Cork oak endophytic fungi as potential biocontrol agents against biscogniauxia mediterranea and diplodia corticola

An increase in cork oak diseases caused by Biscogniauxia mediterranea and Diplodia corticola has been reported in the last decade. Due to the high socio-economic and ecologic importance of this plant species in the Mediterranean Basin, the search for preventive or treatment measures to control these diseases is an urgent need. Fungal endophytes were recovered from cork oak trees with different disease severity levels, using culture-dependent methods. The results showed a higher number of potential pathogens than beneficial fungi such as cork oak endophytes, even in healthy plants. The antagonist potential of a selection of eight cork oak fungal endophytes was tested against B. mediterranea and D. corticola by dual-plate assays. The tested endophytes were more efficient in inhibiting D. corticola than B. mediterranea growth, but Simplicillium aogashimaense, Fimetariella rabenhorstii, Chaetomium sp. and Alternaria alternata revealed a high potential to inhibit the growth of both. Simplicillium aogashimaense caused macroscopic and microscopic mycelial/hyphal deformations and presented promising results in controlling both phytopathogens’ growth in vitro. The evaluation of the antagonistic potential of non-volatile and volatile compounds also revealed that A. alternata compounds could be further explored for inhibiting both pathogens. These findings provide valuable knowledge that can be further explored in in vivo assays to find a suitable biocontrol agent for these cork oak diseases.This work was supported by FEDER funds through COMPETE (Programa Operacional Factores de Competitividade) and by national funds by FCT (Fundacao para a Ciencia e a Tecnologia) in the framework of the project POCI-01-0145-FEDER-028635, as well as for financial support to BioISI (UIDB/04046/2020) and CIMO (UID/AGR/00690/2020) by national funds by FCT/MCTES/PIDDAC. D. Costa thanks FCT for PhD grant SFRH/BD/120516/2016

Phylogenetic analysis and genetic diversity of the xylariaceous ascomycete Biscogniauxia mediterranea from cork oak forests in different bioclimates

Cork oak is a tree species with ecological importance that contributes to economic and social development in the Mediterranean region. Cork oak decline is a major concern for forest sustainability and has negative impacts on cork oak growth and production. This event has been increasingly reported in the last decades and seems to be related with climate changes. Biscogniauxia mediterranea is an endophytic fungus of healthy cork oak trees that turns into a pathogen in trees weaken by environmental stress. Understanding the drivers of B. mediterranea populations diversity and differentiation is expected to allow a better control of cork oak decline and preserve forest sustainability. Endophyte isolates from different cork oak forests were identified as B. mediterranea and their genetic diversity was evaluated using phylogenetic and microsatellite-primed PCR analyses. Genetic diversity and variability of this fungus was correlated with environmental/phytosanitary conditions present in forests/trees from which isolates were collected. High genetic diversity and variability was found in B. mediterranea populations obtained from different forests, suggesting some degree of isolation by distance. Bioclimate was the most significant effect that explained the genetic variability of B. mediterranea, rather than precipitation or temperature intensities alone or disease symptoms. These findings bring new implications for the changing climate to cork oak forests sustainability, cork production and quality.This work was supported by FEDER funds through COMPETE (Programa Operacional Factores de Competitividade) and by national funds by FCT (Fundação para a Ciência e a Tecnologia) in the framework of the project POCI-01-0145-FEDER-028635, as well as for financial support to BioISI (UIDB/04046/2020), CBMA (UIDB/04050/2020), and CIMO (UID/AGR/00690/2020) by national funds by FCT/MCTES/PIDDAC. D. Costa thanks FCT for PhD grant SFRH/BD/120516/2016.info:eu-repo/semantics/publishedVersio

Olive fungal epiphytic communities are affected by their maturation stage

The phyllosphere comprises the aerial parts of plants and is colonized by a great diversity of microorganisms, either growing inside (as endophytes) or on the surface (as epiphytes) of plant tissues. The factors that structure the diversity of epiphytes and the importance of these microorganisms for host plant protection have been less studied when compared to the case of endophytes. In this work, the epiphytic fungal communities from fruits of the olive tree (olives) in different maturation stages (green and semi-ripened), obtained from different olive orchard managements (integrated and organic production) and from distinct cultivars displaying different susceptibilities to olive anthracnose (Cobrançosa and Madural), are compared by using a metabarcoding approach. We discuss whether such differences in host resistance against anthracnose depend on both the fungal taxa or fungal community composition. A total of 1565 amplicon sequence variants (ASVs) were obtained, mainly belonging to the Ascomycota phylum and Saccharomycetes class. Although significant differences on epiphytic fungal richness were observed among olives obtained in different production systems and maturation stages, these factors in addition to host cultivar did not influence the composition of the epiphytes. Despite these results, a co-inertia analysis showed that Aureobasidium spp. and Sporocadaceae spp. were positively associated with the green olives of the cv. Madural produced under integrated production, while Saccharomycetales spp. (Kluyveromyces, Candida, Kazachstania and Saccharomyces) were positively associated with the semi-ripened olives of the cv. Cobrançosa obtained from organic production. The discriminant power of these fungi, some of them recognized as biocontrol agents, suggest that they might be important in conferring differences on host plant susceptibility to anthracnose.This research was funded by FEDER funds through COMPETE (Programa Operacional Factores de Competitividade) and by national funds by FCT (Fundação para a Ciência e a Tecnologia) in the framework of the projects PTDC/ASP-PLA/31133/2017, as well as BioISI (UIDB/04046/2020), CBMA (UIDB/04050/2020) and Mountain Research Center—CIMO (UIDB/00690/2020).info:eu-repo/semantics/publishedVersio

Bacteria could help ectomycorrhizae establishment under climate variations

Rhizosphere microbiome is one of the main sources of plant protection against drought. Beneficial symbiotic microorganisms, such as ectomycorrhizal fungi (ECMF) and mycorrhiza helper bacteria (MHB), interact with each other for increasing or maintaining host plant fitness. This mutual support benefits all three partners and comprises a natural system for drought acclimation in plants. Cork oak (Quercus suber L.) tolerance to drought scenarios is widely known, but adaptation to climate changes has been a challenge for forest sustainability protection. In this work, ECMF and MHB communities from cork oak forests were cross-linked and correlated with climates. Cenococcum, Russula and Tuber were the most abundant ECMF capable of interacting with MHB (ECMF~MHB) genera in cork oak stands, while Bacillus, Burkholderia and Streptomyces were the most conspicuous MHB. Integrating all microbial data, two consortia Lactarius/Bacillaceae and Russula/Burkholderaceae have singled out but revealed a negative interaction with each other. Russula/Burkholderaceae might have an important role for cork oak forest sustainability in arid environments, which will be complemented by the lower drought adaptation of competitive Lactarius/Bacillaceae. These microbial consortia could play an essential role on cork oak forest resilience to upcoming climatic changes.This work was supported by FEDER funds through COMPETE (Programa Operacional Factores de Competitividade) and by national funds by FCT (Fundação para a Ciência e a Tecnologia) in the framework of the projects SuberControl (PTDC/ASP-SIL/28635/2017), BioISI (UIDB/04046/2020) and CIMO (UIDB/00690/2020).info:eu-repo/semantics/publishedVersio

Improvements in the enzymatic degradation of textile dyes using ionic-liquid-based surfactants

The intensive use of water containing dyes by the textile industry, and consequently the contamination of soils and water, represents serious environmental concerns. Amongst the several processes applied in the treatment of textile effluents, biological-based processes, if designed to be cost-effective and ecofriendly, are promising alternatives to decolorize textile effluents. In this work we investigate and propose the novel use of ionic liquids (ILs) with surfactant characteristics to improve the degradation of the largely used and highly hydrophobic textile dye indigo carmine (IC) by laccase. An initial screening on the activity of laccase in aqueous solutions of twelve surfactant-based ILs from three different families, namely tetraalkylammonium- and imidazolium-based cationic surfactants and cholinium-based anionic surfactants, at different concentrations, was carried out. Significant improvements in the activity of laccase were observed with decyltrimethylammonium bromide, [N10111]Br, and 1-decyl-3-methylimidazolium chloride, [C10mim]Cl, at 75 mM (above the critical micellar concentration of each IL). These ILs were then investigated in aqueous solutions to simultaneously encapsulate laccase and IC for the in situ enzymatic biodegradation of the dye. The use of ILs remarkably increases the degradation rate of the dye and decolorization efficiency; a degradation efficiency of IC of 82% is attained in 0.5 h using aqueous solutions of [N10111]Br, whereas without IL only 6% of IC is degraded. Furthermore, 93% of the dye decolorization was achieved with [N10111]Br. The overall gathered results show that it is possible to significantly improve the degradation of hydrophobic dyes by enzymes using appropriate surfactant-based ILs, while foreseeing the use of the treated water by the same textile industries in new dyeing steps and thus contributing to a substantial decrease of the economic input and environmental footprint of these industries.publishe

Advances achieved by ionic-liquid-based materials as alternative supports and purification platforms for proteins and enzymes

Ionic liquids (ILs) have been applied in several fields in which enzymes and proteins play a noteworthy role, for instance in biorefinery, biotechnology, and pharmaceutical sciences, among others. Despite their use as solvents and co-solvents, their combination with materials for protein- and enzyme-based applications has raised significant attention in the past few years. Among them, significant advances were brought by supported ionic liquids (SILs), in which ILs are introduced to modify the surface and properties of materials, e.g., as ligands when covalently bond or when physiosorbed. SILs have been mainly investigated as alternative supports for enzymes in biocatalysis and as new supports in preparative liquid chromatography for the purification of high-value proteins and enzymes. In this manuscript, we provide an overview on the most relevant advances by using SILs as supports for enzymes and as purification platforms for a variety of proteins and enzymes. The interaction mechanisms occurring between proteins and SILs/ILs are highlighted, allowing the design of efficient processes involving SILs. The work developed is discussed in light of the respective development phase and innovation level of the applied technologies. Advantages and disadvantages are identified, as well as the missing links to pave their use in relevant applications.publishe

Illuminating Olea europaea L. endophyte fungal community

A wide array of fungal endophytes is known to inhabit plant tissues and were recently recognized as essential for plant health. A better description of the scarcely known endophyte microbiota in olive tree phyllosphere is the first step for elucidating the microbial interactions that lead to olive disease establishment. In this work, the fungal endophytic community of the phyllosphere of different olive tree cultivars (Cobrançosa, Galega vulgar, Madural, Picual, Verdeal Transmontana) is revealed by using a metabarcoding strategy targeting ITS1 barcode. A total of 460 OTUs were obtained, increasing the broad view of fungal endophytes inhabiting the olive tree phyllosphere, in particular yeast endophytes. New endophytes were persistently found in all cultivar tissues. Different olive tree cultivars depicted distinct endophyte communities. Olive cultivars exhibited dissimilar amounts of fungi with distinct ecological functions, which could explain at least in part their differential susceptibility/tolerance to olive diseases.This work was supported by FEDER funds through COMPETE (Programa Operacional Factores de Competitividade) and by national funds by FCT (Fundaç˜ao para a Ciˆencia e a Tecnologia) in the framework of the projects EXCL/AGR-PRO/0591/2012, PTDC/AGR-PRO/4354/2012, and PTDC/ASP-PLA/31133/2017, as well as BioISI (UIDB/04046/2020) and Mountain Research Center - CIMO (UIDB/00690/2020).info:eu-repo/semantics/publishedVersio

Morfometria do trato vocal por ressonância magnética: simulação de padrões patológicos articulatórios

Introdução - A análise da forma ou morfometria de estruturas anatómicas, como o trato vocal, pode ser efetuada a partir de imagens bidimensionais (2D) como de aquisições volumétricas (3D) de ressonância magnética (RM). Esta técnica de imagem tem vindo a ter uma utilização crescente no estudo da produção da fala. Objetivos - Demonstrar como pode ser efetuada a morfometria do trato vocal a partir da imagem por ressonância magnética e ainda apresentar padrões anatómicos normais durante a produção das vogais [i a u] e dois padrões articulatórios patológicos em contexto simulado. Métodos - As imagens consideradas foram recolhidas a partir de aquisições 2D (Turbo Spin-eco) e 3D (Flash Gradiente-Eco) de RM em quatro sujeitos durante a produção das vogais em estudo; adicionalmente procedeu-se à avaliação de duas perturbações articulatórias usando o mesmo protocolo de RM. A morfometria do trato vocal foi extraída com recurso a técnicas manuais (para extração de cinco medidas articulatórias) e automáticas (para determinação de volumes) de processamento e análise de imagem. Resultados - Foi possível analisar todo o trato vocal, incluindo a posição e a forma dos articuladores, tendo por base cinco medidas descritivas do posicionamento destes órgãos durante a produção das vogais. A determinação destas medições permitiu identificar quais as estratégias mais comummente adotadas na produção de cada som, nomeadamente a postura articulatória e a variação de cada medida para cada um dos sujeitos em estudo. No contexto de voz falada intersujeitos, foi notória a variabilidade nos volumes estimados do trato vocal para cada som e, em especial, o aumento do volume do trato vocal na perturbação articulatória de sigmatismo. Conclusão - A imagem por RM é, sem dúvida, uma técnica promissora no estudo da fala, inócua, não-invasiva e que fornece informação fiável da morfometria do trato vocal.Introduction - The shape or morphologic analysis of anatomical structures, such as the vocal tract can be performed from two-dimensional (2D) or volumetric acquisitions (3D) of magnetic resonance imaging (MRI). This imaging technique has had an increasing use in the study of speech production. Objectives - To determine a method to perform the morphometric analysis of the vocal tract from magnetic resonance imaging; to present anatomical patterns during the normal speech production of some vowels and two pathological articulatory disorders in simulated context. Methods - The image data was collected from 2D (Turbo Spin Echo) and 3D (Flash Gradient Echo) acquisitions of MRI of four subjects during the production of three vowels; in addition, two articulatory disorders were assessed using this imaging protocol. The morphology of the vocal tract was extracted using manual and automatic techniques of image processing and analysis. Results - Based on five articulatory measurements, it was possible to study the entire vocal tract during vowel production, including the position and shape of the articulators involved. Based on these measurements, it was possible to identify the strategies most commonly adopted in the production of each sound, including the articulatory posture and the modification of each measure for the subjects under study. Concerning the voices of the different speakers, the variability in the assessed volumes of the vocal tract for each sound was found, and in particular, the increased vocal tract volume in the articulatory disorder - the sigmatism. Conclusion - MRI is a promising technique for speech production studies, safe, non-invasive and that provides reliable information concerning the morphometric analysis of the vocal tract

Influence of Prolonged Wearing of Unstable Shoes on Upright Standing Postural Control

To study the influence of prolonged wearing of unstable shoes on standing postural control in prolonged standing workers. The participants were divided into two groups: one wore unstable shoes while the other wore conventional shoes for 8 weeks. Stabilometry parameters related to centre of pressure (CoP), rambling (RM) and trembling (TR) as well as the total agonist/antagonist muscle activity, antagonist co-activation and reciprocal activation were evaluated during upright standing, before and after the 8 week period. In both moments, the subjects were evaluated wearing the unstable shoes and in barefoot. The unstable shoe condition presented increased CoP displacement related variables and decreased co-activation command compared to barefoot before and after the intervention. The prolonged wearing of unstable shoes led to: (1) reduction of medial-lateral CoP root mean square and area; (2) decreased anteroposterior RM displacement; (3) increased anteroposterior RM mean velocity and mediolateral RM displacement; (4) decreased anteroposterior TR RMS; and (5) increased thigh antagonist co-activation in the unstable shoe condition. The unstable shoe condition is associated to a higher destabilizing effect that leads to a selection of more efficient and accurate postural commands compared to barefoot. Prolonged wearing of unstable shoes provides increased effectiveness and performance of the postural control system, while wearing of unstable shoes in upright standing, that are reflected by changes in CoP related variables and by a reorganization of postural control commands

Usefulness of neck ultrasonography in the follow-up of patients with differentiated thyroid cancer

Neck ultrasonography (US) is recommended for the assessment of all patients with thyroid carcinoma after initial therapy, since even low-risk patients with undetectable stimulated thyroglobulin (Tg) may present cervical metastases. In the case of these metastases, US is the most sensitive method and is superior to whole-body 131I scanning. Cervical lymph nodes with a diameter > 5 mm presenting thin calcifications and/or cystic degeneration have almost always a malignant etiology. In the absence of these characteristics, a round shape and the absence of an echogenic hilum are suspicious findings, whereas elongated lymph nodes with a visible echogenic hilum are considered benign. Doppler flow analysis helps with the differential diagnosis, usually revealing peripheral or mixed hypervascularization in malignant cases. In the presence of suspicious lymph nodes upon US, fine-needle aspiration cytology and measurement of Tg in the needle lavage fluid are useful and complementary exams for the definition of the etiology, with the combination of the two methods showing elevated sensitivity and 100% specificity. US is also useful before thyroidectomy, even contributing in some cases to modify the surgical planning, and before ablation for the measurement of thyroid remnants and detection of persistent lymph node metastases. Another application of this imaging method is to guide the injection of ethanol (sclerotherapy) or the introduction of electrodes for radiofrequency ablation in selected cases of isolated lymph node metastases as an alternative to traditional therapies.A ultra-sonografia (US) cervical é recomendada na avaliação de todos pacientes com carcinoma de tireóide após a terapia inicial, pois mesmo indivíduos de baixo risco com tireoglobulina (Tg) estimulada indetectável podem apresentar metástases cervicais. Para estas metástases, a US é o método mais sensível, superior à pesquisa de corpo inteiro (PCI) com 131I. Linfonodos cervicais com diâmetro > 5 mm com calcificações finas e/ou degeneração cística quase sempre são de etiologia maligna. Na ausência destas características, o formato arredondado e a ausência do hilo ecogênico são achados suspeitos, enquanto linfonodos alongados e com hilo ecogênico visível são considerados benignos. A avaliação do fluxo, através do doppler, auxilia no diagnóstico diferencial, usualmente revelando hipervascularização periférica ou mista nos casos malignos. Na presença de linfonodos suspeitos na US, a avaliação citológica do material obtido através da punção aspirativa por agulha fina (PAAF) e a dosagem da Tg, obtida do lavado da agulha, são testes úteis e complementares para definir a etiologia, com elevada sensibilidade quando combinados e especificidade de 100%. A US também é útil antes da tiroidectomia, auxiliando e até, em alguns casos, modificando o planejamento cirúrgico; e antes da ablação, para mensuração dos remanescentes tireoidianos e pesquisa de metástases linfonodais persistentes. Outra aplicação desse método de imagem é guiar a injeção de etanol (escleroterapia) ou a introdução de eletrodos para ablação com radiofreqüência em casos selecionados de metástases linfonodais isoladas, como alternativa às terapias convencionais.Santa Casa de Belo Horizonte Serviço de EndocrinologiaInstituto Alpha de GastroenterologiaUniversidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Departamento de MedicinaUNIFESP, EPM, Depto. de MedicinaSciEL