Los hongos entomopatógenos y sus extractos en el control de la mosca del olivo Bactrocera oleae (Diptera: Tephritidae)

La mosca del olivo Bactrocera oleae es el fitófago más destructivo para el olivar por atacar al fruto, parte cosechable del cultivo, lo que origina graves pérdidas de cantidad de la producción, pero más grave, también en su calidad, donde este díptero constituye la principal amenaza biótica. Hoy en día el control de esta mosca es un verdadero desafío, tanto por la escasez de materias activas insecticidas disponibles, como por las primeras citas de aparición de resistencia a las mismas. Además, los efectos negativos de los insecticidas químicos sobre el medioambiente y los seres vivos, que han impulsado la normativa reguladora de la Unión Europea que emana a todos los estados miembros, promulgan los métodos no químicos de control de plagas, con énfasis en los bioinsecticidas. Dentro de éllos, los hongos entomopatógenos presentan gran potencial para ser usados dentro de los programas de control integrado de la mosca del olivo. Su presencia natural en los ecosistemas de olivar, su modo de acción por contacto, y su capacidad de secretar nuevas moléculas con actividad insecticida son factores que les sitúan en primera línea mundial para el desarrollo de estrategias alternativas de control para la mosca del olivo

Production of Microsclerotia by Metarhizium sp., and Factors Affecting Their Survival, Germination, and Conidial Yield

Microsclerotia (MS) produced by some species of Metarhizium can be used as active ingredients in mycoinsecticides for the control of soil-dwelling stages of geophilic pests. In this study, the MS production potential of two Metarhizium brunneum strains and one M. robertsii strain was evaluated. The three strains were able to produce MS in liquid fermentation, yielding between 4.0 × 106 (M. robertsii EAMa 01/158-Su strain) and 1.0 × 107 (M. brunneum EAMa 01/58-Su strain) infective propagules (CFU) per gram of MS. The EAMa 01/58-Su strain was selected for further investigation into the effects of key abiotic factors on their survival and conidial yield. The MS were demonstrated to be stable at different storage temperatures (−80, −18, and 4 °C), with a shelf-life up to one year. The best temperature for MS storage was −80 °C, ensuring good viability of MS for up to one year (4.9 × 1010 CFU/g MS). Moreover, soil texture significantly affected CFU production by MS; sandy soils were the best driver of infective propagule production. Finally, the best combination of soil temperature and humidity for MS germination was 22.7 °C and 7.3% (wt./wt.), with no significant effect of UV-B exposure time on MS viability. These results provide key insights into the handling and storage of MS, and for decision making on MS dosage and timing of application

Entomopathogenic Fungi-Mediated Solubilization and Induction of Fe Related Genes in Melon and Cucumber Plants

Endophytic insect pathogenic fungi have a multifunctional lifestyle; in addition to its well-known function as biocontrol agents, it may also help plants respond to other biotic and abiotic stresses, such as iron (Fe) deficiency. This study explores M. brunneum EAMa 01/58-Su strain attributes for Fe acquisition. Firstly, direct attributes include siderophore exudation (in vitro assay) and Fe content in shoots and in the substrate (in vivo assay) were evaluated for three strains of Beauveria bassiana and Metarhizium bruneum. The M. brunneum EAMa 01/58-Su strain showed a great ability to exudate iron siderophores (58.4% surface siderophores exudation) and provided higher Fe content in both dry matter and substrate compared to the control and was therefore selected for further research to unravel the possible induction of Fe deficiency responses, Ferric Reductase Activity (FRA), and relative expression of Fe acquisition genes by qRT-PCR in melon and cucumber plants.. In addition, root priming by M. brunneum EAMa 01/58-Su strain elicited Fe deficiency responses at transcriptional level. Our results show an early up-regulation (24, 48 or 72 h post inoculation) of the Fe acquisition genes FRO1, FRO2, IRT1, HA1, and FIT as well as the FRA. These results highlight the mechanisms involved in the Fe acquisition as mediated by IPF M. brunneum EAMa 01/58-Su strain

Optimizing wheat seed treatment with entomopathogenic fungi for improving plant growth at early development stages

Aim of study: Entomopathogenic fungi (EPF) are biocontrol agents, plant growth promoters, and increase tolerance to biotic-abiotic stresses. In this study we investigated the factors associated to the application method, which are crucial for the interaction between the fungus and the host plant at initial crop growth stages. Area of study: The study was performed in Cordoba (Spain) Material and methods: Three experiments were performed to investigate: (i) the effect of different concentrations of the surfactant Tween® 80 (0, 0.5, 1, 5, and 10%) on wheat seed coating with conidia of Metarhizium brunneum and seed and conidia viability; (ii) the performance of wheat seedlings at first growth stages after their inoculation with Beauveria bassiana or M. brunneum via seed coating or soil drenching; and (iii) the role of soil sterilization and seed disinfection on leaf concentration of chlorophyll (SPAD) and B. bassiana or M. brunneum colonization. Main results: Tween® 80 concentration linearly improved seed coating (up to 127%) without altering wheat seeds and fungal conidia germination. Seedling length of inoculated plants was significantly increased with B. bassiana and M. brunneum (67% and 46%, respectively) via seed coating. Seed disinfection was key to achieve an enhancement in wheat SPAD (10-18%) with B. bassiana or M. brunneum concerning Control, that combined with sterilization of soil showed the highest endophyte colonization rates (up to 83.3% with both fungi) Research highlights: The surfactant concentration, application method, seed disinfection, and soil sterilization are key parameters to improve the potential benefits on the EPF-plant relationship

Soil drenching with entomopathogenic fungi for control of the soil-dwelling life stages and adults of the same generation of Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae)

Four Beauveria bassiana and three Metarhizium brunneum isolates were evaluated, as soil drenches, against Spodoptera littoralis prepupae. Treatment efficacy was determined by assessing total mortality during development from prepupae through to pupae and adults; mortality and sub-lethal effects on reproduction were also quantified for adults emerging from surviving prepupae/pupae. All isolates were pathogenic but overall mortality varied between 31.7 and 83.3% (0% for control); average survival time was 7.5–10.5 days (14.0 days for control). From 1.7–15.0% of adults emerging from surviving prepupae/pupae were deformed (0% in control). Contact with fungal suspensions as prepupae/pupae caused a significant reduction in fecundity of emerging adult females (15–58.9%), and a significant reduction in egg viability (6.8–28.4%) compared with controls. Two isolates were selected for virulence evaluation against S. littoralis prepupae. The LC50s were 1.7 × 107 and 1.8 × 107 conidia ml−1 and the median survival times were 7 and 6 days for isolates EAMa 01/58-Su and EAMb 09/01-Su, respectively. Destruxin A was present in pupae developing from prepupae treated with isolates EAMa 01/58-Su (0.010 ± 0.002 µg pupae−1) and EAMb 09/01-Su (0.015 ± 0.003 µg pupae−1). The use of entomopathogenic fungi as soil drenches could be a key component of S. littoralis IPM strategies due to direct reductions in the number of soil-dwelling life stages and, also, the significant reduction in reproductive potential of surviving adults

Persistence of Metarhizium brunneum (Ascomycota: Hypocreales) in the Soil Is Affected by Formulation Type as Shown by Strain-Specific DNA Markers

The genus Metarhizium has an increasingly important role in the development of Integrated Pest Control against Tephritid fruit flies in aerial sprays targeting adults and soil treatments targeting preimaginals. Indeed, the soil is considered the main habitat and reservoir of Metarhizium spp., which may be a plant-beneficial microorganism due to its lifestyle as an endophyte and/or rhizosphere-competent fungus. This key role of Metarhizium spp. for eco-sustainable agriculture highlights the priority of developing proper monitoring tools not only to follow the presence of the fungus in the soil and to correlate it with its performance against Tephritid preimaginals but also for risk assessment studies for patenting and registering biocontrol strains. The present study aimed at understanding the population dynamics of M. brunneum strain EAMb 09/01-Su, which is a candidate strain for olive fruit fly Bactrocera oleae (Rossi, 1790) preimaginal control in the soil, when applied to the soil at the field using different formulations and propagules. For this, strain-specific DNA markers were developed and used to track the levels of EAMb 09/01-Su in the soil of 4 field trials. The fungus persists over 250 days in the soil, and the levels of the fungus remained higher when applied as an oil-dispersion formulation than when applied as a wettable powder or encapsulated microsclerotia. Peak concentrations of EAMb 09/01-Su depend on the exogenous input and weakly on environmental conditions. These results will help us to optimize the application patterns and perform accurate risk assessments during further development of this and other entomopathogenic fungus-based bioinsecticides

Direct and indirect effects of two endophytic entomopathogenic fungi on survival and feeding behaviour of meadow spittlebug Philaenus spumarius

Beyond Xylella, Integrated Management Strategies for Mitigating Xylella fastidiosa Impact in Europe (BeXyl) (Grant Agreement 101060593). Partner/Coordinador principal: Blanca B. Landa del Castillo, Investigadora Científica del Instituto de Agricultura Sostenible (IAS-CSIC).The most effective way to contain spread of the plant pathogen Xylella fastidiosa in Europe is to manage populations of its main insect vector, Philaenus spumarius L. (Hemiptera: Aphrophoridae). The action of some entomopathogenic endophytic fungi (EEF) on insect pests, directly through contact or indirectly by internal colonization of host plant tissues, may offer an excellent control option within integrated pest management (IPM) programmes. This study evaluated the impact of two EEF strains as control agents of P. spumarius (Metarhizium brunneum strain EAMa 01/58-Su and Beauveria bassiana strain EABb 01/33-Su [Hypocreales: Clavicipitaceae and Cordycipitaceae, respectively]); both strains were known to be excellent control agents of many other serious chewing and sap-sucking agricultural pests. First, the pathogenicity of strains applied directly to P. spumarius adults was evaluated, resulting in 50.0% and 32.0% mortality and a mean survival time of 4.7 and 6.2 days for M. brunneum and B. bassiana, respectively. Furthermore, colonization of Sonchus oleraceus by EEFs was observed after both soil and foliar applications. Greatest colonization (91.0%) was observed in plants treated with foliar applications of M. brunneum. Based on these results, we evaluated the feeding behaviour of P. spumarius on S. oleraceus plants previously colonized by M. brunneum using the ‘Electrical Penetration Graph’ technique. P. spumarius fed in a similar way on EEF-colonized and non-colonized S. oleraceus plants, with the exception that insects feeding on colonized plants reached xylem vessels faster than insects feeding on non-colonized plants. In addition, significantly higher mortality of P. spumarius was observed when feeding on plants treated by M. brunneum compared with the control. The duration of xylem ingestion events indicates that P. spumarius preferred to feed on basal leaves of the plant compared with apical leaves. Our results indicate strong potential for strain EAMa 01/58-Su of M. brunneum to be used as part of an X. fastidiosa IPM programme.This present study was supported by: grant PID2019-103844RB-I00 from the Spanish Ministry of Science and Innovation, Spain; by the Project BeXyl (Beyond Xylella, Integrated Management Strategies for Mitigating Xylella fastidiosa impact in Europe - Grant ID No 101060593) from the European Union’s Horizon Action ‘Food, Bioeconomy Natural Resources, Agriculture and Environment’ Programme; and by grant FP19-XYLELLA from IMIDRA. Also, this publication is part of the project TED2021-132458B-100, funded by MCIN/AEI/10.13039/501100011033 and by the European Union "NextGenerationEU"/PRTR. Recovery, Transformation, and Resilience Plan - Funded by the European Union - NextGenerationEU. Also, we acknowledge financial support from the Spanish Ministry of Science and Innovation; and the Spanish State Research Agency, through the Severo Ochoa and María de Maeztu Program for Centers and Units of Excellence in R&D (Ref. CEX2019-000968-M).With funding from the Spanish government through the "Severo Ochoa Centre of Excellence" accreditation (CEX2019-000968-M).Peer reviewe

Persistence of Metarhizium brunneum (Ascomycota: Hypocreales) in the Soil Is Affected by Formulation Type as Shown by Strain-Specific DNA Markers

The genus Metarhizium has an increasingly important role in the development of Integrated Pest Control against Tephritid fruit flies in aerial sprays targeting adults and soil treatments targeting preimaginals. Indeed, the soil is considered the main habitat and reservoir of Metarhizium spp., which may be a plant-beneficial microorganism due to its lifestyle as an endophyte and/or rhizosphere-competent fungus. This key role of Metarhizium spp. for eco-sustainable agriculture highlights the priority of developing proper monitoring tools not only to follow the presence of the fungus in the soil and to correlate it with its performance against Tephritid preimaginals but also for risk assessment studies for patenting and registering biocontrol strains. The present study aimed at understanding the population dynamics of M. brunneum strain EAMb 09/01-Su, which is a candidate strain for olive fruit fly Bactrocera oleae (Rossi, 1790) preimaginal control in the soil, when applied to the soil at the field using different formulations and propagules. For this, strain-specific DNA markers were developed and used to track the levels of EAMb 09/01-Su in the soil of 4 field trials. The fungus persists over 250 days in the soil, and the levels of the fungus remained higher when applied as an oil-dispersion formulation than when applied as a wettable powder or encapsulated microsclerotia. Peak concentrations of EAMb 09/01-Su depend on the exogenous input and weakly on environmental conditions. These results will help us to optimize the application patterns and perform accurate risk assessments during further development of this and other entomopathogenic fungus-based bioinsecticides

Metarhizium brunneum (Ascomycota; Hypocreales) Treatments Targeting Olive Fly in the Soil for Sustainable Crop Production

Soil treatments with Metarhizium brunneum EAMa 01/58-Su strain conducted in both Northern and Southern Spain reduced the olive fly (Bactrocera oleae) population density emerging from the soil during spring up to 70% in treated plots compared with controls. A model to determine the influence of rainfall on the conidial wash into different soil types was developed, with most of the conidia retained at the first 5 cm, regardless of soil type, with relative percentages of conidia recovered ranging between 56 and 95%. Furthermore, the possible effect of UV-B exposure time on the pathogenicity of this strain against B. oleae adults coming from surviving preimaginals and carrying conidia from the soil at adult emergence was also evaluated. The UV-B irradiance has no significant effect on M. brunneum EAMa 01/58-Su pathogenicity with B. oleae adult mortalities of 93, 90, 79, and 77% after 0, 2, 4, and 6 of UV-B irradiance exposure, respectively. In a next step for the use of these M. brunneum EAMa 01/58-Sun soil treatments within a B. oleae IPM strategy, its possible effect of on the B. oleae cosmopolitan parasitoid Psyttalia concolor, its compatibility with the herbicide oxyfluorfen 24% commonly used in olive orchards and the possible presence of the fungus in the olive oil resulting from olives previously placed in contact with the fungus were investigated. Only the highest conidial concentration (1 × 108 conidia ml−) caused significant P. concolor adult mortality (22%) with enduing mycosis in 13% of the cadavers. There were no fungal propagules in olive oil samples resulting from olives previously contaminated by EAMa 01/58-Su conidia. Finally, the strain was demonstrated to be compatible with herbicide since the soil application of the fungus reduced the B. oleae population density up to 50% even when it was mixed with the herbicide in the same tank. The fungal inoculum reached basal levels 4 months after treatments (1.6 × 103 conidia g soil−1). These results reveal both the efficacy and environmental and food safety of this B. oleae control method, protecting olive groves and improving olive oil quality without negative effects on the natural enemy P. concolor

Impact of the COVID-19 pandemic on patients with paediatric cancer in low-income, middle-income and high-income countries: a multicentre, international, observational cohort study

OBJECTIVES: Paediatric cancer is a leading cause of death for children. Children in low-income and middle-income countries (LMICs) were four times more likely to die than children in high-income countries (HICs). This study aimed to test the hypothesis that the COVID-19 pandemic had affected the delivery of healthcare services worldwide, and exacerbated the disparity in paediatric cancer outcomes between LMICs and HICs. DESIGN: A multicentre, international, collaborative cohort study. SETTING: 91 hospitals and cancer centres in 39 countries providing cancer treatment to paediatric patients between March and December 2020. PARTICIPANTS: Patients were included if they were under the age of 18 years, and newly diagnosed with or undergoing active cancer treatment for Acute lymphoblastic leukaemia, non-Hodgkin's lymphoma, Hodgkin lymphoma, Wilms' tumour, sarcoma, retinoblastoma, gliomas, medulloblastomas or neuroblastomas, in keeping with the WHO Global Initiative for Childhood Cancer. MAIN OUTCOME MEASURE: All-cause mortality at 30 days and 90 days. RESULTS: 1660 patients were recruited. 219 children had changes to their treatment due to the pandemic. Patients in LMICs were primarily affected (n=182/219, 83.1%). Relative to patients with paediatric cancer in HICs, patients with paediatric cancer in LMICs had 12.1 (95% CI 2.93 to 50.3) and 7.9 (95% CI 3.2 to 19.7) times the odds of death at 30 days and 90 days, respectively, after presentation during the COVID-19 pandemic (p<0.001). After adjusting for confounders, patients with paediatric cancer in LMICs had 15.6 (95% CI 3.7 to 65.8) times the odds of death at 30 days (p<0.001). CONCLUSIONS: The COVID-19 pandemic has affected paediatric oncology service provision. It has disproportionately affected patients in LMICs, highlighting and compounding existing disparities in healthcare systems globally that need addressing urgently. However, many patients with paediatric cancer continued to receive their normal standard of care. This speaks to the adaptability and resilience of healthcare systems and healthcare workers globally