Male Spicule Research Articles

Aphelenchoides japonicus n. sp. (Tylenchina: Aphelenchoididae) found in the rhizosphere of Camellia sasanqua and Juniperus chinensis from Japan

Summary Aphelenchoides japonicus n. sp., isolated in the rhizosphere of Camellia sasanqua and Juniperus chinensis from Japan, is described. It is characterised by three lateral incisures, and a slender stylet 8-10 μm long with obvious round basal swellings, an excretory pore located at the level of the nerve ring or slightly posterior, an oblong spermatheca, containing ellipse sperm cells in a single row, but sometimes with spheroid sperm cells in 2-3 rows, a vagina not sclerotised and the vulva having slightly protruding lips and lacking a flap. The post-vulval uterine sac (PUS) is well developed, occupying ca 26.2% (21.5-31.4) of the vulva to anus distance. The male spicules are smoothly arcuate, the condylus very short and rounded, a small rostrum sharply pointed, the dorsal limb 21.6 (19.1-23.8) μm long, lacking a hook-like structure at the distal tip. The tail of both sexes is conical, with a trimucronate process, giving a wide warty mucron-like structure or a star-like appearance. The new species belongs to Group 3 of the Aphelenchoides species sensu Shahina. Phylogenetic analysis based on 18S and 28S D2-D3 of rDNA, along with ITS and mtCOI confirmed its status as a new species.

Morphological and molecular characterisation of Bursaphelenchus parasaudi n. sp. (Tylenchina: Aphelenchoididae) found in loblolly pine logs from the USA

Summary Bursaphelenchus parasaudi n. sp., isolated in Ningbo, P.R. China, from loblolly pine logs imported from the USA is described. It is characterised by a slim body (a = 32-43), lateral field with four lines, excretory pore located at the level of metacorpus, vulva at 76% of total body length, a distinct vulval flap in lateral view, post-uterine branch ca half to two-thirds of the vulva to anus distance long, female tail conical, slightly ventrally curved, with a finely rounded terminus, male spicules with weakly developed rostrum and condylus and cross-striped in their central part, a single precloacal papilla and three pairs of ventro-sublateral papillae. It belongs to the africanus group of the genus Bursaphelenchus and is most similar to B. saudi. Detailed phylogenetic analysis based on the partial 18S region, ITS and 28S D2-D3 region confirmed the status of this nematode as a new species.

Redescription of Raphidascaris (Ichthyascaris) vicentei (Nematoda: Raphidascarididae) Parasitizing Lutjanid Fishes (Actinopterygii: Lutjanidae) from the Northeast Coast of Brazil, Including First Observation Using SEM.

In this study, 143 lutjanid fishes representing four different species from the Northeast coast off Brazil were analyzed for parasites. The aim of the present study is to provide a detailed redescription of R. (I.) vicentei, collected from lutjanid fishes in Brazil, including the first observation of the species using scanning electron microscopy (SEM). Nematodes found clarified in a crescent glycerin series, were photographed and measured (in mm), and some specimens destined for SEM. The Illustrations were made using an optical microscope with a drawing tube. Nematodes found were identified as Raphidascaris (Ichthyascaris) vicentei, infecting 6.29% of the examined fish. Detailed morphological analysis revealed the following characteristics observed for the first time in this species: labial papillae, narrow lateral alae and the detailed structure of male spicules. These contributions help filling the gaps in the literature and expand the knowledge of this parasite morphology. The presence of R. (I.) vicentei in four lutjanid species may indicates low host specificity, highlighting its adaptability to different hosts. Furthermore, new host records are given. This research provides the most comprehensive morphological description of R. (I.) vicentei to date, generating crucial insights for a better understanding of the parasitic biodiversity off the Brazilian Northeastern coast of Brazil, as well as establishing foundation for future investigations on the ecology and impacts of these parasites in fish populations from this region.

Forty years later: a new Andean stream treefrog of the genus Hyloscirtus (Anura: Hylidae) from Ecuador, with comments on arm hypertrophy in the H. larinopygion group.

We present the description of a novel treefrog species inhabiting the Andean streams in southeastern Ecuador that has been erroneously identified as Hyloscirtus lindae for four decades. The new species is closely related to H. tapichalaca and is part of the southern clade of the H. larinopygion group, which comprises five species confined to the southeastern Andes of Ecuador to the northeastern Andes of Peru. It is diagnosed from its close relatives by a unique combination of characteristics, including hypertrophied forelimbs in males, a pronounced, curved, and protruding spine-shaped prepollex, a substantial supracloacal flap, supratympanic fold, digital discs colored in shades of orange-red or bright-red, and the concealed surfaces of limbs displaying a bluish-gray hue and dorsal spicules in males. We briefly explore the implications of this discovery for the evolution of arm morphology in the H. larinopygion group. Our findings underscore the continued importance of periodically reviewing historical specimens, leading to unexpected discoveries; once again confirming the importance of natural history museums and their custodian biological collections.

Morphological and Molecular Characterization of a New Root-Knot Nematode, Meloidogyne limonae n. sp. (Nematoda: Meloidogynidae), Parasitizing Lemon in China.

Root-knot nematodes of the genus Meloidogyne parasitize the roots of thousands of plants and can cause severe damage and yield losses. Here, we report a new root-knot nematode, Meloidogyne limonae n. sp., parasitizing "lemon" (Citrus limon) in Hainan Province, South China. Lemon trees infected by the root-knot nematode showed poor-quality lemons, chlorosis of foliage, weak growth, and numerous root galls with white females and egg masses protruding outside. Phylogenetic trees of sequences within the ribosomal and mitochondrial DNA demonstrated that this species differs clearly from other previously described root-knot nematodes. Morphologically, the new species is characterized by an oval-shaped perineal pattern and the lateral field marked by a ridge of cuticle on one or both sides; the dorsal arch is low, with fine to coarse, smooth cuticle striae; the vulva slit is centrally located at the unstriated area; the spicules of males are arcuate and curved ventrally; the gubernaculum is distinct and curved; the labial disc of second-stage juveniles is prominent and dumbbell shaped; stylet knobs are oval and sloping backwardly; pharyngeal glands are not filling the body cavity and overlapping the intestine ventrally; and the conical tail is gradually tapering. Phylogenetic trees based on the ITS1-5.8S-ITS2, D2-D3 of the 28S rDNA, and COI and COII-16S rRNA genes of the mtDNA showed that M. limonae n. sp. belongs to an undescribed root-knot nematode lineage that is separated from other species with the resemblance in morphology, such as M. floridensis, M. hispanica, M. acronea, and M. paranaensis.

Phylogenetic evidence of a possible Trichuris globulosa species complex in Arabian camels from Kuwait.

During a 1-year study, Trichuris adults were obtained after necropsy of Arabian camels (Camelus dromedarius) from a slaughterhouse in Kuwait. Morphological and molecular identification was performed to confirm the identity of the Trichuris specimens obtained from C. dromedarius. Fifteen male Trichuris specimens were selected, and molecular identification was performed using mitochondrial cytochrome c oxidase subunit I, 12S ribosomal RNA, 16S ribosomal RNA genes and the nuclear internal transcribed spacer 2 (ITS2) region. Through phylogenetic analysis, 2 distinct groups were obtained using the mitochondrial genes, where group 1 showed a close relationship to Trichuris globulosa while group 2 showed a close relationship to Trichuris ovis, providing molecular evidence of a possible T. globulosa species complex. Additionally, the nuclear ITS2 region did not provide enough resolution to distinguish between the 2 groups of Trichuris specimens. Observation of morphological characters revealed variations in the shape of the male spicule sheath, where specimens present either a globular posteriorly truncated swelling or the absence of posteriorly truncated swelling. Moreover, the variations in male spicule sheath does not corroborate with the results of molecular data, suggesting the limited use of this character for identification of T. globulosa. In conclusion, molecular analysis suggests a possible species complex in T. globulosa, with the mitochondrial genetic markers successfully differentiating between the 2 groups. The limited use of the male spicule sheath as a diagnostic character for identification of T. globulosa is suggested.

A revision of the trichostrongylid nematode Cooperia Ransom, 1907, from deer game: recent integrative research confirms the existence of the ancient host-specific species Cooperia ventricosa (Rudolphi, 1809).

The trichostrongylid roundworms of the genus Cooperia, which are important in veterinary medicine, currently comprise 19 valid species that parasitize the small intestine of both free-living and domestic ruminants. Only four Cooperia spp. have been reported in Europe, namely C. oncophora, C. punctata, C. curticei and C. pectinata. In 2018-2022, 25 red deer (Cervus elaphus) and 30 sika deer (Cervus nippon) of both sexes and various ages from several remote locations in the Czech Republic were parasitologically examined. Intestinal nematodes of the genus Cooperia were found only in two northern regions. Using the globally recognized key book on trichostrongylid nematodes, they were preliminarily identified as C. pectinata. However, a molecular analysis of cox2 and ITS rDNA gene sequences revealed that Cooperia sp. parasitizing Czech deer is a separate taxon that is more closely related to C. oncophora than to C. pectinata. A subsequent morphological analysis and literature survey confirmed the independence of deer Cooperia sp., which is similar but not identical to bovid C. pectinata. Previous long-term correct identifications of bovid C. pectinata and misidentifications of deer Cooperia species were caused by a fundamental error in the key book mentioned above. Interestingly, the ancient trichostrongylid nematode Strongylus ventricosus from the type host red deer (Cervus elaphus) shot near Greifswald (Germany) was described by Rudolphi in 1809. Rudolphi's type material (one male and four females) was deposited in the Museum für Naturkunde (Berlin). Later, the ancient species S. ventricosus was taken as a synonym for various Cooperia spp. Our current re-examination of the type male indicated that there is a relatively good agreement with our new material from Czech deer regarding the most important characteristics of S. ventricosus (i.e., the shape and size of the male spicules); however, Rudolphi's type material is in rather poor condition. The suggested resurrection of the deer Cooperia sp. in this study as Cooperia ventricosa (Rudolphi, 1809) requires verification by collecting and analyzing new nematode material from the type locality near Greifswald.

Mating sub-behaviours in Bursaphelenchus xylophilus on agar and evidence for female cooperation

Summary The pine wood nematode, Bursaphelenchus xylophilus, is the cause of pine wilt disease and reproduces by copulation within host pine trees. Female mate selection in B. xylophilus mating behaviour has been reported but definitive evidence is lacking. Furthermore, while the mating behaviour of the free-living nematode Caenorhabditis elegans on agar has been well understood, the mating behaviour of B. xylophilus on agar has not been studied in detail, and the differences in their characteristics remain unclear. Here, we addressed these knowledge gaps by observing B. xylophilus mating behaviour on agar. We identified six primary mating sub-behaviours in B. xylophilus males. Comparative analysis with C. elegans revealed distinct differences in the sequence of mating stages. Notably, B. xylophilus males actively searched for the female’s vulva during the wrapping stage, whereas females exhibited back-and-forth locomotion to align the male spicule with the vulva location. Additionally, we conducted mating assays using anaesthetised female nematodes to evaluate the female’s role in the mating behaviours of B. xylophilus. The mating duration increased significantly when females were anaesthetised, and the probabilities of reaching the wrapping and vulva location stages were reduced. Therefore, B. xylophilus females actively cooperate in mating behaviours. Our findings highlight unique aspects of mating behaviours in B. xylophilus and provide insights concerning differences in mating behaviours between B. xylophilus and C. elegans.

Cosmocerca goroensis n. sp. (Nematoda: Cosmocercidae) from South Africa and its phylogenetic relationships with other cosmocercids based on partial 28S sequences.

Nematodes of the genus Cosmocerca are commonly found in various amphibians in South Africa and in most cases are identified as C. ornata. However, after detailed morphological studies and molecular approaches, three new species of the genus were recently described from three different frogs in South Africa. In present study, we describe another new species - Cosmocerca goroensis parasitising the Northern Pygmy Toad Poyntonophrynus fenoulheti in Soutpansberg mountains, Limpopo province, South Africa. The new species is characterised by prominent sex dimorphism, wide lateral alae, numerous somatic papillae in both sexes, and wide triangularly shaped gubernaculum and simple prominent spicules in males. Cosmocerca goroensis n. sp. distinguished from congeners, previously reported in Southern Africa by the shape of the gubernaculum and arrangement of somatic papillae in males. Morphological differences were confirmed by molecular analysis based on fragments of the 28S gene. Phylogenetic analysis based on the 28S gene fragments, including C. goroensis n. sp. and newly obtained sequence of C. ornata from Pelophylax lessonae from Ukraine, supported previously known data of closer relationships between species of Cosmocerca and Aplectana and more distant with Cosmocercoides spp.

Molecular Characterization and Functional Analysis of GPCR Gene Bx-srh-1 in Pinewood Nematode (Bursaphelenchus xylophilus)

Bursaphelenchus xylophilus, also known as the pine wood nematode (PWD), which causes pine wilt disease (PWD), is one of the most devastating diseases affecting pine forests globally. G protein-coupled receptors (GPCRs) are crucial in many biological processes and serve as privileged points of communication between cells and the surrounding environment. α-pinene has been found to play a crucial role in combating the infection, colonization, and early stages of pathogenesis caused by B. xylophilus. In this study, we investigated the molecular characteristics and biological functions of the GPCR gene Bx-srh-1 in B. xylophilus. Fluorescence in situ hybridization (FISH) was performed to determine the spatial expression patterns of Bx-srh-1 in B. xylophilus. The results indicated that Bx-srh-1 is expressed in the intestine and subcutaneous tissues of J2 and J3 juveniles and in the spicules of adult males and vulvae of adult females. RNA interference (RNAi) was used to analyze Bx-srh-1 gene function, and we examined the expression patterns of Bx-srh-1 in B. xylophilus under α-pinene stress. The RNA interference indicated that Bx-srh-1 was involved in the reproductive ability and pathogenicity of B. xylophilus; the expression levels of Bx-srh-1 significantly increased after the exposure to α-pinene for 12 h, and they peaked at 48 h. Silencing Bx-srh-1 may therefore lead to a reduction in B. xylophilus reproduction and pathogenicity. These results demonstrate that Bx-srh-1 is related to the feeding behavior, reproduction, pathogenicity, and resistance to α-pinene process of B. xylophilus.

Steinernema indicum n. sp., a new entomopathogenic nematode (Nematoda: Steinernematidae) from India

Summary A new species of entomopathogenic nematode, Steinernema indicum n. sp., was isolated from a coconut palm (Cocos nucifera) field in the district of Udupi, Karnataka, India. The infective juveniles (IJ) of S. indicum n. sp. possess a body length of 1043 (869-1172) μm. The lateral fields are formed by eight equally spaced and developed ridges (nine incisures) in the mid-body region, giving the formula 3, 8, 7, 5. The excretory pore is located posterior to the mid-pharynx region (D% = 63), and the hyaline region occupies ca half of the tail length. The first-generation males are characterised by very short spicules 66 (60-70) μm long. The male spicules are moderately curved with a sharp tip and are golden-brown in colour with the manubrium elongate and with a length to width ratio of 1.92:1. There are 11 pairs of genital papillae plus a single midventral papilla located anterior to the cloacal region. The first-generation male of S. indicum n. sp. is characterised by the presence of a single dorsal postcloacal papilla, a typical diagnostic character that has not been reported from any other steinernematid species. The first-generation male tail has a short mucron present in ca 40% of specimens examined and a second-generation male tail showing a short mucron present in ca 65% of male specimens examined. The first and second-generation females possess a slightly protruding post-anal swelling. The new species is further characterised by sequences of the internal transcribed spacer (ITS) and partial 28S regions (D2-D3) of the ribosomal DNA (rDNA). Phylogenetic analyses confirm that S. indicum n. sp. belongs to the glaseri-group and, based on both ITS and D2D3 genes, showed that S. indicum n. sp. is a member of the karii clade.

Morphologic and Genetic Characterization of Protospirura canariensis n. sp. (Nematoda, Spiruridae), a Parasite of the Black Rat Rattus rattus (Rodentia, Muridae) from El Hierro Island (Canary Archipelago, Spain)

A new spirurid nematode, Protospirura canariensis n. sp., a parasite of the black rat Rattus rattus Linnaeus, 1758 (Rodentia: Muridae), in El Hierro Island (Canary Archipelago, Spain), was described by means of light (LM) and scanning electron microscopy (SEM). The most discriminating characteristics between the new species and the remaining species of the genus Protospirura were observed to be the following: (a) the number of tooth-like outgrowths in the sub-median and lateral lobes of the pseudolabia, both in males and females (2 and 4, respectively); (b) the size of the right and left spicules in males (643–715 µm and 309–412 µm, respectively); and (c) the numbers and arrangements of the cloacal papillae in males. The new species has a total of 17 cloacal papillae (4 large and pedunculated pairs of precloacal papillae, an unpaired precloacal papilla and 4 pairs of postcloacal papillae). The arrangement of the postcloacal papillae are as follows: the first pair are large, pedunculated and placed near the posterior edge of cloaca; the three remaining postcloacal pairs are grouped and located near the posterior tip. In the latter group, the papillae in the first pair are large and pedunculated. The parasitized hosts and their geographical distributions were also useful criteria in distinguishing P. canariensis n. sp. from the remaining species of the genus Protospirura. In addition, the cytochrome c oxidase subunit I (cox1) sequence of the new species was obtained and compared with the available data of related species.

Cylicospirura felineus in Puma (Herpailurus yagouaroundi) in southern Brazil - Macroscopic and microscopic aspects

Parasites of the Cylicospirura genus are known to affect domestic and wild carnivores by causing the formation of nodules within the gastric mucosa and pyloric region of parasitized animals. C. felineus is a specimen of this genus that has been reported in Brazil in animal species such as Leopardus geoffroyi and Puma concolor. However, as far as the authors of the present study are aware, reports of C. felineus in jaguarundis (Puma yagouaroundi) do not yet exist. This wild feline species native to the Americas is found only in a few countries - and it is possibly extinct in others, such as the United States. In Brazil, the P. yagouaroundi species is classified as vulnerable by government environmental agencies. Consequently, identifying the diseases and their relevance to this species is of utmost importance for P. yagouaroundi's preservation. This study aims to describe the histopathological lesions found in jaguarundis parasitized by the genus Cylicospirura, and to identify the parasitic species observed in these animals. Eight specimens of P. yagouaroundi necropsied between 2010 and 2020 in the Laboratory of Veterinary Pathology of the Federal University of Paraná (UFPR) in Palotina, Brazil, were studied. Seven specimens were wildlife roadkills, and one was from the Municipal Zoo of Cascavel, Paraná, Brazil. Seven of the necropsied animals presented - in the greater curvature of the stomach, near the cardia - one or more parasitic nodules of 0.5 to 3.0cm in diameter that contained several intralesional reddish filiform parasites. Histologically, these nodules were characterized as transmural parasitic granulomas surrounded by reaction with deposition of dense connective tissue. The parasitological evaluations by light and scanning electron microscopy identified morphological C. felineus' characteristics through the presence of unequal spicules in males, the position of the vulva, and location of the pedunculated papillae. Despite the gastric lesion caused by the parasite, the apparent low pathogenicity to the gastrointestinal system of the evaluated animals, massive infections or large nodules can lead to death - other members of the Spirocercidae family are known to cause organ rupture, esophageal obstructions, and mesenchymal neoplasms. Therefore, C. felineus should be included in the list of gastric parasites with pathogenic potential in the species P. yagouaroundi.

Description of Laimaphelenchus shanxiensis sp. n. (Rhabditida: Aphelenchoididae) from Shanxi province, P.R. China

Summary Laimaphelenchus shanxiensis sp. n. was recovered from the bark samples of Chinese pine tree (Pinus tabuliformis) in Shanxi province, China. The new species is characterised by having 888-1049 μm long females, the cephalic region with six labial sectors divided by ribs, stylet 11-13 μm long, the excretory pore located at the same level with, or slightly posterior to nerve ring, lateral field with four incisures, post-vulval uterine sac 54-83 μm long, vulva with a well-developed anterior flap, tail having a single stalk at terminus ending to a flat structure with 4-7 finger-like appendages in scanning electron microscopy; male spicules 20-22 μm long and two pairs of caudal papillae present. In addition, molecular phylogenetic trees inferred from the partial small subunit (SSU) and D2-D3 segments of large subunit (LSU) of rRNA genes, revealed that this nematode belongs to the Laimaphelenchus and is different from all currently sequenced species of Laimaphelenchus for their SSU and LSU markers.

Ditylenchus rafiqi n. sp. from pomegranate (Punica granatum L.) from India with a tabular key for the species known from India

Summary Ditylenchus rafiqi n. sp. was recovered from the rhizosphere of pomegranate in New Delhi (India), and it is characterised by morphological, morphometrical and molecular methods. The new species is characterised by having a body length of 553-901 μm, delicate stylet (7-10 μm long), longer conus length (5.0-5.5 μm in females and 4.5-5.5 μm in males) with small round knobs sloping backward, seven lateral lines at the mid-body, median bulb well developed with thickening of lumen walls, a post-uterine sac well developed (27-47 μm), tail long, elongate conoid with an almost round terminus (45-74 μm) and male spicule of 15-17.5 μm, gubernaculum simple, leptoderan type of bursa enveloping the tail almost >50% of its tail length. A detailed study based on morphological and morphometrical methods clearly demarcates that D. rafiqi n. sp. differs from other known species of Ditylenchus. The molecular characterisation was done using D2-D3 of 28S rRNA and ITS rRNA markers. Molecular characterisation shows that the new species is closely related to D. acutus, D. gilanicus (D2-D3 marker), and D. medicaginis (ITS rRNA markers). Based on phylogenetic analyses using D2-D3 of LSU and ITS rRNA markers, D. rafiqi n. sp. forms an intermediate clade between the D. triformis and D. dipsaci groups.

Pseudaphelenchus chunanensis n. sp. (Tylenchina: Aphelenchoididae) found in Pinus massoniana from China

Summary Pseudaphelenchus chunanensis n. sp. was isolated in branches of Pinus massoniana from eastern China. The new species is characterised by very short males (402 (360-452) μm) and females (470 (397-540) μm), three incisures in the lateral field, slender stylet with small but conspicuous basal knobs, excretory pore located at the level of anterior margin of the metacorpus or more anterior. Male spicules paired, thorn-like, separate, condylus truncate, sometimes broadly rounded, rostrum conical or triangular with bluntly pointed tip, calomus smoothly tapering together with lamina, thin and strongly arcuate in middle then tapering towards bluntly pointed distal tip, cucullus absent. A long and narrow bursa surrounds the entire tail but inconspicuous in most males. Conical tail bluntly rounded but sometimes has a short mucron. Female tail conical, smoothly tapering and slightly ventrally bent in distal part of tail, terminus fine or bluntly pointed. It is typologically different from six known Pseudaphelenchus species, and detailed phylogenetic analysis based on partial 18S ribosomal DNA (rDNA), internal transcribed spacer (ITS) and the D2-D3 region of 28S rDNA sequences further confirm this nematode as a new species.

Description of Ruehmaphelenchus kuroshioi n. sp. and R. interjectus n. sp. (Tylenchomorpha: Aphelenchoididae) isolated from ambrosia beetles, Euwallacea spp. (Scolytinae), from Japan

Summary Two Ruehmaphelenchus species were isolated from the ambrosia beetles Euwallacea kuroshio and E. interjectus, collected from Okinawa and Kagoshima, Japan, respectively. Both nematodes were successfully cultured on a lawn of the grey mould Botrytis cinerea. Morphological observation and molecular characterisation revealed that both species are undescribed; therefore, the nematodes are described herein as R. kuroshioi n. sp. and R. interjectus n. sp. Phylogenetically, both species are closely related to R. formosanus, R. fujiensis, R. digitulus and R. asiaticus. Ruehmaphelenchus kuroshioi n. sp. is characterised by a conical male and female tail, which has a terminal projection (mucron), small male spicule with a weakly-developed triangular condylus, dorsal and ventral limbs that are connected to each other by three different tissues, and clearly separated distal pairs of genital papillae (P3 and P4). However, R. kuroshioi n. sp. can be distinguished from its close relatives by a combination of male and female tail characters and its distinctive phylogenetic status. Ruehmaphelenchus interjectus n. sp. is almost identical to R. formosanus, sharing characteristic lateral spikes of the spicule; they form a well-supported phylogenetic clade. However, R. interjectus n. sp. is distinguished from R. formosanus by morphometric values and ribosomal RNA, i.e., a 6-bp difference from the ca 1.7 kb molecular sequence of the 18S ribosomal RNA of R. formosanus, which warrants independent species status.

Bursaphelenchus taedae sp. n. (Tylenchina: Aphelenchoididae) – a new species of the eggersi-group in Loblolly pine logs from the USA

Summary Bursaphelenchus taedae sp. n. of the eggersi-group was detected in loblolly pine logs from the USA together with B. antoniae. It is characterised by a relatively small stylet with basal swellings, a lateral field with three lines, and the excretory pore located at the level of the nerve ring. The female has a very small extension of the anterior vulval lip over the vulva (= a ‘vulval flap’), a long PUS extending for 40.1-67.8% of vulva to anus distance, and hook-like tail conical, gradually tapering to a finely rounded or broadly rounded terminus. The male spicules are 17-22 μm long in chord, only slightly ventrally curved, condylus short, truncate, slightly dorsally bent to dorsally hooked, rostrum ca 3-4 μm long, close to the proximal spicule end, without cucullus. Seven genital papillae present. Bursaphelenchus taedae sp. n. can be distinguished from other species of the eggersi-group by morphological and molecular characters.

Description of Ruehmaphelenchus americanum n. sp. (Nematoda: Aphelenchoididae) found in Loblolly pine from the USA

Summary Ruehmaphelenchus americanum n. sp., isolated from southern yellow pine (Pinus taedae L.) from the USA is described and figured. It is characterised by a relatively stout body (a = 30 for females and males), three lines in the lateral field, both oocytes and spermatocytes arranged in two rows, male spicules relatively small (14-18 μm) with weakly developed condylus and rostrum, short tail with a bluntly pointed tip, seven papilliform genital papillae present, female vulva positioned at ca 82% of body length, vulval lips slightly protruding, post-uterine branch extending two-thirds of vulva to anus distance, tail cylindrical, ca two anal body diam. long, terminus forming a spike-like projection or mucron, 7.6-12.2 μm long, with pointed tip. The new species can be separated from 11 known species (except R. thailandae) by male genital papillae arrangement (the second and third pair adjacent vs separated). Detailed phylogenetic analysis based on 18S and 28S D2-D3 region ribosomal RNA (rRNA) sequences has confirmed the status of this nematode as a new species.

Detection and identification of Ektaphelenchoides diversislocis sp. n. isolated from Pinus woodlands of China and Japan

Summary Understanding the occurrence and distribution of parasitic nematodes is crucial – some are economic pests, and some are important predators of their insect associates. In our recent nematode inventory survey, two populations of an ektaphelechid nematode were detected in the branches of Pinus trees; later the same nematode population was detected in plant quarantine examinations of wood packaging material imported from Japan. The species was processed and identified as Ektaphelenchoides diversislocis sp. n. The new species can be characterised by having three lateral lines, stylet long and tripartite, excretory pore at the level of nerve ring, comparatively longer post-vulval uterine sac, indistinct rectum and anus. Female posterior body region conical, gradually narrowing, like a mucron or filiform. Male spicule with well-developed condylus, triangular rostrum, and cucullus absent. The species is morphologically and molecularly close to E. compsi. The new species was characterised with near full-length 18S, 28S D2-D3 regions, and internal transcribed spacer (ITS) rRNA genes and mitochondrial cytochrome oxidase I (COI) gene sequences. The species within the Ektaphelenchoides genus are considered to have predatory roles against insect associates; therefore, the discovery of E. diversislocis sp. n. from local and foreign woodlands warrants increased sampling and research attention.