First Report of Banana bract mosaic virus in 'Cavendish' Banana in Ecuador.

Banana bract mosaic virus (BBrMV), a member of the genus Potyvirus, family Potyviridae, is the causal agent of bract mosaic disease. The disorder has been considered a serious constraint to banana and plantain production in India and the Philippines, where the virus was first identified (3). To date, the presence of BBrMV has been reported only in a few banana-growing countries in Asia (3). In the Americas, BBrMV has been detected by ELISA tests in Colombia only (1). The efficient spread of BBrMV through aphids and vegetative material increases the quarantine risk and requires strict measures to prevent entrance of the virus to new areas. In Ecuador-the world's number one banana exporter-the banana industry represents the main agricultural income source. Thus, early detection of banana pathogens is a priority. In June of 2012, mosaic symptoms in bracts and bunch distortion of 'Cavendish' banana were observed in a commercial field in the province of Guayas, Ecuador. Leaves from 35 symptomatic plants were tested for Cucumber mosaic virus (CMV), Banana streak virus (BSV), and BBrMV using double antibody sandwich ELISA kits from Adgen (Scotland, UK). Twenty-one plants tested positive for BBrMV but not for CMV or BSV. In order to confirm the ELISA results, fresh or lyophilized leaf extracts were used for immunocapture reverse transcription (IC-RT)-PCR. In addition, total RNA was extracted from the ELISA-positive samples and subjected to RT-PCR. The RT reactions were done using both random and oligo dT primers. Several sets of primers, flanking conserved regions of the virus coat protein (CP), have been used for PCR-detection of BBrMV (2,3,4). The Ecuadorian BBrMV isolate was successfully detected by three primer sets with reported amplification products of 324, 280, and 260 nucleotides long, respectively (3,4). Amplification products of the expected size were purified and sequenced. All the nucleotide sequences obtained from 20 PCR-positive symptomatic plants were 100% identical between each other. However, 99% identity was observed when PCR products from the Ecuadorian isolate were compared with the corresponding fragment of a BBrMV isolate from the Philippines (NCBI Accession No. DQ851496.1). PCR products of the Ecuadorian isolate, amplified by the different CP primers described above, were assembled into a 408-bp fragment and deposited in the NCBI GenBank (KC247746). Further testing confirmed the presence of BBrMV in symptomatic plants from four different provinces. To our knowledge, this is the first report of BBrMV in Ecuador and the first BBrMV partial nucleotide sequence reported from the Americas. It is worth mentioning that primer set Bract 1/Bract 2, which amplifies a 604-bp product (2), was not effective in detecting the Ecuadorian isolate. It is hypothesized that nucleotide variation at the reverse primer site is the cause of the lack of amplification with this primer set, since the forward primer is part of the sequenced product and no variation was found. Sequencing of the entire CP region is underway to conduct phylogenetic analysis and determine genetic relationships across several other BBrMV isolates. References: (1) J. J. Alarcon et al. Agron 14:65, 2006. (2) M. F. Bateson and J. L. Dale. Arch. Virol 140:515, 1995. (3) E. M. Dassanayake. Ann. Sri Lanka Dept. Agric. 3:19, 2001. (4) M. L. Iskra-Caruana et al. J. Virol. Methods 153:223, 2008.

First Report of Raspberry bushy dwarf virus in Andean Blackberry (Rubus glaucus) in Central Ecuador.

During the past two decades, several viruses have been identified from Rubus spp. in wild and commercial plantings around the world (2). In Ecuador, approximately 14 tons of blackberries are produced each year from an estimated area of 5,500 ha. In 2012, a preliminary survey was conducted to determine the presence of RNA viruses in Rubus glaucus, the most prevalent blackberry in Ecuador. Fifteen plants showing leaf mottling and severe mosaic were leaf-sampled from each of five different fields in Azuay Province. A total of 12 pooled samples of 20 g were obtained from the collected symptomatic tissue and used for dsRNA extraction using a cellulose-based protocol for detection of RNA viruses in plants (3). Three dsRNA segments of approximately 5 kbp, 2 kbp, and 900 bp were observed from all 12 dsRNA preparations. The dsRNA was heat-denatured and used as template for the generation of cDNA library using the universal random primer 5'-GCCGGAGCTCTGCAGAATTCNNNNNN-3', for reverse transcription (RT), and the anchor primer 5'-GCCGGAGCTCTGCAGAATTC-3'for PCR as described (1). The PCR products were cloned using a StrataClone Kit (Agilent, CA) and sequenced (Macrogen, Korea). Sequence analysis revealed the presence of Raspberry bushy dwarf virus (RBDV), a pollen-borne Idaeovirus naturally found in several Rubus spp. worldwide. Approximately 120 RBDV sequences obtained from the Ecuadorean isolate were assembled into two contigs belonging to RNA1 and RNA2. Both sequences were re-confirmed by RT-PCR using specific primers. Partial sequences were assigned GenBank Accessions KC315894, KC315893, and KC315892 for the replicase, MP and CP, respectively. Furthermore, BLAST searches showed that the nucleotide sequence corresponding to the replicase was 95% similar to an isolate from the resistance breaking R15 strain (S51557.1), whereas the MP and CP nucleotide sequences were up to 98% similar to a Slovenian isolate (EU796088.1). Primers designed to amplify a 427-bp portion of the CP were used to detect RBDV from four blackberry plantings in two distant production areas: Ambato in Tungurahua Province and Paute in Azuay Province. Leaf mottling and severe mosaic was observed in 90% of blackberry fields in those two locations. Leaf samples (n = 90) were randomly collected from both symptomatic and asymptomatic plants in each location. In Ambato, RBDV was detected in 50% and 40% of symptomatic and asymptomatic plants, respectively. In Paute, RBDV was present in 70% of symptomatic plants and 29% of asymptomatic plants. The presence of RBDV in asymptomatic plants suggests the virus might not be the sole causal agent of the disorder. Further studies are needed to determine the role of RBDV in the observed symptoms, since virus complexes responsible for increased severity of symptoms have been commonly reported in Rubus spp. (4). R. glaucus is native to the tropical highlands (from Ecuador to Mexico) and differs from blackberries commercially grown in the United States and Europe. Therefore, RBDV-induced symptoms reported in blackberry grown in the United States and Europe may not be extrapolated to the Andes berry. To the best of our knowledge, this is the first report of RBDV from blackberry in Ecuador. References: (1) P. Froussard. Nucleic Acids Res. 20:2900, 1992. (2) R. R. Martin et al. Plant Dis. 97:168, 2013. (3). T. J. Morris and J. A. Dodds. Phytopathology 69:854. 1979. (4) D. F. Quito-Avila et al. J. Virol. Methods 179:38, 2012.

In-hospital mortality associated factors in elderly patients with invasive mechanical ventilation in the emergency department.

AIMS: To identify factors associated with in-hospital mortality, to estimate the intubation rate and to describe in-hospital mortality in patients over 65 years old with invasive mechanical ventilation (IMV) in the emergency department (ED). METHODS: Retrospective cohort study of patients over 65 years old, who were intubated in an ED of a high complexity hospital between 2016 and 2018. Demographic data, comorbidities, and severity scores on admission were described. Bivariate and multivariate analyses were performed with logistic regression according to mortality and possible confounders. RESULTS: A total of 285 patients with a mean age of 80 years required IMV in the emergency department, for a median of 3 days, and with a mean APACHE II score of 20 points of severity. The IMV rate was .48% (95% CI .43-.54), and 55.44% (158) died. Mortality-associated factors after age and sex adjustment were stroke (OR 2.13; 95% CI 1.21-3.76), chronic kidney failure, (OR 4.,38; 95% CI 1.91-10.04), Charlson index (OR 1.19; 95% CI 1.02-1.38), APACHE II score (OR 1.07; 95% CI 1.02-1.12), and SOFA score (OR 1.14; 95% CI 1.03-1.27). DISCUSSION: Our IMV rate was lower than that stated by Johnson et al. in the United States in 2018 (.59%). In-hospital mortality in our study exceeded that predicted by the APACHE II score (40%) and SOFA (33%). However it was consistent with that reported by Lieberman et al. in Israel and Esteban et al. in the United States. CONCLUSIONS: Although the IMV rate was low in the ED, more than half the patients died during hospitalization. Pre-existing cerebrovascular and renal diseases and high results in the comorbidities index and severity scores on admission were independent factors associated with in-hospital mortality.

[Acute lumbalgia frequency and treatment at a private hospital emergency plant]. / Frecuencia de lumbalgia aguda y su tratamiento en la central de emergencias de un hospital privado.

INTRODUCTION: Acute back pain (LA) is a considerable problem in any care setting. The Emergency Department (ED) intended to provide immediate, timely and effective care. Our objective was to estimate the prevalence and describe the usual care process. MATERIAL AND METHODS: To estimate prevalence, a random sample was included of 5,333 consultations admitted in ED at hospital XX between January and December 2016. LA cases were defined according to the diagnosis at arrival. A retrospective cohort with all LA cases was used to describe complementary studies and therapeutic behaviors established in ED or within the subsequent 30 days outpatient follow-up. RESULTS: LA represented 4.37% (95% CI: 3.83-4.95%) of consultations. A total of 1,096 cases of LA were included: mean age of 53.26 ± 19.85 years, 55.93% (613) female sex and few comorbidities. The median attention time was 3.28 hours and 6.20% was admitted. 70% (778) received treatment in CEA, being the most used administration routes: intravenous (61.86%), and intramuscular (20.62%). The analgesics used were: 55.38% corticosteroids, 55.02% NSAIDs, 34.85% opioids. The complementary studies requested during ED or until 30 days after discharge, were: 52.19% laboratory, 47.17% radiography, 28.38% RM, 10.77% tomography, 9.12% interventionism. CONCLUSIONS: This study provides epidemiological data of LA in Argentina and demonstrates the need to promote the rational use of resources.

INTRODUCCIÓN: La lumbalgia aguda (LA) es un problema en diferentes ámbitos de atención. La Central de Emergencias de Adultos (CEA) intenta brindar atención inmediata, especializada y eficaz. Nuestro objetivo fue estimar la prevalencia y describir el proceso de atención habitual. MATERIAL Y MÉTODOS: Para la estimación de prevalencia, se incluyó una muestra aleatoria de 5,333 consultas admitidas en CEA de un solo hospital entre Enero y Diciembre de 2016. Se definió como numerador a los casos de LA según diagnóstico de triaje al ingreso. Una cohorte retrospectiva con todos los casos de LA se utilizó para describir estudios complementarios y conductas terapéuticas instaurados en CEA, con seguimiento ambulatorio posterior. RESULTADOS: La lumbalgia aguda representó 4.37% (IC 95%: 3.83-4.95%) de las consultas. Se incluyeron 1,096 casos de LA: edad media de 53.26 ± 19.85 años, 55.93% (613) sexo femenino y escasas comorbilidades. La mediana de tiempo de atención fue de 3.28 horas y se internó 6.20%, 70% (778) recibió tratamiento en CEA, las vías de administración más utilizadas fueron: endovenosa (61.86%) e intramuscular (20.62%). Los analgésicos utilizados fueron: 55.38% corticoides, 55.02% AINE, 34.85% opiáceos. Los estudios complementarios solicitados en CEA o hasta los 30 días postegreso fueron: 52.19% laboratorio, 47.17% radiografía/s, 28.38% resonancia/s, 10.77% tomografía/s, 9.12% intervencionismo/s. CONCLUSIONES: Este estudio aporta datos epidemiológicos sobre LA y evidencia la necesidad de fomentar el uso racional de los recursos.

[Assessment of an educational intervention regarding blood cultures as process indicators in an emergency department in Argentina]. / Evaluación del impacto de una intervención educativa en indicadores de gestión de un servicio médico de urgencias de Argentina.

OBJECTIVE: The aim of this study was to describe 2 process indicators related to taking blood cultures (BC) in an Adult Emergency Department of a tertiary university hospital in Buenos Aires,and to describe the changes after a series of educational activities for health professionals was implemented during May 2016 as regards the appropriate indication of BC and the proper collection technique. MATERIALS AND METHODS: A retrospective cohort study was designed to assess its effectiveness, which consecutively included all patients admitted during 2015-2016. The BC request rate was used as a process indicator, and the percentage of contaminated BCs and the true positives rate were used as quality indicators. Both were measured monthly and prospectively during the period of study. RESULTS: The annual adjusted rate of BC requests was 4.9% (95% CI 4.8-5) in 2015 and 2.9% (95% CI 2.8-2.9) in 2016. The rate of false positive (contaminated) BCs was 4.5% in 2015 and 4.3% after the educational intervention. The true positive BCs were 8.3% in 2015 and 12% post-intervention. CONCLUSIONS: These findings prove how important and effective the educational interventions are.