Multi-modal adaptor-clathrin contacts drive coated vesicle assembly.

Clathrin-coated pits are formed by the recognition of membrane and cargo by the AP2 complex and the subsequent recruitment of clathrin triskelia. A role for AP2 in coated-pit assembly beyond initial clathrin recruitment has not been explored. Clathrin binds the ß2 subunit of AP2, and several binding sites have been identified, but our structural knowledge of these interactions is incomplete and their functional importance during endocytosis is unclear. Here, we analysed the cryo-EM structure of clathrin cages assembled in the presence of ß2 hinge-appendage (ß2HA). We find that the ß2-appendage binds in at least two positions in the cage, demonstrating that multi-modal binding is a fundamental property of clathrin-AP2 interactions. In one position, ß2-appendage cross-links two adjacent terminal domains from different triskelia. Functional analysis of ß2HA-clathrin interactions reveals that endocytosis requires two clathrin interaction sites: a clathrin-box motif on the hinge and the "sandwich site" on the appendage. We propose that ß2-appendage binding to more than one triskelion is a key feature of the system and likely explains why assembly is driven by AP2.

A systematic review of self-report measures of generalized shame.

OBJECTIVE: Shame is a transdiagnostic emotion of strong clinical and research interest. Yet, there is a lack of consensus on the definition and varying methods employed across self-report measures, potentially affecting our ability to accurately study shame and examine whether clinical interventions to alter shame are effective. This paper offers a systematic review of self-report measures of generalized shame. METHODS: PubMed, PsycInfo, and Web of Science were searched. Studies were included when they were available in English and the primary aim was to evaluate measurement properties of scales or subscales designed to measure generalized shame in adults. RESULTS: Thirty-six papers examining 19 scales were identified, with measures of trait shame more common than state shame. Construct validity, internal consistency, and structural validity were relative strengths. Development and content validity studies were lacking and suffered from low methodological quality. CONCLUSIONS: All measures evaluated needed additional research to meet criteria for recommended use.

Patterns of host tree use within a lineage of saproxlic snout-less weevils (Coleoptera: Curculionidae: Scolytinae: Scolytini).

The influence of plants in the diversification of herbivorous insects, specifically those that utilize moribund and dead hosts, is little explored. Host shifts are expected because the effectiveness of toxic secondary chemicals is lessened by decay of dead plants. Feeding on dead plants also releases herbivorous insect lineages from diversifying within a particular plant lineage. Thus, phylogenetic constraints on the herbivorous insect lineage imposed by the host plants are diminished and repeated patterns of species diversification in an association with unrelated host trees is hypothesized (i.e., taxon cycle). Scolytini, a diverse weevil tribe, specialize on many different dead and moribund plant taxa as a source of food. These species and their hosts offer an opportunity to examine the association between dead host plants and the extent of phylogenetic constraints. A phylogeny of the Scolytini was reconstructed with likelihood and Bayesian analyses of DNA sequence data from nuclear (28S, CAD, ArgK) and mitochondrial (COI) genes. Ancestral host usage and geography was reconstructed using likelihood criteria and conservation of host use was tested. Results supported a monophyletic Scolytini, Ceratolepis, Loganius, and a paraphyletic Scolytus, Camptocerus and Cnemonyx. Diversification of the Scolytini generally occurred well after their host taxa diversified and suggests a sequential evolution of host use. In this scenario the beetle imposes little selection pressure on the tree but the tree provides a platform for beetle evolution. Major changes in host tree use occurred during periods of global cooling associated with changes in beetle biogeography. Diversification of beetles occurred on common and widespread hosts and there was likely a single origination of conifer-feeding from angiosperm-feeding species during the early Pliocene and a radiation of beetle species from the Palearctic to the Nearctic. Overall, the observed patterns of Scolytini host use are conserved and are similar to those expected in a taxon pulse diversification. That is, after a host switch to an unrelated tree, the beetles diversify within the host plant lineage. The need to locate an ephemeral food resource, i.e., a dying tree, likely maintains host specificity once a host shift occurs. These findings suggest that characteristics of dead and moribund host plants (e.g. secondary chemicals) influence the diversification of these saproxlic weevils despite the reduction of selection pressures.

Differential Diagnosis for the Painful Tingling Arm.

ABSTRACT: The painful tingling arm is a common presenting complaint for the musculoskeletal physician. The differential diagnosis for upper-extremity pain associated with paresthesias will be the focus of this review. Symptoms are often neurologic in etiology, originating from the spinal cord, nerve root(s), brachial plexus, or peripheral nerve(s). Localizing the pathology starts with a comprehensive understanding of neuromuscular anatomy. It also is imperative to understand the function of these respective structures. The differential diagnosis can be narrowed with a thorough history, including an assessment of sport-specific risk factors, along with a comprehensive physical examination and functional assessment. It is important to determine the sensory distribution of the patient's symptoms. If weakness also is present, the affected muscles must be identified. While the diagnosis can often be made clinically, electrodiagnostics, magnetic resonance imaging, and ultrasound can be used as needed for confirmation and more specific localization. Nonneurologic structures also may be causative or contributory to the patient's symptoms and also should be considered.

Validation of self-reported help-seeking, and measurement of the patient interval, for cancer symptoms: an observational study to inform methodological challenges in symptomatic presentation research.

BACKGROUND: To improve earlier presentation with potential symptoms of cancer, accurate data are needed on how people respond to these symptoms. It is currently unclear how self-reported medical help-seeking for symptoms associated with cancer by people from the community correspond to what is recorded in their general practice records, or how well the patient interval (time from symptom onset to first presentation to a health-professional) can be estimated from patient records. METHOD: Data from two studies that reviewed general practice electronic records of residents in Scotland, (i) the 'Useful Study': respondents to a general population survey who reported experiencing symptoms potentially associated with one of four common cancers (breast, colorectal, lung and upper gastro-intestinal) and (ii) the 'Detect Cancer Early' programme: cancer patients with one of the same four cancers. Survey respondents' self-reported help-seeking (yes/no) was corroborated; Cohen's Kappa assessed level of agreement. Combined data on the patient interval were evaluated using descriptive analysis. RESULTS: 'Useful Study' respondents' self-report of help-seeking showed exact correspondence with general practice electronic records in 72% of cases (n = 136, kappa 0.453, moderate agreement). Between both studies, 1269 patient records from 35 general practices were reviewed. The patient interval could not be determined in 44% (n = 809) of symptoms presented by these individuals. CONCLUSIONS: Patient self-report of help-seeking for symptoms potentially associated with cancer offer a reasonably accurate method to research responses to these symptoms. Incomplete patient interval data suggest routine general practice records are unreliable for measuring this important part of the patient's symptom journey.

Characterization of a novel method for the production of single-span membrane proteins in Escherichia coli.

The large-scale production and isolation of recombinant protein is a central element of the biotechnology industry and many of the products have proved extremely beneficial for therapeutic medicine. Escherichia coli is the microorganism of choice for the expression of heterologous proteins for therapeutic application, and a range of high-value proteins have been targeted to the periplasm using the well characterized Sec protein export pathway. More recently, the ability of the second mainstream protein export system, the twin-arginine translocase, to transport fully-folded proteins into the periplasm of not only E. coli, but also other Gram-negative bacteria, has captured the interest of the biotechnology industry. In this study, we have used a novel approach to block the export of a heterologous Tat substrate in the later stages of the export process, and thereby generate a single-span membrane protein with the soluble domain positioned on the periplasmic side of the inner membrane. Biochemical and immuno-electron microscopy approaches were used to investigate the export of human growth hormone by the twin-arginine translocase, and the generation of a single-span membrane-embedded variant. This is the first time that a bonafide biotechnologically relevant protein has been exported by this machinery and visualized directly in this manner. The data presented here demonstrate a novel method for the production of single-span membrane proteins in E. coli.

Phylogenomics clarifies repeated evolutionary origins of inbreeding and fungus farming in bark beetles (Curculionidae, Scolytinae).

Bark and ambrosia beetles (Curculionidae, Scolytinae) display a conspicuous diversity of unusual genetic and ecological attributes and behaviors. Reconstructing the evolution of Scolytinae, particularly the large and ecologically significant tribe Cryphalini (pygmy borers), has long been problematic. These challenges have not adequately been addressed using morphological characters, and previous research has used only DNA sequence data from small numbers of genes. Through a combination of anchored hybrid enrichment, low-coverage draft genomes, and transcriptomes, we addressed these challenges by amassing a large molecular phylogenetic dataset for bark and ambrosia beetles. The resulting DNA sequence data from 251 protein coding genes (114,276 bp of nucleotide sequence data) support inference of the first robust phylogeny of Scolytinae, with a special focus on the species rich tribe Cryphalini and its close relatives. Key strategies, including inbreeding mating systems and fungus farming, evolved repeatedly across Scolytinae. We confirm 12 of 16 hypothesized origins of fungus farming, 6 of 8 origins of inbreeding polygyny and at least 11 independent origins of a super-generalist host range. These three innovations are statistically correlated, but their appearance within lineages was not necessarily simultaneous. Additionally, the evolution of extreme host plant generalism often preceded, rather than succeeded, fungus farming. Of the high-diversity tribes of Scolytinae, only Xyleborini is monophyletic, Corthylini is paraphyletic and Cryphalini is highly polyphyletic. Cryphalini sensu stricto is part of a clade containing the genera Hypothenemus, Cryphalus and Trypophloeus, and the tribe Xyloterini. Stegomerus and Cryptocarenus (Cryphalini) are part of a clade otherwise containing all Corthylini. Several other genera, including Ernoporus and Scolytogenes (Cryphalini), make up a distantly related clade. Several of the genera of Cryphalini are also intermixed. For example, Cryphalus and Hypocryphalus are intermingled, as well as Ernoporicus, Ptilopodius and Scolytogenes. Our data are consistent with widespread polyphyly and paraphyly across Scolytinae and within Cryphalini, and provides new insights into the evolution of inbreeding mating systems and fungus farming in the species rich and ecologically significant weevil subfamily Scolytinae.

TatA complexes exhibit a marked change in organisation in response to expression of the TatBC complex.

The twin-arginine translocation (Tat) system is an integral membrane protein complex that accomplishes the remarkable feat of transporting large, fully folded polypeptides across the inner membrane of bacteria, into the periplasm. In Escherichia coli, Tat comprises three membrane proteins: TatA, TatB and TatC. How these proteins arrange themselves in the inner membrane to permit passage of Tat substrates, whilst maintaining membrane integrity, is still poorly understood. TatA is the most abundant component of this complex and facilitates assembly of the transport mechanism. We have utilised immunogold labelling in combination with array tomography to gain insight into the localisation and distribution of the TatA protein in E. coli cells. We show that TatA exhibits a uniform distribution throughout the inner membrane of E. coli and that altering the expression of TatBC shows a previously uncharacterised distribution of TatA in the inner membrane. Array tomography was used to provide our first insight into this altered distribution of TatA in three-dimensional space, revealing that this protein forms linear clusters in the inner membrane of E. coli upon increased expression of TatBC. This is the first indication that TatA organisation in the inner membrane alters in response to changes in Tat subunit stoichiometry.

Does emergency presentation of cancer represent poor performance in primary care? Insights from a novel analysis of linked primary and secondary care data.

BACKGROUND: People diagnosed with cancer following emergency presentation have poorer short-term survival. To what extent this signifies a missed opportunity for earlier diagnosis in primary care remains unclear as little detailed data exist on the patient/general practitioner interaction beforehand. METHODS: Analysis of primary care and regional data for 1802 cancer patients from Northeast Scotland. Adjusted odds ratios (OR) and 95% confidence intervals (CIs) for patient and GP practice predictors of emergency presentation. Qualitative context coding of primary care interaction before emergency presentation. RESULTS: Emergency presentations equalled 20% (n=365). Twenty-eight per cent had no relevant prior GP contact. Of those with prior GP contact 30% were admitted while waiting to be seen in secondary care, and 19% were missed opportunities for earlier diagnosis. Associated predictors: no prior GP contact (OR=3.89; CI 95% 2.14-7.09); having lung (OR=23.24; 95% CI 7.92-68.21), colorectal (OR=18.49; CI 95% 6.60-51.82) and upper GI cancer (OR=18.97; CI 95% 6.08-59.23); ethnicity (OR=2.78; CI 95% 1.27-6.06). CONCLUSIONS: Our novel approach has revealed that emergency cancer presentation is more complex than previously thought. Patient delay, prolonged referral pathways and missed opportunities by GPs all contribute, but emergency presentation can also represent effective care. Resources should be used proportionately to raise public and GP awareness and improve post-referral pathways.

Protein transport by the bacterial Tat pathway.

The twin-arginine translocation (Tat) system accomplishes the remarkable feat of translocating large - even dimeric - proteins across tightly sealed energy-transducing membranes. All of the available evidence indicates that it is unique in terms of both structure and mechanism; however its very nature has hindered efforts to probe the core translocation events. At the heart of the problem is the fact that two large sub-complexes are believed to coalesce to form the active translocon, and 'capturing' this translocation event has been too difficult. Nevertheless, studies on the individual components have come a long way in recent years, and structural studies have reached the point where educated guesses can be made concerning the most interesting aspects of Tat. In this article we review these studies and the emerging ideas in this field. This article is part of a Special Issue entitled: Protein trafficking and secretion in bacteria. Guest Editors: Anastassios Economou and Ross Dalbey.

History of the Exotic Ambrosia Beetles Euwallacea interjectus and Euwallacea validus (Coleoptera: Curculionidae: Xyleborini) in the United States.

Exotic insects are constantly intercepted at U.S. ports-of-entry. Of these, wood-boring beetles, particularly xyleborine ambrosia beetles, are sometimes missed during port inspections and become established in the United States. Euwallacea validus (Eichhoff) and Euwallacea interjectus (Blandford) are morphologically similar Asian ambrosia beetle species that vary by their fungal associates and their potential to cause economic damage. Euwallacea validus and E. interjectus were first discovered in New York (1975) and Hawaii (1976), respectively. Euwallacea validus was collected multiple times from widely separated localities and is assumed to have spread throughout the eastern United States. The discovery of E. interjectus in Florida (2011) and Texas (2011) prompted our review of the E. validus specimens because of the potential misidentification of the species. In addition, using mitochondrial cytochrome oxidase I (COI) DNA data and phylogenetic analysis, we tested the hypothesis that multiple introductions account for the U.S. populations of E. interjectus and E. validus. Our review of 7,184 specimens revealed an earlier introduction to the mainland for E. interjectus, which was first collected from Louisiana in 1984. This species is distributed in the South while E. validus occurs in the North with a known area of syntopy in northeastern Georgia. The extent of the syntopy within the United States is unknown and further investigation is required. Phylogenetic analysis of 24 E. interjectus and 20 E. validus individuals resolved clades that associated with each species and gross geographic provenance. Four well-supported clades represented E. interjectus which included the following localities: 1) Hawaii and Thailand; 2) Vietnam, Taiwan, and Texas; 3) Okinawa (Japan); and 4) Japan and several southern U.S. states. One clade comprised all E. validus specimens from Japan and the mainland United States. Four and two haplotypes were found for the E. interjectus and E. validus specimens, respectively, in mainland United States. Except for the Texas specimen, the haplotypes differed by one nucleotide. The relationship of the haplotypes and their sequence similarity suggested that the provenance of E. validus and the majority of E. interjectus haplotypes was Japan while the Texas haplotype originated later and from a location near Taiwan. Given the high nucleotide sequence difference between the Hawaiian and Thai haplotypes, the exact origin of the Hawaiian E. interjectus is unknown but likely Southeast Asia. A broader investigation including more SE Asian individuals will help to further explain the introduction of E. interjectus into Hawaii and Texas.

Coptoborus ochromactonus, n. sp. (Coleoptera: Curculionidae: Scolytinae), an emerging pest of cultivated balsa (Malvales: Malvaceae) in Ecuador.

A new species of xyleborine ambrosia beetle has been found to attack balsa, Ochroma pyramidale (Cavanilles ex Lamarck) Urban, in Ecuador. Coptoborus ochromactonus Smith & Cognato is described and its biology is reported. Large-scale surveys were conducted between 2006 and 2009, and observational studies were carried out between 2010 and 2013 in Ecuadorian commercial plantations to determine life history and host preference characteristics. C. ochromactonus attacked balsa between 1.5 and 3 yr in age. Successful attacks were more prevalent in smaller diameter trees and unhealthy trees. In general, attacks and beetle-caused mortality were more prevalent during the dry summer months when trees were under more moisture and light stress. Fungal mycelia were consistently observed coating beetle galleries and are likely the true damaging agent to balsa trees.

A new widely distributed invasive alien species of Amasa ambrosia beetles (Coleoptera: Curculionidae: Scolytinae: Xyleborini).

Amasa parviseta Knek & Smith, new species is described from Australia, Brazil, Uruguay, France and Spain. The species is native to Australia and appears to have spread widely in association with introduced Eucalyptus species.

Extracellular vesicles from GABAergic but not glutamatergic neurons protect against neurological dysfunction following cranial irradiation.

Cranial irradiation used to control brain malignancies invariably leads to progressive and debilitating declines in cognition. Clinical efforts implementing hippocampal avoidance and NMDAR antagonism, have sought to minimize dose to radiosensitive neurogenic regions while normalizing excitatory/inhibitory (E/I) tone. Results of these trials have yielded only marginal benefits to cognition, prompting current studies to evaluate the potential of systemic extracellular vesicle (EV) therapy to restore neurocognitive functionality in the irradiated brain. Here we tested the hypothesis that EVs derived from inhibitory but not excitatory neuronal cultures would prove beneficial to cognition and associated pathology. Rats subjected to a clinically relevant, fractionated cranial irradiation paradigm were given multiple injections of either GABAergic- or glutamatergic-derived EV and subjected to behavioral testing. Rats treated with GABAergic but not glutamatergic EVs showed significant improvements on hippocampal- and cortical-dependent behavioral tasks. While each treatment enhanced levels of the neurotrophic factors BDNF and GDNF, only GABAergic EVs preserved granule cell neuron dendritic spine density. Additional studies conducted with GABAergic EVs, confirmed significant benefits on amygdala-dependent behavior and modest changes in synaptic plasticity as measured by long-term potentiation. These data point to a potentially more efficacious approach for resolving radiation-induced neurological deficits, possibly through a mechanism able to restore homeostatic E/I balance.

A reevaluation of the use of rhizome scars to age plants of Trillium erectum (Melanthiaceae)1.

PREMISE OF STUDY: Herbaceous perennials are important long-lived plants in North American forests. Trillium has been used as a model organism to examine the effects of ecological processes on age structure in herbaceous forest perennials. Here, the methods of aging Trillium rhizomes are critically examined. • METHODS: Rhizomes of seedlings, single-bract plants, three-bract nonflowering plants, and flowering plants of Trillium erectum were examined. The patterns of cataphyll and scape scars on rhizomes were examined with respect to demographic category. • KEY RESULTS: Trillium rhizomes produce two cataphyll scars per year on single-bract, three-bract nonflowering, and flowering plants. Scape scars were only evident on rhizomes of three-leaved nonflowering and flowering plants. The percentage of intact rhizomes ranged from 10-67% for three-bract nonflowering plants, and 0-51% for flowering plants. Rhizomes in all demographic categories had evidence of recessing tissues from the oldest portion of the rhizome indicating that accurate age estimates are not possible on many plants. • CONCLUSIONS: Accepted methods of aging Trillium rhizomes have significant drawbacks. The primary problem is that rhizomes rot from the oldest portions in all demographic categories. A second problem is that plants producing multiple scape scars in a given year could mistakenly be counted as multiple years. Finally, confusing terminology and misrepresentations in Trillium literature suggests that many previous studies did not correctly determine age. Given the challenges of aging Trillium, we suggest that future studies use rhizome aging to study early demographic stages only and the ecological processes that influence their growth.

New species, taxonomic changes and newly recorded species of Webbia Hopkins, ambrosia beetles from Thailand and neighbouring countries (Coleoptera: Curculionidae: Scolytinae: Xyleborini).

Four new species, Webbia aculeata Sittichaya, Smith & Beaver sp. nov., Webbia granulosa Sittichaya, Smith & Beaver sp. nov., Webbia planicauda Beaver, Sittichaya & Smith sp. nov., and Webbia spinosulcata Sittichaya, Smith & Beaver sp. nov., are described. Arixyleborus dissimilis (Eggers, 1923), and Arixyleborus orbiculatus (Eggers, 1923) are transferred to Webbia. Two new synonyms and a new combination are proposed: Webbia dissimilis (Eggers, 1923) comb. nov. (=Webbia costulatula Schedl, 1953 syn. nov.); Webbia hatanakai Browne, 1986 (=Webbia turbinata Maiti & Saha, 1986 syn. nov.). A lectotype is designated for Webbia orbiculata (Eggers, 1923) comb. res.. Three species, Webbia bituberculata (Browne, 1977), Webbia subuculae (Browne, 1962), and Webbia suturalis Browne, 1955 are recorded in Thailand for the first time. With the inclusion of the Webbia species described and recorded herein, the diversity of Webbia is increased to 41 species. A key is given to the 16 species recorded from Thailand.

New species and newly recorded species of Anisandrus Ferrari, 1867 ambrosia beetles from Thailand (Coleoptera, Curculionidae, Scolytinae, Xyleborini).

Five new species, Anisandrusmontanussp. nov., A.phithakpasp. nov., A.tanaosisp. nov., A.tritonsp. nov., and A.uniseriatussp. nov. are described from Thailand. Anisandruscarinensis (Eggers, 1923) is reported from Thailand for the first time and A.apicalis is removed from the Thai fauna. With the inclusion of the species described and recorded here, the diversity of Anisandrus is increased to 40 species, of which 11 occur in Thailand. A synoptic list and a key to the Anisandrus of Thailand are presented.

Social anxiety and MDMA-assisted therapy investigation: a novel clinical trial protocol.

Background: Social anxiety disorder (SAD) is a serious and prevalent psychiatric condition that heavily impacts social functioning and quality of life. Though efficacious treatments exist for SAD, remission rates remain elevated and a significant portion of those affected do not access effective treatment, suggesting the need for additional evidence-based treatment options. This paper presents a protocol for an open-label pilot study of MDMA-assisted therapy (MDMA-AT) for social anxiety disorder. The study aims to assess preliminary treatment outcomes, feasibility and safety, and psychological and physiological processes of change in the treatment of SAD with MDMA-AT. A secondary aim includes the development of a treatment manual for MDMA-AT for SAD. Method: The outlined protocol is a randomized, open-label delayed treatment study. We will recruit 20 participants who meet criteria with moderate-to-severe social anxiety disorder (SAD) of the generalized subtype. Participants will be randomly assigned to an immediate treatment (n = 10) or delayed treatment condition (n = 10). Those in the immediate treatment condition will proceed immediately to active MDMA-AT consisting of three preparation sessions, two medicine sessions in which they receive oral doses of MDMA, and six integration sessions over approximately a 16-week period. The delayed treatment condition will receive the same intervention after a 16-week delay. Our primary outcome is SAD symptom reduction as measured by the Liebowitz Social Anxiety Scale administered by blinded raters at post-treatment and 6 month follow up. Secondary outcomes include changes in functional impairment, feasibility and safety measures, and novel therapeutic processes of change including shame and shame-related coping, belongingness, self-concealment, and self-compassion at post-treatment. Exploratory outcomes are also discussed. Discussion: The results of this pilot trial advance the field's understanding of the acceptability and potential effectiveness of MDMA-AT for social anxiety disorder and provide an overview of relevant therapeutic mechanisms unique to SAD. We hope findings from this protocol will inform the design of subsequent larger-scale randomized controlled trials (RCT) examining the efficacy of MDMA-AT for SAD. Clinical trial registration: https://clinicaltrials.gov/, NCT05138068.

Bradycardic Arrest in a Child with Complex Congenital Heart Disease Due to Sugammadex Administration.

The neuromuscular blocking drugs rocuronium and vecuronium are often used during general anesthesia. These drugs temporarily paralyze the patient and thus both facilitate placement of an endotracheal tube and prevent any patient movement during surgery. Reversal of neuromuscular blockade is necessary at the end of surgery to avoid postoperative weakness and adverse respiratory events in the recovery room. Neostigmine, the traditional reversal agent, may not completely restore muscle strength. Sugammadex is a reversal agent that is more effective and quicker acting than neostigmine. In adults, sugammadex administration has rarely been associated with bradycardia and cardiac arrest. In healthy children, the bradycardia that occurs after sugammadex administration is benign and does not require intervention. There is 1 case report of a 10- to 15-second bradycardic arrest after sugammadex administration to a 10-year-old child with heart disease. The present case report describes an 8-month-old child with complex congenital heart disease who experienced a 10-minute bradycardic arrest after the administration of sugammadex. Pediatric anesthesiologists should be aware that sugammadex administration to children with heart disease may cause hemodynamically significant bradycardia.

Fusarium kuroshium is the primary fungal symbiont of an ambrosia beetle, Euwallacea fornicatus, and can kill mango tree in Japan.

This study identifies fungi associated with Euwallacea fornicatus and determines whether these fungal species play the role of primary symbiont. E. fornicatus adults that emerged from the branches of infested trees in Okinawa main island, Japan, were collected and used to isolate fungi. Fusarium kuroshium and Penicillium citrinum were the most dominant fungal associates of females and males, respectively. F. kuroshium was much more frequently isolated from the head, including mycangia (fungus-carrying organs), of females than any other body parts. We inoculated healthy mango saplings with F. kuroshium or F. decemcellulare, both of which were symbionts of E. fornicatus females infesting mango trees. F. kuroshium decreased leaf stomatal conductance and rate of xylem sap-conduction area and increased length and area of xylem discoloration of the saplings, thereby weakening and killing some. These results suggest that F. kuroshium, a mycangial fungus of E. fornicatus, inhibits water flow in mango trees. This study is the first to report that F. kuroshium causes wilt disease in mango trees and that it is a primary fungal symbiont of E. fornicatus.