Chitosan-based bioactive formulations for the control of powdery mildew in viticulture

Grapevine is highly susceptible to fungal diseases, whose incidence and severity increase due to climate change. The present work focuses on the assessment of eight combinations of natural products with chitosan oligomers with fungicidal capacity that may be effective in the integrated control of powdery mildew, in compliance with Article 14 of the European Directive 2009/128/EC. Their efficacy was evaluated in field conditions against natural infections, in a plot with high disease pressure during a growing season (assaying both foliar or root application), and against overwintering inoculums (chasmothecia) through in vitro tests. In addition, their possible biostimulant capacities were evaluated based on harvest yields. Treatments based on chitosan oligomers in combination with secondary metabolites of Streptomyces spp. and chitosan oligomers combined with hydrolyzed gluten showed the best results in terms of disease control. Given the high efficacy of these formulations, comparable to that of conventional antifungals, they constitute an interesting alternative for the control of this disease whose treatment can, in some cases, represent almost half of the production costs

Transformación vía Agrobacterium tumefaciens para inducir tolerancia a la podredumbre blanca del aguacate

Comunicación presentada en el VII World Avocado Congress, celebrado en Cairns (Australia) del 5 al 9 de septiembre de 2011.[EN] One of the most important limiting factors for avocado production in Spain is the disease caused by the fungus Rosellinia necatrix . Genetic manipulation could be useful for the introduction of fungal resistance traits into this crop. A n efficient Agrobacterium - mediated transformation protocol for avocado using AGL1 Agrobacterium strain and somatic embryos as the target material has been established by our group, although embryo conversion rate into plants needs to be improved. For that reason, we are using the strawberry, another Rosellinia necatrix ́s host, as model species to test the effect of several transgenes (two of fungal origin, chit 42 chitinase and β - 1,3 - glucanase from Trichoderma harzianum , and one of plant origin, At NPR1), on inducing tolerance to this fungus. Strawberry transformation with the β - 1,3 - glucanase gene has allowed the selection of two lines, β6 and β10, with enhanced tolerance to R. necatrix while no positive results were obtained following transformation with the chit - 42 gene. In relation to the At NPR1 gene more than 30 independent transgenic lines have been obtained whose tolerance to R. necatrix is currently under evaluation. Concerning avocado transformation, more than 10 independent transgenic lines (derive d from an embryogenic line of an immature Duke7 zygotic embryo) have been obtained with At NPR1 gene. Plants have been recovered from one line and efforts are underway to recover plants from other lines following micrografting of the transgenic sprouted sho ots onto in vitro germinated seedlings.[ES] Uno de los factores limitantes de la producción de aguacate en España es la enfermedad causada por el hongo R. necatrix. La manipulación genética podría ser de utilidad para introducir caracteres de resistencia en este cultivo. Se ha establecido un sistema eficiente de transformación en aguacate usando la cepa de Agrobacterium AGL1 y células embriogénicas como diana, sin embargo, la conversión en plantas de los embriones transgénicos necesita ser mejorada. Por esta razón, estamos utilizando la fresa, otro huésped de R. necatrix, como especie modelo para testar el efecto de varios transgenes (2 de origen fúngico, la quitinasa chit-42 y la β-1,3-glucanasa de Trichoderma harzianum, y uno derivado de plantas, AtNPR1), en la inducción de tolerancia a este patógeno tras la transformación de esta especie. La transformación de fresa con el gen de β -1,3-glucanasa ha permitido la selección de dos líneas, β6 y β10, con mayor tolerancia a R. necatrix, mientras que no se han obtenido resultados positivos con el gen chit-42. En relación con el gen AtNPR1, se han obtenido más de 30 líneas transgénicas independientes, cuya tolerancia frente a R. necatrix se está evaluando en la actualidad. En relación con la transformación de aguacate, más de 10 líneas transgénicas independientes (derivadas de una línea embriogénica obtenida a partir de un embrión zigótico inmaduro del cv. Duke 7) se han obtenido con el gen AtNPR1. Se han recuperado plantas de una línea y actualmente se está intentando recuperar plantas de otras líneas mediante microinjerto de los embriones transgénicos germinados.Este trabajo se ha realizado en el marco del proyecto AGL2008 - 05453 - C02 - 01/AGR.Peer Reviewe

Catalogue of parasites and diseases of the common cockle Cerastoderma edule

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Integrated flow cytometry and sequencing to reconstruct evolutionary patterns from dysplasia to acute myeloid leukemia

Clonal evolution in acute myeloid leukemia (AML) originates long before diagnosis and is a dynamic process that may affect survival. However, it remains uninvestigated during routine diagnostic workup. We hypothesized that the mutational status of bone marrow dysplastic cells and leukemic blasts, analyzed at the onset of AML using integrated multidimensional flow cytometry (MFC) immunophenotyping and sorting (FACS) with next-generation sequencing (NGS), could reconstruct leukemogenesis. Dysplastic cells were detected by MFC in 285 of 348 (82%) newly-diagnosed AML patients. Presence of dysplasia according to MFC and WHO criteria had no prognostic value in the elderly. NGS of dysplastic cells and blasts isolated at diagnosis identified three evolutionary patterns: stable (n=12/21), branching (n=4/21) and clonal evolution (n=5/21). In patients achieving complete response, integrated MFC and FACS with NGS showed persistent measurable residual disease (MRD) in phenotypically normal cell types, as well as the acquisition of genetic traits associated with treatment resistance. Furthermore, whole-exome sequencing of dysplastic and leukemic cells at diagnosis and of MRD uncovered different clonal involvement in dysplastic myelo-erythropoiesis, leukemic transformation and chemoresistance. Altogether, we showed that it is possible to reconstruct leukemogenesis in approximately 80% of newly diagnosed AML patients, using techniques other than single-cell multiomics.ACKNOWLEDGEMENTS: The authors acknowledge the patients, caregivers, and the biobank of the University of Navarra. This work was supported by grants from the Área de Oncología del Instituto de Salud Carlos III, Centro de Investigacion Biom ´ edica en ´ Red (CIBER-ONC) (CB16/12/00369, CB16/12/00233, CB16/12/ 00489, and CB16/12/00284), Instituto de Salud Carlos III/Subdireccion General de Investigaci ´ on Sanitaria (FIS numbers PI16/ ´ 01661, PI16/00517, and PI19/01518), and the Plan de Investigacion´ de la Universidad de Navarra (PIUNA 2014-18). This work was supported internationally by the Cancer Research UK, FCAECC, and AIRC under the Accelerator Award Program (EDITOR)

Integrated flow cytometry and sequencing to reconstruct evolutionary patterns from dysplasia to acute myeloid leukemia

Clonal evolution in acute myeloid leukemia (AML) originates long before diagnosis and is a dynamic process that may affect survival. However, it remains uninvestigated during routine diagnostic workups. We hypothesized that the mutational status of bone marrow dysplastic cells and leukemic blasts, analyzed at the onset of AML using integrated multidimensional flow cytometry (MFC) immunophenotyping and fluorescence-activated cell sorting (FACS) with next-generation sequencing (NGS), could reconstruct leukemogenesis. Dysplastic cells were detected by MFC in 285 of 348 (82%) newly diagnosed patients with AML. Presence of dysplasia according to MFC and World Health Organization criteria had no prognostic value in older adults. NGS of dysplastic cells and blasts isolated at diagnosis identified 3 evolutionary patterns: stable (n = 12 of 21), branching (n = 4 of 21), and clonal evolution (n = 5 of 21). In patients achieving complete response (CR), integrated MFC and FACS with NGS showed persistent measurable residual disease (MRD) in phenotypically normal cell types, as well as the acquisition of genetic traits associated with treatment resistance. Furthermore, whole-exome sequencing of dysplastic and leukemic cells at diagnosis and of MRD uncovered different clonal involvement in dysplastic myelo-erythropoiesis, leukemic transformation, and chemoresistance. Altogether, we showed that it is possible to reconstruct leukemogenesis in ∼80% of patients with newly diagnosed AML, using techniques other than single-cell multiomics.This work was supported by grants from the Área de Oncología del Instituto de Salud Carlos III, Centro de Investigación Biomédica en Red (CIBER-ONC) (CB16/12/00369, CB16/12/00233, CB16/12/00489, and CB16/12/00284), Instituto de Salud Carlos III/Subdirección General de Investigación Sanitaria (FIS numbers PI16/01661, PI16/00517, and PI19/01518), and the Plan de Investigación de la Universidad de Navarra (PIUNA 2014-18). This work was supported internationally by the Cancer Research UK, FCAECC, and AIRC under the Accelerator Award Program (EDITOR)

Monitoring emergence of SARS-CoV-2 B.1.1.7 Variant through the Spanish National SARS-CoV-2 Wastewater Surveillance System (VATar COVID-19) from December 2020 to March 2021

Background Since its first identification in the United Kingdom in late 2020, the highly transmissible B.1.1.7 variant of SARS-CoV-2, become dominant in several European countries raising great concern. Aim The aim of this study was to develop a duplex real-time RT-qPCR assay to detect, discriminate and quantitate SARS-CoV-2 variants containing one of its mutation signatures, the ΔHV69/70 deletion, to trace the community circulation of the B.1.1.7 variant in Spain through the Spanish National SARS-CoV-2 Wastewater Surveillance System (VATar COVID-19). Results B.1.1.7 variant was first detected in sewage from the Southern city of Málaga (Andalucía) in week 20_52, and multiple introductions during Christmas holidays were inferred in different parts of the country, earlier than clinical epidemiological reporting by the local authorities. Wastewater-based B.1.1.7 tracking showed a good correlation with clinical data and provided information at the local level. Data from WWTPs which reached B.1.1.7 prevalences higher than 90% for ≥ 2 consecutive weeks showed that 8.1±1.8 weeks were required for B.1.1.7 to become dominant. Conclusion The study highlights the applicability of RT-qPCR-based strategies to track specific mutations of variants of concern (VOCs) as soon as they are identified by clinical sequencing, and its integration into existing wastewater surveillance programs, as a cost-effective approach to complement clinical testing during the COVID-19 pandemic.This work was partially supported by the COVID-19 wastewater surveillance project (VATar COVID19), funded by the Spanish Ministry for the Ecological Transition and the Demographic Challenge of and the Spanish Ministry of Health; grants from CSIC (202070E101) and MICINN co-founded by AEI FEDER, UE (AGL2017-82909); grant ED431C 2018/18 from the Conselleria de Educacion, Universidade e Formacion Profesional, Xunta de Galicia (Spain); Direccio General de Recerca i Innovacio en Salut (DGRIS) Catalan Health Ministry Generalitat de Catalunya through Vall de Hebron Research Institute (VHIR), and Centro para el Desarrollo Tecnologico Industrial (CDTI) from the Spanish Ministry of Economy and Business, grant number IDI-20200297. Pilar Truchado is holding a Ramon y Cajal contract from the Ministerio de Ciencia e Innovacion. Adan Martinez is holding a predoctoral fellowship FI_SDUR from Generalitat de Catalunya. We gratefully acknowledge all the staff involved in the VATar COVID-19 project, working with sample collection and logistics. The authors are grateful to Promega Corporation (Madison, US) for technical advice, and thank Andrea Lopez de Mota for her technical support.N

Monitoring Emergence of the SARS-CoV-2 B.1.1.7 Variant through the Spanish National SARS-CoV-2 Wastewater Surveillance System (VATar COVID-19)

Since its first identification in the United Kingdom in late 2020, the highly transmissible B.1.1.7 variant of SARS-CoV-2 has become dominant in several countries raising great concern. We developed a duplex real-time RT-qPCR assay to detect, discriminate, and quantitate SARS-CoV-2 variants containing one of its mutation signatures, the ΔHV69/70 deletion, and used it to trace the community circulation of the B.1.1.7 variant in Spain through the Spanish National SARS-CoV-2 Wastewater Surveillance System (VATar COVID-19). The B.1.1.7 variant was detected earlier than clinical epidemiological reporting by the local authorities, first in the southern city of Málaga (Andalucía) in week 20_52 (year_week), and multiple introductions during Christmas holidays were inferred in different parts of the country. Wastewater-based B.1.1.7 tracking showed a good correlation with clinical data and provided information at the local level. Data from wastewater treatment plants, which reached B.1.1.7 prevalences higher than 90% for ≥2 consecutive weeks showed that 8.1 ± 2.0 weeks were required for B.1.1.7 to become dominant. The study highlights the applicability of RT-qPCR-based strategies to track specific mutations of variants of concern as soon as they are identified by clinical sequencing and their integration into existing wastewater surveillance programs, as a cost-effective approach to complement clinical testing during the COVID-19 pandemic.This work was partially supported by the COVID-19 wastewater surveillance project (VATar COVID19), funded by the Spanish Ministry for the Ecological Transition and the Demographic Challenge and the Spanish Ministry of Health, grants from CSIC (202070E101) and MICINN cofounded by AEI FEDER, UE (AGL2017-82909), grant ED431C 2018/18 from the Consellería de Educación, Universidade e Formación Profesional, Xunta de Galicia (Spain), Direcció General de Recerca i Innovació en Salut (DGRIS) Catalan Health Ministry Generalitat de Catalunya through Vall d’Hebron Research Institute (VHIR), and Centro para el Desarrollo Tecnológico Industrial (CDTI) from the Spanish Ministry of Economy and Business, grant number IDI-20200297. P.T. is holding a Ramón y Cajal contract from the Ministerio de Ciencia e Innovación. A.M. is holding a predoctoral fellowship FI_SDUR from Generalitat de Catalunya. We gratefully acknowledge all the staff involved in the VATar COVID-19 project, working with sample collection and logistics. The authors are grateful to Promega Corporation (Madison, US) for technical advice and thank Andrea Lopez de Mota for her technical support.Peer reviewe

Atlas des parasites et maladies de la coque commune Cerastoderma edule

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Inventario de parasitos e enfermidades do berberecho Cerastoderma edule

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Catálogo de parasitas e patologias do berbigão-vulgar Cerastoderma edule

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