Efeito do plasma frio atmosférico em revestimento de cera de carnaúba na inativação de Colletotrichum brevisporum

Cera de carnaúba é utilizada, com sucesso, em revestimento de frutos por formar uma barreira contra a perda de umidade, troca de gases e ação antimicrobiana. No presente trabalho, pretende-se melhorar esta última propriedade através do tratamento por plasma. Plasma produzido por descarga de barreira dielétrica (DBD), usando pulsos de voltagem de 45 kV, repetidos na frequência de 405 Hz, durante 9 min, foi aplicado diretamente sobre esporos de fungo Colletotrichum brevisporum (4x104 conidia.cm-1) ou sobre superfície dos revestimentos de cera. Verificou-se que o revestimento, após tratado por plasma, apresentou resistência antifúngica equivalente àquela existente quando aplicada diretamente sobre os fungos. Este resultado foi justificado com base na literatura, em que o tratamento com plasma em lipídios estimula a oxidação das cadeias lipídicas, resultando na formação de compostos carboxílicos, que possuem propriedades antimicrobianas. Palavras-chave: Plasma DBD, eletrodos coplanares, antifúngico, atmosférico, inativação

Identification and pathogenicity of Macrophomina species collected from weeds in melon fields in Northeastern Brazil

"This is the peer reviewed version of the following article: Negreiros, AMP, Sales Júnior, R, León, M, et al. Identification and pathogenicity of Macrophomina species collected from weeds in melon fields in Northeastern Brazil. J Phytopathol. 2019; 167: 326 337. , which has been published in final form at https://doi.org/10.1111/jph.12801. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving."[EN] In this work, a collection of 94 Macrophomina isolates obtained from roots of two weed species, Trianthema portulacastrum and Boerhavia diffusa, collected during surveys conducted during 2015 and 2016 in melon production fields in Northeastern Brazil, were characterized by using molecular techniques. Phylogenetic analysis of the EF1-alpha gene allowed the identification of 32 isolates as M. phaseolina and 62 isolates as M. pseudophaseolina. Results of a pathogenicity test performed on melon seedlings of the cv. "Gladial" revealed that all M. phaseolina isolates inoculated were able to cause disease to melon seedlings, but only some M. pseudophaseolina isolates were able to infect them. This study represents the first report of M. pseudophaseolina in both T. portulacastrum and B. diffusa weeds, which are prevalent in the main Brazilian melon producing and exporting regions. Information about the biology and epidemiology of M. pseudophaseolina is scarce because of its recent description; thus, further research is needed for a better understanding of this fungus as a potentially emerging pathogen of melon and other crops.Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior-Brazil (CAPES); Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)Negreiros, AMP.; Sales Junior, R.; León Santana, M.; de Assis Melo N.J.; Michereff, S.; de Queiroz Ambrósio M.M.; De Sousa Medeiros, H.... (2019). Identification and pathogenicity of Macrophomina species collected from weeds in melon fields in Northeastern Brazil. Journal of Phytopathology. 167(6):326-337. https://doi.org/10.1111/jph.12801S3263371676Agustí-Brisach, C., Gramaje, D., León, M., García-Jiménez, J., & Armengol, J. (2011). Evaluation of Vineyard Weeds as Potential Hosts of Black-Foot and Petri Disease Pathogens. Plant Disease, 95(7), 803-810. doi:10.1094/pdis-12-10-0888A. C. Alfenas R. Mafia G. Métodos em fitopatologia 2016 Ed. UFV Universidade Federal de Viçosa Viçosa Brasil 516Ambrósio, M. M. Q., Dantas, A. C. A., Martínez-Perez, E., Medeiros, A. C., Nunes, G. H. S., & Picó, M. B. (2015). Screening a variable germplasm collection of Cucumis melo L. for seedling resistance to Macrophomina phaseolina. Euphytica, 206(2), 287-300. doi:10.1007/s10681-015-1452-xAnuário Anuário ‐ Anuário Brasileiro da Fruticultura 2018 2018 Ed. Gazeta Santa Cruz Santa Cruz do Sul Brazil 88Baird, R. E., & Brock, J. H. (1999). First Report of Macrophomina phaseolina on Cotton (Gossypium hirsutum) in Georgia. Plant Disease, 83(5), 487-487. doi:10.1094/pdis.1999.83.5.487bBaird, R. E., Watson, C. E., & Scruggs, M. (2003). Relative Longevity of Macrophomina phaseolina and Associated Mycobiota on Residual Soybean Roots in Soil. Plant Disease, 87(5), 563-566. doi:10.1094/pdis.2003.87.5.563Carbone, I., & Kohn, L. M. (1999). A Method for Designing Primer Sets for Speciation Studies in Filamentous Ascomycetes. Mycologia, 91(3), 553. doi:10.2307/3761358Chaves, A. L. R., Braun, M. R., Eiras, M., Colariccio, A., & Galleti, S. R. (2003). Erigeron bonariensis: hospedeira alternativa do Lettuce mosaic virus no Brasil. Fitopatologia Brasileira, 28(3), 307-311. doi:10.1590/s0100-41582003000300014Claudino, M. R., & Soares, D. J. (2014). Pathogenicity and aggressiveness of Macrophomina phaseolina isolates to castor (Ricinus communis). Tropical Plant Pathology, 39(6), 453-456. doi:10.1590/s1982-56762014000600006Cohen, R., Omari, N., Porat, A., & Edelstein, M. (2012). Management of Macrophomina wilt in melons using grafting or fungicide soil application: Pathological, horticultural and economical aspects. Crop Protection, 35, 58-63. doi:10.1016/j.cropro.2011.12.015FAOSTAT(2018). FAO statistical databases food and agriculture organization of the United Nations.http://www.fao.org/faostat/en/#home.Farr D. F. &Rossman A. Y.(2018). Fungal Databases. National Fungus Collections ARS USDA: U.S.https://nt.ars-grin.gov/fungaldatabases/.Fuhlbohm, M. J., Ryley, M. J., & Aitken, E. A. B. (2012). New weed hosts of Macrophomina phaseolina in Australia. Australasian Plant Disease Notes, 7(1), 193-195. doi:10.1007/s13314-012-0082-6Funnell-Harris, D. L., O’Neill, P. M., Sattler, S. E., & Yerka, M. K. (2016). Response of Sweet Sorghum Lines to Stalk Pathogens Fusarium thapsinum and Macrophomina phaseolina. Plant Disease, 100(5), 896-903. doi:10.1094/pdis-09-15-1050-reIBGE(2018). Instituto Brasileiro de Geografia e Estatística.https://sidra.ibge.gov.br/home/pms/brasil.Jacob, C. J., Krarup, C., Díaz, G. A., & Latorre, B. A. (2013). A Severe Outbreak of Charcoal Rot in Cantaloupe Melon Caused by Macrophomina phaseolina in Chile. Plant Disease, 97(1), 141-141. doi:10.1094/pdis-06-12-0588-pdnKumar, S., Stecher, G., & Tamura, K. (2016). MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Molecular Biology and Evolution, 33(7), 1870-1874. doi:10.1093/molbev/msw054Machado, A. R., Pinho, D. B., & Pereira, O. L. (2014). Phylogeny, identification and pathogenicity of the Botryosphaeriaceae associated with collar and root rot of the biofuel plant Jatropha curcas in Brazil, with a description of new species of Lasiodiplodia. Fungal Diversity, 67(1), 231-247. doi:10.1007/s13225-013-0274-1Machado, A. R., Pinho, D. B., Soares, D. J., Gomes, A. A. M., & Pereira, O. L. (2018). Bayesian analyses of five gene regions reveal a new phylogenetic species of Macrophomina associated with charcoal rot on oilseed crops in Brazil. European Journal of Plant Pathology, 153(1), 89-100. doi:10.1007/s10658-018-1545-1Medeiros, A. C., Melo, D. R. M. de, Ambrósio, M. M. de Q., Nunes, G. H. de S., & Costa, J. M. da. (2015). Métodos de inoculação de Rhizoctonia solani e Macrophomina phaseolina em meloeiro (Cucumis melo). Summa Phytopathologica, 41(4), 281-286. doi:10.1590/0100-5405/2083Miller M. A. Pfeiffer W. &Schwartz T.(2012). The CIPRES science gateway: enabling high‐impact science for phylogenetics researchers with limited resources. In: Proceedings of the 1st Conference of the Extreme Science and Engineering Discovery Environment: Bridging from the extreme to the campus and beyond (pp.39).Chicago IL.Mir, Z. R., Singh, P. K., Zaidi, P. H., Vinayan, M. T., Sharma, S. S., Krishna, M. K., … Nair, S. K. (2018). Genetic analysis of resistance to post flowering stalk rot in tropical germplasm of maize ( Zea mays L.). Crop Protection, 106, 42-49. doi:10.1016/j.cropro.2017.12.004Mbaye, N., Mame, P. S., Ndiaga, C., & Ibrahima, N. (2015). Is the recently described Macrophomina pseudophaseolina pathogenically different from Macrophomina phaseolina? African Journal of Microbiology Research, 9(45), 2232-2238. doi:10.5897/ajmr2015.7742Nylander J. A. A.(2004). MrModeltest V2. Program Distributed by the Author: Evolutionary Biology Centre Uppsala University Sweden.Reuveni, R., Krikun, J., Nachmias, A., & Shlevin, E. (1982). The role ofMacrophomina phaseolina in a collapse of melon plants in Israel. Phytoparasitica, 10(1), 51-56. doi:10.1007/bf02981892Ronquist, F., Teslenko, M., van der Mark, P., Ayres, D. L., Darling, A., Höhna, S., … Huelsenbeck, J. P. (2012). MrBayes 3.2: Efficient Bayesian Phylogenetic Inference and Model Choice Across a Large Model Space. Systematic Biology, 61(3), 539-542. doi:10.1093/sysbio/sys029Rusuku, G., Buruchara, R. A., Gatabazi, M., & Pastor-Corrales, M. A. (1997). Occurrence and Distribution in Rwanda of Soilborne Fungi Pathogenic to the Common Bean. Plant Disease, 81(5), 445-449. doi:10.1094/pdis.1997.81.5.445Sales Junior, R., Oliveira, O. F. de, Medeiros, É. V. de, Guimarães, I. M., Correia, K. C., & Michereff, S. J. (2012). Ervas daninhas como hospedeiras alternativas de patógenos causadores do colapso do meloeiro. Revista Ciência Agronômica, 43(1), 195-198. doi:10.1590/s1806-66902012000100024Short, G. E. (1980). Survival ofMacrophomina phaseolinain Soil and in Residue of Soybean. Phytopathology, 70(1), 13. doi:10.1094/phyto-70-13Francisco, de A. S. e S., & Carlos, A. V. de A. (2016). The Assistat Software Version 7.7 and its use in the analysis of experimental data. African Journal of Agricultural Research, 11(39), 3733-3740. doi:10.5897/ajar2016.11522Stamatakis, A. (2014). RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics, 30(9), 1312-1313. doi:10.1093/bioinformatics/btu033Thompson, J. D., Higgins, D. G., & Gibson, T. J. (1994). CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research, 22(22), 4673-4680. doi:10.1093/nar/22.22.4673Wrather, J. A., Anderson, T. R., Arsyad, D. M., Gai, J., Ploper, L. D., Porta-Puglia, A., … Yorinori, J. T. (1997). Soybean Disease Loss Estimates for the Top 10 Soybean Producing Countries in 1994. Plant Disease, 81(1), 107-110. doi:10.1094/pdis.1997.81.1.107Wrather, J. A., Anderson, T. R., Arsyad, D. M., Tan, Y., Ploper, L. D., Porta-Puglia, A., … Yorinori, J. T. (2001). Soybean disease loss estimates for the top ten soybean-producing counries in 1998. Canadian Journal of Plant Pathology, 23(2), 115-121. doi:10.1080/0706066010950691

Recolhimento de embalagens vazias de agrotóxicos no Rio Grande do Norte de 2006 a 2014

Pesticides are substances used to control unwanted biological agents agricultural production, and improper disposal of its packaging can contaminate the environment, and endanger public health. Thus, this study aims to quantify the volume of packaging types that have been processed and sent for incineration and recycling by Empties Receiving Center of Pesticide Mossoro - RN in the years 2006 to 2014. In this period were received at central a total of 488.76 tons of empty pesticide containers and by the end of 2014, 450.53 t of this material were processed and transported to recycling or incineration, totaling approximately 92% of packages received by the Central. Packaging of High Density Polyethylene and cardboard were those that represented the highest percentage of processing and withdrawal of Central 40.36 and 28.10%, respectively. In contrast, smaller percentages were taken from the steel and to the lids of containers 2.16 and 4.03%, respectively. Contaminated or not cleanable packaging accounted for 11.44% of the total amount processed.Os agrotóxicos são substâncias utilizadas para o controle de agentes biológicos não desejáveis a produção agrícola, e o descarte inadequado de suas embalagens podem contaminar o meio ambiente e ainda colocar em risco a saúde pública. Sendo assim, este trabalho tem por objetivo quantificar o volume de tipos de embalagens que foram processadas e encaminhadas para incineração e reciclagem pela Central de Recebimento de Embalagens Vazias de Agrotóxicos de Mossoró – RN nos anos de 2006 a 2014. Neste período deram entrada na central um total de 488.76 t de embalagens vazias de agrotóxicos e até o final de 2014, 450.53 t desse material foram processados e transportados para recicladoras ou incineradoras, totalizando aproximadamente 92% das embalagens recebidas pela Central. As embalagens de Polietileno de Alta Densidade e papelão foram as que representaram o maior percentual de processamento e retirada da Central 40,36 e 28,10%, respectivamente. Em contrapartida, os menores valores percentuais de retirada foram para o aço e as tampas das embalagens 2,16 e 4,03%, respectivamente. As embalagens contaminadas ou não laváveis representaram 11,44% do valor total processado

Monosporascus cannonballus: AGENTE CAUSAL DO COLAPSO OU MORTE SÚBITA DO MELOEIRO

O fungo Monosporascus cannonballus destaca-se como um dos principais fitopatógenos envolvidos no Colapso ou Morte Súbita do meloeiro. Trata-se de um fitopatógeno habitante natural do solo, termófilo e bem adaptado às condições áridas e semi-áridas, e por isso, responsável por grandes perdas, limitando o cultivo de melão em diversas regiões do mundo. Inúmeras medidas de controle têm sido estudadas, entretanto, estas ainda são pouco eficazes. Apesar da importância desta síndrome, poucos são os estudos sobre o potágeno. Desta forma, informações epidemiológicas da doença constituem importantes aspectos no desenvolvimento de estratégias para seu manejo integrado

Efeito de doses de mycosim tri-ton no cultivo do melão e da melancia conduzido em dois solos naturalmente infestados

This study investigated the behavior of plants of watermelon and melon with different doses of mycorrhiza in two types of naturally infested soil in greenhouse conditions. The variables analyzed were: plant height, diameter of the hypocotyl and root fresh weight. Statistical analysis showed a positive effect between types of soil texture, and is considered a sandy texture frankly the best one. For tests with watermelon no statistical difference regarding doses of mycorrhiza and its interaction with the two soil types, however with melon statistical difference regarding doses of mycorrhiza in relation to root fresh weight.Este trabalho objetivou estudar o comportamento de plantas de melancia e de melão a diferentes doses de micorriza, em dois tipos de solo naturalmente infestados, em condições de casa de vegetação. As variáveis analisadas foram: altura da planta, diâmetro do hipocótilo e peso fresco de raízes. A análise estatística evidenciou efeito positivo entre os tipos de textura de solo, sendo considerada a textura franca arenoso a melhor delas. Para os testes com melancia não houve diferença estatística quanto às doses de micorriza e sua interação com os dois tipos de solo, no entanto com melão houve diferença estatística quanto às doses de micorriza com relação ao peso fresco de raiz

Monosporascus cannonballus: agente causal do colapso ou morte súbita do meloeiro

The fungus Monosporascus cannonballus stands out as one of the main pathogens involved in the Melon Collapse or Sudden Wilt. This is a pathogen natural inhabitant of soil, thermophilic and well adapted to arid and semi-arid conditions, and therefore, responsible for major losses, limiting the cultivation of melons in different regions of the world. Several control measures have been studied, however, these are not still very effective. Despite the importance of this syndrome, there are few studies about the pathogen. Thus, informations about the disease epidemiology are an important aspects in developing strategies for integrated management.O fungo Monosporascus cannonballus destaca-se como um dos principais fitopatógenos envolvidos no Colapso ou Morte Súbita do meloeiro. Trata-se de um fitopatógeno habitante natural do solo, termófilo e bem adaptado às condições áridas e semi-áridas, e por isso, responsável por grandes perdas, limitando o cultivo de melão em diversas regiões do mundo. Inúmeras medidas de controle têm sido estudadas, entretanto, estas ainda são pouco eficazes. Apesar da importância desta síndrome, poucos são os estudos sobre o potágeno. Desta forma, informações epidemiológicas da doença constituem importantes aspectos no desenvolvimento de estratégias para seu manejo integrado.El hongo Monosporascus cannonballus se destaca como uno de los principales patógenos implicados en el colapso del melón o la muerte súbita. Este es un habitante natural del patógeno del suelo, termofílicas, y bien adaptados a zonas áridas y semiáridas, y por lo tanto responsables de grandes pérdidas al restringir el cultivo de melones en distintas regiones del mundo. varias medidas de control han sido estudiados, sin embargo, estos siguen siendo muy eficaces. A pesar de la importancia de este síndrome, hay pocos estudios sobre el potágeno. Así, la información epidemiológica de la enfermedad, son aspectos importantes en el desarrollo de estrategias para la gestión integrada

Thrips collected in watermelon crops in the semiarid of Rio Grande do Norte, Brazil

The aim of this research was to report the occurrence of two species of thrips in watermelon crops in the semiarid region of Rio Grande do Norte, Brazil. From August to September 2011, we performed weekly sampling of thrips in a commercial watermelon production area with 10,000m2, using 20 Moericke traps. We captured a total of 431 thrips, belonging to Frankliniella schultzei (Trybom) (Thripidae) and Haplothrips gowdeyi (Franklin) (Phlaeothripidae). This is the first report of H. gowdeyi in watermelon crops in Brazi

Design and Fabrication of a Biomimetic Smart Material for Electrochemical Detection of Carbendazim Pesticides in Real Samples with Enhanced Selectivity

Agricultural products are vitally important for sustaining life on earth and their production has notably grown over the years worldwide in general and in Brazil particularly. Elevating agricultural practices consequently leads to a proportionate increase in the usage of pesticides that are crucially important for enhanced crop yield and protection. These compounds have been employed excessively in alarming concentrations, causing the contamination of soil, water, and air. Additionally, they pose serious threats to human health. The current study introduces an innovative tool for producing appropriate materials coupled with an electrochemical sensor designed to measure carbendazim levels. The sensor is developed using a molecularly imprinted polymer (MIP) mounted on a glassy carbon electrode. This electrode is equipped with multi-walled carbon nanotubes (MWCNTs) for improved performance. The combined system demonstrates promising potential for accurately quantifying carbendazim. The morphological characteristics of the synthesized materials were investigated using field emission scanning electron microscopy (FESEM) and the Fourier-transform infrared (FTIR) technique. The analytical curve was drawn using the electrochemical method in the range of 2 to 20 ppm while for HPLC 2–12 ppm; the results are presented as the maximum adsorption capacity of the MIP (82.4%) when compared with NIP (41%) using the HPLC method. The analysis conducted using differential pulse voltammetry (DPV) yielded a limit of detection (LOD) of 1.0 ppm and a repeatability of 5.08% (n = 10). The results obtained from the analysis of selectivity demonstrated that the proposed electrochemical sensor is remarkably efficient for the quantitative assessment of carbendazim, even in the presence of another interferent. The sensor was successfully tested for river water samples for carbendazim detection, and recovery rates ranging from 94 to 101% were obtained for HPLC and 94 to 104% for the electrochemical method. The results obtained show that the proposed electrochemical technique is viable for the application and quantitative determination of carbendazim in any medium

EFEITO DE FLUAZINAM NO CONTROLE Monosporascus cannonballus, AGENTE CAUSAL DO DECLÍNIO DE RAMAS EM MELOEIRO

The use of fungicides is a key measure of control used to halt the vine decline in melon by Monosporascus cannonballus. This study aimed to ascertain the effectiveness of the active principles fluazinam and methyl thiophanate in control of M. cannonballus in melon. Melon plants (yellow type) were grown in pot with soil naturally infested with the pathogen. The soil was originated from areas with a history of vine decline in melons. It was used a design in DIC with 10 treatments and 5 replications. The treatments consisted of increasing doses of these active principles alone or in combination. Were utilized two controls: autoclaved soil and naturally infested soil. The parameters analyzed were fresh weight of roots, damage to hypocotyl in primary and secondary roots. The analysis of variance indicated that all treatments that showed the presence of active principle fluazinam differ significantly from other treatments. In addition, the analysis did not indicate differences among the doses used in the experiment. Thus, it appears that this active principle can be recommended at its lowest dose to control the disease once it is properly registered to this crop. It was found phytotoxic effects of the active principle methyl thiophanate in melon plants during this experiment

Eficiência de Bacillus subtilis e B. pumilus no controle de Podosphaera xanthii em meloeiro

This study aimed to prove the efficiency of Bacillus subtilis and B. pumilus on the control of muskmelon powdery mildew (Podosphaera xanthii). The essay was installed in a commercial field in the city of Mossoró- RN, during the period of December/2008 to January/2009. The experimental design was randomized blocks with nine treatments and four replications, being the treatments characterized by doses of the following biological and chemical products: Serenade (B. subtilis) (1.34 g.L -1 ) at doses of 2.5, 5.0 and 7.5 mL.L-1 ; Sonata (B. pumilus) (1.38 g.L-1 ) at doses of 2.5, 5.0 and 7.5 mL.L-1 ; Score® (difenoconazole) (250 g.L-1 ) at a dose of 0.3 mL.L-1 ; Amistar® (azoxystrobin) (500 g.kg-1 ) at a dose of 0.21 g.L -1 . The witness did not receive any treatment. Aplications were done every 7 days, for five weeks and the evaluations were done every 7 days, for four weeks, being the first done on the thirst week after the experiment implantation. The evaluations were based in the note scale ranging from 0 (zero) to 5 (five), where “0” corresponds to no infected plant and “5” to infection superior to 40% of leaf area. The biological products Serenade at dose of 750 mL.100L-1 and Sonata at doses of 250 and 500 mL.100L-1 , applied into preventive spraying, controlled satisfactorily powdery mildew on melon when compared to the standard products Amistar® and Score® , used in commercial scale, being recommended for the biological control of this pathogen in melon culture.Este trabalho objetivou comprovar a eficiência de Bacillus subtilis e B. pumilus no controle do oídio do meloeiro (Podosphaera xanthii). O experimento foi instalado em campo comercial da cultura no município de Mossoró- RN, durante o período de dezembro de 2008 a janeiro de 2009. O delineamento experimental utilizado foi em blocos ao acaso com nove tratamentos e quatro repetições, sendo os tratamentos caracterizados por concentrações dos seguintes produtos biológicos e químicos: Serenade (B. subtilis) (1,34 g.L-1 ) nas doses de 2,5; 5,0 e 7,5 mL.L -1 ; Sonata (B. pumilus) (1,38 g.L-1 ) nas doses de 2,5; 5,0 e 7,5 mL.L -1 ; Score® (difenoconazole) (250 g.L-1 ) na dose de 0,3 mL.L -1 ; Amistar® (azoxistrobina) (500 g.kg-1 ) na dose de 0,21 g.L -1 . A testemunha não recebeu tratamento. As aplicações foram realizadas a cada sete dias, durante cinco semanas e as avaliações foram realizadas semanalmente, durante quatro semanas, sendo a primeira realizada na terceira semana após a implantação do experimento. As avaliações foram baseadas na escala de notas variando de zero a cinco, onde zero corresponde a planta não infectada e cinco à infecção superior a 40% da área foliar. Os produtos biológicos Serenade na dose 7,5 mL.L -1 e Sonata, nas doses 2,5 e 7,5 mL.L -1 , aplicados em pulverização preventiva, controlaram satisfatoriamente o oídio em meloeiro quando comparado aos produtos padrões Amistar® e Score® . Os mesmos, quando devidamente registrados para a cultura, podem ser recomendados para o controle do oídio