Abstract Anopheles calderoni considered a potential malaria vector, was only reported in Colombia during the last decade. Due to its recent report and the confusion with closely related species, little is known about the actual distribution and epidemiological importance of this species. In this work, the distribution and entomological parameters of A. calderoni were determined in several localities in Colombia. Anopheles specimens were collected in 10 departments from various regions in Colombia between 2009 and 2015. We characterized the ITS2 region for the collected specimens and performed a bioinformatic analysis to predict its taxonomic identification using a molecular test. We also determined the natural infection by Plasmodium and calculated entomological parameters. A. calderoni was registered in 4 departments in west Colombia. The highest biting rate (14.7 bites per night) was detected in Tumaco, SW Colombia, where a specimen was found to be infected by Plasmodium falciparum. The bionformatic analysis generated an ITS2 pattern that allows differentiation of this species. The presence of A. calderoni in regions with ecological diverse characteristics suggests that this species tolerates varied environments, and its infection with Plasmodium parasites indicates its role as a vector of local importance.

Resumen Anopheles calderoni, considerado vector potencial de malaria, fue reportado en la última década en Colombia. Debido a su registro reciente y a la confusión con especies cercanas, se conoce poco sobre su distribución real e importancia epidemiológica. En este trabajo se determinó la distribución y parámetros entomológicos de A. calderoni en diversas localidades de Colombia. Se recolectaron especímenes de Anopheles en 10 departamentos de varias regiones de Colombia, entre 2009 y 2015. Se caracterizó la región ITS2 para los individuos recolectados y se realizó un análisis bioinformático para predecir su identificación taxonómica mediante una prueba molecular. Se determinó la infección natural por Plasmodium y se calcularon parámetros entomológicos. Se registraron especímenes de A. calderoni en 4 departamentos del oeste colombiano. La tasa de picadura más alta (14.7 picaduras por noche) se presentó en Tumaco, en el sureste de Colombia, donde se detectó un ejemplar infectado por Plasmodium falciparum. El análisis bioinformático produjo patrones de ITS2 que permiten diferenciar a A. calderoni. La presencia de esta especie en regiones con características ecológicas diversas, sugiere que tolera diferentes ambientes y su infección con parásitos Plasmodium, indica su posible papel como vector de importancia local.

Resumen Introducción. En Colombia hay tres especies de mosquitos Anopheles implicadas como vectores primarios en la transmisión de la malaria o paludismo; sin embargo, el rol local de algunas especies de Anopheles aún debe determinarse. Objetivo. Determinar la abundancia, la composición y la infección natural de mosquitos anofelinos con Plasmodium spp. en dos regiones endémicas de malaria en Colombia. Materiales y métodos. Se recolectaron mosquitos del género Anopheles usando los métodos de recolección con cebo humano y en reposo en corrales de ganado vacuno, en nueve localidades de dos regiones endémicas para malaria en Colombia. Los especímenes se identificaron morfológicamente y se confirmaron por reacción en cadena de la polimerasa (PCR) de los polimorfismos en la longitud de los fragmentos de restricción (Restriction Fragment Length Polymorphism, RFLP) en el espaciador intergénico ribosómico nuclear 2 (Internal Transcribed Spacer, ITS-2) (PCR-RFLPITS2). Los especímenes se procesaron y analizaron mediante ELISA y PCR anidada basada en la subunidad pequeña del ARN ribosómico (small subunit ribosomal RNA, ssrRNA) para determinar la infección por Plasmodium. Resultados. Se recolectaron 1.963 mosquitos Anopheles correspondientes a nueve especies. Anopheles nuneztovari fue la especie predominante (53,5 %), seguida por A. darlingi (34,5 %), A. triannulatus s.l. (6 %) y por otras especies (˜5,9 %). Tres especies se encontraron naturalmente infectadas con Plasmodium spp.: A. darlingi, A. nuneztovari y A. triannulatus s.l. Conclusiones. La infección natural de A. darlingi y A. nuneztovari indica que estos vectores primarios siguen siendo actores principales en la transmisión de malaria en las localidades estudiadas de los departamentos del Valle del Cauca y Chocó. Además, el espécimen A. triannulatus s.l. infectado, recolectado en corrales de animales de la localidad estudiada en el departamento de Córdoba, indica que existe la necesidad de estudios futuros para establecer la importancia epidemiológica de esta especie dada su abundancia y comportamiento antropofílico oportunista.

Abstract Introduction: In Colombia there are three Anopheles species implicated in malaria transmission as primary vectors; however, the local role of some Anopheles species must still be defined. Objective: To determine the abundance, composition and natural infection rates for Anopheles mosquitoes with Plasmodium spp. in two malaria-endemic regions of Colombia. Materials and methods: Anopheles mosquitoes were collected using the human-landing catches and while resting in livestock corrals in nine localities of two malaria-endemic regions of Colombia. Mosquitoes were morphologically identified and confirmed by PCR-RFLP-ITS2. Identified mosquitoes were processed and tested for Plasmodium parasite infection by ELISA and ssrRNA-based nested PCR. Results: We collected 1,963 Anopheles mosquitoes corresponding to nine species. The most abundant species were Anopheles nuneztovari (53.5%) and A. darlingi (34.5%), followed by A. triannulatus s.l. (6%), and other species (˜5.9%). Three species were naturally infected with Plasmodium spp.: A. darlingi, A. nuneztovari and A. triannulatus s.l. Conclusions: Natural infection of A. darlingi and A. nuneztovari indicate that these malaria vectors continue to be effective carriers of Plasmodium in the localities under study in Valle del Cauca and Chocó. Additionally, the infected A. triannulatus s.l. collected in livestock corrals in the locality of the department of Córdoba suggests the need for further studies to define the epidemiological importance of this species given its abundance and opportunistic anthropophilic behavior.

Abstract: Anopheles calderoni, a species recorded in Colombia and Ecuador in 2010, is considered a malaria vector in Peru and a potential vector in the Colombian Pacific Coast. Given the recent report of A. calderoni, little is known on its morphological and genetic variability in the Colombian territory. As a first approach, this study evaluated wing morphometric variation on A. calderoni of 2 localities from western Colombia, Guadalajara de Buga and San Andrés de Tumaco. Eight morphometric characters of the wing were measured for 76 females (Buga n = 36, Tumaco n = 40) that were compared using univariate and multivariate analyzes. Pairwise comparison of wing character measures between the 2 populations showed statistically significant differences (p < .05). Discriminant analysis based on these characters, assigned with low accuracy (< 60%) each specimen to its original geographical population. Wing size differences between these A. calderoni populations may be due to micro-environmental characteristics of each locality rather than to genetic differences. The knowledge on A. calderoni population structure is relevant for the design of targeted vector control strategies.

Resumen: Anopheles calderoni, una especie registrada para Colombia y Ecuador en el 2010, es considerada un vector de malaria en Perú y un vector potencial en la costa pacífica colombiana. Dado el reciente registro de A. calderoni, poco se conoce acerca de su variabilidad morfológica y genética en el territorio colombiano. Como una primera aproximación, en este estudio se evaluó la variabilidad morfométrica alar de A. calderoni de 2 localidades del occidente colombiano, Guadalajara de Buga y San Andrés de Tumaco. Se midieron 8 variables morfométricas del ala de 76 hembras (Buga n = 36, Tumaco n = 40) y se compararon mediante análisis univariado y multivariado. La comparación pareada de las medidas de los caracteres del ala entre las 2 poblaciones reveló diferencias estadísticamente significativas (p < 0.05). El análisis discriminante, con base en estos caracteres, asignó con una baja precisión (< 60%) los especímenes a su población geográfica original. Las diferencias en el tamaño alar entre las poblaciones de A. calderoni pueden deberse a características microambientales de cada localidad más que a diferencias genéticas. El conocimiento de la estructura poblacional de A. calderoni es relevante para el diseño de estrategias de control vectorial dirigidas.

Bacillus cereus es una bacteria genéticamente diversa que se encuentra comúnmente en el ambiente. Contamina los alimentos afectando la salud humana, al ingerir el microorganismo y/o sus toxinas, la emética o las enterotoxinas. En Colombia son escasos los reportes de intoxicación por B. cereus y se estima que hay un gran subregistro. Por lo anterior, se recomienda aumentar la vigilancia de este patógeno y realizar estudios sobre aspectos relevantes que permitan aplicar medidas de control para disminuir las intoxicaciones por B. cereus. El objetivo de esta revisión bibliográfica es presentar información actualizada sobre B. cereus, que incluye aspectos de su biología, taxonomía, toxinas, alimentos que contamina y metodologías para detectar, prevenir y controlar este microorganismo. La información presentada es de utilidad para el público en general, especialmente personas vinculadas al sector de alimentos, inocuidad alimentaria y control de procesos.

Bacillus cereus is a genetically diverse bacterium commonly found in the environment. It contaminates food, thus affecting human health upon ingestion of the microorganism and/or its toxins, the emetic or enterotoxins. In Colombia, reports of intoxication by B. cereus are scarce and under-registration is presumed. Because of this, it is recommended to increase surveillance of this pathogen and to develop studies on relevant aspects that allow the application of control measures to reduce intoxications by B. cereus. The aim of this review is to present current information on B. cereus, including aspects of its biology, taxonomy, toxins, food that it contaminates and methodologies for the detection, prevention and control of this microorganism. This information is useful for the general public, especially people involved with the food sector, food safety and process control.

Bacillus cereus é uma bactéria geneticamente diversa normalmente encontrada no ambiente. Contamina os alimentos dos humanos e estes acabam prejudicados quando ingerem o microorganismo e/ou suas toxinas, a emética ou as enterotoxinas. Na Colômbia existem poucos relatórios de intoxicação por B. cereus, mas se estima um grande sub-registro. Essa estimação faz com que seja recomendável aumentar a vigilância deste patógeno e estudar aspetos relevantes que permitam aplicar medidas de controle, para diminuir as intoxicações por B. cereus. O objetivo desta revisão bibliográfica é apresentar informação atualizada sobre a B. cereus, incluindo aspectos da sua biologia, taxonomia, toxinas, alimentos que pode contaminar e metodologias para detectar, prevenir e controlar este microorganismo. Esta informação é útil para o público geral, especialmente para pessoas ligadas ao setor dos alimentos, à segurança alimentar e ao controle de processos

Resumen Introducción y objetivo: La caries dental es una de las enfermedades infecciosas de la cavidad oral más comunes en el mundo. El objetivo de este estudio fue determinar la prevalencia de Streptococcus mutansy sus genotipos en muestras de saliva de niños de 6 y 7 años, con y sin caries dental. Materiales y métodos: Cuarenta y siete (47) muestras de saliva fueron obtenidas de niños de 6-7 años, seleccionados aleatoriamente de una escuela pública en Medellín, Colombia. Se realizaron evaluaciones orales para determinar el índice COP-D (número de dientes permanentes cariados, obturados y perdidos). Los aislamientos fueron identificados mediante PCR para diferenciación de S. sobrinusy S. mutans, API-20STREP y análisis de ARNr 16S. Se determinaron mediante PCR los genotipos c e, fy ken las cepas identificadas como S. mutans. Resultados: La prevalencia de S. mutansfue 14,9%. De las 47 muestras de saliva, 57,4% (27) correspondieron a niños con caries dental, y 8,5 % (4), fueron positivas para S. mutans genotipo c, 2,1 % (1 cada una) genotipo f genotipo ky genotipo cy k, respectivamente. Conclusión: La prevalencia de S. mutansfue menor que la reportada en estudios previos de niños colombianos con características demográficas similares. Se encontró presencia de genotipos c, fy kde S. mutansen niños con caries dental, pero no en el grupo de niños sin caries dental. Este es el primer estudio en reportar la presencia del genotipo ken Colombia. Es necesario realizar más investigaciones en nuestro país para aclarar la relación de S. mutans genotipo k con caries dental y enfermedad cardiovascular

Abstract Introduction and objective: Tooth decay is one of the most prevalent infectious diseases of the oral cavity. The aim of this study was to determine the prevalence of Streptoccocus mutans and its genotypes in saliva samples from 6 and 7 year old children with and without dental caries. Materials and methods: Forty seven saliva samples were obtained from 6-7 year-old children, randomly selected from a Public School in Medellin, Colombia. Oral examinations in order to determine the DMF-T Index (Decay, Missing and Filling Teeth) were carried out. Isolates were identified by the Streptoccocus sobrinus and Streptococcus mutans PCR, API 20STREP and 16S rRNA sequence analysis. Genotypes c, e, f and k were detected by PCR. Results: Prevalence of S. mutanswas 14.9%. Of the 47 saliva samples, 57.4% (27) corresponded to children with dental caries, and 8.5% (4) were positive for S. mutans genotype c, 2.1% (1 each) genotype f genotype k, and genotype cand k, respectively. Conclusion: Prevalence of S. mutanswas lower than in previous studies of Colombian children with similar demographic characteristics. Streptoccocusmutans'genotype c, fand kwere found in children with caries but not in the group without caries. This is the first report of S. mutans genotype kin Colombia; this genotype requires further study to clarify its relation with dental caries and cardiovascular disease in Colombia

Bacillus cereus es un contaminante de alimentos conocido por ser patogénico para los humanos, causando síndromes de vómito y diarrea. En este estudio se evaluó la presencia de B. cereus toxigénicos utilizando PCR múltiple directamente en complementos dietarios para niños y en almidón de yuca colectados en Medellín, Colombia. De 75 muestras de complemento dietario para niños, 70,7% estuvieron contaminadas con B. cereus toxigénicos y se detectaron cuatro diferentes consorcios toxigénicos: I: nheA, hblC, cytK (9,8%), II: nheA, hblC (2%), III: hblC, cytK (41.2%), IV: hblC (47%). De 75 muestras de almidón de yuca, 44% estuvieron contaminadas con B. cereus toxigénicos y se determinaron cuatro diferentes consorcios toxigénicos: I: nheA, hblC, cytK (48.5%), II: nheA, hblC, cytK, cesB (3%), III: hblC, cytK (30,3%), IV: hblC (18.2%). En general, en los complementos dietarios para niños sólo se detectaron consorcios enterotoxigénicos, mientras que en el almidón los consorcios enterotoxigénicos predominaron sobre el emético. La PCR múltiple fue de utilidad para detectar contaminación con B. cereus toxigénicos permitiendo la detección directa y simultánea de todos los genes tóxicos en los alimentos. Este estudio es el primero en Colombia en evaluar B. cereus toxigénicos y proporciona información importante para la evaluación de riesgos microbiológicos en los alimentos pulverizados.

Bacillus cereus is a food contaminant and a known human pathogen that can cause emetic and diarrheal syndromes. In this study we evaluated the presence of toxigenic B. cereus by multiplex PCR directly in dietary complement for children and cassava starch samples collected on Medellin, Colombia. Of 75 dietary complement for children samples evaluated, 70.7% were contaminated with toxigenic B. cereus and four different toxigenic consortia were detected: I: nheA, hblC, cytK (9.8%), II: nheA, hblC (2%), III: hblC, cytK (41.2%), IV: hblC (47%). Of 75 cassava starch samples, 44% were contaminated with toxigenic B. cereus and four different toxigenic consortia were determined: I: nheA, hblC, cytK (48.5%), II: nheA, hblC, cytK, cesB (3%), III: hblC, cytK (30.3%), IV: hblC (18.2%). In general, in dietary complement for children only enterotoxigenic consortia were detected while in cassava starch the enterotoxigenic consortia predominated over the emetic. Multiplex PCR was useful to detect toxigenic B. cereus contamination allowing direct and simultaneous detection of all toxin genes in foods. This study is the first in Colombia to evaluate toxigenic B. cereus, providing information of importance for microbiological risk evaluation in dried foods.

Bacillus cereus é um contaminante de alimentos e é conhecido por ser patogénico nos seres humanos ocasionando síndromes de vômitos e diarreia. Neste estudo foi avaliada a presença de B. cereus toxigênicos por PCR multiplex diretamente em complementos da dieta para crianças e amido de mandioca, em amostras coletadas em Medellín, na Colômbia. De 75 amostras dos complementos da dieta para crianças, 70,7% estiveram contaminadas com B. cereus toxigênicos e foram detectados quatro diferentes consórcios: I: nheA, hblC, cytK (9,8%), II: nheA, hblC (2%), III: hblC, cytK (41,2%), IV: hblC (47%). De 75 amostras de amido de mandioca, 44% estiveram contaminadas com B. cereus toxigênicos e quatro consórcios diferentes foram determinados: I: nheA, hblC, cytK (48,5%), II: nheA, hblC, cytK, cesB (3%) III: hblC, cytK (30,3%), IV: hblC (18,2%). Em geral, nos complementos da dieta para crianças foram detectados apenas consórcios enterotoxigênicos, não obstante no amido os consórcios enterotoxigênicos predominaram sobre o emético. A PCR multiplex foi útil para detectar contaminação com B. cereus toxigênico permitindo a detecção direta e simultânea de todos os genes tóxicos em alimentos. Este estudo é o primeiro na Colômbia em avaliar B. cereus toxigênico e providencia informação importante para a avaliação de riscos microbiológicos em alimentos pulverizados.

Virulence and antibiotic resistance are significant determinants of the types of infections caused by Staphylococcus aureus and paediatric groups remain among the most commonly affected populations. The goal of this study was to characterise virulence genes of methicillin-susceptible S. aureus (MSSA) and methicillin-resistant S. aureus (MRSA) strains isolated from a paediatric population of a Colombian University Hospital during 2009. Sixty MSSA and MRSA isolates were obtained from paediatric patients between zero-14 years. We identified the genes encoding virulence factors, which included Panton-Valentine leucocidine (PVL), staphylococcal enterotoxins A-E, exfoliative toxins A and B and toxic shock syndrome toxin 1. Typing of the staphylococcal chromosome cassette mec (SCCmec) was performed in MRSA strains. The virulence genes were more diverse and frequent in MSSA than in MRSA isolates (83% vs. 73%). MRSA strains harboured SCCmec types IVc (60%), I (30%), IVa (7%) and V (3%). SCCmec type IVc isolates frequently carried the PVL encoding genes and harboured virulence determinants resembling susceptible strains while SCCmec type I isolates were often negative. PVL was not exclusive to skin and soft tissue infections. As previously suggested, these differences in the distribution of virulence factor genes may be due to the fitness cost associated with methicillin resistance.

To elucidate the Anopheles nuneztovari s.l. taxonomic status at a microgeographic level in four malaria endemic localities from Antioquia and Córdoba, Colombia, fragments of the cytochrome oxidase subunit I (COI) and the white gene were used. The COI analysis showed low genetic differentiation with fixation index (F ST) levels between -0.02-0.137 and Nm values between 3-∞, indicating the presence of high gene flow among An. nuneztovari s.l. populations from the four localities. The COI network showed a single most common haplotype, type 1 (n = 55), present in all localities, as the likely ancestral haplotype. Analysis of the white gene showed that An. nuneztovari s.l. populations from both departments grouped with haplotypes 19 and 20, which are part of lineage 3 reported previously. The results of the present study suggest that An. nuneztovari s.l. is a single taxon in the area of the present study.

Se optimizó un protocolo para la extracción de ADN a partir de mosquitos anofelinos, evaluando varios tiempos de incubación de la muestra con el acetato de potasio (AcK). Los resultados evidenciaron que la mayor concentración de ADN y frecuencia de amplificación se obtuvieron al utilizar el tratamiento de 1 h, lo que sugiere que es posible reducir el tiempo de incubación con AcK sin afectar la cantidad y calidad del ADN extraído.

A DNA extraction protocol for anopheline mosquitoes was optimized by the evaluation of various incubation times of the sample with Potassium acetate (KAc). The results showed that the highest DNA concentration and amplification frequency were obtained with the 1 h treatment, which suggests that is possible to reduce the incubation time with KAc without affecting the amount and quality of the DNA extracted.

An understanding of the taxonomic status and vector distribution of anophelines is crucial in controlling malaria. Previous phylogenetic analyses have supported the description of six species of the Neotropical malaria vector Anopheles (Nyssorhynchus) albitarsis s.l. (Diptera: Culicidae): An. albitarsis, Anopheles deaneorum, Anopheles marajoara, Anopheles oryzalimnetes, Anopheles janconnae and An. albitarsis F. To evaluate the taxonomic status of An. albitarsis s.l. mosquitoes collected in various localities in the Colombian Caribbean region, specimens were analyzed using the complete mitochondrial DNA cytochrome oxidase I (COI) gene, the ribosomal DNA (rDNA) internal transcribed spacer 2 (ITS2) region and partial nuclear DNA white gene sequences. Phylogenetic analyses of the COI gene sequences detected a new lineage closely related to An. janconnae in the Caribbean region of Colombia and determined its position relative to the other members of the complex. However, the ITS2 and white gene sequences lacked sufficient resolution to support a new lineage closely related to An. janconnae or the An. janconnae clade. The possible involvement of this new lineage in malaria transmission in Colombia remains unknown, but its phylogenetic closeness to An. janconnae, which has been implicated in local malaria transmission in Brazil, is intriguing.

El Grupo Oswaldoi, subgénero Nyssorhynchus (Diptera: Culicidae), incluye 16 especies. Algunas de estas especies presentan una alta variabilidad morfológica intraespecífica y similitud interespecie; lo anterior puede ocasionar dificultad en la identificación de la hembra al usar las claves morfológicas, como ocurre con Anopheles nuneztovari, considerado vector primario de malaria en Colombia. En este trabajo se comparó la utilidad de cinco claves morfológicas para la identificación de especímenes A. nuneztovari recolectados en la localidad de Puerto Anchica, Montelibano, Córdoba, Colombia. Se analizaron tres relaciones morfométricas y se confirmó la identificación morfológica utilizando la técnica PCR-RFLP basada en secuencias ITS2. El análisis de 41 hembras usando las claves, mostró que los especímenes A. nuneztovari presentaron sobrelapamiento morfológico con otras especies del Grupo Oswaldoi, como: A. rangeli, A. oswaldoi, A. evansae y A. benarrochi. El análisis molecular confirmó que todos los especímenes corresponden a A. nuneztovari. Dado que el uso de claves morfológicas continúa siendo la estrategia de elección para la identificación de anofelinos, los resultados sugieren que en caso de especies problemáticas, es conveniente confirmar la identificación mediante herramientas moleculares desarrolladas con el respaldo de la identificación de estadios inmaduros.

The Oswaldoi Group, subgenus Nyssorhynchus (Diptera: Culicidae), includes 16 species. Some of these species exhibit high intraspecific morphological variability and interspecific similarity; this can lead to difficulty in the identification of females by using morphological keys, as is the case with Anopheles nuneztovari, considered a primary malaria vector in Colombia. In this study we compared the usefulness of five morphological keys for the identification of A. nuneztovari specimens collected in the locality of Puerto Anchica, Montelibano, Córdoba, Colombia. Three morphometric ratios were analyzed and morphological identification results were confirmed using the PCR-RFLP technique based on ITS2 sequences. The analysis of 41 females using the keys showed that A. nuneztovari specimens presented overlap with other species of the Oswaldoi Group, such as A. rangeli, A. oswaldoi, A. evansae and A. benarrochi. Molecular analysis confirmed that all of the specimens corresponded to A. nuneztovari. Because the use of morphological keys continue to be a strategy of choice for the identification of anophelines, the results suggest that in the case of problematic species, it is convenient to confirm identification using molecular methods developed with the support of the identification of immature stages.

Malaria is a serious health problem in the states of Córdoba and Antioquia, Northwestern Colombia, where 64.4% of total Colombian cases were reported in 2007. Because little entomological information is available in this region, the aim of this work was to identify the Anopheles species composition and natural infectivity of mosquitoes distributed in seven localities with highest malaria transmission. A total of 1,768 Anopheles mosquitoes were collected using human landing catches from March 2007-July 2008. Ten species were identified; overall, Anopheles nuneztovari s.l. was the most widespread (62%) and showed the highest average human biting rates. There were six other species of the Nyssorhynchus subgenus: Anopheles albimanus (11.6%), Anopheles darlingi (9.8%), Anopheles braziliensis (6.6%), Anopheles triannulatus s.l. (3.5%), Anopheles albitarsis s.l. and Anopheles oswaldoi s.l. at < 1%; and three of the Anopheles subgenus: Anopheles punctimacula, Anopheles pseudopunctipennis s.l. and Anopheles neomaculipalpusat < 1% each. Two species from Córdoba, An. nuneztovari and An. darlingi, were found to be naturally infected by Plasmodium vivax VK247, as determined by ELISA and confirmed by nested PCR. All species were active indoors and outdoors. These results provide basic information for targeted vector control strategies in these localities.