Complete Genome Sequences of Two Salmonella enterica Lytic Phages, NBSal006 and NBSal007.

The use of bacteriophages as antimicrobial agents represents a promising alternative for the control of pathogenic bacteria. Here, we present the complete genome sequences of two novel Salmonella enterica lytic bacteriophages, NBSal006 and NBSal007, candidates for Salmonella biocontrol.

Extracción y evaluación de compuestos antivirales de Chondracanthus chamissoi y Chlorella peruviana contra el virus dengue serotipo 2 / Extraction and antiviral activity assessment of compounds from Chondracanthus chamissoi and Chlorella peruviana against serotype 2 dengue virus

RESUMEN Objetivo: extraer y evaluar la actividad antiviral de los compuestos de Chondracanthus chamissoi y Chlorella peruviana contra DENV-2 en células Vero-76. Materiales y métodos: se extrajeron el carragenano de Chondracanthus chamissoi, los carbohidratos solubles de Chondracanthus chamissoi y Chlorella peruviana y se realizó la prueba de toxicidad en células VERO-76 y la evaluación de la actividad antiviral. Resultados: se obtuvieron carragenanos de la fase de esporofito y gametofito de Chondracanthus chamissoi, los mismos que fueron identificados, mediante infrarrojo, como k-carragenano. Por cromatografía se identificaron nueve azúcares (ribosa, xilosa, arabinosa, fructuosa, manosa, galactosa, sucrosa, maltosa y lactosa) en la muestra de carbohidratos solubles de Chondracanthus chamissoi fase gametofito y cuatro azucares (glucosa, sucrosa, maltosa y lactosa) en la de Chlorella peruviana. Los compuestos de Chondracanthus chamissoi y la solución de carbohidratos solubles de Chlorella peruviana no presentaron efecto citotóxico; los carbohidratos del extracto crudo de Chlorella peruviana sí los tuvieron. Todas las fracciones del extracto crudo de Chondracanthus chamissoi fase gametofítica fueron positivas por la prueba de reducción del número de placas(50) a la dilución 1:5. El k-carragenano de Chondracanthus chamissoi en ambas fases y los extractos crudos de carbohidratos solubles de Chondracanthus chamissoi, Chlorella peruviana y la solución de carbohidratos solubles de Chlorella peruviana inhibieron el crecimiento del virus dengue, pero no los carbohidratos del extracto crudo de la Chlorella peruviana. Conclusiones: los compuestos obtenidos de Chondracanthus chamissoi y Chlorella peruviana presentan actividad antiviral contra DENV-2 por lo cual es necesario continuar los estudios del potencial antiviral de estos compuestos fraccionados y purificados.

ABSTRACT Objective: to extract and determine the antiviral activity of compounds from Chondracanthus chamissoi and Chlorella peruviana against DENV-2 in Vero-76 cells. Materials and methods: carrageenan from Chondracanthus chamissoi and soluble carbohydrates from Chlorella peruviana were extracted, and toxicity tests in VERO-76 cells and antiviral activity were determined. Results: carrageenan from Chondracanthus chamissoi sporophyte and gametophyte phases were obtained, the compound was identified as k-carrageenan using infrared light. Nine sugars (ribose, xylose, arabinose, fructose, mannose, galactose, sucrose, maltose, and lactose) were identified using chromatography in the soluble carbohydrate mix from Chondracanthus chamissoi, and four sugars (glucose, sucrose, maltose, and lactose) from Chlorella peruviana were identified. Compounds from Chondracanthus chamissoi and Chlorella peruviana soluble carbohydrate solutions did not show any cytotoxic effect, carbohydrates from the raw extract from Chlorella peruviana did have this effect. All fractions from the raw extract from the gametophyte phase from Chondracanthus chamissoi were positive in the plate reduction test at 1:5 dilution. K-carrageenan from Chondracanthus chamissoi in both phases, as well as the crude extracts of soluble carbohydrates from Chondracanthus chamissoi and Chlorella peruviana as well as the soluble carbohydrate solution from Chlorella peruviana inhibited growth of dengue virus, but that was not the case with the carbohydrates of the raw extract from Chlorella peruviana. Conclusions: compounds obtained from Chondracanthus chamissoi and Chlorella peruviana show antiviral activity against DENV-2, so it is necessary to continue studies aiming to determine the antiviral potential of these fractioned and purified compounds.

[Dengue in Peru: a quarter century after its reemergence]. / Dengue en el Perú: a un cuarto de siglo de su reemergencia.

A health problem each time more frequent and dispersed in tropical and subtropical areas of the world, including Peru where it entered in 1990, is dengue. It is produced by the dengue virus with four serotypes and transmitted by Aedes aegypti, a vector that coexists with humans and whose presence is favored by deficient sanitary, social and economic conditions. Manifestations of severe forms of the disease such as shock and bleeding, are related to the frequent co-circulation of the four serotypes and the emergence of new genotypes such as American/Asian serotype 2. The new classification of the disease by WHO as dengue with or without warning signs and severe dengue, is contributing to more timely diagnosis and treatment, enabling reductions in mortality. Of note is the need to highlight the surveillance of acute febrile illness and Aedes indices that contribute to a timely diagnosis and guide vector control measures through sanitary education and environmental management with community and intersectoral participation, in a creative manner according to ecological niches. An alternative for complementary prevention would be vaccination using tetravalent vaccines whose safety and efficacy must be guaranteed before its use in the population under the framework of comprehensive strategies.

Dengue en el Perú: a un cuarto de siglo de su reemergencia / Dengue in Peru: a quarter century after its reemergence

Un problema sanitario cada vez más frecuente y disperso en áreas tropicales y subtropicales del mundo, incluido el Perú donde ingresó en 1990, es el dengue. Es producido por el virus del dengue con cuatro serotipos y transmitido por el Aedes aegypti, vector que convive con los humanos y cuya presencia es favorecida por deficientes condiciones sanitarias, sociales y económicas. Manifestaciones de formas severas de la enfermedad como el choque y sangrado, están relacionadas con la frecuente cocirculación de los cuatro serotipos y la aparición de nuevos genotipos como el americano/asiático del serotipo 2. La nueva clasificación de la enfermedad por la OMS como dengue con o sin signos de alarma y dengue grave, está contribuyendo a un diagnóstico y tratamiento más oportunos, permitiendo reducir la letalidad. Debe destacarse la necesidad de la vigilancia del síndrome febril y los índices aédicos que contribuyan a un diagnóstico oportuno y orienten las medidas de control vectorial mediante educación sanitaria y manejo ambiental con participación comunitaria e intersectorial, de manera creativa según los nichos ecológicos. Una alternativa de prevención complementaria sería la vacunación utilizando vacunas tetravalentes cuya seguridad y eficacia deben estar garantizadas antes de su uso poblacional en el marco de estrategias integrales.

A health problem each time more frequent and dispersed in tropical and subtropical areas of the world, including Peru where it entered in 1990, is dengue. It is produced by the dengue virus with four serotypes and transmitted by Aedes aegypti, a vector that coexists with humans and whose presence is favored by deficient sanitary, social and economic conditions. Manifestations of severe forms of the disease such as shock and bleeding, are related to the frequent co-circulation of the four serotypes and the emergence of new genotypes such as American / Asian serotype 2. The new classification of the disease by WHO as dengue with or without warning signs and severe dengue, is contributing to more timely diagnosis and treatment, enabling reductions in mortality. Of note is the need to highlight the surveillance of acute febrile illness and Aedes indices that contribute to a timely diagnosis and guide vector control measures through sanitary education and environmental management with community and intersectoral participation, in a creative manner according to ecological niches. An alternative for complementary prevention would be vaccination using tetravalent vaccines whose safety and efficacy must be guaranteed before its use in the population under the framework of comprehensive strategies.

[Genetic variability of Aedes aegypti determined by mitochondrial gene ND4 analysis in eleven endemic areas for dengue in Peru]. / Variabilidad genética del Aedes aegypti determinada mediante el análisis del gen mitocondrial ND4 en once áreas endémicas para dengue en el Perú

In order to establish the genetic variability of Aedes aegypti determined by the analysis of the MT-ND4 gene, in eleven endemic regions for dengue in Peru, 51 samples of Ae. Aegypti were tested. The genetic variability was determined through the amplification and sequencing of a fragment of 336 base-pairs of MT ND4, the analysis of intra-specific phylogeny was conducted with the Network Ver. 4.6.10 program; and the phylogenetic analysis, with the Neighbor Joining distance method. The presence of five haplotypes of Ae. Aegypti grouped in two lineages was identified: the first one includes haplotypes 1, 3 and 5, and the second one comprises haplotypes 2 and 4. The geographic distribution of each of the haplotypes found is also shown. It is concluded that this variability is caused by the active migration of this vector and the human activity-mediated passive migration.

Variabilidad genética del Aedes aegypti determinada mediante el análisis del gen mitocondrial Nd4 en once áreas endémicas para dengue en el Perú / Genetic variability of Aedes Aegypti determined by mitochondrial gene ND4 analysis in eleven endemic areas for dengue in Peru

Con el objetivo de establecer la variabilidad genética de Aedes aegypti determinada por el análisis del gen mitocondrial ND4, se analizaron 51 especímenes de Ae. aegypti en once regiones endémicas para dengue en el Perú. La variabilidad genética se determinó mediante la amplificación y secuenciación de un fragmento de 336 pares de bases del gen mitocondrial ND4. El análisis de filogenia intraespecífica se realizó con el programa Network Ver. 4.6.10; y el análisis filogenético, con el método de distancia Neighbor Joining. Se identificó la presencia de cinco haplotipos de Ae. aegypti agrupados en dos linajes: el primero agrupa a los haplotipos 1, 3 y 5 y el segundo agrupa los haplotipos 2 y 4, se muestra además la distribución geográfica de cada uno de los haplotipos encontrados. Se concluye que esta variabilidad se debe tanto a la migración activa de este vector como a la migración pasiva mediada por la actividad humana.

In order to establish the genetic variability of Aedes aegypti determined by the analysis of the MT-ND4 gene, in eleven endemic regions for dengue in Peru, 51 samples of Ae. Aegypti were tested. The genetic variability was determined through the amplification and sequencing of a fragment of 336 base-pairs of MT ND4, the analysis of intra-specific phylogeny was conducted with the Network Ver. 4.6.10 program; and the phylogenetic analysis, with the Neighbor Joining distance method. The presence of five haplotypes of Ae. Aegypti grouped in two lineages was identified: the first one includes haplotypes 1, 3 and 5, and the second one comprises haplotypes 2 and 4. The geographic distribution of each of the haplotypes found is also shown. It is concluded that this variability is caused by the active migration of this vector and the human activity-mediated passive migration.

[Hantavirus pulmonary syndrome (Rio Mamore virus) in the Peruvian Amazon region]. / Síndrome pulmonar por Hantavirus (Virus Río Mamoré) en la Amazonía Peruana.

Hantavirus infection is a viral zoonotic infection borne by rodents which most letal form clinical is the Hantavirus Pulmonary Syndrome (SPH, Spanish abbreviation). The Mamore River variant originates in South America and was found in rodents without any association to human diseases. Two cases of SPH were identified in the Peruvian Amazon region in November 2011. In both cases, a molecular diagnostic testing was conducted by the Instituto Nacional de Salud from Peru. A phylogenetic analysis of a viral genome fragment and a histopathological evaluation were conducted. Both patients developed adult respiratory distress syndrome and refractory shock. A patient died and another one recovered 12 days later.

Síndrome pulmonar por Hantavirus (Virus Río Mamoré) en la Amazonía Peruana / Hantavirus pulmonary syndrome (Rio Mamore Virus) in the Peruvian Amazon region

La hantavirosis es una infección viral zoonótica transmitida por roedores cuya forma clínica más letal es el síndrome pulmonar por Hantavirus (SPH). La variante río Mamoré es autóctona de Sudamérica y fue descrita en roedores sin asociarla a enfermedad en humanos. Se presenta dos casos de SPH causados por hantavirus río Mamoré en la Amazonía peruana en noviembre de 2011. En ambos casos, el diagnóstico confirmatorio fue molecular, efectuados en el Instituto Nacional de Salud de Perú. Se realizó análisis filogenético del fragmento de genoma viral y la evaluación histopatológica. Ambos pacientes evolucionaron a síndrome de distrés respiratorio del adulto y estado de choque refractario. Un paciente falleció y el otro se recuperó a los doce días.

Hantavirus infection is a viral zoonotic infection borne by rodents which most letal form clinical is the Hantavirus Pulmonary Syndrome (SPH, Spanish abbreviation). The Mamore River variant originates in South America and was found in rodents without any association to human diseases. Two cases of SPH were identified in the Peruvian Amazon region in November 2011. In both cases, a molecular diagnostic testing was conducted by the Instituto Nacional de Salud from Peru. A phylogenetic analysis of a viral genome fragment and a histopathological evaluation were conducted. Both patients developed adult respiratory distress syndrome and refractory shock. A patient died and another one recovered 12 days later.

Síndrome pulmonar por Hantavirus (Virus Río Mamoré) en la Amazonía Peruana / Hantavirus pulmonary syndrome (rio mamore virus) in the peruvian amazon region

La hantavirosis es una infección viral zoonótica transmitida por roedores cuya forma clínica más letal es el síndrome pulmonar por Hantavirus (SPH). La variante río Mamoré es autóctona de Sudamérica y fue descrita en roedores sin asociarla a enfermedad en humanos. Se presenta dos casos de SPH causados por Hantavirus río Mamoré en la Amazonía peruana en noviembre de 2011. En ambos casos, el diagnóstico confirmatorio fue molecular, efectuados en el Instituto Nacional de Salud de Perú. Se realizó análisis filogenético del fragmento de genoma viral y la evaluación histopatológica. Ambos pacientes evolucionaron a síndrome de distrés respiratorio del adulto y estado de choque refractario. Un paciente falleció y el otro se recuperó a los doce días(AU)

[Circulation of a different lineage of dengue virus serotype 2 American / Asian genotype in the Peruvian Amazon, 2010]. / Circulación de un linaje diferente del virus dengue 2 genotipo América / Asia en la región amazónica de Perú, 2010.

Our objective was to determine the genotype of the dengue virus type 2 (DENV-2) that circulated in the Amazon region of Peru between November 2010 and January 2011. We analyzed eight samples collected during dengue surveillance activities in the cities of Iquitos, Yurimaguas, Trujillo, Tarapoto and Lima between November 2010 and January 2011 that were sent to Insitituto Nacional de Salud. The viruses were isolated in C6/36 HT cell line. Viral RNA was extracted and the serotype (RT - PCR multiplex) and genotype (RT-Nested PCR of the region E/NS1) were determined. Finally, the E/ NS1 amplicons were sequenced and analyzed by phylogeny. The phylogenetic analysis revealed the introduction of a different lineage which entered in Peru by the end of 2010. These isolates found in Iquitos and other cities in Peru are closely related to DENV-2 isolates that circulated in Brazil during 2007 and 2008, associated with severe dengue cases and deaths. In conclusion, we detected the introduction of a different lineage of DENV-2 America / Asia genotype in Peru that could be associated with the presence of more severe cases.

[Clinical characteristics of patients admitted to hospital "Cesar Garayar García", Iquitos during the dengue epidemic, January-February 2011]. / Características clínicas de pacientes internados en el Hospital de Apoyo de Iquitos "César Garayar García" durante la epidemia de dengue, enero-febrero de 2011.

We conducted a clinical description of confirmed dengue cases admitted to the Hospital " César Garayar García" in Iquitos from January 25 to February 5, 2011. According to current WHO classification, major warning signs leading to hospitalization were: abdominal pain (28/28) and persistent vomiting (15/28), the causes of severity were: hypotension (9 / 13), severe bleeding (6 / 13) and plasma extravasation with respiratory distress (4 / 13). The median patient age was 22 years, however, 8 / 13 patients classified as severe dengue were under 16 years. In conclusion, unlike previous epidemics, we found a high involvement of young people and severity in the clinical presentation predominating a high frequency of shock (hypotension) and vascular leakage, which could be related to the circulation of an emerging variant DENV-2 which is more virulent.

Circulación de un linaje diferente del virus dengue 2 genotipo América / Asia en la región amazónica de Perú, 2010 / Circulation of a different lineage of dengue virus serotype 2 American / Asian genotype in the Peruvian amazon, 2010

El objetivo del estudio fue determinar el genotipo del virus dengue tipo 2 (DENV-2) que circuló en la región Amazónica de Perú entre noviembre de 2010 y enero de 2011. Se analizaron ocho muestras de pacientes captados durante la vigilancia para dengue en las ciudades de Iquitos, Yurimaguas, Trujillo, Tarapoto y Lima entre noviembre de 2010 y enero de 2011 que fueron remitidas al Instituto Nacional de Salud. Se realizó el aislamiento viral en la línea C6/36 HT y la extracción del ARN viral. Se aplicaron técnicas de biología molecular para establecer el serotipo (RT - PCR múltiple) y genotipo (RT-Nested PCR de la región E/NS1) seguidas de secuenciación y análisis filogenético. El análisis filogenético reveló la introducción de un linaje diferente que ingresó a Perú a finales del 2010. Estos aislamientos encontrados en Iquitos y otras ciudades de Perú están muy relacionados con aislamientos de DENV-2 que circularon en Brasil durante el 2007 y 2008 asociados con casos de dengue grave y muertes. En conclusión se detectó la introducción de un linaje diferente del DENV-2 genotipo América/Asia en Perú que podría estar asociado con la presencia de casos más graves de dengue.

Our objective was to determine the genotype of the dengue virus type 2 (DENV-2) that circulated in the Amazon region of Peru between November 2010 and January 2011. We analyzed eight samples collected during dengue surveillance activities in the cities of Iquitos, Yurimaguas, Trujillo, Tarapoto and Lima between November 2010 and January 2011 that were sent to Insitituto Nacional de Salud. The viruses were isolated in C6/36 HT cell line. Viral RNA was extracted and the serotype (RT - PCR multiplex) and genotype (RT-Nested PCR of the region E/NS1) were determined. Finally, the E/ NS1 amplicons were sequenced and analyzed by phylogeny. The phylogenetic analysis revealed the introduction of a different lineage which entered in Peru by the end of 2010. These isolates found in Iquitos and other cities in Peru are closely related to DENV-2 isolates that circulated in Brazil during 2007 and 2008, associated with severe dengue cases and deaths. In conclusion, we detected the introduction of a different lineage of DENV-2 America / Asia genotype in Peru that could be associated with the presence of more severe cases.

Características clínicas de pacientes internados en el Hospital de Apoyo de Iquitos "César Garayar García" durante la epidemia de dengue, enero-febrero de 2011 / Clinical characteristics of patients admitted to hospital "Cesar Garayar García", Iquitos during the dengue epidemic, January-February 2011

Se realizó una descripción clínica de pacientes con dengue confirmado internados en el Hospital de Apoyo de Iquitos "César Garayar García" desde el 25 de enero al 05 de febrero de 2011. Según la actual clasificación de la OMS, los principales signos de alarma que motivaron hospitalización fueron: dolor abdominal (28/28) y vómitos persistentes (15/28); las causas de gravedad fueron: hipotensión (9/13), sangrado grave (6/13) y extravasación de plasma con dificultad respiratoria (4/13). La mediana de edad de los pacientes fue 22 años, sin embargo, 8/13 pacientes clasificados como dengue grave fueron menores de 16 años. En conclusión, se evidencia una mayor afectación de la población joven y gravedad en la presentación clínica a diferencia de epidemias anteriores, predominando el shock (hipotensión) por extravasación vascular, lo cual podría estar relacionado con la circulación de una variante emergente del DENV-2 con mayor virulencia.

We conducted a clinical description of confirmed dengue cases admitted to the Hospital " César Garayar García" in Iquitos from January 25 to February 5, 2011. According to current WHO classification, major warning signs leading to hospitalization were: abdominal pain (28/28) and persistent vomiting (15/28), the causes of severity were: hypotension (9 / 13), severe bleeding (6 / 13) and plasma extravasation with respiratory distress (4 / 13). The median patient age was 22 years, however, 8 / 13 patients classified as severe dengue were under 16 years. In conclusion, unlike previous epidemics, we found a high involvement of young people and severity in the clinical presentation predominating a high frequency of shock (hypotension) and vascular leakage, which could be related to the circulation of an emerging variant DENV-2 which is more virulent.

Circulación de un linaje diferente del virus dengue 2 genotipo América / Asia en la región amazónica de Perú, 2010 / Circulation of a different lineage of dengue virus serotype 2 American / Asian genotype in the Peruvian amazon, 2010

El objetivo del estudio fue determinar el genotipo del virus dengue tipo 2 (DENV-2) que circuló en la región Amazónica de Perú entre noviembre de 2010 y enero de 2011. Se analizaron ocho muestras de pacientes captados durante la vigilancia para dengue en las ciudades de Iquitos, Yurimaguas, Trujillo, Tarapoto y Lima entre noviembre de 2010 y enero de 2011 que fueron remitidas al Instituto Nacional de Salud. Se realizó el aislamiento viral en la línea C6/36 HT y la extracción del ARN viral. Se aplicaron técnicas de biología molecular para establecer el serotipo (RT - PCR múltiple) y genotipo (RT-Nested PCR de la región E/NS1) seguidas de secuenciación y análisis filogenético. El análisis filogenético reveló la introducción de un linaje diferente que ingresó a Perú a finales del 2010. Estos aislamientos encontrados en Iquitos y otras ciudades de Perú están muy relacionados con aislamientos de DENV-2 que circularon en Brasil durante el 2007 y 2008 asociados con casos de dengue grave y muertes. En conclusión se detectó la introducción de un linaje diferente del DENV-2 genotipo América/Asia en Perú que podría estar asociado con la presencia de casos más graves de dengue.(AU)

Our objective was to determine the genotype of the dengue virus type 2 (DENV-2) that circulated in the Amazon region of Peru between November 2010 and January 2011. We analyzed eight samples collected during dengue surveillance activities in the cities of Iquitos, Yurimaguas, Trujillo, Tarapoto and Lima between November 2010 and January 2011 that were sent to Insitituto Nacional de Salud. The viruses were isolated in C6/36 HT cell line. Viral RNA was extracted and the serotype (RT - PCR multiplex) and genotype (RT-Nested PCR of the region E/NS1) were determined. Finally, the E/ NS1 amplicons were sequenced and analyzed by phylogeny. The phylogenetic analysis revealed the introduction of a different lineage which entered in Peru by the end of 2010. These isolates found in Iquitos and other cities in Peru are closely related to DENV-2 isolates that circulated in Brazil during 2007 and 2008, associated with severe dengue cases and deaths. In conclusion, we detected the introduction of a different lineage of DENV-2 America / Asia genotype in Peru that could be associated with the presence of more severe cases.(AU)