Severe Outbreak of Dry Core Rot in Apple Fruits cv. Fuji Caused by Kalmusia variispora During Pre-harvest in Maule Region, Chile.

Apple (Malus × domestica) is an important fruit crop in Chile, with a cultivated area of 32,313 ha concentrated (63%) in Maule Region (35°25' S). Unusual core rot on 'Fuji' apples was observed at harvest in a commercial orchard in Curicó, Maule Region, with an incidence ranged between 22 to 35% in 2018 and 2019. Previously, in 2017, an incidence of 30% was estimated on 'Fuji' fruits destined to the Asian market. Internal decay symptoms consisted of dry, corky light to dark-brown tissue, within the seed locules initially. In moderate to severe cases, the necrotic lesion progresses deeper into the mesoderm. External symptoms were quite subtle, and typically, the disease goes unnoticed. However, infected fruit ripen earlier. Small pieces (2-3 mm) from the internal lesion margin of symptomatic apples (n = 50) were placed on Potato Dextrose Agar (PDA) (2%) and incubated at 20°C for 10 days. Pure cultures (n = 41) were obtained and transferred to Malt Extract Agar (MEA) (2%). Colonies on MEA produced an even to slight undulating buff margin with white woolly aerial mycelium, and immerse ochreous in the center, changing gray to olivaceous aerial mycelium with age. On the underside, colonies were umber and buff in the center and margin, respectively. After 10 days, numerous densely aggregate dark-brown mature pycnidia were observed. Aseptate conidia were subglobose to cylindrical, straight, and sometimes curved with rounded at both ends, that was initially hyaline to pale olive, thin, smooth wall with mean dimensions of (2.9-) 3.4 (-4.4) x (1.5-) 1.8 (2.2) µm (n=50). Based on morphology, the fungus was identified as Kalmusia variispora (Verkley et al. 2014). The internal transcribed space (ITS), portion of ß-tubulin (TUB), and large subunits of the nuclear ribosomal RNA (LSU) loci were used for molecular identification, using primers ITS4/ITS5, Bt2a/Bt2b, and LR0R/LR5 (Ariyawansa et al., 2014). BLAST searches indicated 100% identity with K. variispora (ex-type CBS 121517). The maximum parsimony phylogenetic analysis placed Chilean isolates in the K. variispora clade. Sequences were deposited in GenBank (OL711706 to OL711709, OL739499 to OL739502 and OL711710 to OL711713 for ITS, TUB and LSU, respectively). Pathogenicity tests were conducted using four K. variispora isolates. 'Fuji' apples (n = 20) were surface disinfested (75% ethanol, 30 s) and then wounded and inoculated with conidial suspension (50 L of 106 conidia/mL) deposited in the middle and into the core region using a sterile fine-tipped micropipette. Additionally, 20 one-year dormant rooted cuttings 'Fuji' and 'Cripps Pink' were pruned and immediately inoculated on the pruning wound. An equal number of apples and rooted cuttings treated with sterile water were used as controls. The experiments were repeated once. All inoculated fruits developed lateral lesions (22 to 37 mm) and dry core rot (18 to 36 mm) symptoms identical to those described in the original outbreak, after 20 days at 20°C in a commercial packing box. The inoculated cuttings produced canker lesions of 10 to 21 mm in length, and dieback symptoms were observed after 3 months. No symptoms were observed on the negative controls. Koch's postulates were fulfilled by 100% reisolating K. variispora. Previously, Alternaria spp. have been reported as the primary pathogen associated with moldy core and dry core rot of apples worldwide (McLeod et al., 2014) and in Chile (Elfar et al., 2018). However, Kalmusia spp. have been associated with dry core rot in apples (McLeod et al., 2014) and have been isolated from canker symptoms on apples in Chile (Díaz et al. 2021). To our knowledge, this is the first report of a severe outbreak of K. variispora causing dry core rot in apples in Chile and worldwide.

Long-term cryopreservation of Lentinus crinitus strains by wheat grain technique.

Lentinus crinitus (Basidiomycota: Polyporales) is a saprophytic fungus with biotechnological importance described more than 20 years ago. However, there are few studies on the long-term preservation of this basidiomycete. Cryopreservation is a long-term storage technique that reduces the metabolic activity of microorganisms, but its success depends on the adjustment of the freezing process, the cryoprotectants, and the protective substrates for each species. This study aimed to assess the mycelial viability and genetic stability of L. crinitus strains cryopreserved at -86 °C for two years by the wheat grain technique using different cryoprotectants and freezing methods. Three strains of L. crinitus (U9-1, U13-5, and U15-12) were subjected to different concentrations and types of cryoprotectants (dimethyl sulfoxide, glycerol, glucose, and sucrose), freezing methods such as immediate freezing from 25 to -86 °C and progressing freezing from 25 to -86 °C in a freezing container with isopropyl alcohol to control the rate of cell freezing at -1 °C min-1, protective substrate (wheat grain and 2% malt extract agar), and cryopreservation period (1, 6, 12, and 24 months). After thawing, samples were evaluated for mycelial viability, time to mycelial recovery, mycelial stability, and genetic stability of the fungus. All techniques achieved effective cryopreservation at -86 °C, mainly with the wheat grain technique. All cryoprotectants (3.5% glycerol, 1.5% dimethyl sulfoxide, 25% sucrose, and 5% glucose), freezing methods (immediate and gradual), and protective substrate (wheat grain and malt extract agar) were effective for cryopreservation of the three L.crinitus strains in an ultra-low temperature freezer for two years. Mycelial viability, mycelial stability, and genetic stability of the fungus were not affected after two-year cryopreservation, evidencing the robustness of the long-term cryopreservation technique and the fungus.

Desafios na gestão em saúde frente a pandemia de covid-19: relato de experiência / Challenges in health management in front of the COVID-19 pandemic: experience report

Objetivo: descrever a experiência vivenciada pela gerência de enfermagem durante a pandemia de COVID-19 em uma instituição de saúde não referenciada para a doença. Métodos: estudo descritivo, do tipo relato de experiência, que apresenta os desafios enfrentados e atividades desenvolvidas pela gerência de Enfermagem em uma instituição pública de saúde do estado do Ceará de atenção secundária, não referenciada para a COVID-19, no período de março a maio de 2020. Síntese de dados: Planejamento e organização do serviço: realizou-se na unidade de saúde, treinamento prévio para diminuir riscos de contaminação, afastou-se grupos de risco, estabeleceu-se fluxos para situações pontuais de atendimento a doença, cirurgias eletivas foram canceladas por decretos estaduais. Desafios enfrentados nos processos de trabalho da equipe: porcentagem relevante de profissionais adquiriram a doença, a saúde mental dos profissionais também foi consideravelmente afetada, as escalas de serviço previstas não puderam ser cumpridas em sua totalidade e situações de estresse perante atendimentos inesperados a COVID-19 foram percebidos. Conclusão: conclui-se que, mesmo se tratando de um hospital secundário não referenciado para a COVID-19, independente de estratégias pré-estabelecidas pela gerência de Enfermagem, houve dificuldades na adaptação dos profissionais ao período de pandemia e ocorreram situações que exigiram rapidez e preparo para intervenções.

Objective: to describe the experience of nursing management during the COVID-19 pandemic in a health institution not referred to the disease. Method: a descriptive, experience report type study that presents the challenges faced and activities developed by the nursing management in a public health institution in the state of Ceará for secondary care, not referenced to COVID-19, in the period from March to May 2020. Data synthesis: Planning and organization of the service: training was carried out at the health unit, prior training to reduce risks of contamination, risk groups were removed, flows were established for specific situations of care for the disease, elective surgeries were canceled for state decrees. Challenges faced in the team's work processes: a relevant percentage of professionals acquired the disease, the mental health of the professionals was also considerably affected, the scheduled service schedules could not be fully complied with and situations of stress due to unexpected visits to COVID-19 were perceived. Conclusion: it is concluded that, even though it is a secondary hospital not referenced to COVID-19, regardless of pre-established strategies by the Nursing management, there were difficulties in adapting professionals to the pandemic period and situations that required speed and preparation for interventions.