Description and molecular characterization of Trachactinolaimus persicus sp. n. from Iran (Nematoda, Dorylaimida, Actinolaimidae), with new insights into the taxonomy and evolutionary relationships of the genus.

A new species of the free-living nematode genus Trachactinolaimus, collected in natural habitats of northern Iran, is described, including morphological and molecular (28S-rDNA) data. Trachactinolaimus persicus sp. n. is characterized by its 1.95-2.44 mm long body, lip region weakly offset by depression and 18-20 µm wide, odontostyle 25-27 µm long, neck 540-636 µm long, pharyngeal expansion occupying one-half of the total neck length, bipartite uterus 2.6-3.9 body diameters long, vulva (V = 49-53) pore-like, tail long and filiform in both sexes (174-223 µm, c = 10.0-13.4, c' = 5.9-7.0 in females, 165-196 µm, c = 10.7-13.8, c' = 4.6-5.8 in males), spicules 68-75 µm long, and 12-14 almost contiguous ventromedian supplements with hiatus. Molecular data supports the monophyly of the genus and the hypothesis that Dorylaimidae are the sister group of actinolaims. The taxonomy of Trachactinolaimus is updated, including diagnosis, list of species, key to their identification, and a compendium of their main morphometrics. Dominiactinolaimus is regarded as its junior synonym.

Description of Acromoldavicus xerophilus n. sp. (Nematoda, Rhabditida, Elaphonematidae) from the southern Iberian Peninsula, including a key to species of the genus.

A new species of the genus Acromoldavicus is described from coastal sand dunes and sandy soil in the southeast of the Iberian Peninsula. Acromoldavicus xerophilus n. sp. is characterized by its 557-700 µm body length, cuticle tessellated, lip region with three pairs of expanded lips bearing a large labial expansion, primary axils bearing guard processes with two different morphology, secondary axils lacking guard processes, stoma short and tubular with prostegostom bearing prominent rhabdia directed towards the stoma lumen, female reproductive system monodelphic-prodelphic, post-vulval sac 0.6-0.9 times body diameter, rectum very large, female tail short with biacute terminus and males unknown. The description, light micrographs, scanning electron microscope images, illustrations, and molecular analyses are provided. Molecular analyses (based on 18S and 28S rDNA) revealed its relationship with some species of the genera Cephalobus (18S tree), Nothacrobeles, Paracrobeles, and Spinocephalus (28S tree). Keys to species identification of this genus are also included.

Integrative taxonomy and phylogenetic relationships between representatives of genera within Merliniinae (Nematoda: Merliniidae), with new data on fourteen known and one unknown species.

The application of integrative taxonomic approaches is useful to species delineation based on a combination of distinct types of characters, here morphological features and ribosomal DNA sequences. In this study, we surveyed ectoparasitic nematodes of the subfamily Merliniinae in cultivated and natural environments in Iran. Results of morphological and morphometrical studies, light and scanning electron microscopic observations, and molecular analyses allowed us the identification of fourteen known and one unknown species including representatives of the genera Amplimerlinius (five species), Geocenamus (one species), Merlinius (three species), Nagelus (two species), Paramerlinius (one species), Scutylenchus (two species), and Telomerlinius (one species). The unknown species, Scutylenchus sp., characterized by having 35-50 incisures at mid-body; lateral field with 6 longitudinal incisures; lip region slightly offset by a constriction, flattened at front end; bearing 5-7 annuli; cephalic framework not refractive; stylet robust, 18.3-27 µm long; post anal intestinal sac absent; tail elongate conical, dorsally convex, with 24 (19-28) annuli in ventral side, ending to a smooth terminus and males common; spicules 24.5-31 µm long. The phylogenetic analyses were carried out using molecular data from nuclear ribosomal DNA (rDNA) genes viz. D2-D3 expansion segments of the large ribosomal subunit (28S rRNA), partial small ribosomal subunit (18S rRNA), and internal transcribed spacer (ITS). The molecular variability of D2-D3 expansion segments of the 28S rRNA and partial 18S rRNA was low in this family in comparison to the ITS region, which could be a more helpful molecular marker in species and genus identification.

The enigmatic genus Loofilaimus (Nematoda, Dorylaimida, Loofilaimidae) revisited 25 years after, with remarkable new insights into its phylogeny.

The evolutionary relationships of the nematode genus Loofilaimus are explored with an integrative approach, combining morphological and molecular (28S-rDNA) data. Never recorded since its original description in 1998, the finding of fresh specimens of its type and only species, L. phialistoma, allowed us to obtain SEM observations and sequencing, both for the first time, resulting in relevant aspects to elucidate its phylogeny. Morphologically, the genus is characterized by two autapomorphies affecting its lip region and pharynx. Molecular study revealed that it represents a very restricted evolutionary trend within Dorylaimida. The clade (Nygolaimina + (Loofilaimus + Dorylaimina)) is well supported. Loofilaimidae is accepted as a separate and valid family, which should also include Bertzuckermania.

Morphological and molecular characterization of Metaxonchium magnum sp. n. from Iran (Nematoda, Dorylaimida, Belondiridae), with new insights into the phylogeny of the genus.

A new species of the genus Metaxonchium is described from a natural habitat in Iran. Metaxonchium magnum sp. n. is characterized by its 3.62-4.65 mm long body, lip region cap-like and offset by constriction and 13-16 µm wide, odontostyle fusiform and 14-17 µm long, neck 1016-1359 µm long, both parts of the pharynx separated by a short isthmus-like narrowing, pharyngeal expansion occupying 74.2 (73-77)% of total neck length in females and 70.4 (66-72)% in males, female genital system mono-opistho-ovarian, didelphic, anterior genital branch a large uterine sac with a small terminal mass occupying 7-14% of body length, posterior uterus long and tripartite with a Z-like differentiation, V = 50-52, caudal region short and rounded (24-41 µm, c = 99-161, c' = 0.5-0.7), spicules 90-105 µm long and 10-13 spaced ventromedian supplements with hiatus. Analysis of D2-D3 28S rDNA sequences of the new species suggests that Metaxonchium might not be a monophyletic taxon, a matter that should be confirmed after future research.

Redescription of Diastolaimus grossus (Truskova & Eroshenko, 1977) Andrássy, 1984 (Rhabditida, Chambersiellidae) from Europe and comments on its phylogenetic position.

Two populations of the species Diastolaimus grossus have been obtained from bark of trees in Bosnia and Herzegovina and the Czech Republic. The species is described and characterized in detail using morphological techniques (light and electron scanning microscopy) and morphometrical (Gower General Similarity coefficient of morphological characters) and molecular analyses, including phylogenetic analysis of all related and already sequenced species of the family Chambersiellidae. Morphological and molecular analyses, based on 18S and 28S ribosomal DNA sequences, show that the family Chambersiellidae is polyphyletic, being the subfamily Chambersiellinae related with Cephalobomorpha and Tylenchomorpha, and the subfamily Macrolaiminae is located into Panagrolaimomorpha. The genus Diastolaimus, previously belonging to Macrolaiminae, is transferred to Chambersiellinae. Diastolaimus mexicanus is proposed as a junior synonym of D. grossus.

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

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

Redescription and synonymization of Oscheius citri Tabassum, Shahina, Nasira and Erum, 2016 (Rhabditida, Rhabditidae) from India and its taxonomical consequences.

A population of a nematode species belonging to the genus Oscheius was isolated in western Uttar Pradesh, India. Morphological and morphometrical studies on this species showed its high similarity with six species described previously from Pakistan (Oscheius citri, O. cobbi, O. cynodonti, O. esculentus, O. punctatus and O. sacchari). The molecular analysis of the ITS1-5.8S-ITS2 rDNA sequences of the Indian population and the six species described from Pakistan showed that all the sequences are almost identical. Thus, based on morphological and molecular characteristics, all of the six above-mentioned Pakistani species and Indian strain do not differ from each other, hence can be considered synonyms. The correct name for this taxon is the first described species O. citri. Additionally, the phylogenetic analysis of the 18S rDNA and the 28S rDNA sequences showed that Oscheius citri is sister to the clade formed by O. chongmingensis and O. rugaoensis from China. The high similarity of morphological and morphometric characteristics of O. citri and other species, O. maqbooli, O. nadarajani, O. niazii, O. shamimi and O. siddiqii, suggest their conspecificity; however, lack of molecular data for these species does not allow this hypothesis to be tested.

Morphological and molecular characterization of Labronema montanum sp. n. (Dorylaimida, Dorylaimidae) from Spain.

A new species of the genus Labronema, collected in a natural mountain habitat of the southern Iberian Peninsula, in Spain, is studied, including its morphological and morphometric characterization, SEM observations, and D2-D3 28S-rRNA sequences. Labronema montanum sp. n. is distinguishable by its 1.56 to 2.08 mm long body, with lip region being offset by constriction and 19 to 24 µm broad, odontostyle 21 to 29 µm long, neck 417 to 551 µm long, pharyngeal expansion 205 to 272 µm long, the presence of three cardiac lobes at the pharyngo-intestinal junction, a long and tripartite uterus, longitudinal vulva (V = 57-60), a short and rounded caudal region (19-35 µm, c = 56-86, c = 0.5-0.8), spicules 65 to 76 µm long, and 20 to 25 nearly contiguous ventromedian supplements with hiatus.A new species of the genus Labronema, collected in a natural mountain habitat of the southern Iberian Peninsula, in Spain, is studied, including its morphological and morphometric characterization, SEM observations, and D2-D3 28S-rRNA sequences. Labronema montanum sp. n. is distinguishable by its 1.56 to 2.08 mm long body, with lip region being offset by constriction and 19 to 24 µm broad, odontostyle 21 to 29 µm long, neck 417 to 551 µm long, pharyngeal expansion 205 to 272 µm long, the presence of three cardiac lobes at the pharyngo-intestinal junction, a long and tripartite uterus, longitudinal vulva (V = 57-60), a short and rounded caudal region (19­35 µm, c = 56-86, c = 0.5-0.8), spicules 65 to 76 µm long, and 20 to 25 nearly contiguous ventromedian supplements with hiatus.

Morphological and molecular characterization of Lenonchium zanjanense sp. n. (Dorylaimida, Nordiidae) from Iran, with new insights into the phylogeny of the genus and a revision of its taxonomy.

The new species Lenonchium zanjanense sp. n. is described from a natural habitat of Zanjan province, Iran, including line, light microscopy and scanning electron microscopy illustrations and a molecular (18S, 28S) study. It is characterized by its 3.50-4.51 mm long body, rounded lip region, continuous and 13.5-15.5 µm broad, odontostyle 21-24 µm long, neck 362-490 µm long, double guiding ring, pharyngeal expansion 190-285 µm long, female genital system didelphic-amphidelphic, uterus simple and 185-320 µm long or 3.4-5.9 times the corresponding body diameter, vulva nearly equatorial (V = 45-53), tail conical-elongated to filiform (90-165 µm, c = 23-43, c' = 2.4-5.3) with three or four mucro-like projections at the tip, spicules 58-64 µm long and 16-21 contiguous ventromedian supplements ending at the level of the anterior end of the spicules. The taxonomy of the genus is updated, with an emended diagnosis, list of species, key to their identification and a compendium of their main morphometrics. Lenonchium asterocaudatum is regarded as identical and a junior synonym of L. denticaudatum. New insights into the phylogeny of the group are also provided, and the classification of Lenonchium within Nordiidae is seriously questioned.

Redescription of Macrolaimus canadensis Sanwal, 1960 and M. ruehmi Andrássy, 1966 (Nematoda, Rhabditida, Chambersiellidae), and new data on M. crucis Maupas, 1900.

Three species belonging to the genus Macrolaimus, namely M. canadensis, M. crucis and M. ruehmi, have been obtained from areas of natural vegetation in Bosnia and Herzegovina, Corsica (France), the Czech Republic and Spain. These three species are characterized by the body length of males and females, stomatal proportion of the gymnostom and cheilostom, excretory pore position, postvulval uterine sac length, male and female tail length and morphology, and the length and morphology of the spicules and gubernacula. The occurrence of M. crucis in Spain has, as a result of this study, now also been expanded to a larger area of the southern Iberian Peninsula. Morphological and morphometrical analyses showed that M. canadensis and M. ruehmi are very similar, sharing apomorphic characters. In contrast, M. crucis has plesiomorphic characters. Description, measurements and illustrations are provided for these three species.

Morphological and molecular characterization of Tarantobelus arachnicida gen. n., sp. n. (Nematoda, Rhabditida, Brevibuccidae), a parasitic nematode of tarantulas.

A new genus and new species, Tarantobelus arachnicida, was found in the oral opening of tarantula spiders bred in captivity in Poland. The new species is characterized by having a small body (0.77-0.95 mm long in females and 0.66-0.84 mm in males), cuticle poorly annulated by transverse incisures, lateral field inconspicuous, lips separated with small cuticular flaps topping each lip, stoma panagrolaimoid with gymnostom well developed with robust and refringent rhabdia, pharynx panagrolaimoid with isthmus slightly longer than the basal bulb, intestine with cardiac (anterior) and rectal (posterior) areas with narrower walls. Mature females with intestinal cells including needle crystal packs, excretory pore at isthmus level, female reproductive system panagrolaimoid with post-vulval sac 0.4-0.8 times the length of the corresponding body diameter and having very thick walls, vulva very prominent, female rectum 0.8-1.3 times the length of the anal body diameter, female tail conical with acute tip with phasmids at 58-62% of its length. Male tail conical with long and thin mucro, spicules ventrad bent having rounded manubrium and thick gubernaculum. Description, measurements and illustrations of the new species are provided. Molecular analyses show its relationship with Brevibucca and Cuticonema. On the other hand, Medibulla and its corresponding subfamily Medibullinae, previously in Osstellidae, are transferred to Panagrolaimidae, being Shahnematinae, the junior synonym of Medibullinae. Indocephalobus, recently proposed and located in the family Panagrolaimidae, is considered a junior synonym of Diplogastrellus (Diplogasteromorpha), and its only species, I. zebrae, is considered a junior synonym of D. gracilis. In addition, a key to identification of panagrolaimoid genera is included.

Molecular and morphological characterization of Belondira coomansi n. sp. (Nematoda: Dorylaimida, Belondiridae) from Iran.

A new species belonging to the genus Belondira is described from natural areas in Iran. Belondira coomansi n. sp. is characterized by its general size, the dimensions and appearance of its lip region, presence of distinct labial and post-labial sclerotization, the length of the odontostyle and its inconspicuous lumen and aperture, the length of the neck and its pharyngeal expansion, the reduction of the female anterior genital branch to a simple uterine sac, a very short posterior uterus, the anterior position of the vulva, the length and shape of the caudal region with distinctly thick cuticle at its tip, the length of the spicules, and the presence of only one pair of ventromedian supplements. The new species is close to Belondira brevibulba, B. sacchari, B. tenuidens and B. thornei, and it is compared to them. Molecular characterization (D2-D3 expansion segments of the rRNA large subunit) of the new species is also provided, representing only the second species of this genus for which any DNA sequence data are available.

A low-cost technique to manufacture a container to process meiofauna for scanning electron microscopy.

An easy and low-cost method to elaborate a container to dehydrate nematodes and other meiofauna in order to process them for scanning electron microscopy (SEM) is presented. Illustrations of its elaboration, step by step, are included. In addition, a brief methodology to process meiofauna, especially nematodes and kinorhynchs, and illustrations are provided. With this methodology it is possible to easily introduce the specimens, to lock them in a closed chamber allowing the infiltration of fluids and gases (ethanol, acetone, carbon dioxide) but avoiding losing the specimens. After using this meiofauna basket for SEM the results are efficient. Examples of nematode and kinorhynch SEM pictures obtained using this methodology are also included.

Nematodes of the Order Rhabditida from Andalucía Oriental, Spain. The Genera Protorhabditis (Osche, 1952) Dougherty, 1953 and DiploscapterCobb, 1913, with Description of P. spiculocrestata sp. n. and a Species Protorhabditis Key.

A new species of the genus Protorhabditis is described from natural areas in the SE Iberian Peninsula. Protorhabditis spiculocrestata sp. n. is distinguished by its body length 387-707 mum in females and 375-546 mum in males, lip very low and flattened, stoma 14-22 mum long, female tail conical-elongate (48-100 mum, c = 6.4-8.3, c' = 4.8-7.5), phasmid near anus, male tail conical (20-27 mum, c = 18.3-22.3, c' = 1.4-1.5), bursa peloderan closed anteriorly and bears eight papillae (1+2+1+1+3), spicules 23-26 mum long, and gubernaculum 10-16 mum long. Diploscapter coronatus is also presented. Description, measurements and illustrations, including SEM photographs, are provided. A key to species of Protorhabditis is also given as well a compendium of their measurements.

Description and SEM Observations of Stegelletina coprophila sp.n. (Nematoda: Rhabditida) from Caves of Andalucía Oriental, Spain.

A new species of the genus Stegelletina is described from caves of Andalucía Oriental (SE Iberian Peninsula). Stegelletina coprophila sp. n. is characterized by its body 386 to 536 mum long in females and 391 and 521 mum in males, lateral field with three incisures, lips bearing four tines, labial probolae 4 to 6 mum long and bifurcate, pharyngeal corpus 1.5 to 2.9 times isthmus length, spermatheca 18 to 43 mum long, postvulvar sac 0.5 to 1.6 times the corresponding body diameter long, female tail conical (24-34 mum, c = 14.2-17.2, c' = 1.7-2.3), male tail conical (26-35 mum, c = 12.1-17.1, c' = 1.4-1.8), spicules 20 to 25 mum long, and gubernaculum 10 to 13 mum long. Descriptions, measurements and illustrations, including SEM photographs, are provided for the species.

Nematodes of the order rhabditida from andalucía oriental, Spain. The genera nothacrobeles allen &noffsinger, 1971 and zeldia thorne, 1937.

A new species of the genus Nothacrobeles is described from natural areas (a salt lake) in the Southeast Iberian Peninsula. Nothacrobeles lanceolatus sp. n. is characterized by its body length, two rows of cuticular punctations per annulus, labial probolae bifurcate with divergent prongs, pharyngeal corpus 2.4 to 3.5 times isthmus length, spermatheca length, postuterine sac 0.5 to 1.1 times the corresponding body diameter ratio, female tail conical and bearing a spindle-shaped or conical mucro with acute terminus, phasmid at 8 to 17 microm posterior to the anus, male tail conical with acute mucro, spicules length, and gubernaculum length. In addition, Nothacrobeles cf. lunensis and Zeldia punctata are studied. Cervidellus capricornis is transferred to genus Nothacrobeles. A key to species of Nothacrobeles is also provided.

Lipoprotein(a) levels and risk of coronary heart disease in men. The lipid Research Clinics Coronary Primary Prevention Trial.

OBJECTIVE: To examine the relationship between elevated levels of lipoprotein(a) [Lp(a)] and coronary heart disease (CHD) risk in a prospective study. DESIGN: Nested case-control study. The cohort consisted of participants in the Lipid Research Clinics Coronary Primary Prevention Trial. SETTING: Lipid research clinics. PARTICIPANTS: The Lipid Research Clinics Coronary Primary Prevention Trial participants (n = 3806) were men, aged 35 to 59 years, with plasma cholesterol levels of 6.85 mmol/L (265 mg/dL) or greater, low-density lipoprotein cholesterol levels of 4.91 mmol/L (190 mg/dL) or greater, and triglyceride levels less than 3.39 mmol/L. Subjects were randomly assigned to either cholestyramine or placebo treatment. The Lp(a) levels were measured in plasma samples obtained prior to randomization in 233 cases (participants who developed CHD in the course of the study) and 390 matched CHD-free controls. A total of 96.95% of the subjects were white, 2.25% were black, and 0.80% were of other race. MAIN OUTCOME MEASURE: Coronary heart disease (either fatal or nonfatal) events during a follow-up of 7 to 10 years. RESULTS: The Lp(a) levels were significantly higher (21%) in cases than in controls (23.7 mg/dL [0.59 mmol/L] and 19.5 mg/dL [0.49 mmol/L], respectively; P < .02). This difference was still statistically significant (P < .01) after controlling for age, body mass index, cigarette smoking, blood pressure, low-density lipoprotein cholesterol level, and high-density lipoprotein cholesterol level. When subjects were divided by treatment, both cholestyramine-treated and placebo-treated CHD subjects had Lp(a) levels 20% to 22% greater than their matched controls. However, possibly because of smaller sample sizes, these differences were no longer statistically significant. CONCLUSIONS: Our data are consistent with the concept that an elevated Lp(a) level is an independent risk factor for CHD in hypercholesterolemic white men.