Description and phylogenetic relationships of a new genus of Planoceridae (Polycladida, Acotylea) from Shimoda, Japan

We establish a new genus of planocerid polyclad, Heteroplanocera gen. nov., based on a new species, H. katoi sp. nov., collected at a depth of 245 m off the coast of the Izu Peninsula, West Pacific, Japan. Heteroplanocera gen. nov. is distinguished from other planocerid genera by the following characteristics: (i) a pair of nuchal tentacles, (ii) a muscular bulb, (iii) a pair of spermiducal bulbs (instead of a seminal vesicle), (iv) a free prostatic vesicle, (v) a pair of accessory organs with teeth, (vi) a Lang's vesicle, and (vii) the lack of a bursa copulatrix. We provide a partial sequence of the cytochrome c oxidase subunit I (COI) gene (712 bp) as a DNA barcode of the new species. Furthermore, we estimate the phylogenetic position of Heteroplanocera gen. nov. among Planoceridae based on partial sequences of COI and 16S, 18S, and 28S ribosomal DNA. Based on our findings, we discuss the evolution and function of the (i) vaginal cuticularized structure and (ii) accessory organs among planocerid flatworms.

Description of a new species Coomansus batxatensis (Mononchida, Mononchidae) from Vietnam, with an updated key to species

Coomansus batxatensis sp. nov., recorded from Vietnam, is described and illustrated and its phylogenetic relationship within the Mononchida is analysed. The molecular data (18S and 28S ribosomal DNA) are provided for the new species. The new species is characterized by small body size (body length, L = 0.7–0.9 mm); buccal cavity sub-rectangular in shape, flattened at base, 21–24 × 12–13 μm or 1.9 (1.7–2.0) times as long as wide; posterior position of dorsal tooth apex (59–63% from the base of buccal cavity); pars refringens vaginae with faint and small (2.5 × 1.7 μm) teardrop-shaped pieces, short pars distalis vaginae; and males with short spicules (50–51.5 μm) with rounded head and conical blade part. The new species is close to Coomansus parvus but differs from it by the smaller buccal cavity size, more posterior position of the dorsal tooth apex, longer tail and presence of males. An updated identification key to Coomansus species and a compendium of all the species known are presented.

Comparative study of four known species of the genus Acrobeles von Linstow, 1877 (Nematoda, Cephalobidae) with ‘single’ and ‘double’ cuticle from coastal dunes in Spain

The nematode genus Acrobeles is composed of two morphological groups distinguished by the presence (‘double’ cuticle) or absence (‘single’ cuticle) of the refringent inner layer of the cuticle. In the present study, four species of this genus, two with ‘single’ cuticle (Acrobeles ciliatus and Acrobeles cylindricus) and two with ‘double’ (Acrobeles aenigmaticus and Acrobeles complexus) are studied from coastal dunes in Spain. This study provides detailed morphological and morphometrical analyses for the four species, while molecular analysis, based on 18S and 28S ribosomal DNA, is provided for A. complexus. The four species are studied with scanning electron microscopy, which is obtained for the first time for A. cylindricus. These analyses revealed morphological and molecular differentiations between both groups, appearing as two related monophyletic entities. The subgenera Acrobeles and Seleborca, formerly considered as separate genera, are proposed to accommodate both groups.

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.

Characterization of Nothacrobeles lanceolatus Abolafia & Peña-Santiago, 2003 (Rhabditida, Cephalobidae), a typical species of coastal dunes from Spain

The nematode Nothacrobeles lanceolatus was described in arid soils from Cabo de Gata-Níjar Natural Park, province of Almería (Spain). Since then this species has not been found again. In this study, new data on the distribution and morphological, morphometrical and molecular characteristics (based on 18S and 28S ribosomal DNA) of N. lanceolatus is provided. This species is characterized by having a tessellated cuticle, with its inner layer bearing two rows of cuticular punctations per annulus, lips quadrangular with eight pinnae, labial probolae bifurcate with divergent prongs surrounded by pinnae, pharynx with corpus 2.5–4.0 times isthmus length, post-vulval uterine sac 1.4–2.2 times the corresponding body diameter, and female and male tails conical, bearing a conical mucro with acute terminus. The phylogenetic position of this and related species of the genus Nothacrobeles is discussed. Additionally, bacteria (Pasteuria) and fungi (Catenaria, Nematocida) feeding on N. lanceolatus are showed including light microscopy and scanning electron microscopy illustrations.

Presence and Genetic Identity of Symbiodiniaceae in the Bioeroding Sponge Genera Cliona and Spheciospongia (Clionaidae) in the Spermonde Archipelago (SW Sulawesi), Indonesia

Members of the family Symbiodiniaceae form symbiotic relationships with several metazoan groups on coral reefs, most notably scleractinian corals. However, despite their importance to the health of coral reefs, their relationship with other host organisms such as bioeroding sponges (Clionaidae) is still relatively understudied. In this study we investigate the presence and identity of Symbiodiniaceae in Clionaidae species in Indonesia and evaluate findings related to the evolution and ecology of the host-symbiont relationship. Clionaidae were collected throughout the Spermonde Archipelago in Indonesia. Morphological and molecular techniques were used to identify the sponge host (28S ribosomal DNA) and their Symbiodiniaceae symbionts (ITS2). Seven Clionaidae species were found, of which four species contained Symbiodiniaceae. Cliona aff. orientalis, Cliona thomasi and Spheciospongia maeandrina were host to Cladocopium, while Spheciospongia digitata contained Durusdinium and Freudenthalidium. In the remaining species: Cliona sp., Cliona utricularis and Spheciospongia trincomaliensis no evidence of the presence of Symbiodiniaceae was found. Our results provide the first record of Symbiodiniaceae in the sponge genus Spheciospongia. Additionally, we provide the first findings of Freudenthalidium and first molecular evidence of Durusdinium in bioeroding sponges. Our results indicate coevolution between Spheciospongia digitata, Spheciospongia maeandrina and their symbionts. We discuss that the diversity of Symbiodiniaceae within bioeroding sponges is likely far greater than currently reported in literature. Considering the threat bioeroding sponges can pose to the health of coral reefs, it is crucial to understand Symbiodiniaceae diversity within Clionaidae and their effect on the functioning of Clionaidae species. We propose that the identity of the symbiont species is mostly related to the host species, but we did observe a potential case of environmental adaptation related to environmental stressors.

Coltriciella multipileata (Agaricomycetes, Hymenochaetaceae), a new species from Mexico, related to ectomycorrhizal lineages

Coltriciella multipileata is described here as a new species from Mexico. The species grow on soils in open areas at the vicinity of living trees of Pinus patula, and its known only from the Parque Recreativo Los Colomos, Jalisco. The phylogenetic analysis based on partial nuclear 28S ribosomal DNA sequences, recovered Coltriciella multipileata as sister group with a specimen reported as ectomycorrhizal, and closely related with other two species that are considered saprophitic. According with our preliminary analysis of character states reconstruction, the ectomycorrhizal condition appeared early in the Coltriciella clade, with a high rate of transitions between ectomycorrhizal and saprophitic conditions. A key for species similar to Coltriciella multipileata is presented.

From qualitative to quantitative insect metabarcoding: an in tandem multilocus mosquito identification methodology

In the era of emergence and re-emergence of vector-borne diseases, a high throughput trap-based insect monitoring is essential for the identification of invasive species, study of mosquito populations and risk assessment of disease outbreaks. Insect DNA metabarcoding technology has emerged as a highly promising methodology for unbiased and large-scale surveillance. Despite significant attempts to introduce DNA metabarcoding in mosquito or other insect surveillance qualitative and quantitative metabarcoding remains a challenge. In the present study, we have developed a methodology of in-tandem identification and quantification using cytochrome oxidase subunit I (COI) combined with a secondary multilocus identification and quantification involving three loci of 28S ribosomal DNA. The presented methodology was able to identify individual species in pools of mosquitoes with 95.94% accuracy and resolve with high accuracy (p = 1, χ2 = 2.55) mosquito population composition providing a technology capable of revolutionizing mosquito surveillance through metabarcoding. The methodology, given the respective dataset, has the potential to be applied to various small animal populations.

A survey of entomopathogenic nematodes and their symbiotic bacteria in agricultural areas of northern Thailand

Entomopathogenic nematodes (EPNs) Steinernema and Heterorhabditis and their symbiotic bacteria, Xenorhabdus and Photorhabdus, have been successfully used for the control of insect pests. The objectives of this study were to survey the EPNs and symbiotic bacteria in the agricultural areas of the Phitsanulok province, Thailand, and to study the association between the soil parameters and presence of EPNs. We collected 200 soil samples from 40 soil sites in agricultural areas (field crops, horticulture crops and forest). The prevalence of EPNs was 8.0% (16/200). Fifteen of the EPN isolates were molecularly identified (based on 28S ribosomal DNA and internal transcribed spacer regions) as Steinernema siamkayai. Seven isolates of Xenorhabdus stockiae were identified using recombinase A sequencing. Phylogenetic analysis revealed that all the Steinernema and Xenorhabdus isolates were closely related to S. siamkayai (Indian strain) and X. stockiae (Thai strain), respectively. Significantly more EPNs were recovered from loam than from clay. Although the association between soil parameters (pH, temperature and moisture) and the presence of EPNs was not statistically significant, the elevation levels of the soil sites with and without EPNs were found to be different. Moreover, statistical comparisons between the agricultural areas revealed no significant differences. Therefore, we concluded that S. siamkayai is associated with X. stockiae in agricultural areas and that there is no association between the soil parameters of agricultural areas and presence of EPNs, except for soil texture and the elevation. Steinernema siamkayai may be applied as a biocontrol agent in agricultural areas.