its phylogenetic position with other oligochaetes in the group, demonstrating the proximity between the endoparasite D. lutzi and the free-living oligochaete D. superterrenus.