Re-emergence of rabies virus maintained by canid populations in Paraguay.

Paraguay has registered no human cases of rabies since 2004, and the last case in dogs, reported in 2009, was due to a variant maintained in the common vampire bat "Desmodus rotundus". In 2014, a dog was diagnosed as positive for rabies with aggression towards a boy and all required measures of control were successfully adopted. Epidemiological investigation revealed that the dog was not vaccinated and had been attacked by a crab-eating fox, "zorro" (Cerdocyon thous). The sample was diagnosed by the Official Veterinary Service of the Country and sent to the Center on Rabies Research from the University of São Paulo, Brazil, for antigenic and genetic characterization. A second sample from a dog positive for rabies in the same region in 2015 and 11 samples from a rabies outbreak from Asuncion in 1996 were also characterized. The antigenic profile of the samples, AgV2, was compatible with one of the variants maintained by dogs in Latin America. In genetic characterization, the samples segregated in the canine (domestic and wild species)-related group in an independent subgroup that also included samples from Argentina. These results and the epidemiology of the case indicate that even with the control of rabies in domestic animals, the virus can still circulate in wildlife and may be transmitted to domestic animals and humans, demonstrating the importance of continuous and improved surveillance and control of rabies, including in wild species, to prevent outbreaks in controlled areas.

Phylogenetic Diversity of Koala Retrovirus within a Wild Koala Population.

Koala populations are in serious decline across many areas of mainland Australia, with infectious disease a contributing factor. Koala retrovirus (KoRV) is a gammaretrovirus present in most wild koala populations and captive colonies. Five subtypes of KoRV (A to E) have been identified based on amino acid sequence divergence in a hypervariable region of the receptor binding domain of the envelope protein. However, analysis of viral genetic diversity has been conducted primarily on KoRV in captive koalas housed in zoos in Japan, the United States, and Germany. Wild koalas within Australia have not been comparably assessed. Here we report a detailed analysis of KoRV genetic diversity in samples collected from 18 wild koalas from southeast Queensland. By employing deep sequencing we identified 108 novel KoRV envelope sequences and determined their phylogenetic diversity. Genetic diversity in KoRV was abundant and fell into three major groups; two comprised the previously identified subtypes A and B, while the third contained the remaining hypervariable region subtypes (C, D, and E) as well as four hypervariable region subtypes that we newly define here (F, G, H, and I). In addition to the ubiquitous presence of KoRV-A, which may represent an exclusively endogenous variant, subtypes B, D, and F were found to be at high prevalence, while subtypes G, H, and I were present in a smaller number of animals. IMPORTANCE: Koala retrovirus (KoRV) is thought to be a significant contributor to koala disease and population decline across mainland Australia. This study is the first to determine KoRV subtype prevalence among a wild koala population, and it significantly expands the total number of KoRV sequences available, providing a more precise picture of genetic diversity. This understanding of KoRV subtype prevalence and genetic diversity will be important for conservation efforts attempting to limit the spread of KoRV. Furthermore, KoRV is one of the only retroviruses shown to exist in both endogenous (transmitted vertically to offspring in the germ line DNA) and exogenous (horizontally transmitted between infected individuals) forms, a division of fundamental evolutionary importance.

Phylogeography and evolutionary history of rodent-borne hantaviruses.

Hantavirus (Family Bunyaviridae) are mostly associated to rodents and transmitted to man by inhalation of aerosolized infected excreta of these animals. The human infection by hantaviruses can lead to severe diseases such as hemorrhagic fever with renal syndrome (HFRS) in Asia and Europe, and pulmonary syndrome (HPS) in the Americas. To determine the origin, spreading and evolutionary dynamics of rodent-borne hantaviruses, 190 sequences of nucleoprotein (N) of hantaviruses identified in 30 countries, from 1985 to 2010, were retrieved from the GenBank and analyzed using the BEAST program. Our evolutionary analysis indicates that current genetic diversity of N gene of rodent-borne hantaviruses probably was originated around 2000 years ago. Hantavirus harbored by Murinae and Arvicolinae subfamilies, probably, were originated in Asia 500-700 years ago and later spread toward Siberia, Europe, Africa and North America. Hantavirus carried by Neotominae subfamily, probably, emerged 500-600 years ago in Central America and spread toward North America. Finally, hantaviruses associated to Sigmodontinae occurred in Brazil 400 years ago and were, probably, originated from Neotominae-associated virus from northern South America. These data offer subsidies to understand the time-scale and worldwide dissemination dynamics of rodent-borne hantaviruses.

Viral load of human bocavirus-1 in stools from children with viral diarrhoea in Paraguay.

Since their discovery, four species of human bocavirus (HBoV) have been described in patients with respiratory and gastrointestinal diseases. However, a clear causal association between HBoV-1 and gastroenteritis has not been demonstrated. In this study, we describe the detection and quantification of HBoV-1 in stools from children with acute non-bacterial gastroenteritis using quantitative polymerase chain reaction. HBoV-1 genome was detected in 10.6% of stools with frequent association with rotavirus and norovirus. The median of HBoV-1 viral load was 1.88 × 104 genome/ml, lower than previously shown in secretions of patients with respiratory infections, without any obvious association between high viral load and presence of HBoV as single agent. Thus, although HBoV-1 was frequently detected in these patients, there is no clear causal association of this agent with diarrhoea. Indeed, HBoV-1 DNA in stools of patients with gastroenteritis without respiratory symptoms may be a remnant of previous infections or associated with prolonged shedding of virus in the respiratory or digestive tracts.

Prevalence of adverse events in the hospitals of five Latin American countries: results of the 'Iberoamerican Study of Adverse Events' (IBEAS).

BACKGROUND: Interest in patient safety (PS) is growing exponentially, fuelled by epidemiological research unveiling the extent of unsafe care. However, there is little information about the frequency of harm in developing and transitional countries. To address this issue, the authors performed a study known as the Iberoamerican Adverse Event Study, through a collaborative between the governments of Argentina, Colombia, Costa Rica, Mexico and Peru, the Spanish Ministry of Health, Social Policy and Equality, the Pan American Health Organization and the WHO Patient Safety. METHODS: The study used a cross-sectional design, involving 58 hospitals in the five Latin American countries, to measure the point prevalence of patients presenting an adverse event (AE) on the day of observation. All inpatients at the time of the study were included. RESULTS: A total of 11 379 inpatients were surveyed. Of these, 1191 had at least one AE that the reviewer judged to be related to the care received rather than to the underlying conditions. The estimated point prevalence rate was 10.5% (95% CI 9.91 to 11.04), with more than 28% of AE causing disability and another 6% associated with the death of the patient. Almost 60% of AE were considered preventable. CONCLUSIONS: The high rate of prevalent AE found suggests that PS may represent an important public-health issue in the participating hospitals. While new studies may be needed to confirm these results, these may already be useful to inspire new PS-improvement policies in those settings.

Diseño del estudio IBEAS: prevalencia de efectos adversos en hospitales de Latinoamérica / IBEAS design: adverse events prevalence in Latin American hospitals

Objetivos. Describir las características y metodología del estudio IBEAS: prevalencia de efectos adversos en hospitales de Latinoamérica, que persigue realizar una aproximación a la magnitud, trascendencia e impacto de los eventos adversos (EA); identificar áreas y problemas prioritarios de la seguridad del paciente; incrementar la masa crítica de profesionales involucrados en la seguridad del paciente e incorporar a la agenda de los países objetivos y actividades para mejorar la seguridad del paciente. Método. Estudio sobre la seguridad de los pacientes. Ámbito: 35 hospitales de 5 países: Argentina, Colombia, Costa Rica, México y Perú, a través del análisis de los efectos adversos identificados. Diseño de prevalencia utilizando la revisión de la historia clínica. Resultados. Se describen las implicaciones en el uso del diseño transversal en el estudio de EA, tanto en los recursos necesarios como en la validez interna y en la utilidad para la gestión de riesgos sanitarios. Conclusiones. El diseño transversal es eficiente en tiempo y recursos y fácil de realizar, y aunque no permite estudiar la totalidad del episodio de hospitalización, ha demostrado ser capaz para sostener un sistema de vigilancia. Debido a un posible sesgo de supervivencia, los EA que ocasionen un ingreso se verán sobrerrepresentados, y también aquellos relacionados con la infección nosocomial o los que sean difícilmente identificables si no se ve al paciente. La comunicación con el personal de planta (mientras el paciente está hospitalizado) favorece el juicio de la causalidad del efecto adverso y de su evitabilidad(AU)

Objectives. To describe the methodological characteristics of the IBEAS study: adverse events prevalence in Latin American hospitals, with the aim of analysing the magnitude, significance and impact of adverse events (AE); to identify the main problems associated with patient safety AE; to increase the capacity of professionals involved in patient safety; and the setting up of patient safety agendas in the participating countries. Methods. A patient safety study launched in 35 Latin American hospitals through the analysis of AE in 5 countries: Argentina, Colombia, Costa Rica, Mexico and Peru, using a cross-sectional study using a review of clinical records as the main method. Results. The implications of using a cross-sectional design when studying AE are described, in terms of resources required, internal validity and usefulness related to risk management. Conclusions. The cross-sectional design seems an efficient methodology in terms of time and resources spent, as well as being easy to carry out. Although the cross-sectional design does not review the all hospital episodes, it is able to provide a reliable estimate of prevalence and to support a surveillance system. Because of a possible survival bias, it is likely that the AE which led to hospital admissions will be overestimated, as well as the health related infections or those adverse events which are difficult to identify if the patient is not examined (e.g. contusions). Communication with the ward staff (if the patient is still hospitalised) help in finding the causality and their prevention(AU)

[IBEAS design: adverse events prevalence in Latin American hospitals]. / Diseño del estudio IBEAS: prevalencia de efectos adversos en hospitales de Latinoamérica.

OBJECTIVES: To describe the methodological characteristics of the IBEAS study: adverse events prevalence in Latin American hospitals, with the aim of analysing the magnitude, significance and impact of adverse events (AE); to identify the main problems associated with patient safety AE; to increase the capacity of professionals involved in patient safety; and the setting up of patient safety agendas in the participating countries. METHODS: A patient safety study launched in 35 Latin American hospitals through the analysis of AE in 5 countries: Argentina, Colombia, Costa Rica, Mexico and Peru, using a cross-sectional study using a review of clinical records as the main method. RESULTS: The implications of using a cross-sectional design when studying AE are described, in terms of resources required, internal validity and usefulness related to risk management. CONCLUSIONS: The cross-sectional design seems an efficient methodology in terms of time and resources spent, as well as being easy to carry out. Although the cross-sectional design does not review the all hospital episodes, it is able to provide a reliable estimate of prevalence and to support a surveillance system. Because of a possible survival bias, it is likely that the AE which led to hospital admissions will be overestimated, as well as the health related infections or those adverse events which are difficult to identify if the patient is not examined (e.g. contusions). Communication with the ward staff (if the patient is still hospitalised) help in finding the causality and their prevention.

Detección del virus influenza pandémica A (H1N1) en Paraguay 2009, y amplificación de genes hemaglutinina y neuraminidasa / Detection of pandemic influenza A (H1N1) virus in Paraguay in 2009, and amplification of the hemagglutinin and neuraminidase genes

Introducción: El virus de influenza pandémica A (H1N1), cuya circulación se inició en abril del año 2009 en México y Estados Unidos, se constituyó en el último virus pandémico desde los casos detectados en Hong Kong en 1968. El genoma del virus de influenza A está formado por 8 segmentos ARN de cadena simple (polaridad negativa), que codifican para 10 proteínas. Los genes hemaglutinina y neuraminidasa codifican para dos proteínas de superficie y son los utilizados en los análisis de variabilidad genética. Objetivos: a) Detectar la circulación del virus pandémico en pacientes con sospecha clínica de infección por influenza, y b) Diseñar una estrategia para amplificar de forma completa los genes hemaglutinina y neuraminidasa. Materiales y Métodos: Fueron analizados por Real-Time RT-PCR (transcripción reversa y reacción en cadena de la polimerasa en tiempo real) un total de 181 muestras de hisopado faríngeo, colectadas o remitidas al Hospital de Clínicas, del 6 de agosto al 11 de octubre de 2009. Para el diseño de amplificación de los genes hemaglutinina y neuraminidasa, se han utilizado herramientas bioinformáticas y reacción en cadena de la polimerasa. Resultados: Del total de muestras analizadas, 27 (14.9 %) dieron resultado positivo para el nuevo virus pandémico. Por otra parte, la amplificación completa de ambos genes proporcionó los resultados esperados: 1678-pares de bases (pb) para la hemaglutinina, y 1427-pb para la neuraminidasa. Conclusiones: La implementación de esta tecnología de amplificación permitirá posteriormente la secuenciación de estos genes a fin de determinar las variaciones genéticas del virus que podrían tener un impacto en la salud humana.

Introduction: The pandemic influenza A (H1N1) virus, whose circulation was detected in April 2009 in Mexico and the United States, is the latest pandemic virus since the cases reported in Hong Kong in 1968. The genome of the influenza A virus consists of 8 segments of single-stranded RNA of negative polarity, coding for 10 proteins. The hemagglutinin and neuraminidase genes encode for two surface proteins and are used in the analysis of genetic variability. Objectives: a) to detect circulation of the pandemic virus in patients with clinical suspicion of influenza infection and b) design a strategy to fully amplify the hemagglutinin and neuraminidase genes.Materials and Methods: A total of 181 pharyngeal swabs were collected and sent to the Hospital de Clínicas for analysis using Real-Time RT-PCR (reverse transcription and polymerase chain reaction in real time) between 6 August and 11 October 2009. To design the amplification of hemagglutinin and neuraminidase genes, we used bioinformatic tools and polimerase chain reaction. Results: Of the samples analyzed, 27 (14.9%) were positive for the new pandemic virus. Moreover, the complete amplification of both genes provided the expected results: 1678-base pairs (bp) for the hemagglutinin, and 1427-bp for neuraminidase. Conclusions: The use of this technology for amplification will eventually allow sequencing to identify genetic variations of the virus that could have an impact on human health.

Study of external root resorption during orthodontic treatment in root filled teeth compared with their contralateral teeth with vital pulps.

AIM: To determine whether root filled teeth and those with vital pulps exhibit a similar degree of external root resorption (ERR) as a consequence of orthodontic treatment. METHODOLOGY: The study sample consisted of 77 patients, with a mean age of 32.7 +/- 10.7 years, who had one root filled tooth before completion of multiband/bracket orthodontic therapy for at least 1 year. For each patient, digital panoramic radiographs taken before and after orthodontic treatment were used to determine the proportion of external root resorption (PRR), defined as the ratio between the root resorption in the root filled tooth and that in its contralateral tooth with a vital pulp. The student's t-test, anova and logistic regression analysis were used to determine statistical significance. RESULTS: The mean PRR was 1.00 +/- 0.13, indicating that, in the total sample, there were no significant differences in root resorption in the root filled teeth and their contralateral teeth with vital pulps. Multivariate logistic regression analysis suggested that PRR was significantly greater in incisors (P = 0.0014; odds ratio = 6.2885, C.I. 95% = 2.0-19.4), compared to other teeth, and in women (P = 0.0255; odds ratio = 4.2, C.I. 95% = 1.2-14.6), compared to men. CONCLUSIONS: There was no significant difference in the amount or severity of external root resorption during orthodontic movement between root filled teeth and their contralateral teeth with vital pulps.

Sequence and phylogenetic analysis of the VP4 gene of human rotaviruses isolated in Paraguay.

Nucleotide and amino acid analyzes of the VP4 gene of human rotaviruses isolated both in Paraguay and worldwide were carried out in order to increase our knowledge about the complex pattern of evolution of this virus in nature. Paraguayan strains bearing the P[8] genotype were grouped in the lineages P[8]-1, P[8]-2, and P[8]-3. Regardless of the year of detection, all of the G4 and G9 strains were related to lineage P[8]-3, whereas the G1 strains were related to the three lineages detected in Paraguay; this fact reinforces the notion of the existence of constraints within specific populations of rotavirus strains except for the G1 strains. In addition, we propose a phylogenetic classification for the P[4] strains in five different lineages (i.e. P[4]-1 to P[4]-5). The findings presented in this paper reinforce the importance of a continuous surveillance of rotavirus strains in order to predict the possible variants that will circulate in a country, and ultimately improve current vaccination programs.

Productividad científica del Paraguay en el área de biomedicina: un análisisbibliométrico / Scientific productivity of Paraguay in the field of biomedicine: a bibliometric

Para medir la producción científica de un país es usual que se contabilice la cantidad y calidad deartículos científicos publicados por sus investigadores. Con el objetivo de identificar la cantidad depublicaciones de autoras paraguayos indexadas en la base de datos PubMed en los últimos 20 años,determinar su distribución temporal y el aporte de las diferentes instituciones al desarrollo científicodel Paraguay; se buscaron las investigaciones publicadas en el periodo 1986-2005 en PubMed, pormedio de frases unidas por conectores booleanos. El 67% (71/106) de los artículos hallados fueronpublicados en los últimos 10 años (1996-2005), indicando el fortalecimiento de las cienciasbiomédicas. Las tres instituciones paraguayas con mayor número de publicaciones pertenecen a laUniversidad Nacional de Asunción y son el Instituto de Investigaciones en Ciencias de la Salud(IICS), Facultad de Ciencias Médicas(FCM)y Facultad de Ciencias Químicas (FCQ) con el 39,6%,16% y 15% de todas las publicaciones, respectivamente. Es destacable el gran número depublicaciones de las dos primeras instituciones en los últimos 10 años, registrando 76% y 88% desus publicaciones, en revistas indexadas al Medline. Finalmente, 82% de los artículos fueronpublicados en revistas que poseen factor de impacto (intervalo 0,191-4,927), dando un promediode 1,932. Aunque es un valor bajo comparándolo con el de países limítrofes, es aceptable para unpaís en crecimiento científico. Por otra parte, pese al crecimiento lineal del número de laspublicaciones anuales de autores paraguayos, la producción científica en biomedicina en elParaguay es muy baja. Por lo tanto, creemos que es importante fomentarla, ya que la excelenciaeducativa y la salud pública son indispensables para el crecimiento socio-económico del Paraguay

Electroferotipos cortos de rotavirus detectados en adultos con diarrea en Paraguay / Short electropherotypes of rotavirus detected in adults with diarrhea in Paraguay

La presencia de rotavirus en adultos ha sido subestimada por mucho tiempo debido a que la mayor incidencia y patogenicidad se observa en niños menores de 5 años. Con el fin de determinar los electroferotipos circulantes en adultos, se empleó la técnica de PAGE para analizar 23 aislados de rotavirus de individuos adultos (promedio de edad: 42,5 años; rango:24 a 67 años), recolectados entre Septiembre de 2003 a Febrero de 2005 del Laboratorio del Hospital San Roque, Asunción. En las 23 muestras se detectaron 5 electroferotipos diferentes, 19 presentaron alguno de los 3 patrones largos detectados (denominados LA, LB y LC) y 4 presentaron algún patrón corto (SA y SB). Desde 1998 hasta la fecha, nuestro grupo de trabajo detectó un solo caso de rotavirus con patrón corto (aislado en septiembre de 1999) luego de analizar más de 1500 heces de niños con diarrea aguda. Estudios previos en población infantil, han reportado que se necesitan varios años de estudio para identificar rotavirus con patrones cortos en una comunidad, sin embargo nuestros resultados sugieren que además es importante ampliar las variables demográficas de la población de estudio para encontrar más variantes de rotavirus, como en este caso es la aparición de patrones cortos en adultos sin que estos se presenten en niños.

Although rotavirus is recognized as the major ethiologic agent of acute diarrheal disease in children, the role of rotavirus as a pathogen in adults has long been underappreciated. In order to determine the electropherotypes of rotavirus that circulated in adults in Paraguay, 23 rotavirus isolated from patients older than 24 years (mean=42.5 range = 24­67) were analyzed by PAGE. Samples were collected at the Clinical Lab of the San Roque Hospital, Asunción,Paraguay from September 2003 to February 2005. Five different electropherotypes were detected, 19 samples showed long patterns (named LA, LB and LC) and 4 samples a short pattern (named SA and SB). Since 1998, our group has only detected one sample with a short pattern electropherotype (isolated in September 1999) in more than 1500 grounds of children with acute diarrhea. These results showed that it is necessary to study many epidemic years to detect rotavirus with short electropherotypes. However, we found that it is important to increase the demographic variables to find more rotavirus variability in a community, exemplified here by the short electropherotypes detected in adults.

Dosis letal media (LD50) y alteraciones ultraestructurales hepaticas inducidas por Eugenia uniflora, N.V. "Nängapiry" / Median lethal dose (LD50) and hepatic ultrastructural changes induced by Eugenia uniflora, N.V. \"Nängapiry\"

Se determinó la dosis letal media del extracto etanol: aguda 7:3 de hojas de Eugenia uniflora por vía intraperitoneal en ratones Balb-C, evaluando las alteraciones hepáticas por microscopía óptica y electrónica