Sequence analysis of SWEET transporters from trypanosomatids and evaluation of its expression in Trypanosoma cruzi.

Trypanosoma cruzi is an obligate parasite that uses glucose as one of the main resources to maintain its survival and proliferation. In eukaryotic cells glucose transport across membranes is mediated by facilitated transport through a variety of transporters. Herein, genes from the recently described SWEET family of carbohydrate transporters were identified in trypanosomatid parasites, including the medically important species T. cruzi and Leishmania spp. The identified genes have sequences with the typical attributes of known SWEET transporters. The expression of TcSWEET, the gene for the SWEET transporter found in the T. cruzi genome, was evidenced by immunohistochemistry using a polyclonal serum raised against peptides selected from the deduced TcSWEET protein sequence. In Western blot analysis, this α-TcSWEET serum detected proteins within the theoretical molecular mass for TcSWEET (25.8 kDa) in total epimastigote lysates, suggesting its expression at this parasite stage. Additionally, this serum stained epimastigotes at localizations consistent with the cell body and the flagellum. Together, these data suggests that SWEET transporters may contribute to glucose transport in trypanosomatid parasites.

Environmental Surveillance of Polioviruses in Armenia, Colombia before Trivalent Oral Polio Vaccine Cessation.

Although acute flaccid paralysis (AFP) surveillance is the "gold standard" for detecting cases of polio, environmental surveillance can provide supplementary information in the absence of paralytic poliomyelitis cases. This study aimed to detect the introduction and/or circulation of wild poliovirus or vaccine-derived polioviruses (VDPV) in wastewater, covering a significant population of Armenia, Colombia, before trivalent oral polio vaccine (OPV) cessation. Between March and September 2015, 24 wastewater samples were collected from eight study sites in eight communes of Armenia, Colombia. Virus detection and characterization were performed using both cell culture (i.e., RD or L20B cells) and RT-PCR. Polioviruses were isolated in 11 (45.8%) of 24 wastewater samples. All isolates were identified as Sabin strains (type 1 = 9, type 3 = 2) by intratypic differentiation. Type 2 poliovirus was not detected in any of the samples. No wild poliovirus or VDPV was detected among the isolates. Non-polio enterovirus was identified in 8.3% (2/24) of the samples. This study revealed the excretion of Sabin poliovirus from OPV-immunized individuals, as well as the absence of VDPV and wild poliovirus in wastewaters of Armenia, Colombia. This confirms that environmental surveillance is an effective method, as an additional support to AFP surveillance, to monitor poliovirus during the OPV-to-IPV (inactivated polio vaccine) transition period.

Identification, Characterization, Immunolocalization, and Biological Activity of Lucilin Peptide.

Maggots from the Lucilia sp. genus are used for debridement of infected and necrotic wounds. Broad-spectrum antimicrobial activity has been described in the excretion/secretions (ES1) of these larvae. This study identifies the genetic sequence of a cecropin-like antimicrobial peptide from Lucilia eximia. Total RNA was extracted and used for PCR-RACE amplification of a cecropin, the native peptide was immunolocalized in the tissues and secretions of the larvae, and a synthetic analog was used to explore its antimicrobial, cytotoxic, LPS neutralizing and wound-healing activities in vitro. The genetic cDNA sequence of a cecropin-like antimicrobial peptide in L. eximia called "Lucilin" was amplified, corresponding to 63 aa completed protein and 40 aa mature peptide; the structure of the mature peptide was predicted as an α-helix. The peptide was immunolocalized in the salivary glands, fat body, the ES, and hemolymph of the maggots. Lucilin synthetic peptide analog was active against E. coli DH10B with a MIC2 of 7.8 µg/mL, E. coli extended spectrum b-lactamase (ESBL) (MIC: 15.6 µg/mL), and Enterobacter cloacae (MIC: 125 µg/mL), but it was not active against Pseudomonas aeruginosa and Staphylococcus epidermidis; and had no cytotoxic or hemolytic activity. It showed immunomodulatory activity against human peripheral blood mononuclear cells (PBMCs) stimulated with LPS, reducing the TNF-α production when treated at 17 µg/mL and induces cell migration of Hacat at 5 and 50 µg/mL. Lucilin is a cecropin-like peptide from L. eximia with antimicrobial activity against Gram negative bacteria and immunomodulatory activities, decreasing the TNF-α production in PBMCs and inducing cellular migration in human keratinocytes.

[Modulation of high-density lipoprotein and cytokine IL-1ß and IL-6 levels in patients with dengue]. / Modulación de los niveles de lipoproteínas de alta densidad y las citoquinas IL-1ß e IL-6 en pacientes con dengue.

OBJECTIVES: To assess High-Density Lipoprotein (HDL) and pro-inflammatory cytokine levels in patients with dengue classified according to the severity of the clinical presentation. MATERIALS AND METHODS: A descriptive study was carried out with 70 individuals with dengue, classified as no alarm signs (DSSA), with signs of alarm (DCSA), and severe dengue (DG); 15 healthy donors were included as controls (CS). The serum levels of IL-1ß and IL-6 were quantified through ELISA, and the HDL levels through serum by a colorimetric test. In addition, the platelet count and the hematocrit percentage were determined. RESULTS: Levels of IL-1ß and IL-6 were found to be increased in patients with DENV vs. the healthy controls. Said increase was more evident in patients with dengue and in line with the severity of the clinical presentation. The HDL levels were found to be decreased in patients with dengue versus healthy controls. The correlation analysis showed statistically significant associations between HDL and the platelet number, as well as between the hematocrit percentage and the levels of IL-1ß. CONCLUSIONS: The increased IL-1ß and IL-6 and the decreased HDL levels in the most severe clinical stages suggest that both factors are involved in the development of the pathogenesis and severity. The correlations allow the observation of a potential association between HDL and platelet count, which allows us to provide an approach to the possible role of HDL in the pathogenesis of dengue, but it is still necessary to perform additional studies that will allow to state their importance in the course of the DENV infection.

OBJETIVOS: Evaluar los niveles de lipoproteínas de alta densidad (HDL) y citoquinas proinflamatorias en pacientes con dengue clasificados según la gravedad del cuadro clínico. MATERIALES Y MÉTODOS: Se realizó un estudio descriptivo, incluyendo 70 individuos con dengue, clasificados como: sin signos de alarma (DSSA), con signos de alarma (DCSA) y dengue grave (DG); 15 donantes sanos fueron incluidos como controles (CS). Los niveles séricos de IL-1ß e IL-6 fueron cuantificados por ELISA, y los de HDL en suero por ensayo colorimétrico; además se determinó el recuento de plaquetas y el porcentaje de hematocrito. RESULTADOS: Los niveles de IL-1ß e IL-6 se encontraron aumentados en los pacientes con DENV respecto a los controles sanos. Dicho aumento fue más evidente en los pacientes con dengue, de acuerdo con la gravedad del cuadro clínico. Los niveles de HDL se hallaron disminuidos en los pacientes con dengue comparados con los controles sanos. Los análisis de correlación mostraron asociaciones estadísticamente significativas entre HDL y el número de plaquetas; así como entre el porcentaje de hematocrito y los niveles de IL-1ß. CONCLUSIONES: El aumento de IL-1ß e IL-6 y la disminución de HDL en los estadios clínicos más graves sugieren que ambos factores están implicados en el desarrollo de la patogénesis y severidad. Las correlaciones permiten observar una posible asociación entre HDL y el número de plaquetas que permite dar una aproximación al posible papel de las HDL en la patogénesis del dengue, pero aún es necesario realizar estudios adicionales que permitan elucidar su importancia en el curso de la infección por DENV.

Modulación de los niveles de lipoproteínas de alta densidad y las citoquinas IL-1β e IL-6 en pacientes con dengue / Modulation of high-density lipoprotein and cytokine IL-1β and IL-6 levels in patients with dengue

RESUMEN Objetivos. Evaluar los niveles de lipoproteínas de alta densidad (HDL) y citoquinas proinflamatorias en pacientes con dengue clasificados según la gravedad del cuadro clínico. Materiales y métodos. Se realizó un estudio descriptivo, incluyendo 70 individuos con dengue, clasificados como: sin signos de alarma (DSSA), con signos de alarma (DCSA) y dengue grave (DG); 15 donantes sanos fueron incluidos como controles (CS). Los niveles séricos de IL-1β e IL-6 fueron cuantificados por ELISA, y los de HDL en suero por ensayo colorimétrico; además se determinó el recuento de plaquetas y el porcentaje de hematocrito. Resultados. Los niveles de IL-1β e IL-6 se encontraron aumentados en los pacientes con DENV respecto a los controles sanos. Dicho aumento fue más evidente en los pacientes con dengue, de acuerdo con la gravedad del cuadro clínico. Los niveles de HDL se hallaron disminuidos en los pacientes con dengue comparados con los controles sanos. Los análisis de correlación mostraron asociaciones estadísticamente significativas entre HDL y el número de plaquetas; así como entre el porcentaje de hematocrito y los niveles de IL-1β. Conclusiones. El aumento de IL-1β e IL-6 y la disminución de HDL en los estadios clínicos más graves sugieren que ambos factores están implicados en el desarrollo de la patogénesis y severidad. Las correlaciones permiten observar una posible asociación entre HDL y el número de plaquetas que permite dar una aproximación al posible papel de las HDL en la patogénesis del dengue, pero aún es necesario realizar estudios adicionales que permitan elucidar su importancia en el curso de la infección por DENV.

ABSTRACT Objectives. To assess High-Density Lipoprotein (HDL) and pro-inflammatory cytokine levels in patients with dengue classified according to the severity of the clinical presentation. Materials and Methods. A descriptive study was carried out with 70 individuals with dengue, classified as no alarm signs (DSSA), with signs of alarm (DCSA), and severe dengue (DG); 15 healthy donors were included as controls (CS). The serum levels of IL-1β and IL-6 were quantified through ELISA, and the HDL levels through serum by a colorimetric test. In addition, the platelet count and the hematocrit percentage were determined. Results. Levels of IL-1β and IL-6 were found to be increased in patients with DENV vs. the healthy controls. Said increase was more evident in patients with dengue and in line with the severity of the clinical presentation. The HDL levels were found to be decreased in patients with dengue versus healthy controls. The correlation analysis showed statistically significant associations between HDL and the platelet number, as well as between the hematocrit percentage and the levels of IL-1β. Conclusions. The increased IL-1β and IL-6 and the decreased HDL levels in the most severe clinical stages suggest that both factors are involved in the development of the pathogenesis and severity. The correlations allow the observation of a potential association between HDL and platelet count, which allows us to provide an approach to the possible role of HDL in the pathogenesis of dengue, but it is still necessary to perform additional studies that will allow to state their importance in the course of the DENV infection.

[Outbreak of malaria in an indigenous population living in an urban area of Armenia, Colombia, 2012]. / Descripción de un brote epidémico de malaria en una comunidad indígena asentada en la zona urbana de Armenia, Colombia, 2012.

INTRODUCTION: Malaria is the most frequent mosquito-borne parasitic infection in the tropical regions of the world. Due to different factors, including climate change, this disease can emerge or reemerge in different areas in the planet. OBJECTIVE: To describe an autochthonous outbreak of urban malaria in Armenia, Colombia. MATERIALS AND METHODS: After two children consulted a second level hospital located in the south of Armenia with malaria symptoms and their diagnosis was confirmed through the presence of Plasmodium vivax trophozoites detected by a positive thick blood smear, a visit was conducted to the residence of the children. An active search for cases was carried out, the environmental and sociocultural conditions were described and mosquito larvae and pupae were collected and identified. Control measures were implemented and their effectiveness was evaluated as well. RESULTS: The active search in the community allowed the identification of 11 probable cases, three of which were P. vivax malaria cases in children confirmed by blood microscopy. The entomological analysis of mosquito larvae and pupae showed Anopheles spp., Anopheles punctimacula and Anopheles argyritarsis. CONCLUSION: This is the first report of an autochthonous malaria outbreak by P. vivax in the city of Armenia, associated with the subnormal settlement of an indigenous community. The banks of the Quindío river offer the eco-epidemiological conditions that allow the establishment of the reproductive cycle of the malaria mosquito vector.

Descripción de un brote epidémico de malaria en una comunidad indígena asentada en la zona urbana de Armenia, Colombia, 2012 / Outbreak of malaria in an indigenous population living in an urban area of Armenia, Colombia, 2012

Introducción. La malaria o paludismo es la enfermedad parasitaria transmitida por mosquitos vectores más frecuente en las regiones tropicales. Debido a diversos factores, incluido el cambio climático, la enfermedad puede aparecer o reaparecer en diferentes lugares del mundo. Objetivo. Describir un brote autóctono de malaria urbana en la ciudad de Armenia, Colombia. Materiales y métodos. Después de la consulta en un hospital de segundo nivel del sur de Armenia de dos niños con síntomas de malaria, cuyo diagnóstico mediante gota gruesa confirmó la presencia de trofozoítos de Plasmodium vivax, se visitó el sector donde residían para la búsqueda activa de casos. Se describieron las condiciones ambientales y socioculturales del sector y se recolectaron e identificaron entomológicamente larvas, pupas y adultos de mosquitos. Igualmente, se implementaron medidas de control y se evaluó su efectividad. Resultados. La búsqueda activa en la comunidad permitió la identificación de 11 casos probables de malaria, de los cuales se confirmaron por microscopía tres casos de malaria por P. vivax en niños. El análisis entomológico de las larvas y pupas de mosquitos mostró la presencia de Anopheles spp ., Anopheles punctimacula y Anopheles argyritarsis. Conclusiones. Se reportó el primer brote autóctono de malaria por P.vivax en Armenia, asociado con un asentamiento subnormal en una comunidad indígena y se constató cómo en los márgenes del río Quindío se presentan las condiciones eco-epidemiológicas que permiten el ciclo de reproducción del mosquito vector de la malaria.

Introduction: Malaria is the most frequent mosquito-borne parasitic infection in the tropical regions of the world. Due to different factors, including climate change, this disease can emerge or reemerge in different areas in the planet. Objective: To describe an autochthonous outbreak of urban malaria in Armenia, Colombia. Materials and methods: After two children consulted a second level hospital located in the south of Armenia with malaria symptoms and their diagnosis was confirmed through the presence of Plasmodium vivax trophozoites detected by a positive thick blood smear, a visit was conducted to the residence of the children. An active search for cases was carried out, the environmental and sociocultural conditions were described and mosquito larvae and pupae were collected and identified. Control measures were implemented and their effectiveness was evaluated as well. Results: The active search in the community allowed the identification of 11 probable cases, three of which were P. vivax malaria cases in children confirmed by blood microscopy. The entomological analysis of mosquito larvae and pupae showed Anopheles spp., Anopheles punctimacula and Anopheles argyritarsis. Conclusion: This is the first report of an autochthonous malaria outbreak by P. vivax in the city of Armenia, associated with the subnormal settlement of an indigenous community. The banks of the Quindío river offer the eco-epidemiological conditions that allow the establishment of the reproductive cycle of the malaria mosquito vector.

[Absence of poliovirus circulation in Colombian departments with vaccination coverage below 80%]. / Ausencia de circulación de poliovirus en departamentos colombianos con coberturas vacunales inferiores a 80%

This study aims to explore a possible silent circulation of wild and vaccine-derived polioviruses in departments of Colombia with polio vaccination coverage of below 80%. The study collected 52 samples of wastewater concentrated as a result of precipitation with polyethylene glycol and sodium chloride. The viral detection was carried out through isolation and the identification through neutralization of the cytopathic effect, as well as through a conventional polymerase chain reaction following reverse transcription. The isolated polioviruses were characterized by the VP1 gene sequence. In two of the 52 samples, there was a presence of the Sabin type 2 poliovirus with more than 99% sequence similarity with the Sabin type 2 strain polio. Circulation of the nonpolio enterovirus was detected in 17.3% of the samples. The serotypes identified corresponded to coxsackievirus B1, echovirus 30, and echovirus 11. No evidence of the spread of either vaccine-derived poliovirus or wild poliovirus was detected in the departments of Colombia with polio coverage lower than 80%.

Prevalencia de helmintos intestinales en gatos domésticos del departamento del Quindío, Colombia / Prevalence of intestinal helminths in cats in Quindío, Colombia

Introducción. Las enfermedades producidas por helmintos están ampliamente distribuidas en el mundo y muchas de ellas se consideran zoonosis. Los animales de compañía cumplen un papel trascendental en la transmisión a los humanos. Objetivo. Determinar la prevalencia de helmintos intestinales en gatos del departamento del Quindío. Materiales y métodos. Se estudiaron 121 gatos domésticos del departamento del Quindío, de los cuales se registraron los datos de sexo, edad y condición corporal. Se recolectaron heces y se procesaron mediante las técnicas de Ritchie modificada y de Kato-Katz. Resultados. De los 121 gatos, 42,14 % (IC95%: 33,35-50,94) y 45,45 % (IC95%: 36,58-54,32) resultaron parasitados con alguna especie de helminto adulto según la presencia de huevos en sus heces, mediante las técnicas de Ritchie y de Kato-Katz, respectivamente. Toxocara cati fue el parásito más prevalente(Ritchie: 37,2 %; Kato-Katz: 43 %), seguido por Ancylostoma spp. (Ritchie: 7,43 %; Kato-Katz: 5,78 %) y Aelurostrongylus abstrusus (Ritchie: 0,82 %). Sesenta y cinco (53,71 %) gatos eran hembras y 56 (46,28 %) eran machos; la prevalencia de infección fue similar en ambos sexos. Los felinos mayores de 4 años de edad presentaron mayor prevalencia de parásitos (81,8 %), seguidos por los de 1 a 4 años (48,8 %) y, por último, por los menores de un año (28,6 %). Se encontró una buena condición corporal en 77,68 % y este grupo presentó menor frecuencia de helmintos intestinales. Conclusión. La prevalencia de helmintos intestinales en gatos domésticos del departamento del Quindío fue de 43,8 %, lo que hace necesario establecer programas de vigilancia y prevención en la población humana y felina.

Introduction. Diseases caused by helminths are widely distributed in the world and many of them are considered zoonoses in which pets play a major role in transmission to humans. Objective. The prevalence of intestinal helminths was determined in cats in Quindío Province. Materials and methods. One hundred twenty-one cats were characterized --data recorded included sex, age and body condition. Fecal samples were collected and processed using the modified Ritchie and modified Kato-Katz techniques to determine the presence of intestinal helminths. Results. Of the 121 cats, 42.1%, (95% CI: 33.4-50.9) and 45.5% (95% CI: 36.6-54.3) were parasitized with at least one adult helminth species as evidenced by the presence of eggs in their fecal samples. Toxocara cati was the most prevalent parasite (Ritchie: 37.2%, Kato-Katz: 43%), followed by Ancylostoma spp. (Ritchie: 7.4%, Kato-Katz: 5.8%) and Aelurostrongylus abstrusus (Ritchie: 0.82%). Sixty-five cats (53.7%) were females and 56 (46.3%) males; the prevalence of infection was similar in both sexes. Cats older than 4 years had the highest prevalence (81.8%) followed by those aged 1 to 4 years (48.8%) and by those under 1 year (28.6%). The majority of cats, 77.7%, were found to be in good body condition and this group had the lowest frequency of intestinal helminths with both techniques. Conclusion. The prevalence of intestinal helminths in domestic cats in Quindío was 43.8%; it is necessary to establish surveillance and prevention programs in the human and feline populations.

Larvaterapia aplicada a heridas con poca carga de tejido necrótico y caracterización enzimática de la excreción, secreción y hemolinfa de larvas / Effect of maggot therapy on minimally necrotic tissues: characterization of larval enzymatic excretion/secretion

Introducción. Las úlceras crónicas son una afección con un impacto negativo importante en la calidad de vida de los pacientes y en el sistema de salud; la aparición de infecciones y su difícil manejo, así como la presencia de tejido necrótico, afectan el pronóstico de curación. La larvaterapia se presenta como una opción para el desbridamiento y el manejo de infecciones de úlceras crónicas. Objetivo. Evaluar la larvaterapia en heridas con poca carga de tejido necrótico y evaluar las excreciones, secreciones y la hemolinfa de las larvas, respecto a su contenido enzimático. Materiales y métodos. Se reporta una serie de tres casos clínicos con úlceras crónicas y poca carga de tejido necrótico, tratados con larvaterapia, y se evalúa su evolución por los índices PUSH (Pressure Ulcer Scale for Healing) y Wound Bed Score, así como el patrón electroforético y contenido enzimático por zimograma de las excreciones y secreciones, y de la hemolinfa de las larvas. Resultados. Con solo una aplicación de la larvaterapia se evidenció una mejoría del aspecto de la herida y en los puntajes evaluados; en el PUSH hubo una disminución de 2,3 puntos, en promedio, y con el Wound Bed Score, un incremento de 2,7, lo que demuestra una mejoría en ambas escalas. Conclusión. Se encontró una actividad enzimática diversa en su contenido de excreciones y secreciones, con predominio de actividad de la proteasa de tipo serina.

Introduction. Chronic leg ulcers are a burden for the health system and impact quality of life. The infections, the necrotic tissue and the difficult treatment affects the prognosis and healing time. Maggot therapy is presented as an acceptable alternative for the debridement and treatment of this pathology. Objective. The larval therapy was assessed on chronic leg ulcers with little necrotic tissue. Larval excretion and secretion (E/S) was characterized with respect to hemolymph (HL) enzymatic content. Materials and methods. Three patients with chronic leg ulcers and low necrotic tissue were treated with larval therapy and were assessed with the PUSH (pressure ulcer scale for healing) and Wound Bed Score. E/S and HL content was evaluated by SDS PAGE and zymogram. Results. The clinical aspect of the wounds showed improvement, and the scores demonstrated an average decrease of 2.3 for the PUSH and an average increase of 2.7 for the Wound Bed Score. A wide diversity of enzymatic activity in the E/S was demontrated with major activity belonging to serine protease family. Conclusions. Maggot therapy proved an effective treatment in cases with minimal tissue necrosis and can be considered a viable treatment option.

[Effect of maggot therapy on minimally necrotic tissues: characterization of larval enzymatic excretion/secretion]. / Larvaterapia aplicada a heridas con poca carga de tejido necrótico y caracterización enzimática de la excreción, secreción y hemolinfa de larvas.

INTRODUCTION: Chronic leg ulcers are a burden for the health system and impact quality of life. The infections, the necrotic tissue and the difficult treatment affects the prognosis and healing time. Maggot therapy is presented as an acceptable alternative for the debridement and treatment of this pathology. OBJECTIVE: The larval therapy was assessed on chronic leg ulcers with little necrotic tissue. Larval excretion and secretion (E/S) was characterized with respect to hemolymph (HL) enzymatic content. Materials and methods. Three patients with chronic leg ulcers and low necrotic tissue were treated with larval therapy and were assessed with the PUSH (pressure ulcer scale for healing) and Wound Bed Score. E/S and HL content was evaluated by SDS PAGE and zymogram. RESULTS: The clinical aspect of the wounds showed improvement, and the scores demonstrated an average decrease of 2.3 for the PUSH and an average increase of 2.7 for the Wound Bed Score. A wide diversity of enzymatic activity in the E/S was demonstrated with major activity belonging to serine protease family. CONCLUSIONS: Maggot therapy proved an effective treatment in cases with minimal tissue necrosis and can be considered a viable treatment option.

[Prevalence of intestinal helminths in cats in Quindío, Colombia]. / Prevalencia de helmintos intestinales en gatos domésticos del departamento del Quindío, Colombia.

INTRODUCTION: Diseases caused by helminths are widely distributed in the world and many of them are considered zoonoses in which pets play a major role in transmission to humans. OBJECTIVE: The prevalence of intestinal helminths was determined in cats in Quindío Province. MATERIALS AND METHODS: One hundred twenty-one cats were characterized --data recorded included sex, age and body condition. Fecal samples were collected and processed using the modified Ritchie and modified Kato-Katz techniques to determine the presence of intestinal helminths. RESULTS: Of the 121 cats, 42.1%, (95% CI: 33.4-50.9) and 45.5% (95% CI: 36.6-54.3) were parasitized with at least one adult helminth species as evidenced by the presence of eggs in their fecal samples. Toxocara cati was the most prevalent parasite (Ritchie: 37.2%, Kato-Katz: 43%), followed by Ancylostoma spp. (Ritchie: 7.4%, Kato-Katz: 5.8%) and Aelurostrongylus abstrusus (Ritchie: 0.82%). Sixty-five cats (53.7%) were females and 56 (46.3%) males; the prevalence of infection was similar in both sexes. Cats older than 4 years had the highest prevalence (81.8%) followed by those aged 1 to 4 years (48.8%) and by those under 1 year (28.6%). The majority of cats, 77.7%, were found to be in good body condition and this group had the lowest frequency of intestinal helminths with both techniques. CONCLUSION: The prevalence of intestinal helminths in domestic cats in Quindío was 43.8%; it is necessary to establish surveillance and prevention programs in the human and feline populations.

Prevalencia de enterovirus en recién nacidos y lactantes que consultaron a un centro de atención de primer nivel, Armenia, Colombia, 2009 / Prevalence of enterovirus infection in infants in Armenia, Colombia, 2009

Introducción. Los enterovirus están distribuidos por todo el mundo; sin embargo, existe escasa información sobre su circulación en Colombia. Objetivo. Estimar la prevalencia de circulación de enterovirus en niños menores de un año que asistieron a un centro de atención de primer nivel en Armenia, Colombia, en el 2009, e identificar los principales serotipos de enterovirus circulantes. Materiales y métodos. Se tomaron 320 muestras de heces de niños menores de un año de edad. La presencia de enterovirus se determinó mediante transcripción inversa y la reacción en cadena de la polimerasa anidada (RT-N-PCR), empleando iniciadores genéricos de enterovirus. Las muestras que resultaron positivas en la RT-N-PCR, se inocularon en cultivos celulares apropiados para enterovirus. Los aislamientos obtenidos se identificaron por neutralización con la mezcla de sueros de Lim-Benyesh-Melnick. Resultados. Se detectaron enterovirus en 43 de las 320 (13,3 %) muestras de heces mediante RTN-PCR (IC95%: 9,7 a 17,1). Se obtuvo aislamiento viral en 26 de las 43 (60,4 %) muestras de heces positivas por RT-N-PCR. De los 26 aislamientos obtenidos, en 15 se identificó Coxsackie virus B (ocho CVB1, dos CVB2 y cinco CVB5) y 11 echovirus (seis E6 y cinco E30). Conclusiones. La circulación de enterovirus en la población infantil estudiada fue de 13,3 % y los serotipos de enterovirus aislados corresponden con los serotipos de mayor prevalencia global. Los resultados obtenidos indican la factibilidad de emplear la RT-N-PCR como herramienta para vigilar la circulación de enterovirus en muestras de heces.

Introduction. Despite world wide circulation of enteroviruses, little information has accumulated on the circulation of enteroviruses in Colombia. Objective. The prevalence of enterovirus circulation was examined in children under 1 year to identify the most common enterovirus serotypes. Materials and methods. Fecal samples were collected from 320 children under 1 year of age who attended a first-level health center in the city of Armenia, Colombia, in 2009. Enterovirus detection was performed by reverse transcription reaction and nested polymerase chain reaction (RT-N-PCR) using generic enterovirus primers. Samples testing positive in the RT-N-PCR were inoculated into cell cultures susceptible to enterovirus. All isolates were typed by seroneutralization with Lim-Benyesh-Melnick antiserum pools. Results. Overall, enteroviral RNA was detected in 43 of 320 (13.3%; 95% CI: 9.7 to 17.1) fecal samples by RT-N-PCR. Viral isolation was possible in 26 of 43 (60.4%) of the positive samples. Of these, 15 were Coxsackievirus B (eight CVB1, two CVB2, five CVB5) and 11 Echovirus (six E6 and five E30). Conclusions. The enteroviral circulation in a population on newly bornes up to 1 year old was 13.3%;the most frequent enterovirus was the same as those serotypes most commonly isolated in other parts of the world. The use of RT-N-PCR was demonstrably feasible as a tool to monitor the presence of enterovirus in stool samples.

Prevalencia de Mammomonogamus laryngeus (Strongylida: Syngamidae) en ganado bovino de la central de beneficio del municipio de La Tebaida, Quindío, Colombia / Prevalence of Mammomonogamus laryngeus (Strongylida: Syngamidae) nematodes in a bovine slaughterhouse

Introducción. Mammomonogamus laryngeus es un parásito nematodo que suele encontrarse en la laringe de mamíferos del trópico, especialmente en bovinos, ovejas y gatos. Objetivo. Determinar la prevalencia de M. laryngeus en bovinos de la central de beneficios del municipio de La Tebaida, Quindío, Colombia. Materiales y métodos. Para determinar la presencia de infestación por M. laryngeus, se examinó la tráquea y el árbol pulmonar de los animales bovinos sacrificados en la central de sacrificio de La Tebaida, entre diciembre de 2008 y febrero de 2009.Resultados. Se examinaron 500 bovinos. La prevalencia de infestación por M. laryngeus fue de 14,8 % (74); las formas adultas del parásito se localizaron con mayor frecuencia en el repliegue ariteno-epiglótico y en ningún caso en la tráquea o bronquios. Los departamentos del país de donde provenían los animales con mayor prevalencia, fueron: Valle del Cauca (10 %), Quindío (3,6 %) y Caquetá (1,2 %).Conclusiones. El reporte de esta parasitosis en animales bovinos procedentes de otros departamentos, indica la circulación del parásito en Colombia. Por ende, se hace necesario hacer un estudio para definir la prevalencia actual de esta parasitosis en ganado para consumo humano.

Introduction. Mammomonogamus laryngeus is a nematode parasite that is usually found in the larynx of mammals in the tropics, especially in cattle, sheep and cats. Objective. The prevalence of M. laryngeus was determined in cattle brought to an abattoir as an indicator of its prevalence in the general population.Materials and methods. The present study was undertaken in the municipality of La Tebaida, Quindio, Colombia, to determine the presence of M. laryngeus infection in cattle brought to a local abattoir between December and February 2009. The tracheae and lungs of 500 slaughtered cattle were examined. Results. Parasite prevalence was 14.8% (n=74). The majority of worms were located on aryteno-epiglottic folds, but never in the tracheae or bronchi. The infection rate was highest (47%) in 2 to 2.5 year old animals. In infected animals, an average of 6.4 parasite pairs was found, with higher numbers in older animals. The prevalence of infection in cattle was greater than seen in the provinces of Valle del Cauca (10.0%), uindío(3,0%) pr Caquetá (1.2%). Conclusions. Reports of this parasite in cattle indicate its circulation throughout Colombia and recommend surveillance studies where cattle are raised and prepared for human consumption.

Establecimiento de una línea celular primaria a partir de huevos con embrión de Toxocara canis / Establishing a primary cell line from Toxocara canis embryonated eggs

Introducción. Toxocara canis es el segundo nematelminto más prevalente en perros a nivel regional y entre los tres más frecuentes en algunos países de la región. Debido a que la fuente de contaminación es el perro, éste se convierte en un nematodo con gran potencial zoonótico. Por esta razón, consideramos importante disponer de una línea celular de este helminto para el estudio de los aspectos básicos, así como para el desarrollo de técnicas diagnósticas. Objetivo. Obtener una línea celular primaria a partir de huevos con embrión de T. canis. Métodos. Los parásitos se extrajeron del intestino delgado de perros menores de un año. Las células embrionarias se obtuvieron mediante la embriogénesis de los huevos de los nematodos adultos, en cuatro diferentes medios; dos ricos en sustancias nutritivas, el tercero con formol al 1 % y el cuarto con agua destilada. Las células se obtuvieron mediante disociación mecánica de los huevos con embrión mediante la utilización de jeringas 30G. Resultados. El tiempo estimado de obtención de la línea celular fue de 15 días, en los que siete eran utilizados en la embriogénesis de los huevos. Las células respondieron positivamente a los métodos de crioconservación luego de dos días, e inclusive dos meses después, permitiendo fases de replicación de cuatro pases. Conclusiones. Se logró obtener una línea celular de T. canis a partir de huevos con embrión de este helminto. Esta línea celular ayudará al entendimiento de las relaciones patógenas, posibles blancos terapéuticos y para el desarrollo de métodos diagnósticos.

Introduction: Toxocara canis is the second most prevalent nemathelminthes in dogs at regional level and among the three most frequent in some countries in the region. Due to the fact that the dog is the contamination source, it becomes a nematode with a high zoonotic potential, so we consider it important to be able to use the cell line of this helminth to study the basic aspects, as well as the development of diagnostic techniques. Objective: To obtain a primary cell line from embryonated eggs of T.canis. Methods: The parasites were extracted from the small intestines of dogs under one year old. Embryonic cells were obtained by embryogenesis of the eggs secreted by adult worms in four different media; two were rich in nutrients, one was 1% formaldehyde, and the other was distilled water. The cells were obtained by mechanical dissociation of embryonated eggs using 30G needles. Results: The estimated time for obtaining the cell line was fifteen days, from which seven were used for egg embryogenesis. The cells responded positively to the cryopreservation methods after two days or even two months, allowing a replication phase with four passes. Conclusions: We managed to obtain a cell line from T. canis embryonated eggs. This cell line will help the understanding of pathogenic relationships, potential therapeutic targets and for developing diagnostic methods.

[Prevalence of Mammomonogamus laryngeus(Strongylida:Syngamidae) nematodes in a bovine slaughterhouse]. / Prevalencia de Mammomonogamus laryngeus (Strongylida: Syngamidae) en ganado bovino de la central de beneficio del municipio de La Tebaida, Quindío, Colombia.

INTRODUCTION: Mammomonogamus laryngeus is a nematode parasite that is usually found in the larynx of mammals in the tropics, especially in cattle, sheep and cats. OBJECTIVE: The prevalence of M. laryngeus was determined in cattle brought to an abattoir as an indicator of its prevalence in the general population. Materials and methods. The present study was undertaken in the municipality of La Tebaida, Quindio, Colombia, to determine the presence of M. laryngeus infection in cattle brought to a local abattoir between December and February 2009. The tracheae and lungs of 500 slaughtered cattle were examined. RESULTS: Parasite prevalence was 14.8% (n=74). The majority of worms were located on aryteno-epiglottic folds, but never in the tracheae or bronchi. The infection rate was highest (47%) in 2 to 2.5 year old animals. In infected animals, an average of 6.4 parasite pairs was found, with higher numbers in older animals. The prevalence of infection in cattle was greater than seen in the provinces of Valle del Cauca (10.0%), Quindío(3,0%) pr Caquetá (1.2%). CONCLUSIONS: Reports of this parasite in cattle indicate its circulation throughout Colombia and recommend surveillance studies where cattle are raised and prepared for human consumption.

[Prevalence of enterovirus infection in infants in Armenia, Colombia, 2009]. / Prevalencia de enterovirus en recién nacidos y lactantes que consultaron a un centro de atención de primer nivel, Armenia, Colombia, 2009.

INTRODUCTION: Despite world wide circulation of enteroviruses, little information has accumulated on the circulation of enteroviruses in Colombia. OBJECTIVE: The prevalence of enterovirus circulation was examined in children under 1 year to identify the most common enterovirus serotypes. MATERIALS AND METHODS: Fecal samples were collected from 320 children under 1 year of age who attended a first-level health center in the city of Armenia, Colombia, in 2009. Enterovirus detection was performed by reverse transcription reaction and nested polymerase chain reaction (RT-N-PCR) using generic enterovirus primers. Samples testing positive in the RT-N-PCR were inoculated into cell cultures susceptible to enterovirus. All isolates were typed by seroneutralization with Lim-Benyesh-Melnick antiserum pools. RESULTS: Overall, enteroviral RNA was detected in 43 of 320 (13.3%; 95% CI: 9.7 to 17.1) fecal samples by RT-N-PCR. Viral isolation was possible in 26 of 43 (60.4%) of the positive samples. Of these, 15 were Coxsackievirus B (eight CVB1, two CVB2, five CVB5) and 11 Echovirus (six E6 and five E30). CONCLUSIONS: The enteroviral circulation in a population on newly bornes up to 1 year old was 13.3%;the most frequent enterovirus was the same as those serotypes most commonly isolated in other parts of the world. The use of RT-N-PCR was demonstrably feasible as a tool to monitor the presence of enterovirus in stool samples.

Detección de metabolitos secundarios en Ambrosia peruviana Willd y determinación de la actividad antibacteriana y antihelmíntica / Secondary metabolite detection in Ambrosia peruviana Willd and determination of antibacterial and antihelmintic activity

En el conocimiento etnofarmacológico, Ambrosia peruviana es conocida como una planta antiinfecciosa y antihelmíntica. Dados los altos índices de resistencia bacteriana y parasitaria, se realizó la tamización fitoquímica preliminar siguiendo la metodología de Sanabria, y ensayos biológicos frente a aislamientos clínicos bacterianos, parásitos caninos y Artemia salina. Los ensayos de citotoxicidad en A. salina se realizaron por exposición de los adultos a concentraciones variables de los extractos. La actividad antibacteriana se realizó por los métodos de difusión en disco y concentración inhibitoria mínima (CIM). Se determinó el porcentaje de huevos que tenían el embrión del parásito, en el medio de cultivo se mantuvieron en lactato de Ringer con suplemento al 10% de RPMI y 1X ATM. Se determinó el porcentaje de huevos con embrión, o fecundados, liberados en el medio de cultivo con adición de extracto etanólico y acuoso secos de A. peruviana. Los ensayos de la especie vegetal frente a helmintos se realizaron al sumergir los adultos en medio con suplemento de extracto etanólico seco (usando diferentes concentraciones) y con fracciones ricas en alcaloides. Se identificó la presencia de alcaloides, glucósidos cardiotónicos, quinonas, flavonoides, carbohidratos, taninos y saponinas. La concentración letal media para el extracto etanólico seco fue 64,2 µg/ml, mientras que para el extracto acuoso fue de 840,4 µg/ml. Los extractos no presentaron actividad antibacteriana. Los ejemplares adultos de T. canis presentaron disminución de la motilidad frente a extractos secos, mientras que en la fracción de alcaloides murieron luego de 4 horas de exposición. Los extractos de A. peruviana sobre huevos de T. canis permitieron una disminución en el porcentaje de huevos con embrión que no dependía del extracto observado sino de la concentración empleada.

Ambrosia peruviana has been reported as an anti-infective and anti-parasitic plant in the ethno-pharmacological environment. Given the high rates of bacterial and parasitic resistance against commercial drugs recorded, we performed the preliminary phytochemical screening following Sanabria´s method and biological tests against clinical bacterial isolates, dog parasites, and Artemia salina. Cytotoxicity tests in Artemia salina were carried out by exposing adults to extracts of varied concentrations. The antibacterial activity was performed using the disk diffusion method and CIM. Toxocara canis nematodes were kept in Ringer´s lactate supplemented with 10% RPMI and 1X ATM. The embryo generation percentage of eggs released into the culture medium was evaluated adding ethanol and aqueous extracts of dried A. peruviana. The tests of A. peruviana against helminths were performed by immersing adults in a medium supplemented with dried ethanol extract (at various concentrations) and fractions rich in alkaloids. Phytochemical screening allowed preliminary identification of alkaloids, cardiotonic glucosides, quinones, flavonoids, carbohydrates, tannins and saponins. The LC50 for dry ethanol extract was 64.2µg/ml, while for the aqueous extract was 840.4µg/ml. The extracts did not show antibacterial activity. T. canis adults showed motility decrease against dried extracts, not so in alkaloid fraction where they died after a 4h exposure. The use of extracts from A. peruviana over Toxocara canis eggs caused a decrease in the percentage of embryo generation, which did not depend on the extract observed but on the concentration used.