Exploring the use of metabarcoding to reveal eukaryotic associations with mononchids nematodes.

Nematodes play a vital ecological role in soil and marine ecosystems, but there is limited information about their dietary diversity and feeding habits. Due to methodological challenges, the available information is based on inference rather than confirmed observations. The lack of correct dietary requirements also hampers rearing experiments. To achieve insight into the prey of mononchid nematodes, this study employed high-throughput Illumina paired-end sequencing using universal eukaryotic species 18S primers on 10 pooled mononchid nematode species, namely Mylonchulus brachyuris, M. brevicaudatus, Mylonchulus sp., Clarkus parvus, Prionchulus sp. M. hawaiiensis, M. sigmaturellus, M. vulvapapillatus, Anatonchus sp. and Miconchus sp. The results indicate that mononchids are associated with a remarkable diversity of eukaryotes, including fungi, algae, and protists. While the metabarcoding approach, first introduced here for mononchids, proved to be a simple and rapid method, it has several limitations and crucial methodological challenges that should be addressed in future studies. Ultimately, such methods should be able to evaluate the dietary complexity of nematodes and provide a valuable avenue for unraveling the dietary requirements of previously unculturable nematodes. This can contribute to the methodology of understanding their feeding habits and contributions to ecosystem dynamics.

Drivers of salamander extirpation mediated by Batrachochytrium salamandrivorans.

The recent arrival of Batrachochytrium salamandrivorans in Europe was followed by rapid expansion of its geographical distribution and host range, confirming the unprecedented threat that this chytrid fungus poses to western Palaearctic amphibians. Mitigating this hazard requires a thorough understanding of the pathogen's disease ecology that is driving the extinction process. Here, we monitored infection, disease and host population dynamics in a Belgian fire salamander (Salamandra salamandra) population for two years immediately after the first signs of infection. We show that arrival of this chytrid is associated with rapid population collapse without any sign of recovery, largely due to lack of increased resistance in the surviving salamanders and a demographic shift that prevents compensation for mortality. The pathogen adopts a dual transmission strategy, with environmentally resistant non-motile spores in addition to the motile spores identified in its sister species B. dendrobatidis. The fungus retains its virulence not only in water and soil, but also in anurans and less susceptible urodelan species that function as infection reservoirs. The combined characteristics of the disease ecology suggest that further expansion of this fungus will behave as a 'perfect storm' that is able to rapidly extirpate highly susceptible salamander populations across Europe.

Characterization of Hoplolaimus seinhorsti and Hoplolaimus pararobustus (Tylenchina: Hoplolaimidae) from banana, with phylogeny and species delineation in the genus Hoplolaimus.

The morphological and molecular characterisations of two lance nematode species isolated from the rhizosphere of banana, Hoplolaimus seinhorsti and H. pararobustus, are provided based on an integrative study that includes light and scanning electron microscopy, phylogenetic analysis and two tree-based molecular species delimitation methods (GMYC and bPTP). Nineteen new sequences were obtained, including 5 partial 18S rRNA, 6 D2-D3 of 28S rRNA, 1 ITS rRNA and 7 COI mtDNA (the first COI sequences of H. seinhorsti and H. pararobustus), and an updated morphological character comparison of 37 Hoplolaimus species is presented. The tree-based molecular species-delimitation approaches employed gave markedly differing results, and also showed remarkable discrepancies among the investigated genes, although the bPTP output was found to agree well with established morphological species delimitations. Both species-delimitation approaches did, however, provide the same output for the COI mtDNA sequences, and the COI mtDNA gene sequence was also found to correspond better to established morphological species. It is therefore recommended by this paper as representing the most suitable barcode marker for Hoplolaimus species identification. This integrative study also resulted in the corrective reassignment of 17 gene sequences that were previously unidentified or incorrectly classified, as well as concluding that H. pararobustus consists of two cryptic species.

A New Root-Knot Nematode species, Meloidogyne karsseni n. sp. (Nematoda: Meloidogynidae), From Mexico and a Taxonomic Update on M. paranaensis From Guatemala.

A new root-knot nematode (RKN) species, Meloidogyne karsseni n. sp., associated with sweet pepper from Mexico, and a population of M. paranaensis from Guatemala, are described using data from morphological, biochemical (isozyme enzymes), molecular, and phylogenetic analyses. Meloidogyne karsseni n. sp. can be morphologically diagnosed using the combined features of the second-stage juveniles, viz. body length (345 to 422 µm), a conical rounded head region, a post-labial annule lacking transverse striation, a thin stylet 11 to 12 µm long, rounded to oval and backwardly sloping knobs, dorsal gland orifice (DGO) at 5.2 to 6.0 µm from the knobs, a hemizonid just above the secretory-excretory (SE) pore, a tapering tail with finely rounded terminus and one or two very weak constrictions at hyaline tail tip; the female characters viz. oval-to-rounded perineal pattern with coarse striation on lateral sides around the anus, low dorsal arch with finer striations, and distinctly visible lateral lines; and the male characteristics viz. a rounded and continuous head, a post-labial annule without transverse striations, a robust stylet 20 to 24 µm long, rounded-to-oval and slightly backwardly sloping knobs, and a DGO at 2.4 to 2.9 µm from the knobs. In all the studied males of M. paranaensis, a characteristic sclerotization around the duct of SE-pore was also observed for the first time. Sequences of 18S, D2-D3 of 28S, and ITS of rDNA, and cox1 of mtDNA were generated for the two species, and in the phylogenetic trees based on these genes, both species appeared in the tropical RKN species complex clade.

First Reports and Morphological and Molecular Characterization of Pratylenchus delattrei and Quinisulcius capitatus Associated with Chickpea in Ethiopia.

Chickpea (Cicer arietinum L.) is classed among the most important leguminous crops of high economic value in Ethiopia. Two plant-parasitic nematode species, Pratylenchus delattrei and Quinisulcius capitatus, were recovered from chickpea-growing areas in Ethiopia and characterized using molecular and morphological data, including the first scanning electron microscopy data for P. delattrei. New sequences of D2-D3 of 28S, ITS rDNA and mtDNA COI genes have been obtained from these species, providing the first COI sequences for P. delattrei and Q. capitatus, with both species being found for the first time on chickpea in Ethiopia. Furthermore, Pratylenchus delattrei was recovered in Ethiopia for the first time. The information obtained about these nematodes will be crucial to developing effective nematode management plans for future chickpea production.

An Integrated Approach for Synonymization of Rotylenchus rhomboides with R. goodeyi (Nematoda: Hoplolaimidae) Reveals High Intraspecific Mitogenomic Variation.

Rotylenchus is a widely distributed, economically important plant-parasitic nematode group whose species-level identification relies largely on limited morphological characters, including character-based tabular keys and molecular data of ribosomal and mitochondrial genes. In this study, a combined morphological and molecular analysis of three populations of Rotylenchus goodeyi from Belgium, Poland, and the Netherlands revealed important character variations of this species, leading to synonymization of R. rhomboides with R. goodeyi and a high nucleotide variation within cox1 gene sequences in these populations. Additional Illumina sequencing of DNA from individuals of the Dutch population revealed two variants of mitogenomes, each approximately 23 Kb in size, differing by approximately 9% and containing 11 protein-coding genes, 2 ribosomal RNA genes, and as many as 29 transfer RNA genes. In addition to the first representative whole-genome shotgun sequence datasets of the genus Rotylenchus, this study also provides the full-length mitogenome and the ribosomal DNA sequences of R. goodeyi.

Genomic Reconstruction of the Introduction and Diversification of Golden Potato Cyst Nematode Populations in Indonesia.

Potato cyst nematodes (PCNs), the umbrella term for Globodera rostochiensis and G. pallida, coevolved with their Solanaceous hosts in the Andean Mountain region. From there, PCN proliferated worldwide to virtually all potato production areas. PCN is a major factor limiting the potato production in Indonesia. In our survey, only G. rostochiensis was found. Fourteen field populations were collected on Java and Sumatra, and unique variants were called by mapping resequencing data on a G. rostochiensis reference genome. A phylogenetic tree based on 1.4 million unique variants showed a genotypic separation between the outgroup, a Scottish Ro1 population, and all Indonesian populations. This separation was comparable in size with the genotypic distinction between the Javanese and the Sumatran PCN populations. Next, variants within PCN effector gene families SPRYSEC, 1106, 4D06, and venom allergen-like protein (VAL) that all interfere with the host innate immune system were compared. Distinct selective pressures acted on these effector families; while SPRYSECs (4,341 single-nucleotide polymorphisms [SNPs]/insertions or deletions of bases [indels]) behaved like neutral genes, the phylogenetic trees of 1106, 4D06, and VAL proteins (235, 790, and 150 SNPs/indels, respectively) showed deviating topologies. Our data suggest that PCN was introduced on Java not too long after the introduction of potato in the middle of the eighteenth century. Soon thereafter, the pathogen established on Sumatra and started to diversify independently. This scenario was corroborated by diversification patterns of the effector families 1106, 4D06, and VAL. Our data demonstrate how genome resequencing data from a nonindigenous pathogen can be used to reconstruct the introduction and diversification process.

First Report of the Root-Knot Nematode Meloidogyne enterolobii Parasitizing Plantain (Musa spp., AAB) in Nigeria.

Plantain (Musa spp., AAB), an important staple food in Africa with West Africa accounting for 32% of global production, is prone to numerous pests and diseases of which plant-parasitic nematodes are a key concern. This includes root-knot nematodes (RKN; Meloidogyne spp.), which infect the roots, causing them to become galled, deformed and swollen. The nematode Meloidogyne enterolobii is considered a global threat to production of many important agricultural crops due to its extremely virulent and aggressive nature (Philbrick et al. 2020). In 2019, during a survey to identify the diversity of nematodes associated with Musa spp. in Nigeria, RKN females (n = 13) were isolated from a heavily galled root (50-75% galling) from a single plantain cv. Agbagba (Musa spp., AAB) plant in Onne, Rivers State, Nigeria (4°43'08.8"N 7°10'37.5"E). Genomic DNA was extracted from three females and processed individually using worm lysis buffer and proteinase K (Bert et al. 2008). The females were identified as M. enterolobii based on Nad5 mtDNA (Janssen et al. 2016), with GenBank accession no. ON010028, ON010027, ON010026, and were 100% homologous to the M. enterolobii sequences MW965454, KU372358 and KU372359 (Supplementary Figure S1). The sampled plant did not show any specific above-ground symptoms but swellings were apparent on the secondary and tertiary roots, which were associated with RKN females that were embedded in the root tissue. All the life stages were found clustered together in the root cortex, where they created necrotic brown-black lesions. A mean value of 2,604 ± 820 (mean ± standard deviation) males, eggs and second-stage juveniles (J2) were extracted from 5 g of root sub-samples (n = 6) using the Hussey and Barker (1973) NaOCl method. On average 39 females were hand-picked (n = 6) from 5 g fresh root. Pure cultures were established from single egg masses and maintained on RKN-susceptible tomato plants (Solanum lycopersicum cv. Marmande). To conduct Koch's postulates, two-month old plantlets of plantain cv. Agbagba (n = 5) were inoculated with 8000 J2s and eggs (initial population) of M. enterolobii pure cultures in 8 L pots in a screenhouse in Nigeria. Non-inoculated plantlets were included as negative controls. The nematode reproduction factor (RF = final density / initial population) and root damage symptoms were assessed 90 days post-inoculation. All the inoculated plantlets had similar galling symptoms and extensive necrosis as was observed in the field (Supplementary Figure S2), with an average RF = 25.9. No symptoms were observed on control plants. Adult females (n = 2) removed from the roots were identified as M. enterolobii based on Nad5 mtDNA (ON532789, ON532790) confirming that plantain cv. Agbagba is a host of M. enterolobii. In Nigeria, M. enterolobii has been reported to be associated with four plant species belonging to four plant families: Euphorbiaceae (Oyetunde et al. 2022), Cucurbitaceae (Bello et al. 2020), Dioscoreaceae (Kolombia et al. 2016), and Solanaceae (dos Santos et al. 2019). To our knowledge, this is the first report of M. enterolobii on a member of the Musaceae in Nigeria and globally the first report on plantain (Musa spp., AAB). The impact of M. enterolobii on plantain productivity has yet to be determined but given the RF value obtained in the pathogenicity test, plantain is a suitable host. This calls for a comprehensive RKN diversity study to evaluate the geographic spread of M. enterolobii on this important staple food crop in West Africa.

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

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

PPNID: a reference database and molecular identification pipeline for plant-parasitic nematodes.

MOTIVATION: The phylum Nematoda comprises the most cosmopolitan and abundant metazoans on Earth and plant-parasitic nematodes represent one of the most significant nematode groups, causing severe losses in agriculture. Practically, the demands for accurate nematode identification are high for ecological, agricultural, taxonomic and phylogenetic researches. Despite their importance, the morphological diagnosis is often a difficult task due to phenotypic plasticity and the absence of clear diagnostic characters while molecular identification is very difficult due to the problematic database and complex genetic background. RESULTS: The present study attempts to make up for currently available databases by creating a manually-curated database including all up-to-date authentic barcoding sequences. To facilitate the laborious process associated with the interpretation and identification of a given query sequence, we developed an automatic software pipeline for rapid species identification. The incorporated alignment function facilitates the examination of mutation distribution and therefore also reveals nucleotide autapomorphies, which are important in species delimitation. The implementation of genetic distance, plot and maximum likelihood phylogeny analysis provides more powerful optimality criteria than similarity searching and facilitates species delimitation using evolutionary or phylogeny species concepts. The pipeline streamlines several functions to facilitate more precise data analyses, and the subsequent interpretation is easy and straightforward. AVAILABILITY AND IMPLEMENTATION: The pipeline was written in vb.net, developed on Microsoft Visual Studio 2017 and designed to work in any Windows environment. The PPNID is distributed under the GNU General Public License (GPL). The executable file along with tutorials is available at https://github.com/xueqing4083/PPNID. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

'Candidatus Xiphinematincola pachtaicus' gen. nov., sp. nov., an endosymbiotic bacterium associated with nematode species of the genus Xiphinema (Nematoda, Longidoridae).

An intracellular bacterium, strain IAST, 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 IAST was found to be new, being related to the family Burkholderiaceae, class Betaproteobacteria. Fungal endosymbionts Mycoavidus cysteinexigens B1-EBT (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 IAST as a new species, although its obligate intracellular and obligate endosymbiont nature prevented isolation of a definitive type strain. Strain IAST is therefore proposed as representing 'Candidatus Xiphinematincola pachtaicus' gen. nov., sp. nov.

Rotylenchus wimbii n. sp. (Nematoda: Hoplolaimidae) associated with finger millet in Kenya.

Rotylenchus wimbii n. sp. was found associated with finger millet in Kenya and is described based on light microscopy, scanning electron microscopy, and molecular information. Sequence analysis was performed on ITS, 18S, and D2-D3 of 28S of ribosomal DNA and COI of mitochondrial DNA. This new species is characterized by a moderate female body size of 0.6 to 0.8 mm, a continuous hemispherical lip region with four annuli, 3 to 4 irregular blocks on the basal lip annule, absence of longitudinal cuticular striations in anterior region, four lateral lines forming three equal bands which are areolated mainly at pharynx level, a robust stylet of 23 to 27 µm of which 45 to 53% is cone part, and with rounded to sometimes indented knobs, a secretory-excretory pore around level of pharyngo-intestinal junction, didelphic-amphidelphic reproductive system, vulva without distinct epiptygma, indistinct to empty spermatheca, tail usually truncated with 5 to 9 annuli, phasmids located at 7 to 17 annuli anterior to anus, and absence of males. Molecular phylogenies, in combination with species delimitation, supported the distinctiveness of Rotylenchus wimbii n. sp. and revealed some mislabeled Rotylenchus brevicaudatus sequences in GenBank.

Morphological and molecular characterization of Heterodera dunensis n. sp. (Nematoda: Heteroderidae) from Gran Canaria, Canary Islands.

Heterodera dunensis n. sp. from the coastal dunes of Gran Canaria, Canary Islands, is described. This new species belongs to the Schachtii group of Heterodera with ambifenestrate fenestration, presence of prominent bullae, and a strong underbridge of cysts. It is characterized by vermiform second-stage juveniles having a slightly offset, dome-shaped labial region with three annuli, four lateral lines, a relatively long stylet (27-31 µm), short tail (35-45 µm), and 46 to 51% of tail as hyaline portion. Males were not found in the type population. Phylogenetic trees inferred from D2-D3 of 28S, partial ITS, and 18S of ribosomal DNA and COI of mitochondrial DNA sequences indicate a position in the 'Schachtii clade'.

Plant-parasitic nematodes associated with sugarcane in Kilimanjaro, Tanzania.

Morphological and molecular analyses of plant-parasitic nematodes (PPN) from 12 sugarcane plantation sites of Tanganyika Planting Company (TPC) Limited in Kilimanjaro region of Tanzania revealed the presence of six PPN genera, i.e. Helicotylenchus, Hemicycliophora, Pratylenchus, Rotylenchulus, Scutellonema, and Tylenchorhynchus. The genera with the highest densities and present in virtually all samples were Pratylenchus and Rotylenchulus, and the most important species appeared to be R. parvus, P. zeae, T. crassicaudatus, and T. ventrosignatus. A total sequences of 11 partial ITS, 15 D2-D3 of 28S, and 6 partial 18S of rRNA gene, and 7 partial COI gene of mtDNA of these species were obtained in this study. Morphology and molecular data comparisons between the Tanzanian R. parvus and the South African R. parvus indicated that R. parvus is a cryptic species complex. Based on the results of morphological and molecular analyses of T. crassicaudatus and T. agri from China, Haiti, Indonesia, Iran, Niger and the USA, T. agri syn. n. is proposed as a junior synonym of T. crassicaudatus.Morphological and molecular analyses of plant-parasitic nematodes (PPN) from 12 sugarcane plantation sites of Tanganyika Planting Company (TPC) Limited in Kilimanjaro region of Tanzania revealed the presence of six PPN genera, i.e. Helicotylenchus, Hemicycliophora, Pratylenchus, Rotylenchulus, Scutellonema, and Tylenchorhynchus. The genera with the highest densities and present in virtually all samples were Pratylenchus and Rotylenchulus, and the most important species appeared to be R. parvus, P. zeae, T. crassicaudatus, and T. ventrosignatus. A total sequences of 11 partial ITS, 15 D2-D3 of 28S, and 6 partial 18S of rRNA gene, and 7 partial COI gene of mtDNA of these species were obtained in this study. Morphology and molecular data comparisons between the Tanzanian R. parvus and the South African R. parvus indicated that R. parvus is a cryptic species complex. Based on the results of morphological and molecular analyses of T. crassicaudatus and T. agri from China, Haiti, Indonesia, Iran, Niger and the USA, T. agri syn. n. is proposed as a junior synonym of T. crassicaudatus.

First report of Rotylenchulus reniformis infecting turmeric in Vietnam and consequent damage.

Turmeric (Curcuma longa L.) is one of the common medicinal crops of high economical value in Vietnam. A survey in the Central Highlands of Vietnam revealed a turmeric growing area showing serious disease symptoms, including stunting of the plant, yellowing or darkening of the leaf margins and tips, and underdeveloped dry and rotten rhizomes. An inspection for plant pathogens in soil samples from this area revealed a high density and frequency of Rotylenchus reniformis, with a significant relationship between the density of R. reniformis, rhizome weight, and level of plant damage. This study provides the first report of R. reniformis found in parasitic association with turmeric in Vietnam with the support of molecular data and examines its resulting damage.Turmeric (Curcuma longa L.) is one of the common medicinal crops of high economical value in Vietnam. A survey in the Central Highlands of Vietnam revealed a turmeric growing area showing serious disease symptoms, including stunting of the plant, yellowing or darkening of the leaf margins and tips, and underdeveloped dry and rotten rhizomes. An inspection for plant pathogens in soil samples from this area revealed a high density and frequency of Rotylenchus reniformis, with a significant relationship between the density of R. reniformis, rhizome weight, and level of plant damage. This study provides the first report of R. reniformis found in parasitic association with turmeric in Vietnam with the support of molecular data and examines its resulting damage.

Morphological and molecular characterization of Pratylenchus species from Yam (Dioscorea spp.) in West Africa.

The root-lesion nematodes (RLN), Pratylenchus spp., are among the major plant-parasitic nematodes affecting yam (Dioscorea spp.) production in West Africa. The distribution and diversity of RLN species associated with yam was investigated through a soil and tuber survey of the main producing areas in Nigeria and Ghana. Pratylenchus spp. were detected in the yam rhizosphere in 59% of 81 soil samples from Ghana and 39% of 114 soil samples from Nigeria. Pratylenchus spp. were detected in 24 of 400 tubers examined, in combination with root-knot nematodes (Meloidogyne spp.) and their associated damage of galls and crazy roots (79%), and with yam nematode (Scutellonema bradys) and their associated damage of dry rot (17%), although no specific additional symptoms were observed for Pratylenchus spp. Species of Pratylenchus were identified by their morphological features and by sequences of the D2-D3 region of the 28 S rDNA gene and the mitochondrial cytochrome oxidase I gene (COI). Pratylenchus brachyurus was the most frequent RLN species in both the rhizosphere and tubers of yam. Pratylenchus hexincisus was recovered from one tuber collected in Nigeria. While further investigations are required to establish the host status of yam for this nematode, this appears to be the first record of P. hexincisus on yam. The present taxonomical status of P. scribneri and P. hexincisus is discussed.

Phylogeography and Molecular Species Delimitation of Pratylenchus capsici n. sp., a New Root-Lesion Nematode in Israel on Pepper (Capsicum annuum).

Root-lesion nematodes of the genus Pratylenchus parasitize the roots of numerous plants and can cause severe damage and yield loss. Here, we report on a new species, Pratylenchus capsici n. sp., from the Arava rift, Israel, which was characterized by integrative methods, including detailed morphology, molecular phylogeny, population genetics, and phylogeography. This species is widely spread across the Arava rift, causing significant infection in pepper (Capsicum annuum) roots and inhibiting plant growth. Both morphological and molecular species delimitation support the recovered species as a new species. We found high cytochrome oxidase subunit I haplotype diversity, and phylogeography analysis suggests that contemporary gene flow is prevented among different agricultural farms, while population dispersal from weeds (Chenopodium album and Sonchus oleraceus) to pepper occurs on a relatively small scale. Our results suggest that weeds are an important reservoir for the dispersal of P. capsici n. sp., either as the original nematode source or at least in maintaining the population between growing seasons.

First Report of Scutellonema brachyurus (Steiner, 1938) Andrassy, 1958 and Occurrence of Meloidogyne incognita (Kofoid & White, 1919) Chitwood, 1949 in Belgium.

A study of plant-parasitic nematodes in the Botanical garden at Ghent University in Belgium revealed the presence of two tropical nematode species, i.e. Scutellonema brachyurus and Meloidogyne incognita. Scutellonema brachyurus was recovered, only once, for the first time in Belgium from Musa basjoo and is morphologically characterized. M. incognita, forming galls on Hedychium greenii, was recovered in all seasons over three consecutive years and is morphologically and molecularly characterized. Although no unequivocal evidence was found to indicate that these nematodes pose a current threat in Belgium, in the light of climate change, it is crucial to improve our knowledge of potential tropical nematode activity in more Northern countries.

Description of Rotylenchus rhomboides n. sp. and a Belgian population of Rotylenchus buxophilus (Tylenchomorpha: Hoplolaimidae).

During a survey in the Botanical garden of Ghent University, a new species Rotylenchus rhomboides n. sp. and a population of Rotylenchus buxophilus were found. Rotylenchus rhomboides n. sp. is characterized by the presence of a rhomboid-like widening of the mid-ridge of lateral field at the level of vulva, a feature previously unknown within the genus. The population of the new species, composed only by females, has a rounded labial region with 4 to 5 annuli, robust stylet (31-37 µm long), short dorsal esophageal gland (9-19 µm) overlap of the intestine, vulva located slightly posterior to mid-body, and hemispherical or rounded tail shape with large phasmids located 3 to 5 annuli anterior to the level of anus. The hierarchical cluster analysis based on morphological features indicated that the new species closely resembles R. corsicus, R. gracilidens, and R. rugatocuticulatus. The DNA analyses of the D2-D3 of 28S rDNA, ITS rDNA, and COI mtDNA sequences of Rotylenchus rhomboides n. sp. show a close relationship with R. buxophilus, R. goodeyi, R. laurentinus, R. pumilus, and R. incultus, all of which can also be differentiated from the new species by morphological features. The combination of morphological, morphometric, and molecular characteristics confirmed the new species and the first report of R. buxophilus on yam (Dioscorea tokoro) in Belgium.During a survey in the Botanical garden of Ghent University, a new species Rotylenchus rhomboides n. sp. and a population of Rotylenchus buxophilus were found. Rotylenchus rhomboides n. sp. is characterized by the presence of a rhomboid-like widening of the mid-ridge of lateral field at the level of vulva, a feature previously unknown within the genus. The population of the new species, composed only by females, has a rounded labial region with 4 to 5 annuli, robust stylet (31­37 µm long), short dorsal esophageal gland (9­19 µm) overlap of the intestine, vulva located slightly posterior to mid-body, and hemispherical or rounded tail shape with large phasmids located 3 to 5 annuli anterior to the level of anus. The hierarchical cluster analysis based on morphological features indicated that the new species closely resembles R. corsicus, R. gracilidens, and R. rugatocuticulatus. The DNA analyses of the D2-D3 of 28S rDNA, ITS rDNA, and COI mtDNA sequences of Rotylenchus rhomboides n. sp. show a close relationship with R. buxophilus, R. goodeyi, R. laurentinus, R. pumilus, and R. incultus, all of which can also be differentiated from the new species by morphological features. The combination of morphological, morphometric, and molecular characteristics confirmed the new species and the first report of R. buxophilus on yam (Dioscorea tokoro) in Belgium.

Disruption of skin microbiota contributes to salamander disease.

Escalating occurrences of emerging infectious diseases underscore the importance of understanding microbiome-pathogen interactions. The amphibian cutaneous microbiome is widely studied for its potential to mitigate disease-mediated amphibian declines. Other microbial interactions in this system, however, have been largely neglected in the context of disease outbreaks. European fire salamanders have suffered dramatic population crashes as a result of the newly emerged Batrachochytrium salamandrivorans (Bsal). In this paper, we investigate microbial interactions on multiple fronts within this system. We show that wild, healthy fire salamanders maintain complex skin microbiotas containing Bsal-inhibitory members, but these community are present at a remarkably low abundance. Through experimentation, we show that increasing bacterial densities of Bsal-inhibiting bacteria via daily addition slowed disease progression in fire salamanders. Additionally, we find that experimental-Bsal infection elicited subtle changes in the skin microbiome, with selected opportunistic bacteria increasing in relative abundance resulting in septicemic events that coincide with extensive destruction of the epidermis. These results suggest that fire salamander skin, in natural settings, maintains bacterial communities at numbers too low to confer sufficient protection against Bsal, and, in fact, the native skin microbiota can constitute a source of opportunistic bacterial pathogens that contribute to pathogenesis. By shedding light on the complex interaction between the microbiome and a lethal pathogen, these data put the interplay between skin microbiomes and a wildlife disease into a new perspective.