Specimens of nematode belonging to Oscheius genis was isolated through the Galleria bait method from soil collected in a karst cave of Tuscany (Central Italy). Molecular and morphological analyses were performed. Total DNA was extracted from individual nematodes and the mitochondrial COI, the ITS containing region and the 18S rRNA gene were amplified and sequenced. BLAST search at the NCBI discriminate this new taxon, similar to other Oscheius. This species belongs to Dolichura group. Cuticle is finely annulated, stoma is short and cheilorhabdion is simple, not well cuticularized. Female body is almost straight upon fixation, the reproductive system is didelphic and tail is short, conoid with pointed tip. Males are rare and similar to female in general morphology except for smaller size. Male body is straight when heat-killed, testis is single, ventral reflexed. Thei show peloderan bursa , tail short rounded and spicules slender and small. Ingective juveniles are slender withelongate tail and have stoma morphology similar to adult. The nematodes were cultured in Petri dishes on several substrates: Nutrient Agar, Escherichia coli, Botritis cinerea, meat baby food, without satisfactory results. Only Petri dishes method with G. mallonella larvae produced IJs, suggesting the entomopathogenicityof this new taxon.

The wide range of feeding types and the ability to adapt to seasonal succession make nematodes significant indicators of ecological conditions of the soil in which they occur. Soil nematophauna is highly sensitive to any environmental damage and, therefore, the analysis of soil nematode community can be a useful diagnostic tool of soil health changes caused by polluting agents, among which also pesticides. Traditional morpho-taxonomic techniques for analysis of soil nematophauna have been flanked or substituted by more innovative and quicker molecular tools. Effectiveness of morpho-taxonomic and molecular techniques was comparatively evaluated through the analysis of soil nematode community from three selected relatively undisturbed and disturbed sites in Apulia region (Italy). Nematodes for both analyses were extracted from 100g sub-samples of composite soil samples collected at each site. Specimens for morphological analysis were fixed in a 2.5% formaldehyde solution and then identified at family and genus level under an optical microscope. The maturity and trophic diversity indices were determined. Total DNA was extracted from the nematode community of each soil subsample and PCR amplification was performed by using the small subunit (18s) of the ribosomal DNA gene, as diagnostic marker. The 18s rDNA was selected because of the large number of 18s sequences in GenBank, the existence of an 18s-based phylogenetic tree and the conserved nature of this gene to ensure complete phylogenetic coverage of the phylum. Sequence analysis through BLAST allowed to classify most of them at genus level and some of them at species level. Few sequences showed no similarity with those present in the database suggesting that they are new for the scientific community. The maturityand trophic diversity indices were also calculated for genera identified at molecular level. Results confirmed that nematodes are good indicators of soil health, as showing a different level of disturbance for each of the three sampled sites. Both morpho-taxonomic and molecular tecniques showed to be effective, though morpho-taxonomic is more time-consuming and skilfulness-requiring whereas a molecular analysis is largely more expensive.

Aquaporin-8 (AQP8) is a membrane channel permeable to water and ammonia. As AQP8 is expressed inthe inner mitochondrial membrane of several mammalian tissues, we studied the effect of the AQP8expression on the mitochondrial transport of ammonia. Recombinant rat AQP8 was expressed in theyeast Saccharomyces cerevisiae. The presence of AQP8 in the inner membrane of yeast mitochondriawas demonstrated by subcellular fractionation and immunoblotting analysis. The ammonia transportwas determined in isolated mitochondria by stopped flow light scattering using formamide as ammoniaanalog. We found that the presence of AQP8 increased by threefold mitochondrial formamide transport.AQP8-facilitated mitochondrial formamide transport in rat native tissue was confirmed in liver (a mitochondrialAQP8-expressing tissue) vs. brain (a mitochondrial AQP8 non-expressing tissue). Comparativestudies indicated that the AQP8-mediated mitochondrial movement of formamide was markedly higherthan that of water. Together, our data suggest that ammonia diffusional transport is a major function formitochondrial AQP8.

La tutela e la gestione del suolo richiedono grande attenzione, viste le importanti funzioni ambientali, economiche, sociali eculturali che il suolo stesso svolge. I nematodi del suolo possono costituire degli utili bioindicatori dello stato di salute di unsuolo, in quanto sono in grado di rispondere prontamente ai cambiamenti ambientali (stress e inquinamento) e pertantol"analisi della loro distribuzione e attività può essere usata per stabilire lo stato di conservazione di un suolo. Ilbiomonitoraggio della nematofauna può essere condotto utilizzando diversi indici ecologici strettamente correlatiall"arricchimento di materia organica ed ai gruppi trofici dei nematodi presenti nel terreno (Maturity Index e EnrichmentIndex). Nel corso del presente studio sono stati campionati tre diversi siti della regione Puglia, localizzati uno in una riservanaturale, il secondo in prossimità di una centrale elettrica a carbone e il terzo in prossimità di una discarica, e per ciascunodi essi è stata condotta l"analisi della nematofauna presente utilizzando sia approcci morfologici che molecolari. Il DNAtotale della nematofauna presente in ciascun campione è stato estratto e sottoposto ad amplificazione mediante PCR. Il geneper il 18S rRNA è stato utilizzato come marcatore molecolare in quanto in banca dati esistono molte sequenze del 18S, talida coprire l"intero phylum dei nematodi. Sono state determinate circa 100 sequenze per ciascun sito. L"analisi dellesequenze mediante BLAST ha permesso di classificare le sequenze ottenute a livello di genere, alcune delle quali sono stateidentificate anche a livello di specie. Gli indici ecologici sono stati calcolati sia per i dati morfologici che per i datimolecolari rivelando che i risultati dei due approcci sono sovrapponibili. I siti analizzati si sono rivelati più disturbati incorrispondenza della centrale elettrica e della discarica rispetto al sito nella riserva naturale. Questi risultati preliminaridimostrano che i nematodi sono sicuramente dei buoni indicatori dello stato di conservazione di un suolo.

Biocontrol effects of arbuscular mycorrhizal fungi against nematodes have been reported in various plants. Literature data suggest that mycorrhizal symbiosis affects plant-water relationships as well. Moreover, it is well established that water deficit and infection with plant parasitic nematodes represent two environmental stresses with interacting effects under field conditions. Few data are available on the effect of combined mycorrhizae and water stress on the development of nematode feeding sites. We studied the impact of Rizophagus intraradices symbiosis on Meloidogyne incognita and tomato (cv San Marzano nano) interaction, with or without water stress. Plants inoculated or not with R. intraradices, maintained in growth chamber at 25°C, were exposed to mild water stress and subsequently infected with J2s of M. incognita. Galls hand-dissected at 7 and 14 days were processed for light microscopy observations. The analysis performed on cross sections of galls i) with or without mycorrhizae, ii) with water stress and iii) with mycorrhizae and water stress, showed changes in the morphology of galls and nematode feeding sites, affecting density and dimensions of nuclei. The symbiosis with R. intraradices and water stress hampered development and structure of giant cells, showing an effect on the modulation of host plant metabolism. NGS-based analysis of galls transcriptome is under study, to unravel the molecular pathways involved in this multiple interaction. Research partially funded by CNR, Progetto Premiale Aqua.

A new longidorid nematode, Longidorus asiaticusn. sp., is described and illustrated from a population extracted from soil associated with the movement of crape myrtle (Lagerstroemia indica) flowering bonsai trees imported from China into Italy. The new needle nematode is characterised by a small body size (2.74-3.52mm), a bluntly-rounded lip region, ca 12 ?m wide, continuous with body contour,amphidial fovea pocket-shape with posterior end rounded not bilobed, a moderately long and flexible odontostyle ca 85 ?m long, stylet ring located at ca 37 ?m from anterior end, vulva almost equatorial (48-54 %), tail short, about 2/3 of its width, dorsally convex-conoid, with rounded terminus, with a c' ratio ca 0.7, bearing two pairs of caudal pores and male absent. Integrative diagnosis was completed with molecular data obtained using D2-D3 expansion segments of 28S rDNA, ITS-rDNA, and partial 18S-rDNA. The phylogenetic relationships of this species with other Longidorus spp. using D2-D3 expansion segments,ITS and partial 18S-rDNA indicated that L. asiaticus n. sp. clustered together with L. hangzhouensis, Longidorus sp. JH-2014, and L. camelliae: all of them sharing a common Asiatic geographic origin.

Seven tomato cultivars and 9 hybrids coming from a breeding programme, susceptible or resistant to root-knot nematode by carrying the resistance gene Mi-1, were compared by determining free phenol content and catalase (CAT) activity in their leaves. A host suitability test based on the determination of the number of egg masses per root system (EM), after inoculation with an avirulent field population of Meloidogyne incognita, confirmed the response to root-knot nematodes of the positive and negative controls, and revealed that all the hybrids except one were highly resistant to the infection. Catalase activity and free phenols extracted from leaves were detected on 3 different groups of tomato plants: 1) known susceptible cultivars; 2) known resistant cultivars; 3) accessions from the breeding programme. The level of catalase activity of the groups 2 and 3 was similar and approximately 2-fold higher than that of the group 1. Conversely, no consistent difference was found in free phenol content among the 3 groups. Moreover, one pair of susceptible and resistant tomato cultivars was used to check changes of anti-oxidant activities due to nematode infection. Catalase and peroxidase activities were measured in roots 5 days after inoculation with 300 and 600 juveniles (J2) of a M. incognita population, and in uninoculated roots used as controls. Catalase was found to be inhibited and enhanced, respectively, in infected resistant and susceptible roots, with respect to controls. The degree of inhibition was higher in the resistant roots inoculated with 600 J2. Guaiacol peroxidase activity did not change after inoculation of susceptible plants, whereas it was enhanced in infected resistant roots compared with controls. The use of catalase activity as a biochemical marker of Mi-1-mediated resistance in tomato is proposed.

The expression pattern of pathogenesis-related genes PR-1, PR-2 and PR-5, considered as markers for salicylic acid (SA)-dependent systemic acquired resistance (SAR), was examined in the roots and shoots of tomato plants pre-treated with SA and subsequently infected with root-knot nematodes (RKNs) (Meloidogyne incognita). PR-1 was up-regulated in both roots and shoots of SA-treated plants, whereas the expression of PR-5 was enhanced only in roots. The over-expression of PR-1 in the whole plant occurred as soon as 1 day after SA treatment. Up-regulation of the PR-1 gene was considered to be the main marker of SAR elicitation. One day after treatment, plants were inoculated with active juveniles (J2s) of M.incognita. The number of J2s that entered the roots and started to develop was significantly lower in SA-treated than in untreated plants at 5 and 15 days after inoculation. The expression pattern of PR-1, PR-2 and PR-5 was also examined in the roots and shoots of susceptible and Mi-1-carrying resistant tomato plants infected by RKNs. Nematode infection produced a down-regulation of PR genes in both roots and shoots of SA-treated and untreated plants, and in roots of Mi-carrying resistant plants. Moreover, in resistant infected plants, PR gene expression, in particular PR-1 gene expression, was highly induced in shoots. Thus, nematode infection was demonstrated to elicit SAR in shoots of resistant plants. The data presented in this study show that the repression of host defence SA signalling is associated with the successful development of RKNs, and that SA exogenously added as a soil drench is able to trigger a SAR-like response to RKNs in tomato. © 2013 BSPP AND JOHN WILEY & SONS LTD.

Nonalcoholic steatosis is a liver pathology characterized by fat accumulation and severe metabolic alterations involving early mitochondrial impairment and late hepatocyte cell death. However, mitochondrial dysfunction mechanisms remain elusive. Using four models of nonalcoholic steatosis, i.e., livers from patients with fatty liver disease, ob/ob mice, mice fed a high-fat diet, and in vitro models of lipotoxicity, we show that outer mitochondrial membrane permeability is altered and identified a posttranslational modification of voltage-dependent anion channel (VDAC), a membrane channel and NADH oxidase, as a cause of early mitochondrial dysfunction. Thus, in nonalcoholic steatosis VDAC exhibits reduced threonine phosphorylation, which increases the influx of water and calcium into mitochondria, sensitizes the organelle to matrix swelling, depolarization, and cytochrome c release without inducing cell death. This also amplifies VDAC enzymatic and channel activities regulation by calcium and modifies its interaction with proteic partners. Moreover, lipid accumulation triggers a rapid lack of VDAC phosphorylation by glycogen synthase kinase 3 (GSK3). Pharmacological and genetic manipulations proved GSK3 to be responsible for VDAC phosphorylation in normal cells. Notably, VDAC phosphorylation level correlated with steatosis severity in patients. Conclusion: VDAC acts as an early sensor of lipid toxicity and its GSK3-mediated phosphorylation status controls outer mitochondrial membrane permeabilization in hepatosteatosis. (HEPATOLOGY 2013; 57: 93-102)

The Japanese cyst nematode Heterodera elachista was detected parasitizing corn cv Rixxer in Bosco Mesola (Ferrara Province) in Northern Italy. The only previous report of this nematode was in Asia (Japan, China and Iran) attacking upland rice; being this work the first report of this cyst nematode in Europe, and confirmed corn as a new host plant for this species. Integrative morphological and molecular data for this species were obtained using D2-D3 expansion regions of 28S rDNA, ITS1-rDNA, the partial 18S rDNA, the protein-coding mitochondrial gene, cytochrome oxidase c subunit I (COI), and the heat-shock protein 90 (hsp90). Heterodera elachista identified in Northern Italy was morphologically and molecularly clearly separated from other cyst nematodes attacking corn (viz. H. avenae, H. filipjevi, H. delvii, H. oryzae, H. sacchari, H. sorghi, H. zeae, Punctodera chalcoensis, and Vittadera zeaphila) and rice (H. oryzae, H. sacchari). The phylogenetic relationships of H. elachista from Northern Italy with other cyst-nematodes using rDNA and mtDNA showed a separation of the genus Heterodera in various morphospecies groups based on vulval cone structures. The development and parasitic habit of H. elachista on naturally infected corn cv Rixxer confirmed a typical susceptible reaction, including multinucleate syncytial cells in parenchymatic cells. Under greenhouse conditions, H. elachista successfully reproduced on two crops widely used in Northern Italy, such as corn (cv PR 33) and rice (cv Baldo). Considering the limited host-range of this nematode, that include two of the three world's most important crops, special attention is needed for avoiding the dispersal of this nematode into new areas, by movement of soil on equipment, water, and contaminated containers infested soil, or agricultural practices.

One form of liver steatosis, namely Non-Alcoholic Fatty Liver Disease (NAFLD), is a worrisome health problem worldwide characterized by intrahepatic triacylglycerol (TG) overaccumulation. NAFLD is a common feature of metabolic syndrome being often associated with obesity, dyslipidemia and diabetes and mostly closely linked to insulin resistance. The mechanism of NAFLD pathogenesis is object of intense investigation especially regarding complex systems ultimately resulting in excessive TG deposition in hepatocytes. However, scarce is the attention about the relevance of hepatic import of glycerol, the other primary source (as glycerol-3-phosphate) of increased TG in hepatocytes. Obese leptin-deficient (ob/ob) mice, an animal model of NAFLD, were used to evaluate the functional involvement of Aquaporin-9 (AQP9), the major pathway of liver glycerol entry, in hepatosteatosis. By RT-PCR and qPCR, the level of Aqp9 mRNA in the liver of starved obese mice was comparable with the corresponding control lean littermates. By immunoblotting, the AQP9 protein at the hepatocyte sinusoidal plasma membrane of obese mice was markedly lower (33%) than lean mice, a finding fully confirmed by immunohistochemistry. By stopped-flow light scattering, the liver glycerol permeability of ob/ob mice was significantly lower (53%) than lean mice, a finding consistent with both the observed down-regulation of AQP9 protein and increased level of plasma glycerol characterizing obese mice. In summary, our results suggest implication of AQP9 in liver steatosis. The reduction of hepatocyte AQP9 and, consequently, glycerol permeability might be a defensive mechanism to counteract further fat infiltration in liver parenchyma.

Nematodes are widely recognised as bioindicators of the soil environment health. Analysis of soil nematode community is increasingly used to calculate various ecological indices related to enrichment and trophic status of nematofauna. The soil nematode community from three selected relatively undisturbed and disturbed sites in the Apulia region (Italy) was comparatively studied through both morpho-taxonomic and molecular analysis. Nematodes for both analyses were extracted from 100 g sub-samples from composite soil samples collected at each site. Nematodes were fixed in a 2.5% formaldehyde solution and thenidentified at family and genus level under an optical microscope. The maturity and trophic diversity indices were determined. For the molecular study, total DNA was extracted from the nematode community of each soil subsample and PCR amplification was performed by using the small subunit (18S) rDNA, as diagnostic marker, for nematode species discrimination. The 300 sequences available at this moment are still under characterisation. Sequencing of further 18S amplicons is also in progress.

Soil nematodes are organisms that quickly respond to changes (stress and pollutants) in the environment and can be useful ecological indicators of environmental disruption. Since they occur in any environment containing organic carbon, they do not quickly escape from stressful conditions, occupy key positions in soil food webs and can be classified in easily identifiable trophic groups (Bongers and Ferris, 1999). While there are many indices of biological diversity, specific tools have been developed for nematodes, such as the Maturity Index (MI) and the 3 Enrichment lndex (El). These indices are based on an ecological classification where to each taxonomic family is assigned an ecological value that ranges from l (typical families of polluted soils or sediments) to 5 (typical families of soils or sediments). The lower values (1 and 2) belong to colonizer nematodes (c), i.e. opportunists, characterized by a rapid biological cycle and able to quickly invade unstable or polluted habitats. High values (3 to 5) belong to persister nematodes (p), characterized by a slow reproduction rate. Persisters are more sensitive to pollutants and 1 other disturbances than colonizers, therefore MI also serve to measure the impact of mixtures of pollutants and the effect of their complex interactions with biotic and abiotic environment. The aim of this study was to identify the nematofauna, recovered from three different habitats, at morphological and molecular level to provide useful information on the soil features and any possible disturbances by calculating ecological indexes of soil biodiversity

This study reports on the isolation and characterization of four different endoglucanases in the root-lesion nematode Pratylenchus vulnus. The gene structures of two of these, Pv-eng-1 and Pv-eng-2, were fully determined, and Pv-eng-3 and Pv-eng-5 were partially sequenced. Spatial expression of Pv-eng-1, Pv-eng-2 and Pv-eng-5 was examined by in situ hybridization. Pv-eng-1 and Pv-eng-2 transcripts were localized in the subventral oesophageal glands, whilst the Pv-eng-5 transcript was localized in the intestine. Real-time RT-PCR showed that three of the four endoglucanases had the highest transcriptional level in adult males and females, thus demonstrating that adults are also parasitic in P.vulnus. Mixed stages of P.vulnus were treated with double-stranded RNA (dsRNA) of Pv-eng-1 in order to study the effect of gene silencing (RNAi). Silencing Pv-eng-1 by dsRNA targeting of the carbohydrate-binding module (CBM) resulted in a significant reduction (88-98%) of the transcript level, suggesting that P.vulnus is susceptible to RNAi. Furthermore, silencing P.vulnus showed a reduction (54%) in nematode reproduction on carrot minidiscs over a 5week period. These results suggest that silencing of Pv-eng-1 may result in reduction of the ability of the nematode to locate and invade roots and, therefore, to establish and reproduce.

Root-lesion nematodes of the genus Pratylenchus are migratory endoparasites distributed worldwide and regarded as severe constraints of economic crops. Crop damage is often aggravated by their interactions with soilborne fungi and bacteria, resulting in complex diseases which are biological and physiological rather than physical in nature. Morphological identification and species delimitation of these nematodes is still problematic due to their high morphological plasticity, the small number of diagnostic features available at species level, the intraspecific variability of some of these characters and many incomplete descriptions published in the literature. Thus, molecular approach, by using the ribosomal DNA, plays a key role within the taxonomy of this genus. The ITS containing region of eighteen species of Pratylenchus was sequenced and the variation within and between species was assessed. Phylogenetic analyses were conducted by using different tree reconstruction approaches: Bayesian inference (BI), neighbour-joining Log-Det (NJ), maximum parsimony (MP) and maximum likelihood (ML). The alignment revealed small species-specific DNA sequences suitable for the construction of potentially useful species- specific primers or for a more promising approach for DNA barcoding of root-lesion nematodes.

Several juvenile and adult nematodes were isolated after dissection of pupae and adults of the red palm weevil, Rhynchophorus ferrugineus, recovered from an infested Phoenix canariensis Chabaud exemplar in Bari, Italy. Two species of nematodes were recovered, Teratorhabditis synpapillata and Mononchoides macrospiculum n. sp. which is described herein. The mitochondrial cytochrome oxidase I (COI), the ITS-containing region, the 18S rRNA gene (SSU) and the D2-D3 expansion domains of the 28S rRNA gene (LSU) were amplified and sequenced. The new species, M. macrospiculum n. sp., is described at morphological and molecular level. Phylogenetic analyses using SSU and LSU sequences placed M. macrospiculum n. sp. together with M. composticola and M. striatus. The sequences of the Italian population of T. synpapillata are identical to those of T. synpapillata from Japan. This is the first report on the association of M. macrospiculum n. sp. and T. synpapillata with the red palm weevil in Europe.

A new strain of Steinernema carpocapsae (Weiser, 1955) was isolated from soil collected in a lagoon plain in Veneto region (North-East Italy). This new strain was named ItS-CAO1. Molecular and morphological analyses were performed. The ITS region and the 18S rRNA gene were amplified and sequenced. The ITS products were then digested with six restriction enzymes in order to unequivocally identify this species. Nematode virulence was tested against last instar of Galleria mellonella (L.) using different laboratory assays. Insect mortality of this new strain is very high in penetration (100%) and sand column assay (93.3%) and the percentage of penetrating infective juveniles was 57.6 and 42.9, respectively. Larval mortality in one-on-one quality assay was 50% and in exposure time assay it was 50% at 19 minutes. With the results of infectivity assays we can evaluate the possibility to use this new strain in biological control programs.

A nematode survey conducted in 2013 in Algeria, revealed that potato cyst nematodes (PCN) and cereal cyst nematodes (CCN) are widely distributed in several potato and cereal growing regions of the country. Sixteen PCN populations from five localities and five CCN populations from four of these localities were collected and characterized at the morphological and molecular levels. The PCN populations were identified as Globodera rostochiensis and G. pallida occurring separately or in mixed populations. Two species of CCN were detected. Heterodera avenae was found in four localities, whereas H. hordecalis only in one locality in association with H. avenae. The morphological and morphometric identification of PCN and CCN was confirmed by diagnostic ITS-RFLP profiles and sequencing. Phylogenetic analysis of the ITS, D2-D3 expansion domains of the 28S rRNA gene and 18S rRNA gene was made for PCN and CCN populations. Globodera pallida and G. rostochiensis from Algeria show great similarity with European and South American populations. Because of the high divergence among Algerian populations of G. pallida and G. rostochiensis it can be assumed that they were multi-introduced in Algeria. The most divergent population of G. pallida, that formed a well-separated group with some populations from Chile and Peru, suggests a later or independent introduction of this population into Algeria. Heterodera avenae and H. hordecalis formed a well-supported cluster with the corresponding populations.

Additional data on the occurrence and distribution of Xiphinema non-americanum group species in Romania are provided. Xiphinema diversicaudatum, X. index, X. vuittenezi and X. italiae were recovered from vineyards and cherry fruit trees; adults and juvenile stages were described and analysed and the morphology/variability discussed. Multiplex PCR diagnostic test using species-specific primers designed by Wang et al. (2003) yielded amplification products with expected lengths for all screened populations of these four species. Two ribosomal markers (D2-D3 28 LSU rDNA and ITS) were sequenced and ITS RFLP patterns were obtained from two X. vuittenezi populations, which have shown some morphological differences. Comparatively low level of interpopulation genetic dissimilarity (<1 %) was revealed for both markers (for D2D3 - 0.5 %; for ITS - 0.7 %). Both populations of X. vuittenezi studied produced identical ITS-RFLP specific pattern that clearly identify this species.

Several nemamdes, juveniles and adults, were found dissecting some Rhynchophorus ferrugineus pupae and adults from infested Phoenix canariensis exemplar in Bari (Italy). Insect was intact externally but inner tissues were completely liquefied. Nematodes were collected using the water trap method and total DNA was extracted from each individual. The 18s rDNA, the ITS containing region and the mitochondrial cytochrome oxidase I (COI) were amplified md sequenced. ITS-RFLP analysis were also obtained. BLAST search revealed that nucleotides sequences are similar (93%) to Koerneria sp. R81982 (Nematoda: Diplogastidae). Nematodes belonging to Diplogastridae are commonly associated with kts, with different types of association depending on diplogastrid genera. Koerneria spp. are frequently associated with stag and dung hties. Characterization studies are now still in progress for the species identification. Our future purpose is to clarify the kind of association between this specie and the Red Weevil and the eventual role as natural control agent.

Oscheius onirici sp. N. (Nematoda: Rhabditidae) was isolated from a karst cave soil of Central Italy. Molecular and morphological analyses were performed. Total DNA was extracted from individual nematodes and the mitochondrial COI, the ITS containing region, the D2-D3 expansion domains of the 28S rRNA gene and the 18S rRNA gene were amplified and sequenced. BLAST search at NCBI by using all molecular markers revealed that this taxon is similar to Oscheius species. Phylogenetic trees of ITS, 28S and 18S rDNA revealed that O. onirici sp. N. belongs to Dolichura-group. Oscheius onirici sp. N. is characterized by small body size and stoma rhabditoid type. Female reproductive system is amphidelphic. Males are rare with peloderan bursa, spicules slender and small, nine pairs of papillae of different lengths, arranged in a 1+1+1/3+3 pattern. Entomopathogenicity bioassay revealed that this nematode is capable of infecting larvae of Galleria mellonella and Tenebrio molitor.

Endo-1,4-?-glucanases have been found in numerous plant-parasitic nematodes (PPN) and play key roles in nematode--plant interactions. Four ?-1,4-endoglucanase encoding transcripts were cloned and characterised in the root-lesion nematode Pratylenchus vulnus. The P. vulnus endoglucanases show high similarity to other endoglucanases found in other nematodes belonging to glycosyl hydrolase family 5 (GHF5). All deduced proteins from the cloned sequences contained the predicted signal peptide for secretion and three of the four endoglucanases did not contain a carbohydrate-binding module (CBM). Real-time PCR experiments suggested that two of endoglucanase transcripts are expressed through the second-stage juveniles (J2), J3--J4 juveniles, males, and the adult females at different amounts confirming that all life stages are able to penetrate the host plant. In-situ hybridisation showed that both transcripts of endo-1,4-?-glucanases accumulated specifically in the pharyngeal subventral gland cells of all P. vulnus stages, thus suggesting the parasitic behaviour of each life stage. Recent data on these characterised genes will be presented and discussed.

Entomopathogenic nematodes (EPNs) are parasites of soil-dwelling insects that occur in natural and agricultural soils around the world. The current study focuses on the unexplored coastal zone of Lebanon where soil samples were taken in different sites chosen randomly along the coast like beaches, agricultural and herbaceous fields. In total, 350 soil samples were collected, mainly from the southern part of the country. An integrated approach, combining both traditional (morphological) and molecular methods, was used to characterize entomopathogenic nematode species encountered. Two named-species are added to the EPNs catalog in Lebanon from 4 samples out of the total 350 samples isolated: Heterorhabditis indica, reported for the first time in the country (samples AYAB6 and BRA20) and Steinernema feltiae (samples ANFA5 and EDA1). Furthermore, one undescribed potential entomopathogenic nematode belonging to Oscheius genus was recovered. The symbiotic bacteria from S. feltiae and H. indica were also molecularly identified through the use of five gene fragments recA, gyrB, dnaN, gltX and infB. Phylogenetic relationships of entomopathogenic nematodes and their symbiotic bacteria were inferred by using maximum-likelihood analysis. Soil studies were subsequently carried out in order to assess a possible relationship between soil parameters and their effects on EPNs. Results indicate that sandy texture and moisture are key factors for the presence and survival of EPNs in the soil in Lebanon.

A population of Xiphinema barense from wild olive trees in Torre Pozzella, Brindisi province, southern Italy, is described using both morphological and molecular studies and compared with the description of the type specimens. The wild olive nematode population agrees very well with all morphometrics provided in the original description. However, detailed observations of the lumen of the tubular portion of the uterus in paratypes and specimens of the new population revealed a clear pseudo-Z-organ with small granules mixed with crystalloid bodies which were previously undetected. Photomicrographs of adult paratypes, which were lacking in the original description, and of specimens of the new population from wild olive trees are provided. The results of the phylogenetic analyses based on the sequences of the D2-D3 expansion regions of the 28S rRNA gene and ITS rRNA genes confirm the species differentiation and indicate the phylogenetic position of X. barense and its relationship with closely related species.