Vulval Flap Research Articles

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.

Unveiling two novel Ditylenchus (Nematoda: Anguinidae) species inhabiting the growing medium of Rhododendron simsii imported from Japan

Summary The present study describes two new Ditylenchus species, isolated from the growing medium of Rhododendron simsii imported from Japan, at the Ningbo plant quarantine laboratory, Ningbo, P.R. China. Ditylenchus parvicauda n. sp. is an amphimictic species. Females have six incisures in the lateral fields, a medium length slim body, a short delicate stylet with asymmetrical stylet knobs, the secretory-excretory (S-E) pore located posterior to the isthmus and in the anterior region of the basal pharyngeal bulb, a posteriorly located vulva (V = 80.9-83.0), post-uterine sac (PUS) tube-like, and the tail cylindrical, tapering gradually towards finely rounded terminus. Males are similar to females in general morphology and have 10.1-14.9 μm long spicules. The second new species, D. gracicauda n. sp. is characterised by having a long and slender body, the presence of a vulval flap, four lines in the lateral field, a narrow and low lip region not offset, the S-E pore located posterior to the isthmus but not in the anterior region of the basal pharyngeal bulb, a posteriorly located vulva (V = 71.1-76.2), a short post-vulval uterine sac short (4.5-9.2 μm), and the tail slender, tapering gradually towards finely rounded to wedge-shaped terminus. The two new species were also characterised molecularly using 18S, ITS and the D2-D3 expansion segments of 28S rRNA genes. Our phylogenetic analyses showed their independent place among available Ditylenchus species. Several Ditylenchus species are regulated pests, and the introduction of these species into a new environment may result in diseases among native plants or vegetation in the area. Therefore, careful attention must be directed towards newly discovered species, particularly those under quarantine observation.

Ektaphelenchoides factum n. sp. in Pinus taeda from the USA and E. pini in Pinus koraiensis from China

Summary The genus Ektaphelenchoides is a diverse group of nematodes; they are either inhabitants of trees or present in the rhizosphere of economical plants. Members of the genus have ecological relevance in terms of being associated with insects and predators of other nematodes. In the present study, a new Ektaphelenchoides species, E. factum n. sp., is described and illustrated. The new species was isolated from a log sample of Pinus taeda imported from the USA. It is characterised by the lateral field with three incisures, stylet distinctly tripartite. Females having a vulval flap, the anus and rectum indistinct, and the tail (posterior part of body) filiform. Males having arcuate and 21.5 (19.4-22.9) μm long spicules, the lamina of the spicule is smoothly curved to distal end, condylus well developed with a broadly rounded tip, rostrum triangular with finely rounded tip and cucullus absent, tail arcuate and conical with terminus bearing a sharply pointed mucron. The new species is different from other Ektaphelenchoides species in having a vulval flap. Molecular characterisation and phylogenetic studies also supported its unique status. In addition, a Chinese native population of E. pini, detected in Pinus koraiensis branches from Liaoning province, P.R. China, is described.

Characterisation of Paratylenchus cacao n. sp. from Ghana, and P. enigmaticus (Nematoda: Paratylenchinae) from Belgium

Summary Paratylenchus cacao n. sp., a pin nematode associated with cocoa (Theobroma cacao L.) from Ghana, is described herein based on morphology, morphometrics and phylogenetic relationships with other species. Morphologically, P. cacao n. sp. is characterised by a female body length of 174-265 μm, body assuming a ‘C’ or ‘J’ shape after heat relaxation, a flat head, slightly set off lip region, a truncated anterior end, a rigid stylet of 24-31 μm, four lateral lines, vulval region with distinct vulval flaps, and a finely rounded tail with a mucron-like terminus. The findings of the phylogenetic analyses utilising the D2-D3 expansion segments of 28S, ITS of rDNA, and COI of mitochondrial DNA sequences indicate that P. cacao n. sp. is a species closely related to P. leptos. Additionally, based on butterhead lettuce and leek populations from Belgium, the first description of a male, SEM pictures and COI sequences of P. enigmaticus are presented. Both studies exemplify how even potentially important plant-parasitic nematode species can remain unnoticed for a long time. This suggests the need to conduct more nematode diversity studies and underscores the significance of correct nematode identification through the utilisation of tools such as integrative taxonomy.

Detection and identification of Devibursaphelenchus minor sp. n. isolated in the bark from the USA

Summary Bark in an empty container from the USA was inspected in Ningbo port, China, and a Devibursaphelenchus population was detected. It was processed and identified as Devibursaphelenchus minor sp. n. The new species is characterised by the lateral field with three incisures, female stylet tripartite, 13.9 (12.7-14.6) μm long without basal thickenings, secretory-excretory pore at the same level as nerve ring or slightly posterior, vagina not sclerotised, vulval lips slightly protruding and vulval flap absent. Post-vulval uterine sac (PUS) not well developed, very short and obscure, extending for half vulval body diam. long, anus and rectum indistinct or absent. Posterior body region conical elongated, gradually narrowing, slightly ventrally curved, ending with a bluntly pointed terminus. Male rare, the spicules are arcuate, separate, 8.2 μm along the chord line, with well-developed condylus, triangular rostrum with finely rounded tip, cucullus or apophysis absent. Tail conical, dorsally convex, ventrally slightly concave with bluntly pointed terminus, ventral bursa not confirmed. Two pairs of genital papillae present. It differs from all other species in this genus by extremely short body length of females and males. Molecular phylogenetic analyses based upon rRNA D2-D3 sequences confirmed it as a new species.

First Morphological and Molecular Characterization of Paratylenchus vandenbrandei (Rhabditida: Tylenchulidae) in Iran.

Paratylenchus vandenbrandei, has been recovered from the rhizospheric soil of Euphrates poplar (Populus euphratica) in the Karkheh protected area of Khuzestan province, southwestern Iran. The species was identified as P. vandenbrandei by the presence of three lines in the lateral fields; conoid rounded lip region; presence of submedian lobes, a stylet 24.0-28.8 μm long; an excretory pore at the level of the anterior part of the pharyngeal bulb; a round-to-oval spermatheca; presence of vulval flaps; and a conoid tail, with a terminus that is rounded or slightly pointed in some specimens. Males have a conoid tail, with a rounded-to-slightly-pointed terminus. The phylogenetic relationships of the species were reconstructed and investigated using partial sequencing of the D2-D3 expansion segments of large subunits, as well as internal transcribed spacer regions (LSU D2-D3 and ITS rDNA) based on Bayesian inference (BI). P. vandenbrandei has formed a clade with P. neonanus, P. minor, P. nainianus, P. chongqinjensis, P. pedrami, P. baldaccii, P. leptos and P. rostrocaudatus with maximal support (BPP = 1.00). To the best of our knowledge, this is the first report of P. vandenbrandei in Iran and the first molecular characterization of the species worldwide.

Morphometry and Molecular Identification of Haemonchus Cobb, 1898 (Trichostrongylidae: Nematoda) Isolates from Small Ruminants in Tanzania Based on Mitochondrial cox 1 and rRNA-ITS genes.

The genus Haemonchus is the major abomasal parasite of ruminants responsible for substantial economic losses in tropical and temperate regions. This study was conducted to clarify the morphometry and molecular characterisation of Haemonchus species isolated from sheep in Babati district, Tanzania. A total of 486 trichostrongylid nematodes were recovered from five sheep. Of the total worms, 106 nematodes were distinguished by 37 males and 69 females. The asymmetrical length of dorsal ray and the distance of bulb at the apex of spicules were used for identification of males. In females, the linguiform vulvar flap was the most predominant with 33 out of 69 (48%) compared with knobbed morph type which was 25/69 (36%) and smooth morph type with 11/69 (16%). Partial cox1 sequence fragments of Haemonchus contortus isolates showed 98.8%, 99.3%, 99.7%, 99.5%, 99.3%, and 98.4% in male, smooth, knobbed, linguiform A, linguiform B, and linguiform C, respectively; with the average nucleotide divergence ranged from 1.03 to 2.35%. The amplified fragments of ITS-2 genes in knobbed, linguiform A, and smooth morphotypes revealed 99.4%, 98.5%, and 98.3%, respectively. Phylogenetic analysis was evaluated by employing Bayesian inference and maximum-likelihood, and the tree was distinctly separated into three clusters focusing on H. contortus in cluster I within the family Haemonchidae. Genetic drifting, mutation, and modification of the morphological features of the Haemonchus species described to have an impact on the development of drug resistance. Species identification is necessary to understand which species infect animal host. We recommend more studies on the parasites intensity and the strategies for controlling Haemonchus species in Tanzania.

Morphometric and phylogenetic study of Haemonchus isolates in small ruminants from mountainous (Lorestan) and plain (Khuzestan) regions of Iran

Haemonchus species are among ruminants' important blood-feeding and pathogenic nematodes. This study aimed to determine morphometric and phylogenetic characteristics of Haemonchus spp. isolates from sheep and goats in two different regions of Iran. A total of 340 isolates of adult Haemonchus spp. from the animals of Khuzestan and Lorestan provinces were identified using morphometric and molecular characteristics. Differentiating functions (DF) of male Haemonchus spp. as well as vulvar morphology and flap diversity for female Haemonchus spp. were determined. The twenty-four samples of Haemonchus contortus that showed the most phenotypic differences were analyzed for the sequencing of two genes, ITS-2 and ND4. Since the DF was less than 0.63, all male isolates were H. contortus. The results showed significant differences in some morphometric characteristics of males and females of H. contortus in animals of two regions. H. contortus female worms showed smooth (2–20 %), knobbed (32–65 %), and linguiform (20–66 %) forms as the lowest vulvar morphs and most common vulvar flap types. Phylogenetic and sequence analysis of ITS2 and ND4 sequences revealed the presence of five and three genotypes for all the isolates, respectively. Morphometric and molecular findings in the present study confirmed H. contortus as the only species in small ruminants from two regions of Iran. The morphometric indices of male and female isolated worms of H. contortus were significantly different concerning the species of animal (goat and sheep) and the geographical region. This study showed that ND4 is more valuable for studying the diversity and phylogeny of H. contortus than ITS2. In this study, there was no correlation between genotypes of different isolates and morphological characteristics, indicating variations in genotypes did not appear to affect the phenotype of H. contortus.

Paratylenchus Ilicis n. Sp. (Nematoda: Paratylenchinae) Associated with Holly from the Netherlands and New Taxonomical and Phylogenetic Support for the Synonymization of Cacopaurus with Paratylenchus.

Paratylenchus ilicis n. sp. was found associated with holly in the Netherlands and was described based on morphology, morphometrics, rRNA and mitochondrial cox1 genes, phylogenetic relationships with other Paratylenchus species, host information and geographical distribution. This species can be morphologically diagnosed based on its light brown, slightly obese to obese females with tubercles on cuticle, lateral bands widening into an ovoid field around vulva level, stylet length of 70 μm to 100 μm, outstretched to reflexed ovary, rounded sperm-filled spermatheca, vagina opening into a thick-walled rounded space, absence of vulval flaps and vulva at 89% to 95% of body length, very short tail in all life stages, and a characteristic finger-like tail tip in juveniles (J2). The new species is morphologically closest to Cacopaurus pestis but differs based on the absence of a scutellum-like differentiation in the lateral field, the ovaries that can be outstretched or reflexed, and the finger-like tail tip in J2. Furthermore, both species were found to be molecularly distant from one another, found in different habitats, and are associated with different hosts. The high morphological similarity between Cacopaurus and Paratylenchus and our phylogenetic analyses, revealing that the former is embedded within different Paratylenchus clades and thus polyphyletic, provide new evidences for the synonymization of Cacopaurus with Paratylenchus.

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.

SCANNING ELECTRON MICROSCOPIC (SEM) STUDY OF NEMATODE PARASITES OF GOAT IN LUCKNOW DISTRICT UTTAR PRADESH

The goat is a vital milk and meat production source in India and contributes to the national and international economy. Goat production faces multiple challenges; one major problem is parasites leading to high mortality and economic losses. Nematode parasites are major problems to livestock globally, characterized clinically by enteritis, anaemia, emaciation, dehydration, and death. This study revealed the morphological identification of nematode parasites (Haemonchus sp., Oesophagostomum sp., and Trichuris sp.) in goats in the Lucknow region (India). Morphological identification was done by SEM (Scanning Electron Microscope) with the help of Souls in 1982. Scanning electron microscope exposed some species-specific characters, total body length of parasites, posterior region of Haemonchus having well-developed Copulatory Bursa (CB), spicules, Dorsal ray (DR) Anterior region of Haemonchus having cervical papillae (CP), vulval flap, knob, and cervical lamina. Oesophagostomum showed that the collar region on the mouth part and Buccal Capsule (BC) is surrounded by External Corona Radiate (ECR) and Internal Corona Radiate (ICR). Trichuris sp. was showed the anterior end vulva region of the female, and the posterior end of the male was spirally coiled and spicular sheath covered with minute spines (S).

Redescription of Ektaphelenchoides compsi Baujard, 1984 (Tylenchina: Aphelenchoididae) isolated from Pinus massoniana in Fujian Province, China

Summary Ektaphelenchoides compsi is redescribed morphologically with new molecular characterisation. It was isolated from a dead Pinus massoniana tree in Ningde City, Fujian Province, China. Detailed morphology of the spicule, female gonad, hemizonid position, arrangement of male caudal papillae and female tail terminus shape are documented. It is characterised by a lateral field with three lines (forming two bands), tripartite stylet 17.8 (17.0-19.4) μm long without basal thickenings, metacorpus rectangular with anterior 40% granular and posterior part weakly muscular, metacorpal valve slightly posterior to middle of metacorpus, excretory pore at level of nerve ring, vagina with thickened walls and strongly developed muscular bundles, vulval lips slightly protuberant, vulval flap absent, distal region of post-vulval uterine sac appearing as a weakly developed oogonia, anus and rectum indistinct, female posterior part (‘tail’) dorsally convex, conical, terminal region contracted into a bluntly pointed tip. The spicules are arcuate, 15.6 (14.3-16.3) μm along the chord, lamina smoothly curved to distal end, capitulum slightly concave, condylus well-developed with broadly rounded tip and slightly depressed at dorsal end, rostrum triangular with finely rounded tip, cucullus absent, and with seven caudal papillae present. The near full length 18S and 28S D2-D3 regions of rRNA genes sequences were characterised. The phylogenetic analyses revealed that the Fujian population of E. compsi grouped with the Zhejiang population of E. compsi, both being morphologically identical.

Scanning electron microscopy of different vulval structures in a Mexican Haemonchus contortus isolate.

Haemonchus contortus is a parasite species that affects the health and production of grazing small ruminants in different parts of the world. Scanning electron microscopy (SEM) is an important tool for the study of parasites' morphology and taxonomy as it generates images that appear 3D and are generally easier to interpret than optical microscopy images. This study used the SEM to describe the vulval types of adult H. contortus from a Mexican isolate. A total of 14 adult H. contortus females were obtained from two artificially infected goats. Females were fixed and processed by critical point drying and observed with SEM. A collection of SEM images was obtained from these parasites and those images were used to identify the structures previously described by optical microscopy studies. Two different types of vulval structures were described in this Mexican H. contortus isolate: Type 1 (vulval flap), Type 2 (epiptygma). An unusual vulval structure was reported in a single individual. The Type 1 included vulval flaps of different sizes and spatial dispositions, as well as one or more knobs in different positions around the vulva. The Type 2 shows differences in the epiptygma. The present study suggests that the Mexican H. contortus isolate used in donor animals possess intraspecific polymorphism in vulval structures.

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

Summary Ruehmaphelenchus taedae n. sp., isolated from Loblolly pine logs (Pinus taedae L.) from the USA, is described and figured. It is characterised by a relatively slim body (a = 42 and 43 for males and females, respectively), three lines in the lateral field, male spicules relatively small (12-18 μm) with high and dorsally bent condylus and weakly developed rostrum, bursal flap absent, short tail possessing a long terminal spike ending in a bluntly rounded tip and 8.7-13.3 μm long, vulva positioned at ca 83% of body length, vulval flap absent, vulval lips slightly protruding, post-vulval uterine branch extending for less than half of vulva to anus distance, and female tail conoid, ca 3-4 anal body diam. long, with 13.7-18.5 μm terminal projection. The new species can be separated from all other species of the genus by the male tail possessing a long terminal spike and the more anterior excretory pore. Detailed phylogenetic analysis based on 28S D2-D3 region sequences confirmed the status of this nematode as a new species.

Subcutaneous Vulvar Flap Viability Evaluation With Near-Infrared Probe and Indocyanine Green for Vulvar Cancer Reconstructive Surgery: A Feasible Technique.

Introduction: Vulvar cancer is a rare condition affecting older women and accounts for 3–5% of all gynecological cancers. Primary surgical treatment involves the removal of a large amount of tissue for which reconstructive surgery is often necessary with a high rate of postoperative complications. Despite several techniques for the evaluation of vulvar flap viability have been proposed, many methods cannot be performed during surgery and require expensive devices often missing in a gynecological clinic. This study aims to verify the feasibility and the safety of the vulvar flap viability evaluation through a near-infrared endoscopic probe and Indocyanine green (ICG) tracer in a small group of patients and to evaluate long-term vulvar flap outcomes.Methods: Patients with primary vulvar cancer who required surgical treatment and subsequent vulvar flap reconstructive surgery were prospectively included in the study. A 25 mg ICG vial diluted in 20 ml of saline solution was intravenously infused before closing the skin edges of the flaps. All patients were given 0.2 mg/kg body weight of intravenous ICG. After 10–15 min, a near-infrared endoscopic probe was used to evaluate the vulvar flap viability.Results: Of the 18 patients who underwent radical vulvectomy for vulvar cancer during the study period, 15 were included in the analysis. All packaged surgical flaps showed tracer uptake on the surgical margin. No intro-operative complications were recorded neither surgery-related nor to dye infusion. No surgical infection, dehiscence, or necrosis was recorded.Conclusions: Vulvar flap viability assessment using Indocyanine green and a laparoscopic infrared probe is a feasible method. All cases included in the analysis showed a dye uptake on the surgical edge of the flap. Further, prospective studies are needed to confirm the method in clinical practice and to evaluate its superiority over simple subjective clinical evaluation.

First Report of a Sheathoid Nematode (Hemicriconemoides litchi) on Longan in Taiwan.

Longan (Dimocarpus longan Lour.) is a subtropical fruit, widely grown in China, Taiwan, and Southeast Asia. Longan has a high commercial value, US$242,200 of dried longan fruits are exported to the United States from Taiwan every year (Wang et al. 2010). A soil sample from a longan orchard located in Changhua County, Taiwan (24.0162657, 120.529-1457) was collected in June 2019. Sheathoid nematodes were the dominant species in this sample and over 40 adult individuals per 100 g soil were found. In December 2019, sheathoid nematodes were collected again from the same tree and processed for identification by morphological and molecular characteristics. Nematodes were extracted using the modified Baermann funnel method (Wu et al. 2010) for 24 h. The morphometric data from fifteen females with variant tail types were taken. All individuals had a closely sheath, lip region set off with two annuli, stylet frequently slightly dorsally curved with rounded knobs, no vulval flaps, tail narrowing to a broadly rounded terminus, or tapering to a truncate end, anus situated 3-4 annuli posterior to the vulva. The body length = 552 μm (483 to 616 μm), body width at mid-body = 34.02 μm (27.75 to 40.03 μm), a = 15.93 (14.73 to 18.87), b = 4.65 (4.04 to 5.06), V% = 91.57 (86.74 to 92.56), stylet length = 66.66 μm (63.51 to 69.6 μm), tail length = 28.48 μm (23.56 to 37.45 μm), ring number 114 to 130, Rs = 14 to 19, Roes = 22 to 28, RV = 9 to 13, Ran = 5 to 9 and RVan = 3 to 4. Since the stylet length were less than 70μm, they are more fitted to be Hemicriconemoides litchi (Van den Berg et al. 2015). DNA samples extracted using VIAGEN® DirectPCR lysis buffer from single females (n = 10) were processed to amplify the 28S D2-D3 expansion segment and the ITS region using primers sets D2A and D3B, TW81 and AB28, respectively (Van den Berg et al. 2014). The D2-D3 region of the nematodes collected in this study (MT-539384) shared 99% similarity with several H. litchi sequences deposited in the GenBank database (e.g. KP192481, KF856540), and the ITS region (MT556011) also shared 99% similarity with several H. litchi sequences (e.g. KP192482, GQ354786). Therefore, based on morphological, molecular data and phylogenetic relationship analysis (Nguyen et al. 2020) the nematodes from the Longan orchard were determined to be H. litchi. To measure the reproductive factor (Rf), 60 sheathoid nematodes (57 females and three males) recovered from Baermann extraction were inoculated onto root systems of a longan tree in a 15-cm-d pot filled with sterilized soil. One hundred and sixty eight days after inoculation, three 100 cm3 subsamples of soil from the pot were processed as mentioned previously, and the average number was 48/100 cm3 of soil. The final population was approximately 768 nematodes per 1600 cm3 of soil with the Rf value of 12.8 confirming Longan as a host, although no symptoms were observed. The DNA from three individuals in the three subsamples with variant tail types were used to obtain D2-D3 and ITS region sequences, and confirmed the species as H. litchi. Hemicriconemoides spp. is associated with root malformation and nutrient deficiencies on agricultural fruit trees (Milne et al. 1971; McSorley et al. 1980); however, aboveground evidence of damage may not become immediately obvious and the importance of sheathoid nematodes is easily overlooked (Chen et al. 2011). This is the first report of H. litchi as a parasite of longan tree in Taiwan.

Bursaphelenchus paraleoni n. sp. (Tylenchina: Aphelenchoididae) intercepted in Loblolly pine logs imported into China from the USA

Summary Bursaphelenchus paraleoni n. sp. was isolated from Loblolly pine, Pinus taeda, logs from the USA. The new species is characterised by a lateral field with three lines, 13-16 μm long stylet with small basal swellings, and the excretory pore located slightly posterior to the nerve ring. The male spicules are mitten-shaped, the lamina smoothly tapering towards the distal tip. The condylus is broadly rounded, with a small dorsally bent hook, rostrum triangular or conical with finely rounded tip, and cucullus absent. One midventral precloacal papilla, one adcloacal pair and two postcloacal pairs of genital papillae are present. The bursal flap is long and spade-like with the posterior margin truncate or irregular. Females have a small vulval flap 4-6 μm long, a post-vulval uterine sac extending for 34-49% of the vulva to anus distance, a very long tail with slight ventral curvature, mostly filiform and with a finely rounded terminus, occasionally bluntly rounded. Huge variation in the tails of males and females were observed in a culture. It is similar to species of B. eidmanni, B. leoni, and B. silvestris in the leoni-group but detailed phylogenetic analysis based on 18S, ITS and D2-D3 28S sequences confirmed it as a new species.

Bursaphelenchus parayongensis n. sp. (Tylenchina: Aphelenchoididae) found in packaging wood from China

Summary Bursaphelenchus parayongensis n. sp. was isolated from packaging wood of a consignment from China exported to Germany. The new species is characterised by the lateral field with three lines, 9-13 μm long stylet with small basal swellings, and the excretory pore located near the posterior part of metacorpus. The male spicules are mitten-shaped, the lamina smoothly tapers towards the distal tip. The condylus is broadly rounded, with a small dorsally bent hook, rostrum triangular with sharply pointed tip, and without cucullus. One precloacal papilla, one adcloacal pair and two postcloacal pairs of male papillae present, with the posteriormost pair forming ‘gland papillae’. Bursal flap long, spade-like with posterior margin truncate or irregular. Females have a small vulval flap of 2-3 μm long, and a post-vulval uterine sac extending for 55-77% of the vulva to anus distance. Tail cylindrical, mostly (about 70%) with a robust mucron, very short to less than 3 μm, but occasionally tail terminus bluntly rounded, without mucron. It is similar to species in subgroup 3 of the eggersi group including B. carpini, B. clavicauda, B. cryphali and B. yongensis, but morphologically different. Detailed phylogenetic analysis based on partial 18S, ITS and D2-D3 region of 28S sequences has confirmed the status of this nematode as a new species.

Morphometrical and Molecular Characterization of Oesophagostomum columbianum (Chabertiidae: Oesophagostominae) and Haemonchus contortus (Trichostrongylidae: Haemonchinae) Isolated from Goat (Capra hircus) in Sylhet, Bangladesh.

This study was aimed at describing two (2) intestinal nematodes from naturally infected native breed of goats (Capra hircus) in Bangladesh, identified as Oesophagostomum columbianum (Curtice, 1890) Stossich 1899 and Haemonchus contortus (Rudolphi, 1803) Cobb, 1898. The identification was made based on morphometric features and was confirmed by amplifying internal transcribed spacer (ITS) and cytochrome c oxidase (cox1) gene. Well-developed lateral alae, distinct cervical papillae anteriorly to esophageal expansion, and male spicule length (0.73-0.79 mm, n = 2) were characteristically observed in O. columbianum. At the same time, male spicule length (0.40-0.46 mm, n = 2) and position of female vulvar flap (4.30-4.54 mm from posterior end, n = 3) were observed in H. contortus. DNA sequence homology of the ITS and cox1 gene of both specimens revealed the same results, showing similarity to the GenBank sequences of O. columbianum (GenBank No. KC715827; JX188470) and H. contortus (GenBank No. KJ724377; HQ389229). Phylogenetic analysis computed by maximum livelihood (ML) from the ITS nucleotide sequences revealed that the O. columbianum and H. contortus isolates identified in this study were clustered in the same clade with isolates from China and Iran, respectively. This study, for the first time, illustrates the characteristics of O. columbianum and H. contortus in Bangladesh, combining both morphological and molecular data. The universal primer-based polymerase chain reaction (PCR) protocol could be an economical and efficient option for researchers from poor resource settings for precise identification of nematodes. The information generated in this study may contribute to formulating effective control strategies against these nematodes.

Laimaphelenchus africanus n. sp. (Tylenchomorpha: Aphelenchoididae) from South Africa, a morphological and molecular phylogenetic study, with an update to the diagnostics of the genus.

A newly recovered population of the genus Laimaphelenchus from a dead maritime pine wood sample in Potchefstroom, South Africa, representing a new species, named L. africanus n. sp., is herein described and illustrated based on morphological and molecular data. The new species is mainly characterized by the following: 750–987 µm long females; a cephalic region with no disc and six cephalic lobs not divided by ribs; a 10.0–12.5 µm long stylet; four incisures in the lateral field; secretory-excretory pore (SE-pore) at slightly posterior to the nerve ring; vulva with a well-developed anterior flap, vagina with two well-developed sclerotized pieces; post-vulval uterine sac (PUS) 63–125 µm long; tail conical, 30–44 µm long, ventrally curved with a subventral stalk in terminus, lacking tubercles, with six to nine small projections at the tip in scanning electron microscopy (SEM); and rare males with 17 μm long spicules. The new species was morphologically compared to those species of the genus with a stalk in tail terminus, lacking tubercles, a vulval flap and four incisures in the lateral field viz., L. liaoningensis, L. preissii, L. simlaensis, L. sinensis, L. spiflatus, and L. unituberculus. Phylogenetically, the new species was placed into the major Laimaphelenchus clade using partial large subunit ribosomal DNA (LSU rDNA D2-D3) sequences. An overall literature review corroborated the presence of the stalk (currently with two main groups) at the tail end is the main characteristic trait delimiting the genus. A compendium based on the characters of the stalk, presence/absence of a vulval flap in females and number of the lateral lines was also established.