Environmental Transmission of Symbionts in the Mangrove Crabs Aratus pisonii and Minuca rapax: Acquisition of the Bacterial Community through Larval Development to Juvenile Stage.

Aratus pisonii and Minuca rapax are two brachyuran crabs living with bacterial ectosymbionts located on gill lamellae. One previous study has shown that several rod-shaped bacterial morphotypes are present and the community is dominated by Alphaproteobacteria and Bacteroidota. This study aims to identify the mode of transmission of the symbionts to the new host generations and to identify the bacterial community colonizing the gills of juveniles. We tested for the presence of bacteria using PCR with universal primers targeting the 16S rRNA encoding gene from gonads, eggs, and different larval stages either obtained in laboratory conditions or from the field. The presence of bacteria on juvenile gills was also characterized by scanning electron microscopy, and subsequently identified by metabarcoding analysis. Gonads, eggs, and larvae were negative to PCR tests, suggesting that bacteria are not present at these stages in significant densities. On the other hand, juveniles of both species display three rod-shaped bacterial morphotypes on gill lamellae, and sequencing revealed that the community is dominated by Bacteroidota and Alphaproteobacteria on A. pisonii juveniles, and by Alphaprotobacteria, Bacteroidota, and Acidimicrobia on M. rapax juveniles. Despite the fact that juveniles of both species co-occur in the same biotope, no shared bacterial phylotype was identified. However, some of the most abundant bacteria present in adults are also present in juveniles of the same species, suggesting that juvenile-associated communities resemble those of adults. Because some of these bacteria were also found in crab burrow water, we hypothesize that the bacterial community is established gradually during the life of the crab starting from the megalopa stage and involves epibiosis-competent bacteria that occur in the environment.

A new feather mite species of the genus Mycterialges Gaud & Atyeo, 1981 (Acari, Xolalgidae) from the Oriental Stork, Ciconiaboyciana (Ciconiiformes, Ciconiidae) in Korea.

A new feather mite species, Mycterialgesboycianaesp. nov. (Xolalgidae), was identified from the Oriental Stork, Ciconiaboyciana Swinhoe, 1873, in Korea. Males of M.boycianaesp. nov. are distinguished from Mycterialgesmesomorphus Gaud & Atyeo, 1981, in having a single triangular prodorsal shield, sinuous margins of the opisthosoma located between setae e2 and h2 on the hysteronotal shield, an oval-shaped epiandrum without posterior extensions, a shorter tibia + tarsus IV than femoragenu IV, and an absent ambulacral disc of leg IV. Females differ in having a prodorsal shield with a posterior margin that is blunt-angular, and a concave posterior margin of the hysteronotal shield with posterior extensions. This study presents the first record of the feather mite genus Mycterialges in birds of the genus Ciconia (Ciconiidae). Additionally, we determined the phylogenetic relationship among Ingrassiinae using the mitochondrial cytochrome c oxidase subunit (COI).

Population Genomics of Pooled Samples: Unveiling Symbiont Infrapopulation Diversity and Host-Symbiont Coevolution.

Microscopic symbionts represent crucial links in biological communities. However, they present technical challenges in high-throughput sequencing (HTS) studies due to their small size and minimal high-quality DNA yields, hindering our understanding of host-symbiont coevolution at microevolutionary and macroevolutionary scales. One approach to overcome those barriers is to pool multiple individuals from the same infrapopulation (i.e., individual host) and sequence them together (Pool-Seq), but individual-level information is then compromised. To simultaneously address both issues (i.e., minimal DNA yields and loss of individual-level information), we implemented a strategic Pool-Seq approach to assess variation in sequencing performance and categorize genetic diversity (single nucleotide polymorphisms (SNPs)) at both the individual-level and infrapopulation-level for microscopic feather mites. To do so, we collected feathers harboring mites (Proctophyllodidae: Amerodectes protonotaria) from four individual Prothonotary Warblers (Parulidae: Protonotaria citrea). From each of the four hosts (i.e., four mite infrapopulations), we conducted whole-genome sequencing on three extraction pools consisting of different numbers of mites (1 mite, 5 mites, and 20 mites). We found that samples containing pools of multiple mites had more sequencing reads map to the feather mite reference genome than did the samples containing only a single mite. Mite infrapopulations were primarily genetically structured by their associated individual hosts (not pool size) and the majority of SNPs were shared by all pools within an infrapopulation. Together, these results suggest that the patterns observed are driven by evolutionary processes occurring at the infrapopulation level and are not technical signals due to pool size. In total, despite the challenges presented by microscopic symbionts in HTS studies, this work highlights the value of both individual-level and infrapopulation-level sequencing toward our understanding of host-symbiont coevolution at multiple evolutionary scales.

Impact of Wood Age on Termite Microbial Assemblages.

The decomposition of wood and detritus is challenging to most macroscopic organisms due to the recalcitrant nature of lignocellulose. Moreover, woody plants often protect themselves by synthesizing toxic or nocent compounds which infuse their tissues. Termites are essential wood decomposers in warmer terrestrial ecosystems and, as such, they have to cope with high concentrations of plant toxins in wood. In this paper, we evaluated the influence of wood age on the gut microbial (bacterial and fungal) communities associated with the termites Reticulitermes flavipes (Rhinotermitidae) (Kollar, 1837) and Microcerotermes biroi (Termitidae) (Desneux, 1905). We confirmed that the secondary metabolite concentration decreased with wood age. We identified a core microbial consortium maintained in the gut of R. flavipes and M. biroi and found that its diversity and composition were not altered by the wood age. Therefore, the concentration of secondary metabolites had no effect on the termite gut microbiome. We also found that both termite feeding activities and wood age affect the wood microbiome. Whether the increasing relative abundance of microbes with termite activities is beneficial to the termites is unknown and remains to be investigated. IMPORTANCE Termites can feed on wood thanks to their association with their gut microbes. However, the current understanding of termites as holobiont is limited. To our knowledge, no studies comprehensively reveal the influence of wood age on the termite-associated microbial assemblage. The wood of many tree species contains high concentrations of plant toxins that can vary with their age and may influence microbes. Here, we studied the impact of Norway spruce wood of varying ages and terpene concentrations on the microbial communities associated with the termites Reticulitermes flavipes (Rhinotermitidae) and Microcerotermes biroi (Termitidae). We performed a bacterial 16S rRNA and fungal ITS2 metabarcoding study to reveal the microbial communities associated with R. flavipes and M. biroi and their impact on shaping the wood microbiome. We noted that a stable core microbiome in the termites was unaltered by the feeding substrate, while termite activities influenced the wood microbiome, suggesting that plant secondary metabolites have negligible effects on the termite gut microbiome. Hence, our study shed new insights into the termite-associated microbial assemblage under the influence of varying amounts of terpene content in wood and provides a groundwork for future investigations for developing symbiont-mediated termite control measures.

The role of spine length in crustacean and fish associations with echinoids at Los Cabos, Baja California Sur, Mexico / El papel de la longitud de la espina en la asociación de crustáceos y peces con equinoideos en Los Cabos, Baja California Sur, México

Introduction: Echinoids (sea urchins) provide shelter for a variety of facultative or obligatory ectosymbionts. Objective: To evaluate the hypothesis that decapods and fishes prefer to associate with echinoid individuals and species that have longer spines. Methods: We visually studied the frequency of decapod crustaceans and fishes associated with echinoids in shallow water (< 4 m) and deeper water (5-20 m) at Los Cabos, Baja California Sur, Mexico, during 1-6 January 2019. Results: We inspected 1 058 echinoids of six species. Five decapod species associated with three species of echinoids. When compared with other echinoid species, in shallow water, decapods associated 5.1 times more often with the longest-spined echinoid Diadema mexicanum (7.0 times more decapods per individual D. mexicanum); in deeper water, association frequency was similar for all echinoid species. Fourteen fish species associated with four echinoid species. In shallow water, fishes associated 2.6 times more with D. mexicanum (4.5 times more fishes per individual). There was no preferred echinoid species in deeper water. Longer-spined D. mexicanum had more decapods and fishes. Associations were more frequent in shallow water. Multiple individuals and species of decapods and fish often associated together with a single D. mexicanum. The decapod that presumably is Tuleariocaris holthuisi showed a possible obligatory association with one of the equinoids (D. mexicanum); the other decapods and all fish species are facultative associates. Conclusion: Our results support the hypothesis that decapods and fishes associate most frequently with echinoids with the longest spines, presumably to reduce the risk of predation.

Introducción: Los equinoideos (erizos de mar) brindan refugio a una variedad de ectosimbiontes facultativos u obligatorios. Objetivo: Evaluar la hipótesis de que los decápodos y los peces prefieren asociarse con individuos y especies de equinoideos con espinas más largas. Métodos: Estudiamos visualmente la frecuencia de crustáceos decápodos y peces asociados con equinoideos en aguas poco profundas (< 4 m) y aguas más profundas (5-20 m) en Los Cabos, Baja California Sur, México, del 1-6 de enero 2019. Resultados: Examinamos 1 058 equinoideos de seis especies. Cinco especies de decápodos se asociaron con tres especies de equinoideos. Al comparar con otras especies de equinoideos, en aguas poco profundas, los decápodos se asociaron 5.1 veces más frecuentemente con la especie de equinoideo de espinas más largas, Diadema mexicanum (7.0 veces más decápodos por individuo en D. mexicanum); en aguas más profundas, la frecuencia fue similar para todas las especies de equinoideos. Catorce especies de peces se asociaron con 4 especies de equinoideos. En aguas poco profundas, los peces se asociaron 2.6 veces más con D. mexicanum (4.5 veces más peces por individuo). No hubo preferencia por una especie de equinoideo en aguas más profundas. Individuos de D. mexicanum con espinas largas tuvieron más asociación con decápodos y peces. Las asociaciones se dieron con mayor frecuencia en aguas poco profundas. Múltiples individuos y especies de decápodos y peces a menudo se asociaron con un solo D. mexicanum. Un decápodo que presumiblemente es Tuleariocaris holthuisi mostró una posible asociación obligatoria con uno de los equinoideos (D. mexicanum); las otras especies de decápodos y todas las especies de peces presentaron asociaciones facultativas. Conclusión: Nuestros resultados apoyan la hipótesis de que los decápodos y los peces se asociaron con mayor frecuencia con los equinoideos con las espinas más largas, presumiblemente para reducir el riesgo de depredación.

Anaerobic ciliates as a model group for studying symbioses in oxygen-depleted environments.

Anaerobiosis has independently evolved in multiple lineages of ciliates, allowing them to colonize a variety of anoxic and oxygen-depleted habitats. Anaerobic ciliates commonly form symbiotic relationships with various prokaryotes, including methanogenic archaea and members of several bacterial groups. The hypothesized functions of these ecto- and endosymbionts include the symbiont utilizing the ciliate's fermentative end products to increase the host's anaerobic metabolic efficiency, or the symbiont directly providing the host with energy by denitrification or photosynthesis. The host, in turn, may protect the symbiont from competition, the environment, and predation. Despite rapid advances in sampling, molecular, and microscopy methods, as well as the associated broadening of the known diversity of anaerobic ciliates, many aspects of these ciliate symbioses, including host specificity and coevolution, remain largely unexplored. Nevertheless, with the number of comparative genomic and transcriptomic analyses targeting anaerobic ciliates and their symbionts on the rise, insights into the nature of these symbioses and the evolution of the ciliate transition to obligate anaerobiosis continue to deepen. This review summarizes the current body of knowledge regarding the complex nature of symbioses in anaerobic ciliates, the diversity of these symbionts, their role in the evolution of ciliate anaerobiosis and their significance in ecosystem-level processes.

Termites Are Associated with External Species-Specific Bacterial Communities.

All termites have established a wide range of associations with symbiotic microbes in their guts. Some termite species are also associated with microbes that grow in their nests, but the prevalence of these associations remains largely unknown. Here, we studied the bacterial communities associated with the termites and galleries of three wood-feeding termite species by using 16S rRNA gene amplicon sequencing. We found that the compositions of bacterial communities among termite bodies, termite galleries, and control wood fragments devoid of termite activities differ in a species-specific manner. Termite galleries were enriched in bacterial operational taxonomic units (OTUs) belonging to Rhizobiales and Actinobacteria, which were often shared by several termite species. The abundance of several bacterial OTUs, such as Bacillus, Clostridium, Corynebacterium, and Staphylococcus, was reduced in termite galleries. Our results demonstrate that both termite guts and termite galleries harbor unique bacterial communities.IMPORTANCE As is the case for all ecosystem engineers, termites impact their habitat by their activities, potentially affecting bacterial communities. Here, we studied three wood-feeding termite species and found that they influence the composition of the bacterial communities in their surrounding environment. Termite activities have positive effects on Rhizobiales and Actinobacteria abundance and negative effects on the abundance of several ubiquitous genera, such as Bacillus, Clostridium, Corynebacterium, and Staphylococcus Our results demonstrate that termite galleries harbor unique bacterial communities.

Seven new species of Graphilbum from conifers in Norway, Poland, and Russia.

During surveys of insect-associated mycobiomes in Norway, Poland, and Russia, isolates with affinity to Graphilbum (Ophiostomatales, Ascomycota) were recovered. In this study, eight known Graphilbum species as well as the newly collected isolates were compared based on morphology and DNA sequence data for four gene regions. The results revealed seven new species, described here as G. acuminatum, G. carpaticum, G. curvidentis, G. furuicola, G. gorcense, G. interstitiale, and G. sexdentatum. In addition to these species, G. crescericum and G. sparsum were commonly found in Norway. All new species were recovered from conifers in association with bark beetles, cerambycid beetles, and weevils and were morphologically similar, predominantly with pesotum-like asexual morphs. Where sexual morphs were present, these were small ascomata with short necks and rod-shaped ascospores having hyaline sheaths. The results suggest that Graphilbum species are common members of the Ophiostomatales in conifer ecosystems.

Morphology of obligate ectosymbionts reveals Paralaxus gen. nov.: A new circumtropical genus of marine stilbonematine nematodes.

Stilbonematinae are a subfamily of conspicuous marine nematodes, distinguished by a coat of sulphur-oxidizing bacterial ectosymbionts on their cuticle. As most nematodes, the worm hosts have a relatively simple anatomy and few taxonomically informative characters, and this has resulted in numerous taxonomic reassignments and synonymizations. Recent studies using a combination of morphological and molecular traits have helped to improve the taxonomy of Stilbonematinae but also raised questions on the validity of several genera. Here, we describe a new circumtropically distributed genus Paralaxus (Stilbonematinae) with three species: Paralaxus cocos sp. nov., P. bermudensis sp. nov. and P. columbae sp. nov. We used single worm metagenomes to generate host 18S rRNA and cytochrome c oxidase I (COI) as well as symbiont 16S rRNA gene sequences. Intriguingly, COI alignments and primer matching analyses suggest that the COI is not suitable for PCR-based barcoding approaches in Stilbonematinae as the genera have a highly diverse base composition and no conserved primer sites. The phylogenetic analyses of all three gene sets, however, confirm the morphological assignments and support the erection of the new genus Paralaxus as well as corroborate the status of the other stilbonematine genera. Paralaxus most closely resembles the stilbonematine genus Laxus in overlapping sets of diagnostic features but can be distinguished from Laxus by the morphology of the genus-specific symbiont coat. Our re-analyses of key parameters of the symbiont coat morphology as character for all Stilbonematinae genera show that with amended descriptions, including the coat, highly reliable genus assignments can be obtained.

Revealing the metabolic capacity of Streblomastix strix and its bacterial symbionts using single-cell metagenomics.

Lower termites harbor in their hindgut complex microbial communities that are involved in the digestion of cellulose. Among these are protists, which are usually associated with specific bacterial symbionts found on their surface or inside their cells. While these form the foundations of a classic system in symbiosis research, we still know little about the functional basis for most of these relationships. Here, we describe the complex functional relationship between one protist, the oxymonad Streblomastix strix, and its ectosymbiotic bacterial community using single-cell genomics. We generated partial assemblies of the host S. strix genome and Candidatus Ordinivivax streblomastigis, as well as a complex metagenome assembly of at least 8 other Bacteroidetes bacteria confirmed by ribosomal (r)RNA fluorescence in situ hybridization (FISH) to be associated with S. strix. Our data suggest that S. strix is probably not involved in the cellulose digestion, but the bacterial community on its surface secretes a complex array of glycosyl hydrolases, providing them with the ability to degrade cellulose to monomers and fueling the metabolism of S. strix In addition, some of the bacteria can fix nitrogen and can theoretically provide S. strix with essential amino acids and cofactors, which the protist cannot synthesize. On the contrary, most of the bacterial symbionts lack the essential glycolytic enzyme enolase, which may be overcome by the exchange of intermediates with S. strix This study demonstrates the value of the combined single-cell (meta)genomic and FISH approach for studies of complicated symbiotic systems.

The complete mitochondrial genome of the feather mite Trouessartia rubecula Jablonska, 1968 (Astigmata: Analgoidea: Trouessartiidae).

We assembled and annotated the complete mitochondrial genome of Trouessartia rubecula, the first feather mite complete mitochondrial genome from the largest feather mite superfamily Analgoidea (ca. 1150 spp). The mitogenome was composed of 13 protein, 17 tRNA, and 2 rRNA-coding genes and was 14,125 bp in length.

Feather mites of the genera Dubininia and Cacatualges (Acari: Xolalgidae) associated with parrots (Aves: Psittaciformes) of the Old World.

This paper gives a systematic revision of feather mites of the genera Dubininia Vassilev, 1958 and Cacatualges Dabert, Badek and Skoracki, 2007 (Xolalgidae: Ingrassiinae) associated with parrots (Aves: Psittaciformes) of the Old World. Five new species are described: Cacatualges probosciger sp. n. from Probosciger aterrimus (Gmelin) (Cacatuidae) from New Guinea, Dubininia charmosynae sp. n. from Charmosyna pulchella Gray GR (Psittaculidae) from New Guinea, D. micropsittae sp. n. from Micropsitta pusio pusio (Scaltter) (Psittaculidae) from New Guinea, D. nestori sp. n. from Nestor notabilis Gould (Strigopidae) from New Zealand, and D. pezopori sp. n. from Pezoporus wallicus (Kerr) (Psittaculidae) from Tasmania, Australia. Four previously described species of Dubininia are redescribed based on material from type hosts: D. curta (Trouessart, 1885) from Platycercus elegans (Gmelin) (Psittaculidae), D. lorina (Trouessart, 1885) from Lorius domicella (Linnaeus) (Psittaculidae), D. melopsittaci Atyeo and Gaud, 1987 from Melopsittacus undulatus (Shaw) (Psittaculidae), and D. psittacina (Trouessart, 1885) from Strigops harboptilus Gray GR (Strigopidae) from New Zealand. A new diagnosis for the genus Dubininia is provided. A key to all presently known Dubininia species is provided for the first time.

Promiscuous and specific bacterial symbiont acquisition in the amoeboid genus Nuclearia (Opisthokonta).

We isolated 17 strains of the amoeboid genus Nuclearia (Opisthokonta) from five Swiss lakes. Eight of these nucleariid isolates were associated with bacterial endosymbionts and/or ectosymbionts. Amoebae were characterized morphologically and by their 18S rRNA genes. Phylogeny based on molecular data resulted in four established monophyletic branches and two new clusters. A heterogeneous picture emerged by highlighting nucleariids with associated bacteria. Apart from one cluster which consisted of only isolates with and three groups of amoebae without symbionts, we also found mixed clusters. The picture got even more 'blurred' by regarding the phylogeny of symbiotic bacteria. Although seven different bacterial strains could be identified, it seems that we still are only scratching the surface of symbionts' diversity. Furthermore, types of symbioses might be different depending on host species. Strains of Nuclearia thermophila harboured the same endosymbiont even when isolated from different lakes. This pointed to a specific and obligate interaction. However, two isolates of N. delicatula were associated with different endosymbiotic bacteria. Here the symbiont acquisition seemed to be rather promiscuous. This behaviour regarding symbiotic associations is especially remarkable considering the phylogenetic position of these basal opisthokonts.

Feather mites (Acari, Astigmata) associated with birds in an Atlantic Forest fragment in Northeastern Brazil / Ácaros de pena (Acari: Astigmata) associados com aves em um fragmento de Mata Atlântica no Nordeste do Brasil

AbstractThe present study reports associations between feather mites (Astigmata) and birds in an Atlantic Forest fragment in Rio Grande do Norte state, in Brazil. In the laboratory, mites were collected through visual examination of freshly killed birds. Overall, 172 individuals from 38 bird species were examined, between October 2011 and July 2012. The prevalence of feather mites was 80.8%, corresponding to 139 infested individuals distributed into 30 species and 15 families of hosts. Fifteen feather mite taxa could be identified to the species level, sixteen to the genus level and three to the subfamily level, distributed into the families Analgidae, Proctophyllodidae, Psoroptoididae, Pteronyssidae, Xolalgidae, Trouessartiidae, Falculiferidae and Gabuciniidae. Hitherto unknown associations between feather mites and birds were recorded for eleven taxa identified to the species level, and nine taxa were recorded for the first time in Brazil. The number of new geographic records, as well as the hitherto unknown mite-host associations, supports the high estimates of diversity for feather mites of Brazil and show the need for research to increase knowledge of plumicole mites in the Neotropical region.

ResumoO presente estudo reporta associações entre ácaros (Astigmata) e aves em um fragmento de Mata Atlântica no estado do Rio Grande do Norte, Brazil. Em laboratório, ácaros foram coletados através de examinação visual de aves recentemente mortas. No total, 172 indivíduos de 38 espécies de aves foram examinados entre Outubro de 2011 e Julho de 2012. A prevalência de ácaros na comunidade de aves foi de 80,8%, correspondendo a 139 indivíduos infestados distribuídos em 30 espécies e 15 famílias de hospedeiros. Quinze táxons de ácaros de pena foram identificados em nível de espécie, dezesseis em nível de gênero e três em nível de subfamília, distribuídos nas famílias Analgidae, Proctophyllodidae, Psoroptoididae, Pteronyssidae, Xolalgidae, Trouessartiidae, Falculiferidae and Gabuciniidae. Associações ainda não conhecidas entre ácaros e aves foram registradas para onze táxons identificados em nível específico, e nove táxons foram registrados pela primeira vez no Brasil. O número de novos registros geográficos, assim como as associações ácaro-hospedeiro até então desconhecidas, suportam as altas estimativas de diversidade de ácaros de pena e apontam a necessidade de mais pesquisas que ampliem o conhecimento dos ácaros plumícolas da região Neotropical.

Ménage-à-trois: the amoeba Nuclearia sp. from Lake Zurich with its ecto- and endosymbiotic bacteria.

We present a fascinating triad relationship between a eukaryotic amoeba and its two bacterial symbionts. The morphological characteristics of the amoeba allowed for a confident assignment to the genus Nuclearia (Opisthokonta, Nucleariidae), but species identification resulted in an ambiguous result. Sequence analysis indicated an affiliation to the species N. thermophila, however, several morphological features contradict the original description. Amoebal isolates were cultured for several years with their preferred food source, the microcystin-producing harmful cyanobacterium Planktothrix rubescens. Symbioses of the amoeba with ecto- and endosymbiotic bacteria were maintained over this period. Several thousand cells of the ectosymbiont are regularly arranged inside a layer of extracellular polymeric substances produced by the amoeba. The ectosymbiont was identified as Paucibacter toxinivorans (Betaproteobacteria), which was originally isolated by enrichment with microcystins. We found indications that our isolated ectosymbiont indeed contributed to toxin-degradation. The endosymbiont (Gammaproteobacteria, 15-20 bacteria per amoeba) is enclosed in symbiosomes inside the host cytoplasm and represents probably an obligate symbiont. We propose the name "Candidatus Endonucleariobacter rarus" for this bacterium that was neither found free-living nor in a symbiotic association. Nucleariidae are uniquely suited model organisms to study the basic principles of symbioses between opisthokonts and prokaryotes.

Phylogenetic confirmation of the genus Robbea (Nematoda: Desmodoridae, Stilbonematinae) with the description of three new species.

The Stilbonematinae are a monophyletic group of marine nematodes that are characterized by a coat of thiotrophic bacterial symbionts. Among the ten known genera of the Stilbonematinae, the genus Robbea Gerlach 1956 had a problematic taxonomic history of synonymizations and indications of polyphyletic origin. Here we describe three new species of the genus, R. hypermnestra sp. nov., R. ruetzleri sp. nov. and R. agricola sp. nov., using conventional light microscopy, interference contrast microscopy and SEM. We provide 18S rRNA gene sequences of all three species, together with new sequences for the genera Catanema and Leptonemella. Both our morphological analyses as well as our phylogenetic reconstructions corroborate the genus Robbea. In our phylogenetic analysis the three species of the genus Robbea form a distinct clade in the Stilbonematinae radiation and are clearly separated from the clade of the genus Catanema, which has previously been synonymized with Robbea. Surprisingly, in R. hypermnestra sp. nov. all females are intersexes exhibiting male sexual characters. Our extended dataset of Stilbonematinae 18S rRNA genes for the first time allows the identification of the different genera, e.g. in a barcoding approach. http://zoobank.org/urn:lsid:zoobank.org:pub:D37C3F5A-CF2B-40E6-8B09-3C72EEED60B0.