Proteome of urticating setae of Ochrogaster lunifer, a processionary caterpillar of medical and veterinary importance, including primary structures of putative toxins.

Ochrogaster lunifer (Lepidoptera: Notodontidae) is an Australian processionary caterpillar with detachable urticating setae that have a defensive function. These true setae induce inflammation when they contact human skin, and equine foetal loss syndrome if they are accidentally ingested by gravid horses. We used transcriptomics and proteomics to identify proteins and peptides present in and on urticating setae, which may include toxins that contribute to inflammation and/or foetal loss syndromes. This process identified 37 putative toxins, including multiple homologues of the honeybee venom peptide secapin, and proteins with similarity to odorant binding proteins, arylphorins, and the insect immune modulator Diedel. This work identifies candidate molecules that may contribute to the adverse effects of processionary caterpillar setae on human and animal health.

The Database of European Forest Insect and Disease Disturbances: DEFID2.

Insect and disease outbreaks in forests are biotic disturbances that can profoundly alter ecosystem dynamics. In many parts of the world, these disturbance regimes are intensifying as the climate changes and shifts the distribution of species and biomes. As a result, key forest ecosystem services, such as carbon sequestration, regulation of water flows, wood production, protection of soils, and the conservation of biodiversity, could be increasingly compromised. Despite the relevance of these detrimental effects, there are currently no spatially detailed databases that record insect and disease disturbances on forests at the pan-European scale. Here, we present the new Database of European Forest Insect and Disease Disturbances (DEFID2). It comprises over 650,000 harmonized georeferenced records, mapped as polygons or points, of insects and disease disturbances that occurred between 1963 and 2021 in European forests. The records currently span eight different countries and were acquired through diverse methods (e.g., ground surveys, remote sensing techniques). The records in DEFID2 are described by a set of qualitative attributes, including severity and patterns of damage symptoms, agents, host tree species, climate-driven trigger factors, silvicultural practices, and eventual sanitary interventions. They are further complemented with a satellite-based quantitative characterization of the affected forest areas based on Landsat Normalized Burn Ratio time series, and damage metrics derived from them using the LandTrendr spectral-temporal segmentation algorithm (including onset, duration, magnitude, and rate of the disturbance), and possible interactions with windthrow and wildfire events. The DEFID2 database is a novel resource for many large-scale applications dealing with biotic disturbances. It offers a unique contribution to design networks of experiments, improve our understanding of ecological processes underlying biotic forest disturbances, monitor their dynamics, and enhance their representation in land-climate models. Further data sharing is encouraged to extend and improve the DEFID2 database continuously. The database is freely available at https://jeodpp.jrc.ec.europa.eu/ftp/jrc-opendata/FOREST/DISTURBANCES/DEFID2/.

Long-Term Aesthetic Cicatrization Analysis of Lower Eyelid Incision for Orbital Floor Fracture Approach.

Orbital fractures are among the most frequent facial injuries. Of the 3 most widely described approaches in the literature, the lower eyelid approach is the authors' preference. This study retrospectively analyzed the patients treated at the Trauma Center of the Umberto I Hospital, Sapienza University of Rome from January 2010 to December 2020. Inclusion criteria were as follows: diagnosis of pure/impure orbital bone fracture, complete clinical and radiological documentation, and a minimum of 12 months follow-up. Sex, age, etiology, treatment, and associated complications were analyzed using IBM SPSS Statistics. Two internationally validated scales were used for the functional and esthetic long-term evaluation: the Patient and Observer Scar Assessment Scale (POSAS) and the Vancouver one. The scales were compiled by the patient himself and by 3 independent expert observers. Of the 543 patients who underwent surgery in the specified period, 208 fully met the inclusion criteria. One hundred forty-two (68.2%) were males and 66 (31.8%) were females, with a mean age of 40.68 years. The main cause was represented by assaults (33.1%). One hundred seventy-nine patients had a pure orbital fracture (83.8%) and 29 an impure one (16.2%). The most frequent symptoms at the time of diagnosis were diplopia (31.2%), followed by anesthesia of the second trigeminal branch (24.3%). Open reduction with internal fixation was the preferred treatment and proved to be effective in reducing the main signs and symptoms of the fracture in a statistically significant way ( P < 0.05). Long-term esthetic results of the lower eyelid, using the Vancouver and POSASs, were respectively as follows: Vancouver Scar Scale mean total score was 2.41 (range: 0-8), observer POSAS mean total score was 1.83 (range: 1.2-3.9), observer general opinion mean score was 1.66, patient POSAS mean total score was 2.23 (range: 1.33-3.7), and patient general opinion mean score was 2.87. The lower eyelid approach has proven to be reproducible, with a fast-learning curve and a low complication rate. The analysis conducted highlighted an excellent long-term esthetic-functional result. Further studies will be needed to statistically compare the results obtained with other orbital floor surgical approaches.

The Use of Scapular Bone Flaps During Head-and-Neck Reconstruction: Functional, Orthopedic, and Quality of Life Analyses of Flap-Harvest-Related Deficits.

ABSTRACT: The authors sought correlations between harvesting of the scapular bone tip free flap used for head-and-neck reconstruction and any shoulder function deficit, particularly at the level of the rotator cuff. The flap permits reconstruction of large and diverse regions of the head and neck, but long-term harvest morbidity has not been investigated extensively. The authors analyzed the clinical records of all patients who underwent head-and-neck reconstructions using free scapular tip flaps in our department over the past 5 years. The inclusion criteria were complete clinical and radiological documentation and follow-up for at least 12 months. Two populations, a surgical and control population, were analyzed. All patients in both populations underwent the simple shoulder test, an internationally validated self-administered test that highlights functional shoulder deficits, and 3 additional tests at orthopedic visits: the external rotation test, Patte test, and external rotation lag sign test. A head-and-neck cancer-specific questionnaire (University of Washington Quality of Life, version 4) was completed by the surgical patients. All statistical analyses were performed using Software IBM SPSS Statistics (IBM Corp., Armonk, NY).A total of 19 patients met the inclusion criteria: 11 males (57.8%) and 8 females (42.2%) of mean age 60.9 years (range 23-75 years). In addition, 20 age-and sex-matched volunteers with no history of shoulder pathology were evaluated in terms of shoulder functionality. The average score on the simple shoulder test was 10.55 in the healthy population versus 8.31 in the study population ( P < 0.001); in particular, shoulder strength differed between the groups. Similarly, the orthopedic examinations revealed clinically significant differences between the controls and patients on the Patte test and external rotation lag sign test (both P < 0.001), but not the external rotation test ( P > 0.001). The mean University of Washington Quality of Life was 73.39; most patients reported good quality of life (52.6%), followed by very good (15.7%).The free scapular bone tip flap is valuable for reconstruction of head-and-neck defects; the long and reliable vascularpedicle allows rapid patient mobilization. Morbidity was confined to small reductions in shoulder strength and external rotation, which do not affect quality of life.

Spatial orientation of social caterpillars is influenced by polarized light.

Processionary caterpillars of Thaumetopoea pityocampa (in Europe) and Ochrogaster lunifer (in Australia) (Lepidoptera: Notodontidae) form single files of larvae crawling head-to-tail when moving to feeding and pupation sites. We investigated if the processions are guided by polarization vision. The heading orientation of processions could be manipulated with linear polarizing filters held above the leading caterpillar. Exposure to changes in the angle of polarization around the caterpillars resulted in corresponding changes in heading angles. Anatomical analysis indicated specializations for polarization vision of stemma I in both species. Stemma I has a rhabdom with orthogonal and aligned microvilli, and an opaque and rugged surface, which are optimizations for skylight polarization vision, similar to the dorsal rim of adult insects. Stemmata II-VI have a smooth and shiny surface and lobed rhabdoms with non-orthogonal and non-aligned microvilli; they are thus optimized for general vision with minimal polarization sensitivity. Behavioural and anatomical evidence reveal that polarized light cues are important for larval orientation and can be robustly detected with a simple visual system.

Have There Been any Changes in the Epidemiology and Etiology of Maxillofacial Trauma During the COVID-19 Pandemic? An Italian Multicenter Study.

ABSTRACT: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has had a significant impact on people's behavior. The aim of this study has been to evaluate how the SARS-CoV-2 pandemic has impacted the incidence and the features of maxillofacial fractures presented at 6 Italian tertiary centers. Clinical records of all the patients diagnosed for facial fractures between February 23 and May 23, 2019 and 2020 were retrospectively reviewed. Any differences in patient number and characteristics and fracture etiology and site between the 2 groups were then statistically analyzed.There has been a 69.1% decrease in the number of incoming patients during the pandemic. The number of foreign patients has decreased significantly (23.3% versus 9.6%, P = 0.011) while the average age has increased (38.6 versus 45.6 years old, P = 0.01). Specific statistical significant differences for accidental falls (31.8% versus 50.1%, P = 0.005) and sports injuries (16.9% versus 1.4%, P < 0.001) were found. Concerning fracture sites, significant differences have been found in relation to nasal (22.5% versus 11.4%, P = 0.009) and frontal sinus (0.9% versus 4.4%, P = 0.037) fractures. In conclusion, SARS-CoV-2 pandemic has significantly changed the epidemiology and the etiology of facial traumas.

Genetically driven CD39 expression shapes human tumor-infiltrating CD8+ T-cell functions.

In our study, we investigated the role of CD39 on tumor-infiltrating CD8+ T lymphocytes (CD8+ TILs) in colorectal, head and neck and pancreatic cancers. Partially confirming recent observations correlating the CD39 expression with T-cell exhaustion, we demonstrated a divergent functional activity in CD39+ CD8+ TILs. On the one hand, CD39+ CD8+ TILs (as compared to their CD39- counterparts) produced significantly lower IFN-γ and IL-2 amounts, expressed higher PD-1, and inversely correlated with perforin and granzyme B expression. On the other, they displayed a significantly higher proliferative capacity ex vivo that was inversely correlated with the PD-1 expression. Therefore, CD39+ CD8+ TILs, including those co-expressing the CD103 (a marker of T resident memory [TRM] cells), were defined as partially dysfunctional T cells that correlate with tumor patients with initial progression stages. Interestingly, our results identified for the first time a single nucleotide polymorphism (SNP rs10748643 A>G), as a genetic factor associated with CD39 expression in CD8+ TILs. Finally, we demonstrated that compounds inhibiting CD39-related ATPases improved CD39+ CD8+ T-cell effector function ex vivo, and that CD39+ CD8+ TILs displayed effective suppression function in vitro. Overall these data suggest that the SNP analysis may represent a suitable predictor of CD39+ CD8+ T-cell expression in cancer patients, and propose the modulation of CD39 as a new strategy to restore partially exhausted CD8+ TILs.

Performance of Trichopria drosophilae (Hymenoptera: Diapriidae), a Generalist Parasitoid of Drosophila suzukii (Diptera: Drosophilidae), at Low Temperature.

Survival and parasitism activity of Trichopria drosophilae Perkins adults, a cosmopolitan parasitoid of Drosophila spp., were studied under laboratory conditions using five constant temperatures at the lower range known for this enemy, from 4 to 20°C in 4°C increments. Drosophila suzukii Matsumura, an invasive pest of small fruits, was used as a host. Commercially available adult parasitoids were provided with 1) food and D. suzukii pupae; 2) food and no D. suzukii pupae; 3) no food and no pupae. The results show that adult females of T. drosophilae lived longer than males, and both generally benefitted from food supply. The highest level of survival was observed between 8 and 12°C for fed insects, irrespective of whether they were offered host pupae or not. The absence of food led to the highest mortality, but the parasitoid demonstrated considerably resistance to prolonged starvation. Successful parasitism increased steadily with temperature and reached the highest value at 20°C. Conversely, D. suzukii emergence rate was high after exposure of pupae to parasitoids at 4°C, while pupal mortality increased strongly with temperature until 12°C. The findings indicate that T. drosophilae is well adapted to the relatively cold conditions experienced in early spring and in autumn or at high elevations, when the host pupae could be largely available. The long lifespan of the adults and the ability to parasitize the host at low temperature make T. drosophilae potentially useful for the biocontrol of D. suzukii.

A novel veno-arteriovenous extracorporeal membrane oxygenation with double pump for the treatment of Harlequin syndrome.

OBJECTIVES: The Harlequin syndrome is a complication observed in patients receiving peripheral venoarterial extracorporeal membrane oxygenation. This condition is defined as a critical variation in the oxygen saturation between the upper and the lower part of the body deriving from a poor lung function. METHODS: Between July 2018 and November 2019, a total of 60 patients (42 men and 18 women; mean age 57.4 ± 10.0 years; range = 28-71 years) underwent peripheral venoarterial extracorporeal membrane oxygenation in our center. Harlequin syndrome was identified in eight cases (six men and two women; 13.3%) of the 60 venoarterial extracorporeal membrane oxygenation-supported patients. As a result of the Harlequin syndrome, all these patients required conversion to veno-arteriovenous extracorporeal membrane oxygenation. Control and monitoring of the blood flows of the return cannulae were performed using two centrifugal pumps, one for each inlet line, according to the patient requirements to achieve optimum hemodynamic and oxygenation. RESULTS: Mean duration of veno-arteriovenous extracorporeal membrane oxygenation support was 5.3 ± 1.4 days. Seven patients (87.5%) were switched to venovenous extracorporeal membrane oxygenation, and after 13.5 ± 2.7 days, those patients were totally weaned from extracorporeal membrane oxygenation support. One patient (12.5%) had an improvement in the pulmonary function, but the cardiac function was poor. This patient was switched to venoarterial extracorporeal membrane oxygenation, and after 10 days, the patient was completely weaned from extracorporeal membrane oxygenation support. CONCLUSION: The use of a secondary centrifugal pump to manage the blood flow directed to the internal jugular vein, in the veno-arteriovenous extracorporeal membrane oxygenation setup, allows the reduction in the risk of blood clot formation, clotting factor consumption, and pulmonary embolism when compared to the use of an external clamp.

Deciphering the drivers of negative species-genetic diversity correlation in Alpine amphibians.

The evolutionary and ecological importance of neutral and adaptive genetic diversity is widely recognized. Nevertheless, genetic diversity is rarely assessed for conservation planning, which often implicitly assumes a positive correlation between species and genetic diversity. Multiple drivers can cause the co-variation between the genetic diversity of one species and the richness of the whole communities, and explicit tests are needed to identify the processes that can determine species-genetic diversity correlations (SGDCs). Here, we tested whether intrapopulation genetic diversity (at neutral loci) and species richness co-vary in the amphibian communities of a southern Alpine region (Trentino, Italy), using the common frog (Rana temporaria) as focal species for the study of genetic diversity. We also analysed ecological similarity, niche overlap and interspecific interactions between the species, to unravel the processes determining SGDC. The neutral genetic diversity of common frogs was negatively related to species richness. The negative SGDC was probably due to an opposite influence of environmental gradients on the two levels of biodiversity, since the focal species and the other amphibians differ in ecological preferences, particularly in terms of thermal optimum. Conversely, we did not find evidence for a role of interspecific interactions in the negative SGDC. Our findings stress that species richness cannot be used as a universal proxy for genetic diversity, and only combining SGDC with analyses on the determinants of biodiversity can allow to identify the processes determining the relationships between genetic and species diversity.

Processionary Moths and Associated Urtication Risk: Global Change-Driven Effects.

Processionary moths carry urticating setae, which cause health problems in humans and other warm-blooded animals. The pine processionary moth Thaumetopoea pityocampa has responded to global change (climate warming and increased global trade) by extending its distribution range. The subfamily Thaumetopoeinae consists of approximately 100 species. An important question is whether other processionary moth species will similarly respond to these specific dimensions of global change and thus introduce health hazards into new areas. We describe, for the first time, how setae are distributed on different life stages (adult, larva) of major groups within the subfamily. Using the available data, we conclude that there is little evidence that processionary moths as a group will behave like T. pityocampa and expand their distributional range. The health problems caused by setae strongly relate to population density, which may, or may not, be connected to global change.

A total evidence phylogeny for the processionary moths of the genus Thaumetopoea (Lepidoptera: Notodontidae: Thaumetopoeinae).

Processionary moths belong to a group of about 100 species well known for their social behaviour and their urticating systems. The genus Thaumetopoea s.l. includes about 15 species and has been divided into three genera (Helianthocampa, Thaumetopoea s.s., and Traumatocampa) in the last revision, based on key morphological features of the adults and on the host plants of the larvae. We performed a total evidence approach to resolve the phylogeny of the genus Thaumetopoea s.l., analysing all valid taxa included in this group, plus a broad array of close relatives. Thaumetopoea was monophyletic and supported by several apomorphies. Further subclades corroborated by synapomorphies were identified. Our phylogeny suggests that Thaumetopoea must be regarded as a single genus. The mapping of key life history traits on the total evidence tree allowed us to sketch a plausible identikit of the Thaumetopoea ancestor and to track the evolution of the genus. The ancestor originated in the eastern Mediterranean area, and used broadleaved host plants. Subsequently, a switch to conifers occurred, just once, in a large subclade. The ancestor pupated in the soil, like several current species, but in a few taxa this trait was lost, together with the related morphological adaptations.

Rare Complication in Third Maxillary Molar Extraction: Dislocation in Infratemporal Fossa.

AIM: Removal of impacted third maxillary molar is frequently carried out without difficulties and low rate of intraoperative complications. The rare and particularly challenger to manage it is the third molar dislocation into the infratemporal fossa (IF). In this clinical report, the authors present their solution to manage and resolve this particular complication. METHODS: A 28-year-old woman was referred to the emergency rescue unit of the authors' hospital by her dentistry, after the attempt to extract the left impacted maxillary third molar. During the procedure the tooth accidentally dislodged and was lost sight of it. The patient had significant mouth-opening limitation, omolateral mid face swelling and pain. Computer tomography was immediately performed to determine the exact position of the tooth, showing the dental element dislocated into the IF. RESULTS: Considering all of possible complications the best surgical option must guarantee a direct approach and a constant eye contact of the tooth, even in case of further displacement during the procedure, and allow early surgery. The authors used an endoscopic transoral approach through the preexisted access and solved all the issues reducing morbidity. CONCLUSION: Removing tooth from the IF could be burdened by serious risk of bleeding and/or nerve injury. The endoscopic approach provides direct view of the IF reducing morbidity.

Giant "Panfacial" Chordoma Involving Skull Base With Intracranial Invasion.

Chordomas are rare, low-grade, primary malignant bone tumor arising from primitive notochord remnants of the axial skeleton. They rarely occur in an extraosseus craniofacial location such as nasopharynx, nasal septum, paranasal sinuses, and orbits. We report here an extreme case of "panfacial" chordoma involving middle and anterior skull base with intracranial invasion, which reached considerable size because of a delay in diagnosis and treatment. In this case, a wide surgical excision was performed to improve quality of life.

Effects of online tDCS and hf-tRNS on reading performance in children and adolescents with developmental dyslexia: a study protocol for a cross sectional, within-subject, randomized, double-blind, and sham-controlled trial.

Background: Developmental Dyslexia (DD) is a brain-based developmental disorder causing severe reading difficulties. The extensive data on the neurobiology of DD have increased interest in brain-directed approaches, such as transcranial direct current stimulation (tDCS), which have been proposed for DD. While positive outcomes have been observed, results remain heterogeneous. Various methodological approaches have been employed to address this issue. However, no studies have compared the effects of different transcranial electrical stimulation techniques (e.g., tDCS and transcranial random noise stimulation, tRNS), on reading in children and adolescents with DD. Methods: The present within-subject, double-blind, and sham-controlled trial aims to investigate the effects of tDCS and hf-tRNS on reading in children and adolescents with DD. Participants will undergo three conditions with a one-week interval session: (A) single active tDCS session; (B) single active hf-tRNS session; and (C) single sham session (tDCS/hf-tRNS). Left anodal/right cathodal tDCS and bilateral tRNS will be applied over the temporo-parietal regions for 20 min each. Reading measures will be collected before and during each session. Safety and blinding parameters will be recordered. Discussion: We hypothesize that tRNS will demonstrate comparable effectiveness to tDCS in improving reading compared to sham conditions. Additionally, we anticipate that hf-tRNS will exhibit a similar safety profile to tDCS. This study will contribute novel insights into the effectiveness of hf-tRNS, expediting the validation of brain-based treatments for DD.

Extranodal follicular dendritic cell sarcoma of the tonsil: An extremely rare clinical entity.

Follicular dentritic cell sarcomatous neoplasms originate from dendritic cells contained within the lymph nodes. In extranodal location, these neoplasms, are rare clinical entities, and even more so, their location in the head-neck region is extremely rare. Only 17 cases of primary dendritic cell sarcoma of the tonsil are reported in the literature at present. Being such a rare entity, histopathological diagnosis can be complex and requires great expertise and proper immunohistochemical analysis [1]. We present a case of a 48-year-old young man diagnosed with follicular dendritic cell sarcoma of the tonsil who underwent, probably for the first time in the literature, transoral robotic surgical resection.

First report of Coffin-Siris Syndrome with SMARCB1 variant, normal intelligence and mild selective neuropsychological deficits: A case report and literature review.

Background: The SMARCB1 gene encodes a subunit of the BRG1-Associated Factor (BAF) complex, and mutations in this gene have been linked to Coffin-Siris Syndrome (CSS) type 3. CSS is characterized by a range of developmental disabilities, facial dysmorphic features, and feeding difficulties. There's been noted genotype-phenotype correlation in CSS, with cases involving SMARCB1 mutations often exhibiting more severe language impairment and intellectual disability. Method: We conducted a review of reported CSS type 3 cases and presented the first instance of CSS associated with a SMARCB1 variant wherein the patient exhibited normal intelligence and only mild selective neuropsychological deficits. The patient underwent evaluation for feeding challenges, growth delay, and dysmorphic features during their second year of life. Subsequently, CSS diagnosis was confirmed due to a de novo heterozygous c.568C > T (p.Arg190Trp) variant in the SMARCB1 gene. Due to learning difficulties, the patient underwent a comprehensive neuropsychological assessment, which was related to the retrospective reconstruction of her medical and developmental history. Results: The patient demonstrated normal intelligence and adaptive functioning, with specific deficits in arithmetic and selective difficulties in verbal learning and long-term memory. Feeding difficulties and language delay observed in early childhood showed significant improvement over time. Discussion: We discuss this case in relation to previously reported CSS type 3 cases, emphasizing neuropsychological aspects. It's evident that neuropsychological features of CSS can vary among affected individuals, highlighting the importance of personalized support and interventions tailored to specific cognitive and emotional needs by healthcare professionals. Our case suggests avenues for future research to identify specific modifiers of phenotypic expression to explain variability in intellect among patients and pinpoint potential targets for gene therapy.

Seasonality of forest insects: why diapause matters.

Insects have major impacts on forest ecosystems, from herbivory and soil-nutrient cycling to killing trees at a large scale. Forest insects from temperate, tropical, and subtropical regions have evolved strategies to respond to seasonality; for example, by entering diapause, to mitigate adversity and to synchronize lifecycles with favorable periods. Here, we show that distinct functional groups of forest insects; that is, canopy dwellers, trunk-associated species, and soil/litter-inhabiting insects, express a variety of diapause strategies, but do not show systematic differences in diapause strategy depending on functional group. Due to the overall similarities in diapause strategies, we can better estimate the impacts of anthropogenic change on forest insect populations and, consequently, on key ecosystems.

Commodity risk assessment of Tilia cordata and Tilia platyphyllos plants from the UK.

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 Tilia cordata and T. platyphyllos imported from the United Kingdom (UK) as: (a) bundles of budwood/graftwood; (b) 1- to 2-year-old whips, seedlings or transplants; (c) bundles of 1- to 2-year-old cell grown plants; (d) 1- to 7-year-old bare root single plants; and (e) up to 25-year-old single plants in pots, taking into account the available scientific information provided by the UK. A list of pests potentially associated with the commodities was compiled. The relevance of any pest was assessed based on evidence following defined criteria. None of the pests on the list fulfilled all relevant criteria and therefore none were selected for further evaluation. As a result, risk mitigation measures proposed in the technical dossier from the UK were listed, but not further evaluated.

Commodity risk assessment of Corylus avellana plants from the UK.

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 Corylus avellana imported from the United Kingdom (UK) as: (a) bundles of 1- to 2-year old whips or transplants, (b) bundles of 1- to 2-year old cell grown plants, (c) 1- to 7-year old bare root single plants and (d) up to 15-year old single plants in pots, taking into account the available scientific information, including the technical information provided by the UK. All pests associated with the commodity were evaluated against specific criteria for their relevance for this opinion. Two EU quarantine pests, Phytophthora ramorum (non-EU isolates) and Thaumetopoea processionea fulfilled all relevant criteria and were selected for further evaluation. For the selected pests, the risk mitigation measures implemented in the technical dossier from the UK 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 the pest, including uncertainties associated with the assessment. In the assessment of risk, the age of the plants was considered, reasoning that older trees are more likely to be infested mainly due to longer exposure time and larger size. The degree of pest freedom varies among the pests evaluated, with P. ramorum being the pest most frequently expected on the imported plants. The expert knowledge elicitation indicated with 95% certainty that between 9939 and 10,000 of the single plants in pots up to 15-year old will be free from P. ramorum (non-EU isolates).