Molecular Phylogenetic Characterization of Anguina Tritici (Steinbuch, 1799) Filipjev, 1936 (Rhabditida: Anguinidae) on Barley from Iraq.

During a nematological survey in Iraq, in the Bashika area, Ninevah province, an anguinid nematode population was isolated from galls of infected barley plants. The morphological characteristics indicated that the recovered species is identical to Anguina tritici. The barley population of A. tritici was molecularly characterized by sequencing two ribosomal regions (ITS and 18S rRNA genes), and their phylogenetic analyses revealed the newly generated sequences are in sister relation to corresponding sequences of A. tritici from wheat in the Bayesian tree, providing further evidence that the host plant can contribute to the separation of new isolates of plant parasitic nematodes.

Characterisation of Aceria massalongoi and a histopathological study of the leaf galls induced on chaste trees.

The eriophyoid mite Aceria massalongoi (Canestrini) was collected from globoid leaf galls on severely injured chaste trees, Vitex agnus-castus L. (Lamiaceae), in Bari and Bernalda (southern Italy), and on the Ionian island Leukade (Greece). Female, male and nymph were described in detail, following the current morphometric descriptive scheme, supplementing older and incomplete descriptions. Molecular characterization of A. massalongoi from Italy and Greece was conducted by amplifying and sequencing the ribosomal ITS, the D2-D3 expansion domains of the 28S rRNA gene and the mitochondrial COI, for the first time. Phylogenetic trees based on the three molecular markers showed congruent results, confirming that Italian and Greek A. massalongoi populations are the same species that cluster together with some intraspecific variability. Galls, ranging from 0.5 to 2.8 mm in diameter, were randomly distributed on both leaf surfaces, and protruded ca. 1 mm from the leaf surface. Sometimes they were closely aggregated on midrib and leaves, which, consequently, appeared strongly deformed. Close-up observations revealed that gall induction causes hyperplastic proliferation of leaf tissues around the gall chamber hosting mites. The uniserial cell lining inside this chamber provides the nutritional tissue for the mites. All feeding cells contained one or more (frequently 2-3) hypertrophied nuclei and dense granular cytoplasm.

Functional Variation of Two Novel Cellulases, Pv-eng-5 and Pv-eng-8, and the Heat Shock 90 Gene, Pv-hsp-90, in Pratylenchus vulnus and Their Expression in Response to Different Temperature Stress.

Functional characterization of two novel endoglucanase genes, Pv-eng-5 and Pv-eng-8, of the root-lesion nematode Pratylenchus vulnus was carried out. In situ-hybridization experiments revealed that Pv-eng-8 transcript was localized in the pharyngeal glands. Silencing of Pv-eng-5 and Pv-eng-8 resulted in a significant reduction of expression level (52% and 67%, respectively). Furthermore, the silencing of Pv-eng-8 determined a reduction (41%) in nematode reproduction, suggesting that treated nematodes are much less able to process food. Surprisingly, no significant difference on reproduction rate was observed with Pv-eng-5 dsRNA nematodes, suggesting a neofunctionalization of Pv-eng-5 despite the high similarity with nematode endoglucanases. Pratylenchus species are poikilothermic organisms showing close relationships with the environmental temperature. The effects of different temperature ranges revealed that the reproductive potential of P. vulnus increased with increasing temperature from 23 °C to 28 °C, but no reproduction was observed at 33 °C. In real time, increasing temperature from 23 °C to 28 °C the heat shock gene Pv-hsp-90 was differentially expressed in adult stages, while the levels of the effector genes Pv-eng-1 and Pv-eng-8 in females showed no significant differences compared to those observed at 23 °C, only in males Pv-eng-8 level decreased (45%). The upregulation of Pv-hsp-90 in both adult stages suggests a protective mechanism in order to cope with unfavorable environmental conditions.

Occurrence of Sheraphelenchus sucus (Nematoda: Aphelenchoidinae) and Panagrellus sp. (Rhabditida: Panagrolaimidae) Associated with Decaying Pomegranate Fruit in Italy.

Two different nematode species were recovered from pomegranate decaying fruit in two localities in Southern Italy: the mycetophagus nematode Sheraphelenchus sucus and a bacterial feeder nematode belonging to the Panagrolaimidae (Rhabditida) family. Morphometrics of the Italian population of S. sucus closely resemble that of the type population, whereas some differences were found when compared with another population from Iran. Molecular characterization of the Italian S. sucus using the 18S rRNA gene, D2-D3 expansion domains of the 28S rDNA, the ITS region, and the partial mitochondrial COI were carried out. Sequences of the 18S rRNA gene, the D2-D3 domains, and the ITS were analyzed using several methods for inferring phylogeny to reconstruct the relationships among Sheraphelenchus and Bursaphelenchus species. The bacterial feeder Panagrellus sp. was characterized at the molecular level only. The D2-D3 expansion domains and ITS sequences of this Italian panagrolaimid were determined. The D2-D3 sequences of the Italian panagrolaimid showed 99% similarity with the corresponding sequence of Panagrellus sp. associated with Rhynchophorus ferrugineus. This is the first report on the tritrophic association of S. sucus and Rhabditida that uses both insects and pomegranate fruit as hosts.

Glycogen synthase kinase 3-mediated voltage-dependent anion channel phosphorylation controls outer mitochondrial membrane permeability during lipid accumulation.

UNLABELLED: 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.

Commodity risk assessment of Prunus avium plants from United Kingdom.

The European Commission requested the EFSA Panel on Plant Health to prepare and deliver risk assessments for commodities listed in Commission Implementing Regulation (EU) 2018/2019 as 'High risk plants, plant products and other objects'. This Scientific Opinion covers plant health risks posed by plants of Prunus avium possibly grafted on rootstocks of either P. avium, P. canescens, P. cerasus, P. pseudocerasus or their hybrids imported from the UK, taking into account the available scientific information, including the technical information provided by the UK. All pests associated with the commodities were evaluated against specific criteria for their relevance for this opinion. Three quarantine pests Scirtothrips dorsalis, tobacco ringspot virus and tomato ringspot virus), one protected zone EU quarantine pest (Bemisia tabaci (European population), and three non- regulated pests (Colletotrichum aenigma, Eulecanium excrescens and Takahashia japonica) that fulfilled all relevant criteria were selected for further evaluation. The risk mitigation measures proposed in the technical Dossier from the UK were evaluated, taking into account the possible limiting factors. For these pests, expert judgement is given on the likelihood of pest freedom, taking into consideration the risk mitigation measures acting on the pest, including uncertainties associated with the assessment. The degree of pest freedom varies among the pests evaluated, with Colletotrichum aenigma being the pest most frequently expected on the imported potted plants. The Expert Knowledge Elicitation indicated with 95% certainty that between 9971 and 10,000 plants per 10,000 would be free from the above-mentioned fungus.

Commodity risk assessment of plants of 12 selected Prunus species from Moldova.

The European Commission requested the EFSA Panel on Plant Health to prepare and deliver risk assessments for commodities listed in Commission Implementing Regulation (EU) 2018/2019 as 'High-risk plants, plant products and other objects'. This Scientific Opinion covers plant health risks posed by defoliated 1- or 2-year old bare root plants for planting (grafted or not) of 12 Prunus species (Prunus armeniaca, P. avium, P. canescens, P. cerasifera, P. cerasus, P. davidiana, P. domestica, P. dulcis, P. fontanesiana, P. persica, P. salicina, P. tomentosa) imported from Moldova, taking into account the available scientific information, including the technical information provided by the applicant country. The evaluation identified three EU-quarantine pests, Erwinia amylovora (protected zone quarantine pest), Xiphinema rivesi non-EU populations and Xanthomonas arboricola pv. pruni (protected zone quarantine pest), which were selected for further evaluation, based on defined criteria, including their presence in the applicant country. It should be noted that there is uncertainty regarding whether all relevant pests have been identified due to a limited number of scientific publications and pest surveys in Moldova. For the three selected pests, the risk mitigation measures proposed in the technical dossier from Moldova were evaluated taking into account the possible limiting factors. For these pests, an expert judgement is given on the likelihood of pest freedom taking into consideration the risk mitigation measures acting on it, including uncertainties associated with the assessment. The degree of pest freedom varies among the pests evaluated, with Erwinia amylovora being the pest most frequently expected on the imported plants. The Expert Knowledge Elicitation indicated, with 95% certainty, that between 9823 and 10,000 bundles (comprising 10-20 plants per bundle) out of 10,000 bundles would be free from E. amylovora.

Entomopathogenic nematodes and their symbiotic bacteria: from genes to field uses.

The term "microbial control" has been used to describe the use of microbial pathogens (bacteria, viruses, or fungi) or entomopathogenic nematodes (EPNs) to control various insect pest populations. EPNs are among the best biocontrol agents, and major developments in their use have occurred in recent decades, with many surveys having been conducted all over the world to identify EPNs that may have potential in the management of insect pests. For nematodes, the term "entomopathogenic" means "causing disease to insects" and is mainly used in reference to the bacterial symbionts of Steinernema and Heterorhabditis (Xenorhabdus and Photorhabdus, respectively), which cause EPN infectivity. A compendium of our multiannual experiences on EPN surveys and on their collection, identification, characterization, and use in agro-forestry ecosystems is presented here to testify and demonstrate once again that biological control with EPNs is possible and offers many advantages over chemicals, such as end-user safety, minimal damage to natural enemies, and lack of environmental pollution, which are essential conditions for an advanced IPM strategy.

Into the large ears: otitis externa associated with nematodes, mites, and bacteria in Asian elephants (Elephas maximus).

BACKGROUND: The Asian elephant (Elephas maximus), which is an endangered species, harbors several parasites. Among the ectoparasites that it harbors, ear mites of the genus Loxanoetus have the potential to cause external otitis, an inflammation that may also be associated with the presence of other microorganisms. We assessed the relationships between ear mites, nematodes, yeast, bacterial rods, and cocci sampled from the ears of captive Asian elephants in Thailand. In addition, we discuss the possibility that dust-bathing behavior may be triggered by ear mite infestation, and that this in turn may lead to contamination of the ears with soil microorganisms. METHODS: Legally owned captive Asian elephants (n = 64) were sampled. Ear swabs were individually collected from both ears and microscopically examined for the presence of mites, nematodes, yeast, bacterial rods, cocci, and host cells. Mites and nematodes were identified to species level using morphological and molecular methods. RESULTS: Loxanoetus lenae mites were present in 43.8% (n = 28/64) of the animals (19 animals with mites in one ear and nine animals with mites in both ears). Nematodes of the genus Panagrolaimus were detected in 23.4% (n = 15/64) of the animals (10 with nematodes in one ear and five with nematodes in both ears). In adult elephants (Fisher's exact test, P = 0.0278) and female elephants (Fisher's exact test, P = 0.0107), the presence of nematodes in both ears was significantly associated with the presence of mites. In addition, higher categorical burdens of nematodes were also significantly associated with the presence of mites (Fisher's exact test, P = 0.0234) and epithelial cells (Fisher's exact test, P = 0.0108), and marginally significantly associated with bacterial cocci (Fisher's exact test, P = 0.0499). CONCLUSIONS: The presence of L. lenae mites in the ear canals of the Asian elephants was significantly associated with the occurrence of other microorganisms, such as soil nematodes, bacteria and yeasts. The presence of mites in their ears may increase the dust-bathing behavior of elephants which, if confirmed, represents a further paradigmatic example of a parasitic infestation affecting animal behavior.

Commodity risk assessment of Malus sylvestris plants from United Kingdom.

The European Commission requested the EFSA Panel on Plant Health to prepare and deliver risk assessments for commodities listed in Commission Implementing Regulation (EU) 2018/2019 as 'High risk plants, plant products and other objects'. This Scientific Opinion covers plant health risks posed by rooted plants and bundles of bare root plants or rooted cell grown young plants of Malus sylvestris imported from the UK, taking into account the available scientific information, including the technical information provided by the UK. All pests associated with the commodities were evaluated against specific criteria for their relevance for this opinion. Two quarantine pests (tobacco ringspot virus and tomato ringspot virus), one protected zone quarantine pest (Erwinia amylovora) and four non-regulated pests (Colletotrichum aenigma, Meloidogyne mali, Eulecanium excrescens and Takahashia japonica) that fulfilled all relevant criteria were selected for further evaluation. For Erwinia amylovora, special requirements are specified in Commission Implementing Regulation (EU) 2019/2072. Based on the information provided in the dossier, these specific requirements for E. amylovora are met. For the remaining six pests, the risk mitigation measures proposed in the technical Dossier from the UK were evaluated, taking into account the possible limiting factors. For these pests, expert judgement is given on the likelihood of pest freedom, taking into consideration the risk mitigation measures acting on the pest, including uncertainties associated with the assessment. The degree of pest freedom varies among the pests evaluated, with scales (Eulecanium excrescens and Takahashia japonica) being the pests most frequently expected on the imported bundles of bare root plants or rooted cell grown young plants. The expert knowledge elicitation indicated with 95% certainty that between 9,976 and 10,000 bundles (one bundle consisting of 5-15 plants for bare root plants or 25-50 plants for cell grown young plants) per 10,000 would be free from the above-mentioned scales.

Commodity risk assessment of Malus domestica plants from United Kingdom.

The European Commission requested the EFSA Panel on Plant Health to prepare and deliver risk assessments for commodities listed in Commission Implementing Regulation (EU) 2018/2019 as 'High risk plants, plant products and other objects'. This Scientific Opinion covers plant health risks posed by rooted plants in pots, bundles of bare-rooted plants or trees and bundles of budwood and graftwood of Malus domestica imported from the United Kingdom, taking into account the available scientific information, including the technical information provided by the United Kingdom. All pests associated with the commodities were evaluated against specific criteria for their relevance for this opinion. Two quarantine pests (tobacco ringspot virus and tomato ringspot virus), one protected zone quarantine pest (Erwinia amylovora) and four non-regulated pests (Colletotrichum aenigma, Meloidogyne mali, Eulecanium excrescens, Takahashia japonica) that fulfilled all relevant criteria were selected for further evaluation. For E. amylovora, special requirements are specified in Commission Implementing Regulation (EU) 2019/2072. Based on the information provided in the Dossier, the specific requirements for E. amylovora were met. For the remaining six pests, the risk mitigation measures proposed in the technical Dossier from the UK were evaluated considering the possible limiting factors. For the selected pests, expert judgement is given on the likelihood of pest freedom, taking into consideration the risk mitigation measures acting on the pest, including uncertainties associated with the assessment. The degree of pest freedom varies among the pests evaluated, with scales (E. excrescens and T. japonica) being the pests most frequently expected on the imported budwood and graftwood. The Expert Knowledge Elicitation indicated with 95% certainty that between 9,976 and 10,000 bundles (consisting of 50 up to 500 plants each) per 10,000 would be free from the above-mentioned scales.

Commodity risk assessment of Prunus persica and P. dulcis plants from Türkiye.

The European Commission requested the EFSA Panel on Plant Health to prepare and deliver risk assessments for commodities listed in Commission Implementing Regulation (EU) 2018/2019 as 'High risk plants, plant products and other objects'. This Scientific Opinion covers plant health risks posed by plants of Prunus persica and P. dulcis, as budwood/graftwood, rooted or grafted on rootstocks of either P. persica, P. dulcis, P. armeniaca, P. davidiana or their hybrids, imported from Türkiye, taking into account the available scientific information, including the technical information provided by Türkiye. All pests associated with the commodity were evaluated against specific criteria for their relevance for this opinion. Four quarantine pests (peach rosette mosaic virus, tomato ringspot virus, Anoplophora chinensis, Scirtothrips dorsalis) and 14 non-regulated pests (Hoplolaimus galeatus, Lasiodiplodia pseudotheobromae, Neoscytalidium dimidiatum, Neoscytalidium novaehollandiae, Didesmococcus unifasciatus, Euzophera semifuneralis, Lepidosaphes malicola, Lepidosaphes pistaciae, Maconellicoccus hirsutus, Malacosoma parallela, Nipaecoccus viridis, Phenacoccus solenopsis, Pochazia shantungensis, Russellaspis pustulans) that fulfilled all relevant criteria were selected for further evaluation. For these 18 pests, the risk mitigation measures proposed in the technical Dossier from Türkiye were evaluated taking into account the possible limiting factors. For the selected pests, an expert judgement is given on the likelihood of pest freedom taking into consideration the risk mitigation measures acting on the pest, including uncertainties associated with the assessment. The degree of pest freedom varies among the pests evaluated, with fungi from Botryosphaeriaceae family (L. pseudotheobromae, N. dimidiatum and N. novaehollandiae) being the pests most frequently expected on the imported plants. The Expert Knowledge Elicitation indicated with 95% certainty that between 9,813 and 10,000 bundles (consisting of 10 or 25 plants each) per 10,000 would be free from the above-mentioned fungi in the Botryosphaeriaceae family.

Commodity risk assessment of Malus domestica plants from Bosnia and Herzegovina.

The European Commission requested the EFSA Panel on Plant Health to prepare and deliver risk assessments for commodities listed in Commission Implementing Regulation (EU) 2018/2019 as 'High-risk plants, plant products and other objects'. This Scientific Opinion covers plant health risks posed by bare root plants of Malus domestica grafted on rootstocks of either Malus domestica or Malus sylvestris imported from Bosnia and Herzegovina (BIH), taking into account the available scientific information, including the technical information provided by Bosnia and Herzegovina. All pests associated with the commodities were evaluated against specific criteria for their relevance for this opinion. One protected zone quarantine pest, Erwinia amylovora, fulfilled all relevant criteria and was selected for further evaluation. For E. amylovora, special requirements are specified in Commission Implementing Regulation (EU) 2019/2072. Based on the information provided in the Dossier, these specific requirements for E. amylovora are not met.

New occurrence of Meloidogyne graminicola (Nematoda: Meloidogyninae) from rice fields in Italy: Variability and phylogenetic relationships.

Since the first detection of Meloidogyne graminicola in Piedmont, North Italy, in 2016, further inspections for the presence of the rice root-knot nematode were carried out in rice fields of neighboring regions, in accordance with the Italian NPPO (National Plant Protection Organization) to support the official phytosanitary measures, to enable the early detection of the rice pest, and to prevent its spread within the national territory. In 2018, surveys of rice fields in Lombardy region revealed a new occurrence of M. graminicola. In the present study, we confirmed the identification of the rice nematode in Lombardy using the ribosomal ITS region and the mitochondrial COI and COII genes. The sequences and phylogenetic analyses revealed that Lombardy M. graminicola population grouped in all trees in the main cluster containing Meloidogyne species belonging to graminis group, but always in a different subgroup compared to the Piedmont population of M. graminicola. These results clearly suggest that the two Italian populations have been recently and independently introduced and confirm that the geographic origin is not the main factor leading to M. graminicola population variability.

Water Stress Differentially Modulates the Expression of Tomato Cell Wall Metabolism-Related Genes in Meloidogyne incognita Feeding Sites.

Microscopic observations and transcriptomic RNA-Seq analyses were applied to investigate the effect of water stress during the formation of tomato galls formation 1 and 2 weeks after inoculation with the root-knot nematode Meloidogyne incognita. Water stress affected root growth and the nematode ability to mount an efficient parasitism. The effects of water stress on the feeding site development were already observed at 1 week after nematode inoculation, with smaller giant cells, delayed development, and thinner cell walls. These features suggested changes in the expression levels of genes involved in the feeding site formation and maintenance. Gene Ontology (GO) enrichment and expression patterns were used to characterize differentially expressed genes. Water stress modified the expression profile of genes involved in the synthesis, degradation, and remodeling of the cell wall during the development of nematode feeding site. A comparison of gene expression with unstressed galls revealed that water stress intensified the up or downregulation of most genes. However, it particularly influenced the expression pattern of expansin A11 (Solyc04g081870.4.1), expansin-like B1(Solyc08g077910.3.1), a pectin acetylesterase (Solyc08g005800.4.1), and the pectin methylesterase pmeu1 (Solyc03g123630.4.1) which were upregulated in unstressed galls and repressed by water stress, at both sampling times. The expression of most members of the genes involved in cell wall metabolism, i.e., those coding for Csl, fasciclin, and COBRA proteins, were negatively influenced. Interestingly, alteration in the expression profiles of most dirigent protein genes (DIRs) and upregulation of five gene coding for Casparian strip domain protein (CASP)-like proteins were found. Gene expression analysis of galls from water stressed plants allowed us to better understand the molecular basis of M. incognita parasitism in tomato. Specific genes, including those involved in regulation of cellulose synthesis and lignification process, require further study to develop defense strategies against root-knot nematodes.

Commodity risk assessment of Malus domestica plants from Turkey.

The European Commission requested the EFSA Panel on Plant Health to prepare and deliver risk assessments for commodities listed in Commission Implementing Regulation (EU) 2018/2019 as 'High risk plants, plant products and other objects'. This Scientific Opinion covers plant health risks posed by dormant grafted plants, rootstocks, budwood and scions of Malus domestica imported from Turkey, taking into account the available scientific information, including the technical information provided by Turkey. All pests associated with the commodities were evaluated against specific criteria for their relevance for this opinion. Three quarantine pests (Anoplophora chinensis, Lopholeucaspis japonica and tomato ringspot virus), one protected zone quarantine pest (Erwinia amylovora) and eight non-regulated pests (Calepitrimerus baileyi, Cenopalpus irani, Cicadatra persica, Diplodia bulgarica, Hoplolaimus galeatus, Malacosoma parallela, Pratylenchus loosi and Pyrolachnus pyri) that fulfilled all relevant criteria were selected for further evaluation. For E. amylovora, special requirements are specified in Commission Implementing Regulation (EU) 2019/2072. Based on the information provided in the dossier, the specific requirements for E. amylovora were not met. For Anoplophora chinensis, special measures are specified in Commission Implementing Decision (EU) 2012/138. The exporting country does meet the requirement for a certificate regarding plants for planting that originate from Turkish provinces other than Istanbul. For the 10 remaining selected pests, the risk mitigation measures proposed in the technical dossier from Turkey were evaluated taking into account the possible limiting factors. For the selected pests an expert judgement is given on the likelihood of pest freedom taking into consideration the risk mitigation measures acting on the pest, including uncertainties associated with the assessment. The degree of pest freedom varies among the pests evaluated, with D. bulgarica being the pest most frequently expected on the imported plants. The expert knowledge elicitation indicated with 95% certainty that between 9,863 and 10,000 bundles (consisting of 10 or 25 plants each) per 10,000 would be free from D. bulgarica.

Commodity risk assessment of Prunus domestica plants from Ukraine.

The European Commission requested the EFSA Panel on Plant Health to prepare and deliver risk assessments for commodities listed in Commission Implementing Regulation (EU) 2018/2019 as 'High risk plants, plant products and other objects'. This Scientific Opinion covers plant health risks posed by plants of Prunus domestica grafted on Prunus cerasifera imported from Ukraine, taking into account the available scientific information, including the technical information provided by Ukraine. All pests associated with the commodity were evaluated against specific criteria for their relevance for this opinion. One quarantine pest (Lopholeucaspis japonica), two protected zone quarantine pests (Erwinia amylovora and Xanthomonas arboricola pv. pruni) and one non-regulated pest (Eotetranychus prunicola) that fulfilled all relevant criteria were selected for further evaluation. For these four pests, the risk mitigation measures proposed in the technical dossier from Ukraine were evaluated taking into account the possible limiting factors. For the selected pests, an expert judgement is given on the likelihood of pest freedom taking into consideration the risk mitigation measures acting on the pest, including uncertainties associated with the assessment. The degree of pest freedom varies among the pests evaluated, with Xanthomonas arboricola pv. pruni being the pest most frequently expected on the imported plants. The Expert Knowledge Elicitation indicated with 95% certainty that between 9,870 and 10,000 bundles (consisting of 10 plants each) per 10,000 would be free from Xanthomonas arboricola pv. pruni.

Commodity risk assessment of grafted plants of Malus domestica from Moldova.

The European Commission requested the EFSA Panel on Plant Health to prepare and deliver risk assessments for commodities listed in Commission Implementing Regulation (EU) 2018/2019 as 'High risk plants, plant products and other objects'. This Scientific Opinion covers plant health risks posed by defoliated and in dormant phase, grafted bare rooted plants for planting of Malus domestica imported from Moldova, taking into account the available scientific information, including the technical information provided by the applicant country. A list of 1,118 pests potentially associated with the commodity species was compiled. The relevance of these pests was assessed following defined criteria and based on evidence. The EU-quarantine pest Xiphinema rivesi non-EU populations fulfilled these criteria and was selected for further evaluation. For this pest, the risk mitigation measures proposed in the technical dossier from Moldova were evaluated taking into account the possible limiting factors. For this pest, an expert judgement is given on the likelihood of pest freedom taking into consideration the risk mitigation measures acting on it, including uncertainties associated with the assessment. The Expert Knowledge Elicitation indicated, with 95% certainty, that between 9,991 and 10,000 plants per 10,000 would be free of X. rivesi.

Molecular Characterization and Functional Analysis of the Hb-hsp90-1 Gene in Relation to Temperature Changes in Heterorhabditis bacteriophora.

Understanding how entomopathogenic nematodes respond to temperature changes and have adapted to the local environment is crucial to improve their potential as biocontrol agents. In order to improve understanding of Heterorhabditis bacteriophora's potential adaptability to future climate changes, full-length cDNA and the corresponding gene of heat shock protein 90 (Hsp90) were isolated and fully characterized. The reproductive potential of the Apulian strain of H. bacteriophora increased when the temperature rose from 23 to 30°C, but no reproduction was found at 12°C. Expression analyses revealed that Hb-hsp90-1 was differentially expressed in Infective Juveniles (IJs) and adults (hermaphrodites, females and males). Up-regulation of Hb-hsp90-1 was higher during the recovery process in Galleria mellonella larvae than adults, thus confirming the protective role of Hb-hsp90-1 in coping with the host environment. Silencing of Hb-hsp90-1 resulted in a significant reduction (76%) in the expression level. Silenced IJs took longer than untreated nematodes to infect G. mellonella, showing that Hb-hsp90-1 could be also involved in chemosensation. Furthermore, the number of adults and IJs recovered from G. mellonella infected with silenced nematodes and incubated at 30°C was higher than that obtained from G. mellonella infected with untreated nematodes. These data confirm the crucial role of Hb-hsp90-1 allowing acclimation to increased temperatures and modulation of the recovery process.

Pratylenchus vovlasi sp. Nov. (Nematoda: Pratylenchidae) on Raspberries in North Italy with a Morphometrical and Molecular Characterization.

Root-lesion nematode species rank third only to root-knot and cyst nematodes as having the greatest economic impact on crops worldwide. A survey of plant-parasitic nematodes associated with decaying raspberries (Rubus sp.) in northern Italy revealed that root-lesion nematodes were the most frequently occurring species among other phytonematodes. Several Pratylenchus species have been associated with Rubus sp. in Canada (Quebec, British Columbia) and USA (North Carolina, Maryland, New Jersey) including P. penetrans and P. crenatus. In the roots and rhizosphere of symptomatic raspberries, nematodes of two Pratylenchus spp. were detected. Detailed morphometrics of the two root-lesion nematode isolates were consistent with Pratylenchus crenatus and with an undescribed Pratylenchus species. The extracted nematodes were observed and measured as live and fixed materials and subsequently identified by integrative taxonomy (morphometrically and molecularly). The latter species is described herein as Pratylenchus vovlasi sp. nov., resulting morphometrically closest to P. mediterraneus and phylogenetically to P. pratensis. The molecular identification of Pratylenchus vovlasi sp. nov. was carried out by sequencing the ITS region, D2-D3 expansion domains of the 28S rRNA gene and a partial region of the nuclear hsp90 gene. ITS-RFLP and sequence analyses revealed that Pratylenchus vovlasi sp. nov. had species-specific restriction profiles with no corresponding sequences present in the database. The phylogenetic relationships with ITS and D2-D3 sequences placed the Pratylenchus vovlasi sp. nov. in a clade with P. pratensis and P. pseudopratensis. This research confirms the occurrence of cryptic biodiversity within the genus Pratylenchus as well as the need for an integrative approach to the identification of Pratylenchus species.