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Ring nematodes are obligate ectoparasites on crops and natural herbaceous and woody plants, and some species are of economic importance and cause damage to roots of several crops. Recent integrative taxonomical analyses recognized the existence of two cryptic species within the Criconema annuliferum morphotype in Spain. In this study, we corroborated that morphometric, morphological and a multi-locus analysis (including the ribosomal markers D2-D3 expansion segments of 28S rRNA, ITS rRNA, 18S RNA, and the mitochondrial DNA cytochrome oxidase I gene) identified a new lineage clearly separated from C. annuliferum, C. paraannuliferum and C. plesioannuliferum. The new lineage was described herein as Criconema pseudoannuliferum sp. nov., confirming that C. annuliferum species complex species complex comprises a hyper-cryptic species complex. This research analysed soil samples from the rhizosphere of maritime pine (Pinus pinaster Ait.) forests in Bermeja-Crestellina Mountain, located at the western part of Málaga province, southern Spain. The integrative taxonomical analyses revealed the occurrence of a new cryptic species identified using females, males and juveniles with detailed morphology, morphometry and molecular markers, described herein as Criconema pseudoannuliferum sp. nov. All molecular markers (D2-D3, ITS, 18S and COI) were obtained from the same individual that was also used for morphological and morphometric analyses. This research demonstrated the hidden diversity within the C. annuliferum species complex species complex can reach to four lineages under ribosomal and mitochondrial gene markers for one morphospecies group, which includes four species, viz. C. annuliferum, C. paraannuliferum, C. plesioannuliferum, and C. pseudoannuliferum sp. nov. Criconema pseudoannuliferum sp. nov. was detected in moderate soil density in two maritime pine forests (5 and 25 nematodes/500 cm3 of soil) suggesting that does not cause damage to maritime pine. [ABSTRACT FROM AUTHOR]

Paratylenchus species are obligate ectoparasitic nematodes on cultivated and wild herbaceous and woody plants occupying numerous soil categories. Several species may cause damage to several crops (viz. P. dianthus, P. enigmaticus, P. microdorus, P. hamatus and P. epacris on carnation, lettuce, rose and walnut, respectively). This investigation proves and emphasizes the relevance of applying integrative taxonomy for the accurate detection of Paratylenchus species in mountainous wild environments in the Malaga province, Southern Spain. This research analyzed 45 soil samples of maritimus pine and one of green heather in southern Spain and identified fourteen Paratylenchus species, two of them are described herein as new species (P. paraaonli sp. nov., P. plesiostraeleni sp. nov.), six of them were first reports for Spain (P. canchicus, P. nainianus, P. neonanus, P. salubris, Paratylenchus sp. 2 SAS, and P. wuae), and six species (P. caravaquenus, P. microdorus, P. nanus, P. neoamblycephalus, P. sheri, and P. variabilis) have been already reported in Spain. Accordingly, these data increase the biodiversity of pin nematodes in Spain comprising a total of 47 species (33.1% out of 142 total species of this genus). Phylogenetic analyses based on ribosomal and mitochondrial markers (D2-D3, ITS, and partial COI) resulted in a consistent position for the newly described Paratylenchus species in this study (P. plesiostraeleni sp. nov., P. paraaonli sp. nov.). Paratylenchus plesiostraeleni sp. nov. grouped in a separated subclade as unequivocal species from the P. straeleni-complex species (including P. straeleni and P. parastraeleni), and P. paraaonli sp. nov. clustered with P. vitecus, but clearly separate from this species. This study indicates that Paratylenchus species diversity in natural environments may be higher than expected, and this study may help in accurate identifications. [ABSTRACT FROM AUTHOR]

Ring nematodes are obligate ectoparasites on cultivated and wild herbaceous and woody plants, inhabiting many types of soil, but particularly sandy soils. This study explored the morphometrical and molecular diversity of ring nematodes resembling Criconema annuliferum in 222 soil samples from fruit crops in Spain, including almond, apricot, peach and plum, as well as populations from cultivated and wild olives, and common yew. Ring nematodes of the genus Criconema were detected in 12 samples from under Prunus spp. (5.5%), showing a low to moderate nematode soil densities in several localities from southeastern and northeastern Spain. The soil population densities of Criconema associated with Prunus spp. ranged from 1 nematode/500 cm3 of soil in apricot at Sástago (Zaragoza province) to 7950 and 42,491 nematodes/500 cm3 of soil in peach at Ricla and Calasparra (Murcia province), respectively. The integrative taxonomical analyses reveal the presence of two cryptic species identified using females, males (when available), and juveniles with detailed morphology, morphometry, and molecular markers (D2-D3, ITS, 18S, and COI), described herein as Criconema paraannuliferum sp. nov. and Criconema plesioannuliferum sp. nov. All molecular markers from each species were obtained from the same individuals, and these individuals were also used for morphological and morphometric analyses. Criconema paraannuliferum sp. nov. was found in a high soil density in two peach fields (7950 and 42,491 nematodes/500 cm3 of soil) showing the possibility of being pathogenic in some circumstances. [ABSTRACT FROM AUTHOR]

Summary: Specimens of a thin longidorid species collected in Peñalba (Huesca), north-west Spain, were previously described as Paralongidorus iberis. However, we conclude, through scanning electron microscopy and molecular studies on a population from about 15 km from the type locality and on paratype specimens, that this species was originally placed in the wrong genus. Both populations have pore-like amphidial apertures, not slit-like as in Paralongidorus , and the species is therefore transferred to Longidorus. Longidorus iberis n. comb. is regarded as a valid species and is clearly different from closely related species such as L. tabernensis , L. iliturgiensis , L. alvegus and L. indalus in morphometrics and molecular markers. Molecular data are reported for the first time, including the D2-D3 expansion segments of 28S rRNA, ITS1 rRNA, partial 18S rRNA and partial mitochondrial coxI regions. These molecular markers were used for inferring the phylogenetic relationships with other species within Longidorus and Paralongidorus , all clearly separating L. iberis n. comb. from other related taxa and placing the species in the Longidorus clade, rather than with Paralongidorus. [ABSTRACT FROM AUTHOR]

Longidorid nematodes comprise more than 500 species, and Longidorus and Xiphinema are the most diversified, prevalent, and cosmopolitan genera within plant‐parasitic nematodes. This increases the risk of species misidentification. We conducted an integrative morphometric and genetic study on two longidorid species to elucidate the existence of new cases of cryptic speciation within the genera Longidorus and Xiphinema. Detailed morphological, morphometrical, multivariate, and genetic studies were carried out, and mitochondrial and nuclear haploweb analyses were used to differentiate species within the L. iliturgiensis and X. hispanum complexes. Species delimitation using multivariate and haplonet tools of L. iliturgiensis species complex clearly separated L. tabernensis sp. nov. from L. iliturgiensis and L. indalus, and X. subbaetense sp. nov. from X. hispanum and X. adenohystherum. D2‐D3, partial 18S, and partial coxI regions were used for inferring their phylogenetic relationships with other species in each genus. The present study provides new insights into the diversity of Longidorus and Xiphinema species detected in southern Spain, and new evidence of cryptic speciation in both genera. These results support our hypothesis that the biodiversity of Longidoridae in southern Europe is higher than previously supposed and is still not fully clarified. [ABSTRACT FROM AUTHOR]

Three populations of Xiphinema non- americanum group species were detected in agricultural and natural ecosystems, during routine surveys for plant-parasitic nematodes in Spain. Based on morphological and molecular analyses, the species were identified as Xiphinema histriae and Xiphinema lapidosum, being this the first record and molecular characterization of both species in Spain. The morphometrics and morphology of the Spanish populations agree with those of the original description and paratype specimens and the present study provided a first description of the second to fourth juvenile stages of both species. A detailed study on the morphology in the Spanish populations of X. histriae, as well as in paratypes, showed a pseudo- Z-organ with weakly muscularized wall and containing numerous small dense granular bodies, which was different to the original description by Lamberti et al. (1993). This new finding suggests that X. histriae must be considered a member of the morphospecies Group 5 of X. non-americanum. Phylogenetic analysis based on D2 to D3 expansion segments of 28S gene, ITS1 and partial CoxI gene indicated that X. histriae and X. lapidosum are phylogenetically related with other Xiphinema non-americanum group spp. reported from Spain. Considering the pathological and economic importance of this group of nematodes, the combination of morphological characters, measurements, and molecular analysis is crucial for accurate identification of these species. [ABSTRACT FROM AUTHOR]

Root-knot nematodes of the genus Meloidogyne are recognised worldwide as a major production constraint in crops of primary economic importance. Knowledge of their diversity and prevalence, as well as the major environmental and agronomical cues for understanding their distribution in specific areas is of vital importance for designing control measures to reduce significant damage. We provide the first detailed information on the diversity, distribution and levels of Meloidogyne species infecting wild and cultivated olive soils in a wide-region in southern Spain that included 499 sampling sites. Overall Meloidogyne spp. were found in 6.6% of sampled olive plants, with 6.6% and 6.5% for cultivated and wild olive, respectively. We identified five previously described Meloidogyne spp. (Meloidogyne arenaria, M. baetica, M. hapla, M. incognita, M. javanica) and one new species (Meloidogyne oleae sp. nov.) which, characterized using integrative taxonomy, increases the known biodiversity of Meloidogyne spp. affecting olive. Meloidogyne arenaria and M. incognita were only found infecting cultivated olive varieties, while, M. baetica was only found infecting wild olive. Three major parameters drive the distribution of Meloidogyne spp. in cultivated olives in southern Spain, cover vegetation on alley, irrigation and soil texture, but different species respond differently to them. In particular the presence of M. incognita is highly correlated with sandy loamy soils, the presence of M. javanica with irrigated soils and cover vegetation, while the presence of M. arenaria is correlated with the absence of cover vegetation on alley and absence of irrigation. These parameters likely influence the selection of each particular Meloidogyne species from a major dispersal source, such as the rooted plantlets used to establish the orchards. [ABSTRACT FROM AUTHOR]

The genus Xiphinema constitutes a large group of ~275 species of polyphagous, plant-ectoparasitic nematodes that are distributed almost worldwide. Some species affect agricultural crops by feeding directly on root cells as well as by transmitting nepoviruses. Despite their agricultural importance, species discrimination in Xiphinema is difficult, leading to potential misidentification. Integrative taxonomy, based on the combination of molecular analyses and morphology, constitutes a new insight into Xiphinema species identification. In this study we describe two new species of Xiphinema from the Iberian Peninsula (X. macrodora, sp. nov. and X. oleae, sp. nov.) associated with cultivated and wild olive trees. Both species have specific rRNA sequences. Morphologically, Xiphinema macrodora, sp. nov. is characterised by a very long body (7.2-8.7 mm), a very long odontostyle and odontophore (190-206 and 105-120 µm, respectively), and a well developed pseudo-Z-organ, comprising 8 to 12 sclerotised bodies. Xiphinema oleae, sp. nov. is characterised by an odontostyle and an odontophore 136-149 and 65-80 µm long, respectively, and a well developed Z-organ with refractive inclusions (3-5), variable in shape. Additionally, X. macrodora, sp. nov. has the longest body size, and the longest odontostyle and odontophore of any Xiphinema, whereas X. oleae, sp. nov. is the first species with a well developed Z-organ from the Iberian Peninsula. [ABSTRACT FROM AUTHOR]

During a survey for Trichodoridae in cultivated and natural environments, mainly from southern Spain, 7 new Trichodorus species were found. Four of them, T. andalusicus n. sp., Z asturanus n. sp., T. silvestris n. sp. and T. parasilvestris n. sp., belong to the T. lusitanicus morpho-species group characterised in the male by the slightly ventrally curved spicules with a mid-blade constriction with bristles, and in the female by well developed, rounded triangular to quadrangular, vaginal sclerotised pieces. They were mainly differentiated based upon differences in body length, onchiostyle length, spicule shape and number of ventromedian cervical papillae in the male, and size and shape of the vaginal sclerotised pieces in the female. The presence of the T. lusitanicus morpho-species group with 8 species, apparently endemic to the Iberian Peninsula, could point to a recent speciation event. Additionally, 3 new species, T. onubensis n. sp., T. iliplaensis n. sp. and T. paragiennensis n. sp., more closely resembling the morpho-species group of T. sparsus, to which the Spanish species T. giennensis also belongs, were identified. They were differentiated based on body length, length of onchiostyle and position of the secretory-excretory pore and, in the male, by length and shape of the spicules and number of ventromedian cervical papillae and in the female by the vaginal sclerotised pieces. Molecular support to differentiate the new species using the D2-D3 expansion segments of 28S rDNA is also provided. In addition, a new population of T. variabilis from Greece was studied as well as T. lusitanicus type specimens, and a population of T. giennensis and T. lusitanicus were also sequenced. [ABSTRACT FROM AUTHOR]

Rotylenchus vitis n. sp., a new amphimictic species infesting soil and roots of commercial vineyards in southern Spain, is described. Rotylenchus vitis n. sp. is characterised by a truncate lip region with 7-9 annuli and continuous with the body contour, lateral fields areolated at pharyngeal region only, body without longitudinal striations, stylet length of 36-48 μm, vulva position at 47-57%, tail rounded to hemispherical with 13-21 annuli. Morphologically, this species is related to R. cazorlaensis, R. capitatus, R. elegans, R. fabalus, R. iranicus, R. labiodiscus, R. montanus and R. troncapitatus. The results of the phylogenetic analysis based on the sequences of the D2-D3 expansion regions of the 28S and ITS1-rRNA genes confirmed the species differentiation and the close molecular relationship between R. eximius and R. montanus, respectively. [ABSTRACT FROM AUTHOR]

During a recent nematode survey in natural environments of the Los Alcornocales Regional Park narrow valleys, viz., the renowned 'canutos' excavated in the mountains that maintain a humid microclimate, in southern Spain, an amphimictic population of Xiphinema globosum was identified. Morphological and morphometric studies on this population fit the original and previous descriptions and represent the first report from Spain and southern Europe. Molecular characterisation of X. globosum from Spain using D2-D3 expansion regions of 28S rRNA, 18S rRNA and ITS1-rRNA is provided and maximum likelihood and Bayesian inference analysis were used to reconstruct phylogenetic relationships within X. globosum and other Xiphinema species. A supertree solution of the different phylogenetic trees obtained in this study and in other published studies using rDNA genes are presented using the matrix representation parsimony method (MRP) and the most similar supertree method (MSSA). The results revealed a closer phylogenetic relationship of X. globosum with X. diversicaudatum, X. bakeri and with some sequences of unidentified Xiphinema spp. deposited in GenBank. [ABSTRACT FROM AUTHOR]

During recent nematode surveys in a muddy soil around undetermined graminaceous plants in El Rocío, Huelva Province, in southern Spain, and from the rhizosphere of date palm associated with graminaceous vegetation from Abadan, in Khuzestan Province, south-west Iran, populations of Longidorus orientalis were identified. Morphological and morphometrical studies on these populations fit the original description and represent the first report from Spain and Europe. Molecular characterisation of L. orientalis from Spain and Iran, using D2-D3 expansion regions of 28S rRNA, 18S rRNA and ITS-rRNA, is provided. Sequences of the D2-D3 expansion regions and partial 18S genes were analysed using maximum likelihood and Bayesian inference to reconstruct phylogenetic relationships within L. orientalis and other Longidorus species. The results revealed a closer phylogenetic relationship with L. goodeyi for the D2-D3 expansion region and with L. vineacola for the partial 18S region. [ABSTRACT FROM AUTHOR]

A new amphimictic species, Pratylenchus hispaniensis n. sp., parasitising the roots of gum cistus in Andújar (Jaén), southern Spain, is described. The new species is characterised by the presence of numerous males and by the female having a lip region with three annuli, a divided face, a robust stylet (14.5-17.0 μm) with rounded knobs, lateral fields with four lines, V = 80-84, a round spermatheca full of sperm, well developed post-vulval uterine sac and an obliquely truncate tail with irregularly annulated terminus. Morphologically this species is related to P. bhatti, P. kralli, P. mediterraneus, P. pseudofallax and P. thornei. A phenetic study of the 25 most useful diagnostic morphological and allometric characters for Pratylenchus species was done using multivariate factor and linear discriminant analyses. In the factor analysis the first seven factors accounted for 71.1% of the total variance of the characters selected. These factors were related to female tail, pharyngeal overlap, reproductive behaviour, stylet length, L/post-vulval uterine sac ratio, body length and number of lip annuli. Discriminant analysis differentiated Pratylenchus spp. from the three valid species of Zygotylenchus. The results of the phylogenetic analysis based on sequences of the D2-D3 expansion regions of 28S, partial 18S and ITS rRNA genes confirmed the close relationship of P. hispaniensis n. sp. with P. mediterraneus and inferred molecular affinity with P. brzeskii, P. neglectus and P. thornei, in spite of variation in the position of P. hispaniensis n. sp. in the clades. Additional phylogenetic analyses based on the same sets of sequences for P. hispaniensis n. sp., Zygotylenchus guevarai and other Pratylenchidae indicated that Pratylenchus includes several paraphyletic lineages; however, likelihood tests did not reject monophyly of the genus. The inclusion of Pratylenchus, Zygotylenchus, Hirschmanniella, Nacobbus and Apratylenchus in Pratylenchidae was supported. [ABSTRACT FROM AUTHOR]

Paralongidorus litoralis sp. n., a new bisexual species of the genus, is described and illustrated by light microscopy, scanning electron microscopy and molecular studies from specimens collected in a coastal sand dune soil around roots of lentisc (Pistacia lentiscus L.) from Zahara de los Atunes (Cadiz), southern Spain. Paralongidorus litoralis sp. n. is characterised by the large body size (7.5-10.0 mm), a rounded lip region, clearly offset from the body by a collar-like constriction, and bearing a very large stirrup-shaped, amphidial fovea, with conspicuous slit-like aperture, a very long and flexible odontostyle ca 190 μm long, guiding ring located at 35 μm from anterior end, and males with spicules ca 70 μm long. In addition, identification data of a Spanish population of P. paramaximus Heyns, 1965 recovered from sandy soil of a commercial citrus orchard at Alcala de Guadaira (Seville), southern Spain, agree very well with the original description of the species from South Africa. The 18S rRNA and D2 and D3 expansion regions of 28S rRNA gene sequences were obtained for P. litoralis sp. n. and P. paramaximus. Phylogenetic analyses of P. litoralis sp. n. and P. paramaximus rRNA gene sequences and of Longidoridae sequences published in GenBank were done using maximum likelihood and Bayesian inference. In trees generated from the 18S data set Paralongidorus clustered as an external clade from Longidorus, and in trees generated from D2-D3 of 28S dataset Paralongidorus was monophyletic and nested within Longidorus. Maximum likelihood test supported the hypothesis of validity of the Paralongidorus genus. [ABSTRACT FROM AUTHOR]

A new species of awl nematode, Dolichodorus mediterraneus sp. n., is described from southern Spain. The new species is described and illustrated by light microscopy and scanning electron microscopy studies from specimens collected in a sandy soil around roots of cork oak (Quercus suber L.) from Hato Raton, Aznalcázar (Seville), southern Spain. Dolichodorus mediterraneus sp. n. is characterised by a prominent, rounded, lip region, which is offset from the body and has four to six very fine annuli; lateral fields areolated with three lines, ending posterior to anus level; tail 72-122 μm long, abruptly tapering to an acuminate, often spicate, terminus; tail projection 51-79 μ m long; spicules ventrally arcuate, 33-42 μm long; gubernaculum slightly ventrally curved, 14-22 μm long. Scanning electron microscopy observations showed a similar en face pattern to other Dolichodorus spp. Molecular data from the ribosomal large subunit D2-D3 expansion segment of 28S rDNA from paratypes yielded a single fragment of approximately 800 bp. This is the first record of the genus Dolichodorus in Spain and only the second in Europe. A key to species is provided. [ABSTRACT FROM AUTHOR]

Systematic nematode surveys for criconematids in the coastal sand dunes of southern Spain and on the Atlantic coast of Florida, USA, revealed the presence of three populations of sheathoid nematodes identified as Hemicriconemoides promissus, H. ortonwilliamsi and H. wessoni. All of them belong to the fourth group proposed by Decraemer and Geraert (1992) as they are characterised by wide body annuli ca ≥ 5 μm wide and a stylet ca 50 μm long, although all have a distinctly different female tail shape. Comparative SEM studies on H. cocophillus, H. mangiferae, H. brachyurus and H. promissus were done. SEM studies from paratype material and female specimens of a newly discovered population from Spain, confirmed the absence of lateral vulval flaps for both H. promissus populations, justifying the rejection of the synonymy proposed by Germani and Anderson (1991), and the re-establishment of H. promissus as a species distinct from H. brachyurus. The present records of H. promissus and H. ortonwilliamsi extend their geographical distribution from Italy to an additional Mediterranean country and constitute the first reports of both species in Spain. Additional morphometric data and illustrations of a Pensacola, FL, USA, population of H. wessoni, the type species of the genus, are also presented. [ABSTRACT FROM AUTHOR]

SEM studies on a Meloidogyne baetica population provided additional details of the external morphology for female, male and second-stage juveniles. The labial disc in female and male specimens is fused with the medial lips forming a single structure. In second-stage juveniles the lateral lips are triangular with rounded margins. The amphidial opening for all life stages appears oval to rectangular in shape and is located between the labial disc and lateral lips. Lateral fields of male and second-stage juveniles have four incisures irregularly areolated along the entire body. The results of a host-range study for additional natural hosts of M. baetica conducted in wild olive communities growing at Vejer de la Frontera (Cádiz province) in southern Spain are also reported. Apart from the type host, M. baetica was found to infect two natural woody host plants, lentisc (Pistacia lentiscus) and Aristolochia baetica. Hostparasite relationships in these new hosts confirmed the typical susceptible reaction observed in wild and cultivated olives. Similarly, the reproductive fitness, evaluated as the number of eggs per egg mass, was not significantly different in all plant hosts. No infections or galled roots were observed in herbaceous plant species studied and M. baetica must therefore be considered as a parasite of woody plants. [ABSTRACT FROM AUTHOR]

Six crucifer species as potential cover crops, Ethiopian mustard (Brassica carinata line C-101), turnip (Brassica rapa cv. Norfolk), radish (Raphanus sativus), wild rocket (Eruca vesicaria), wild cabbage (Moricandia moricandioides) and white mustard (Sinapis alba), were tested for susceptibility to Meloidogyne arenaria race 2, M. incognita race 1 and M. javanica. Experiments were conducted under glasshouse conditions at 22-28°C for 2 months after inoculation of plants with eggs and second-stage juveniles. All crucifers were infected by Meloidogyne spp. The nematode and crucifer species significantly influenced the severity of root galling and nematode reproduction. Among the plants tested, turnip was the most suitable host for all three Meloidogyne spp. as indicated by severity of root galling and nematode reproduction. The least suitable hosts were wild rocket for M. arenaria, radish for M. incognita and white mustard for M. javanica. The reproduction fitness of M. javanica was greater than that of M. arenaria race 2 and M. incognita race 1 on all plants. [ABSTRACT FROM AUTHOR]

High infection rates of wild olive (Olea europaea sp. sylvestris) feeder roots and soil infestation by a new root-knot nematode were found in sandy soil at Vejer de la Frontera (Cádiz), southern Spain. Morphometric traits and analyses of the nematode esterase electrophoretic pattern as well as of the internal transcribed spacer 1 (ITS1)-5.8S gene and D2-D3 fragment of the 28S gene of rDNA showed that specimens differed clearly from known root-knot nematodes. Studies of host-parasite relationships showed a typical susceptible reaction in naturally infected wild olive plants and in olive planting stocks (cvs. Arbequina and Picual) artificially inoculated with the nematode. However, the nematode did not reproduce in artificially inoculated chickpea, pea, and tomato. Because of the ability of this new nematode to infect wild and cultivated olives only, we suggest the common name, "Mediterranean olive root-knot nematode." The species is herein described and illustrated, and named as Meloidogyne baetica n. sp. The new root-knot nematode can be distinguished from other Meloidogyne spp. by (i) the perineal pattern, which is almost similar to that of M. artiellia, characterized by distinct inner striae forming two distinct longitudinal bands, extending throughout the perineum to just below the vulva; (ii) female excretory pore anterior to the level of stylet knobs, excretory pore distance from anterior end/length of styler ratio extremely small (0.5 to 0.8); and (iii) second-stage juveniles with elongate-conoid tail. Phylogenetic trees derived from maximum parsimony analyses showed that M. baetica is closely related to M. artiellia, the cereal and legume root-knot nematode. [ABSTRACT FROM AUTHOR]

Presents a study that described Hemicriconemoides macrodorus n. sp. from a bisexual population collected in a natural habitat at Santa Elena, Jaén, southern Spain. Method of the study; Results and discussion; Conclusion.

This study delves into the diagnosis of pin nematodes (Paratylenchus spp.) in Spain based on integrative taxonomical approaches using 24 isolates from diverse natural and cultivated environments. Eighteen species were identified using females, males (when available) and juveniles with detailed morphology-morphometry and molecular markers (D2-D3, ITS and COI). Molecular markers were obtained from the same individuals used for morphological and morphometric analyses. The cryptic diversity using an integrative taxonomical approach of the Paratylenchus straeleni-species complex was studied, consisting of an outstanding example of the cryptic diversity within Paratylenchus and including the description of a new species, Paratylenchus parastraeleni sp. nov. Additionally, 17 already known species were identified comprising P. amundseni, P. aciculus, P. baldaccii, P. enigmaticus, P. goodeyi, P. holdemani, P. macrodorus, P. neoamblycephalus, P. pandatus, P. pedrami, P. recisus, P. sheri, P. tateae, P. variabilis, P. veruculatus, P. verus, and P. vitecus. Eight of these species need to be considered as first reports for Spain in this work (viz. P. amundseni, P. aciculus, P. neoamblycephalus, P. pandatus, P. recisus, P. variabilis, P. verus and P. vitecus). Thirty-nine species of Paratylenchus have been reported in Spain from cultivated and natural ecosystems. Although we are aware that nematological efforts on Paratylenchus species in Southern Spain have been higher than that carried out in central and northern part of the country, the present distribution of the genus in Spain, with about 90% of species (35 out of 39 species, and 24 of them confirmed by integrative taxonomy) only reported in Southern Spain, suggest that this part of the country can be considered as a potential hotspot of biodiversity. [ABSTRACT FROM AUTHOR]

Simple Summary: Paratylenchus spp. are vermiform organisms distributed throughout the world that can parasitize a wide variety of cultivated and wild plants. Some species are considered pathogenic in crops; therefore, correct identification is essential to design management strategies. However, the conserved morphology, similarity of morphometric characters and other factors as co-occurrence of more than one species of Paratylenchus in the same soil sample hinder identification to species level. In consequence, this identification should be carried out jointly with morphological, morphometrical and molecular data. The present research aims to provide morphological and molecular characterization of some Paratylenchus species found in Spain and the description of several new species. In previous studies, fifteen species of Paratylenchus, commonly known as pin nematodes, have been reported in Spain. These plant-parasitic nematodes are ectoparasites with a wide host range and global distribution. In this research, 27 populations from twelve Paratylenchus species from 18 municipalities in Spain were studied using morphological, morphometrical and molecular data. This integrative taxonomic approach allowed the identification of twelve species, four of them were considered new undescribed species and eight were already known described. The new species described here are P. caravaquenus sp. nov., P. indalus sp. nov., P. pedrami sp. nov. and P. zurgenerus sp. nov. As for the already known described species, five were considered as first reports for the country, specifically P.enigmaticus, P. hamatus, P. holdemani, P. israelensis, and P. veruculatus, while P. baldaccii, P. goodeyi and P. tenuicaudatus had already been recorded in Spain. This study provides detail morphological and molecular data, including the D2-D3 expansion segments of 28S rRNA, ITS rRNA, and partial mitochondrial COI regions for the identification of different Paratylenchus species found in Spain. These results confirm the extraordinary cryptic diversity in Spain and with examples of morphostatic speciation within the genus Paratylenchus. [ABSTRACT FROM AUTHOR]

The genus Longidorus constitutes a large group of approximately 170 species of plant-ectoparasitic nematodes that are polyphagous and distributed almost worldwide. Some of the species of this genus are vectors of plant viruses. Species discrimination in Longidorus is difficult because the morphology is very conservative, and morphometric characters often overlap, leading to potential misidentification. Integrative taxonomy, based on the combination of molecular analyses with morphology, is a useful and necessary approach in Longidorus species identification. In Spain from 2014 to 2017, we conducted nematode surveys among cultivated and wild plants, from which we identified 13 populations of Longidorus, two of which appeared to represent new species and are described herein as L. iliturgiensis sp. nov. and L. pacensis sp. nov., and 11 populations belonging to eight known species: L. africanus, L. baeticus, L. carpetanensis, L. fasciatus, L. nevesi, L. cf. olegi, L. pini, and L. vallensis. Three species are new geographical records for Spain (L. nevesi, L. cf. olegi, and L. africanus). We report molecular data for L. nevesi, L. cf. olegi, L. carpetanensis and L. pini for the first time. Additionally, we describe the males of L. pini and the juveniles of L. cf. olegi. [ABSTRACT FROM AUTHOR]

• We reveal new insights about the influence of environment in comparison to agricultural management on patterns diversity of PPN in olive ecosystems. • Relatively high levels of shared contributions for space and soil variables. • Effects of agricultural practices on species richness and beta diversity of PPN were less than expected. Numerous studies have documented the distribution of plant and animal communities with respect to spatial structure; however, relatively little is known about the involvement of spatial structures in the diversity of soil organisms such as plant-parasitic nematodes (PPN). Host plants such as olive trees are of particular interest because they host a large number of PPN and have a high economic and cultural importance. In this study, we investigated how different aspects of the environment (i.e. the factors soil, above-ground environment, and agricultural management) and spatial structure shaped the variation of species composition (expressed as beta diversity) and species richness of plant-parasitic nematodes infesting the soil rhizosphere in 376 commercial olive orchards in the south of Spain. We used variation partitioning to assess the relative importance of the unique and shared contributions of the factors describing the environment and spatial structure. To identify sites and species of particular interest, we partitioned beta diversity into local and species contributions. Contrary to our expectation that soil and agricultural management would largely determine the community structure of PPN, more than two-thirds of the variation remained unexplained. Spatial structure and soil were the most important factors shaping species richness and beta diversity. Surprisingly, the effects of agricultural management on species richness were lower than expected, and null [or nonexistent, or nonsignificant] on beta diversity. We found relatively high levels of shared contributions of the different factors, especially in combination with spatial structure, indicating the presence of spatial gradients of the variables describing the environmental factors. Species contributions to beta diversity (SCBD) were positively correlated with nematode prevalence and density range; thus, SCBD could be related to the niche position as reported in other ecosystems. Local contributions to beta diversity (LCBD) were mainly related with habitat filtering mechanisms (e.g. soil physiochemical and agronomic management predictors), suggesting a relationship between nematode total biomass and ecological gradients. Overall, we revealed novel insights into the spatial structure of PPN communities and showed that its beta diversity is less structured by spatial and environmental factors compared to other organism types. [ABSTRACT FROM AUTHOR]

Incidence and nematode population densities of plant-parasitic nematodes were determined in 64 samples of soil and grapevine roots collected from commercial vineyards in southern Spain between October 2003 and May 2005. In addition, a histopathological study was done of root- stock roots naturally infected by root-knot nematodes (Meloidogyne spp.). Nematodes infecting the rootstocks were identified according to conventional procedures, and the Meloidogyne spp. were furthermore identified by sequence characterized amplified region-polymerase chain reaction (SCAR-PCR) and isozyme esterase analyses. The most important plant-parasitic nematodes detected, in order of decreasing frequency of total soil infestation and root infection (percentage of samples), were Mesocriconema xenoplax (34.4%), Meloidogyne incognita (26.6%), Meloidogyne javanica (14.1%), Xiphinema index (12.5%), Xiphinema italiae (10.9%), Pratylenchus vulnus (6.3%), and Meloidogyne arenaria (1.6%). No disease symptoms were observed on aboveground plant parts of the infected grapevines, except for plants in some fields where soil was infested with the virus-vector nematodes X. index and X. italiae. Those grapevines showed a yellow mosaic pattern in leaves early in the growing season and the internode shortening characteristic of infections by Grapevine fanleaf virus. Rootstocks infected by root-knot nematodes (Meloidogyne spp.) showed distorted feeder roots and large- to moderate-sized root galls, present either singly or in clusters. Histopathology of galled roots showed a typical susceptible response to infection by root-knot nematodes: cellular alterations were induced in the cortex, endodermis, pericycle, and vascular system, including giant-cell formation and severe distortion of vascular tissues. Most Meloidogyne egg masses occurred on the surface of the galled root tissues, a position that could facilitate dispersion of the nematode eggs and juveniles and the occurrence of secondary infections. Some of the grapevine rootstocks surveyed in this study (Paulsen 1103, Richter 110, Rupestris du Lot, and SO4) had previously been reported to be resistant to Meloidogyne spp.; however, the population densities of these nematodes found in soil and roots sampled in the present study, as well as the compatible host-parasite relationship revealed by histopathology, indicate a susceptible response to Meloidogyne spp. from southern Spain. [ABSTRACT FROM AUTHOR]

Tumor-necrosis-factor-α inhibitors (anti-TNF-α) are associated with an increased risk of tuberculosis (TB) disease, primarily due to reactivation of latent TB infection (LTBI). We assessed the performance of parallel LTBI screening with tuberculin skin test (TST) and QuantiFERON-TB Gold In-Tube assays (QFT-GIT) before anti-TNF-α treatment in children with immune-mediated inflammatory disorders in a low TB-burden setting. We conducted a multicenter cohort study involving 17 pediatric tertiary centers in Spain. LTBI was defined as the presence of a positive TST and/or QFT-GIT result without clinical or radiological signs of TB disease. A total of 270 patients (median age:11.0 years) were included, mainly with rheumatological (55.9%) or inflammatory bowel disease (34.8%). Twelve patients (4.4%) were diagnosed with TB infection at screening (LTBI, n = 11; TB disease, n = 1). Concordance between TST and QFT-GIT results was moderate (TST+/QFT-GIT+, n = 4; TST-/QFT-GIT+, n = 3; TST+/QFT-GIT-, n = 5; kappa coefficient: 0.48, 95% CI: 0.36-0.60). Indeterminate QFT-GIT results occurred in 10 patients (3.7%) and were associated with young age and elevated C-reactive protein concentrations. Eleven of 12 patients with TB infection uneventfully completed standard LTBI or TB treatment. During a median follow-up period of 6.4 years, only 2 patients developed TB disease (incidence density: 130 (95% CI: 20-440) per 100,000 person-years), both probable de novo infections., Conclusion: A substantial number of patients were diagnosed with LTBI during screening. The dual strategy identified more cases than either of the tests alone, and test agreement was only moderate. Our data show that in children in a low TB prevalence setting, a dual screening strategy with TST and IGRA before anti-TNF-α treatment is effective., What Is Known: • The optimal screening strategy for latent tuberculosis in children with immune-mediated inflammatory disorders remains uncertain. • Children receiving anti-TNF-α drugs are at increased risk of developing severe tuberculosis disease., What Is New: • A dual screening strategy, using TST and an IGRA assay, identified more children with latent tuberculosis than either of the tests alone. • Identification and treatment of latent tuberculosis before initiation of anti-TNF-α therapy averted incident tuberculosis cases., (© 2022. The Author(s).)

This study aimed to evaluate the prevalence and distribution of Paratylenchus species in the main areas of Prunus spp. production in Spain, their ecological constraints, and new molecular tools for the specific identification of major species. Pin nematodes are recognized as plant-parasitic nematodes with a wide host range and global distribution. Therefore, understanding the environmental and agronomic factors involved in their distribution is critical to design environmental and sustainable management strategies to reduce plant damage. A total of 219 sampling sites were surveyed and 12 Paratylenchus species were identified based on an integrative taxonomic approach ( P .  baldaccii , P . enigmaticus , P . goodeyi , P . hamatus , P . holdemani , P . indalus , P . israelensis , P . pedrami , P . tateae , P . tenuicaudatus , P .  veruculatus , and P . zurgenerus ). The most common pin nematode was P . hamatus , followed by P . tenuicaudatus . Nematode abundance was influenced by climatic characteristics, soil chemical properties, and agronomic management practices. Nine explanatory variables were selected as the most strongly associated with Paratylenchus distribution. Specifically, P . tenuicaudatus was significantly correlated with soil chemical characteristics, such as pH and carbon, sulfur, and sodium content, whereas P . goodeyi was closely related to fields with <10 years of almond cultivation. Species-specific PCRs were developed for P . hamatus and P . tenuicaudatus and their validity was evaluated studying the molecular variability of these species and against other Paratylenchus species.

An intracellular bacterium, strain IAS T , was observed to infect several species of the plant-parasitic nematode genus Xiphinema ( Xiphinema astaregiense , Xiphinema incertum , Xiphinema madeirense , Xiphinema pachtaicum , Xiphinema parapachydermum and Xiphinema vallense ). The bacterium could not be recovered on axenic medium. The 16S rRNA gene sequence of IAS T was found to be new, being related to the family Burkholderiaceae, class Betaproteobacteria. Fungal endosymbionts Mycoavidus cysteinexigens B1-EB T (92.9 % sequence identity) and ' Candidatus Glomeribacter gigasporarum' BEG34 (89.8 % identity) are the closest taxa and form a separate phylogenetic clade inside Burkholderiaceae. Other genes ( atpD , lepA and recA ) also separated this species from its closest relatives using a multilocus sequence analysis approach. These genes were obtained using a partial genome of this bacterium. The localization of the bacterium (via light and fluorescence in situ hybridization microscopy) is in the X. pachtaicum females clustered around the developing oocytes, primarily found embedded inside the epithelial wall cells of the ovaries, from where they are dispersed in the intestine. Transmission electron microscopy (TEM) observations supported the presence of bacteria inside the nematode body, where they occupy ovaries and occur inside the intestinal epithelium. Ultrastructural analysis of the bacterium showed cells that appear as mostly irregular, slightly curved rods with rounded ends, 0.8-1.2 µm wide and 2.5-6.0 µm long, possessing a typical Gram-negative cell wall. The peptidoglycan layer is, however, evident only occasionally and not detectable by TEM in most cells. Another irregularly occurring shell surrounding the endosymbiont cells or the cell clusters was also revealed, probably originating from the host cell membrane. Flagella or spore-like cells do not occur and the nucleoid is diffusely distributed throughout the cell. This endosymbiont is transmitted vertically through nematode generations. These results support the proposal of IAS T as a new species, although its obligate intracellular and obligate endosymbiont nature prevented isolation of a definitive type strain. Strain IAS T is therefore proposed as representing ' Candidatus Xiphinematincola pachtaicus' gen. nov., sp. nov.

Reniform nematodes of the genus Rotylenchulus are semi-endoparasites of numerous herbaceous and woody plant species roots and occur largely in regions with temperate, subtropical, and tropical climates. In this study, we provide new records of the nematode Rotylenchulus macrosoma in eight European countries (Czech Republic, France, Germany, Hungary, Italy, Romania, Serbia, and Portugal), in addition to the six Mediterranean countries (Greece, Israel, Jordan, Spain, Syria, and Turkey) where the nematode was previously reported. Four new host species (corn, pea, wheat, and an almond-peach hybrid rootstock) are added to the recorded host species (bean, chickpea, hazelnut, peanut, soybean, and wild and cultivated olive). Molecular analyses based on the cytochrome c oxidase subunit coxI and D2-D3 segments of 28S RNA markers showed high diversity and pronounced genetic structure among populations of Rotylenchulus macrosoma . However, the complexity of phylogeographic patterns in plant-parasitic nematodes may be related to the intrinsic heterogeneity in the distribution of soil organisms, a rare occurrence of a species, or the potential human impact associated with agricultural practices.

Background: Scarce data exist on coronary events following transcatheter aortic valve replacement (TAVR), and no study has determined the factors associated with poorer outcomes in this setting. This study sought to determine the clinical characteristics, outcomes, and prognostic factors of acute coronary syndrome (ACS) events following TAVR., Methods: Multicenter cohort study including a total of 270 patients presenting an ACS after a median time of 12 (interquartile range, 5-17) months post-TAVR. Post-ACS death, myocardial infarction, stroke, and overall major adverse cardiovascular or cerebrovascular events were recorded., Results: The ACS clinical presentation consisted of non-ST-segment-elevation myocardial infarction (STEMI) type 2 (31.9%), non-STEMI type 1 (31.5%), unstable angina (28.5%), and STEMI (8.1%). An invasive strategy was used in 163 patients (60.4%), and a percutaneous coronary intervention was performed in 97 patients (35.9%). Coronary access issues were observed in 2.5% and 2.1% of coronary angiography and percutaneous coronary intervention procedures, respectively. The in-hospital mortality rate was 10.0%, and at a median follow-up of 17 (interquartile range, 5-32) months, the rates of death, stroke, myocardial infarction, and major adverse cardiovascular or cerebrovascular events were 43.0%, 4.1%, 15.2%, and 52.6%, respectively. By multivariable analysis, revascularization at ACS time was associated with a reduction of the risk of all-cause death (hazard ratio, 0.54 [95% CI, 0.36-0.81] P =0.003), whereas STEMI increased the risk of all-cause death (hazard ratio, 2.06 [95% CI, 1.05-4.03] P =0.036) and major adverse cardiovascular or cerebrovascular events (hazard ratio, 1.97 [95% CI, 1.08-3.57] P =0.026)., Conclusions: ACS events in TAVR recipients exhibited specific characteristics (ACS presentation, low use of invasive procedures, coronary access issues) and were associated with a poor prognosis, with a very high in-hospital and late death rate. STEMI and the lack of coronary revascularization determined an increased risk. These results should inform future studies to improve both the prevention and management of ACS post-TAVR.

Olive ( Olea europaea L.) is one of the most important fruit crops in the Mediterranean Basin, because it occupies significant acreage in these countries and often has important cultural heritage and landscape value. This crop can be infected by several Meloidogyne species ( M. javanica , M. arenaria , and M. incognita , among others), and only a few cultivars with some level of resistance to these nematodes have been found. Innovations in intensive olive growing using high planting densities, irrigation, and substantial amounts of fertilizers could increase the nematode population to further damaging levels. To further understand the interactions involved between olive and pathogenic nematodes and in the hope of finding solutions to the agricultural risks, this research aimed to determine the reaction of important olive cultivars in Spain and wild olives to M. javanica infection, including genotypes of the same and other O. europaea subspecies. All olive cultivars tested were good hosts for M. javanica , but high levels of nematode reproduction found in three cultivars (Gordal Sevillana, Hojiblanca, and Manzanilla de Sevilla) were substantially different. In the wild accessions, O. europaea subsp. cerasiformis (genotype W147) and O. europaea subsp. europaea var. sylvestris (genotype W224) were resistant to M. javanica at different levels, with strong resistance in W147 (reproduction factor [Rf] = 0.0003) and moderate resistance in W224 (Rf = 0.79). The defense reaction of W147 to M. javanica showed a strong increase of phenolic compounds but no hypersensitive reaction.

The dagger nematodes Xiphinema pachtaicum and X. index are two of the most widespread and frequently occurring Xiphinema spp. co-infesting vineyards and other crops and natural habitats worldwide. Sexual reproduction is rare in these species. The primary objective of this study was to determine the genetic structure of X. pachtaicum and X. index populations using eight and seven populations, respectively, from different "wine of denomination of origin (D.O.) zones" in Spain and Sardinia (Italy), by studying mitochondrial (cytochrome oxidase c subunit 1 or COI) and nuclear (D2-D3 expansion segments of 28S rDNA) markers. Both Xiphinema spp. showed low intraspecific divergence among COI sequences, ranging from 0.2% (1 base substitution) to 2.3% (10 substitutions) in X. pachtaicum and from 0.2% (1 base substitution) to 0.4% (2 substitutions) in X. index. Population genetic structure was strong for both species. Nevertheless, molecular differences among grapevine-growing areas were not significant, and intrapopulation diversity was very low. It is hypothesized that this genetic homogeneity in the nematode populations reflects their predominant parthenogenetic reproduction mode and low dispersal abilities. Our results also show that X. pachtaicum populations in Spain have possibly been established from two different populations of origin. Results also demonstrated that the two DNA regions studied are suitable diagnostic markers for X. index and X. pachtaicum.

Objectives: To adapt and validate a Spanish-language version (SV) of the National Institutes of Health Stroke Scale (NIHSS) to facilitate its use in Spanish-speaking contexts., Patients and Methods: The methods recommended by the International Quality of Life Assessment Project were followed. Two forward translations and 1 back translation of the NIHSS were developed to ensure lingual and cultural equivalence. A final revised SV-NIHSS was administered by 8 physicians to patients with stroke in 3 clinics in Buenos Aires, Argentina, from September 2003 to December 2003., Results: The study included 102 patients (mean +/- SD age, 73.3+/-6.5 years; 56% women) with stroke (86% ischemic). The SV-NIHSS mean baseline score was 9.78+/-7.04. Interrater reliability was Independently evaluated for 98 patients, showing a high agreement: kappa, 0.77 to 0.99 for the 15 items; interrater correlation coefficient, 0.991 (95% confidence Interval, 0.987-0.994). Intrarater reliability was excellent: kappa, 0.86 to 1.00 for the 15 items; mean intrarater correlation coefficient, 0.994 (95% confidence interval, 0.991-0.996). Construct validity was also adequate; the SV-NIHSS had a negative correlation with baseline Glasgow Coma Scale (Spearman coefficient = -0.574, P < .001) and with Barthel index at 3 months (Spearman coefficient = -0.658, P < .001). Patients with different Rankin scores at 3 months also had significantly different baseline SV-NIHSS scores, from a mean of 4.29+/-2.21 for Rankin score of 0 to a mean of 29.40+/-3.97 for Rankin score of 6 (P < .001)., Conclusion: This study shows that a Spanish-language version of the NIHSS developed with internationally recommended methods is reliable and valid when applied in a Spanish-speaking setting.