Pathogens, reservoirs, and vectors involved in the transmission of vector-borne and zoonotic diseases in a Colombian region.

The ecology of vector-borne diseases (VBDs) is an important system of great complexity, which involves the knowledge about the pathogens and animal species entailed in maintaining transmission cycles in a given locality, including those that act as vectors and reservoirs for the transmitted pathogens. To understand the ecology of some VBDs, we studied vectors, reservoirs, and pathogens of different VBDs, including dengue, leishmaniasis, Chagas disease, malaria, Zika, and chikungunya in the municipality of La Mesa, Cundinamarca, Colombia, a locality close to the capital, Bogotá. Vectors and mammals were sampled in urban and rural areas between May and August 2019. Molecular analyses were performed for the detection of pathogens in mammals and vectors, and of blood-meal sources in insects. Several vectors and mammals collected in this study have been involved in pathogen transmission cycles or may have a potential role in them. The findings of this study suggest that in the municipality of La Mesa, there are both vector and potential reservoir species, which are or could be implicated in the maintenance of the cycles of vector-borne diseases such as leishmaniasis and Chagas disease. Although arbovirus infections, such as dengue, are reported in the municipality, arbovirus presence was not detected. These findings highlight the importance of ongoing surveillance of vectors and associated control operations in La Mesa, of relevance to other locations where vectors and animal hosts also occur.

Altitudinal distribution and species richness of triatomines (Hemiptera:Reduviidae) in Colombia.

BACKGROUND: Chagas disease is considered to be endemic in up to 40% of the territory of Colombia, and to date 27 triatomine species have been reported the country. The purpose of this study was to update the geographical distribution of triatomine species in Colombia and assess the species richness patterns and their altitudinal distribution. METHODS: Occurrence data were compiled between 2007 and 2020, including from reports of entomological surveillance from the Instituto Nacional de Salud (INS), the Centro de Investigaciones en Microbiología y Parasitología Tropical (CIMPAT) at Universidad de Los Andes and a review of the literature. Geographic Information Systems (GIS) were used to describe general species richness patterns of the Triatominae subfamily. To establish the altitudinal distribution of the triatomine species, ranges were obtained from reports with unique elevation values. A generalized linear model was fitted, based on a Poisson distribution, to test the relation between triatomine species richness and Chagas disease cases (2012-2019). RESULTS: An updated geographical and altitudinal distribution for triatomine species in Colombia was established, with 507 municipalities added to the previously known distributions. The greatest triatomine richness in Colombia was found to be concentrated in the northeastern region of the country, extending towards the center to the departments of Arauca, Casanare and Meta. Regarding the altitudinal distribution, the study revealed that the species Rhodnius prolixus and Triatoma dimidiata have the greatest altitudinal ranges. The data also suggest a positive relation between species richness and number of Chagas disease cases reported per department. CONCLUSIONS: Altitudinal ranges for 17 triatomine species found in Colombia are presented. Species richness and species composition patterns are also described, and areas with a higher risk of transmission based on the relation found with Chagas disease cases are highlighted. This updated distribution reveals that Panstrongylus geniculatus is the triatomine with the largest presence by municipalities in Colombia, being reported in 284 municipalities, followed by Rhodnius prolixus in 277 municipalities.

Host selection pattern and flavivirus screening of mosquitoes in a disturbed Colombian rainforest.

Studies on the feeding behavior of hematophagous insects, particularly those of medical importance, are relevant for tracking possible pathogen transmission routes and identifying biases in the choice of vertebrates. We evaluated host selection of blood-feeding mosquitoes in a disturbed forest in the Magdalena Medio valley in Colombia from March 2017 to April 2018, after the introduction of Zika virus to the Americas from the 2015-2016 outbreak. We estimated vertebrate diversity and collected blood-engorged female mosquitoes. Genomic DNA/RNA was extracted from the mosquito's abdomen for vertebrate host identification and pathogen detection. We performed conventional PCR and sequencing, using universal primers targeting vertebrate regions of the eukaryotic mitochondrial genome to determine bloodmeal host. Additionally, we tested for the presence of flaviviruses in all mosquito samples with RT-PCR. Based on the identity and quantity of detected bloodmeals, we performed mosquito-vertebrate interaction network analysis and estimated topology metrics. In total, we collected 292 engorged female mosquitoes representing 20 different species. Bloodmeal analyses identified 26 vertebrate species, the majority of which were mammals (N = 16; 61.5%). No flaviviruses of medical importance were detected from the samples. Although feeding patterns varied, network analyses showed a high degree of specialization by mosquitoes and revealed ecological and phylogenetic relationships among the host community. We conclude that host selection or preference by mosquitoes is species specific.

Entomological characterization of Aedes mosquitoes and arbovirus detection in Ibagué, a Colombian city with co-circulation of Zika, dengue and chikungunya viruses.

BACKGROUND: Dengue, Zika and chikungunya are arboviruses of significant public health importance that are transmitted by Aedes aegypti and Aedes albopictus mosquitoes. In Colombia, where dengue is hyperendemic, and where chikungunya and Zika were introduced in the last decade, more than half of the population lives in areas at risk. The objective of this study was to characterize Aedes spp. vectors and study their natural infection with dengue, Zika and chikungunya in Ibagué, a Colombian city and capital of the department of Tolima, with case reports of simultaneous circulation of these three arboviruses. METHODS: Mosquito collections were carried out monthly between June 2018 and May 2019 in neighborhoods with different levels of socioeconomic status. We used the non-parametric Friedman, Mann-Whitney and Kruskal-Wallis tests to compare mosquito density distributions. We applied logistic regression analyses to identify associations between mosquito density and absence/presence of breeding sites, and the Spearman correlation coefficient to analyze the possible relationship between climatic variables and mosquito density. RESULTS: We collected Ae. aegypti in all sampled neighborhoods and found for the first time Ae. albopictus in the city of Ibagué. A greater abundance of mosquitoes was collected in neighborhoods displaying low compared to high socioeconomic status as well as in the intradomicile compared to the peridomestic space. Female mosquitoes predominated over males, and most of the test females had fed on human blood. In total, four Ae. aegypti pools (3%) were positive for dengue virus (serotype 1) and one pool for chikungunya virus (0.8%). Interestingly, infected females were only collected in neighborhoods of low socioeconomic status, and mostly in the intradomicile space. CONCLUSIONS: We confirmed the co-circulation of dengue (serotype 1) and chikungunya viruses in the Ae. aegypti population in Ibagué. However, Zika virus was not detected in any mosquito sample, 3 years after its introduction into the country. The positivity for dengue and chikungunya viruses, predominance of mosquitoes in the intradomicile space and the high proportion of females fed on humans highlight the high risk for arbovirus transmission in Ibagué, but may also provide an opportunity for establishing effective control strategies.

Spermathecal Filling in Aedes aegypti and Aedes albopictus: Effects of Female and Male Body Sizes and Species.

The mosquitoes Aedes aegypti (L.) (Diptera: Culicidae) and Aedes albopictus (Skuse) (Diptera: Culicidae) are ecologically similar species that have evolved independently in their native ranges, Ae. aegypti in Africa and Ae. albopictus in Asia. Where their invasive ranges overlap, interspecific mating is facilitated by both species swarming to mate around bloodmeal hosts during daylight. Here, we test hypotheses to account for variation in spermathecal filling in females of Ae. aegypti and Ae. albopictus, particularly, the effect of mosquito size on the number of spermathecae containing sperm after intraspecific and interspecifc matings of these two species. Results show that significantly more spermathecae contained sperm in large than in small Ae. albopictus females, but there was no effect of Ae. aegypti female body size on the average number of spermathecae filled. Among inseminated females, there was no effect of male size or cross-type on number of spermathecae with sperm. Differences in the effect of female size on the number of spermathecae that store sperm in Ae. aegypti and Ae. albopictus imply differences in the mating biology of these two species.

[Diagnostic accuracy of three technologies for the diagnosis of multi-drug resistant tuberculosis].

INTRODUCTION: Multi-drug resistant (MDR-TB) and extensively drug-resistant (XDR-TB) tuberculoses are a global public health problem. Their timely detection might reduce the burden of the disease and the economic impact on health systems worldwide. OBJECTIVE: To conduct a literature review of the diagnostic accuracy of three molecular tests to detect multi-drug resistant and extensively drug-resistant tuberculoses. MATERIALS AND METHODS: A systematic literature review following Cochrane methodology was carried out to study the diagnostic accuracy of three molecular tests to detect MDR-TB and XDR-TB in previous studies among immunocompetent population. Articles indexed in Medline and Embase were reviewed starting in 2007. Diagnostic accuracy was reported by sensitivity, specificity, and positive and negative predictive values of each test. RESULTS: In total, 8, 12 and 13 studies were included to assess the diagnostic accuracy of GeneXpert MTB/RIF®, GenoType MTBDRplus® and GenoType MTBDRsl®, respectively. The specificity of GeneXpert MTB/RIF® ranged between 91 and 100%, and its sensitivity between 33.3 and 100%. The sensitivity of GenoType® MTBDRplus® ranged between 88 and 100%. The sensitivity and specificity of GenoType MTBDRsl® to evaluate drug resistance ranged between 56 and 100% and 21 and 100%, respectively. CONCLUSION: The three diagnostic tests evaluated have shown an adequate diagnostic accuracy to detect MDR and XDR tuberculoses.

Precisión diagnóstica de tres pruebas moleculares para detectar la tuberculosis multirresistente / Diagnostic accuracy of three technologies for the diagnosis of multi-drug resistant tuberculosis

Resumen Introducción. La tuberculosis multirresistente (TB-MDR) y la extremadamente resistente (TB-XDR) constituyen un problema de salud pública a nivel mundial. Su detección oportuna permitiría reducir la carga de la enfermedad y su impacto económico en los sistemas de salud. Objetivo. Revisar sistemáticamente la información relacionada con la precisión diagnóstica de tres pruebas moleculares para detectar la tuberculosis multirresistente y la extremadamente resistente. Materiales y métodos. Se hizo una revisión sistemática de la literatura, según los lineamientos de Cochrane, de los estudios en población inmunocompetente relacionados con la precisión diagnóstica de tres pruebas moleculares para detectar la tuberculosis multirresistente y la extremadamente resistente. La búsqueda de los estudios publicados a partir del 2007 se hizo en Medline y Embase. La precisión diagnóstica de las pruebas se estableció con base en los valores máximos y mínimos de sensibilidad y especificidad, y en los valores predictivos positivos y negativos. Resultados. Se detectaron ocho estudios sobre la precisión diagnóstica de la prueba GeneXpert MTB/RIF(r), 12 sobre la de GenoType MTBDRplus(r) y 13 sobre la de GenoType MTBDRsl(r). La especificidad de GeneXpert MTB/RIF(r) osciló entre 91 y 100 % y su sensibilidad, entre 33,3 y 100 %. La sensibilidad de GenoType MTBDRplus(r) varió entre 82 y 100 %, en tanto que la sensibilidad y la especificidad de GenoType(r) MTBDRsl fluctuaron entre 56 y 100 % y 21 y 100 %, respectivamente. Conclusión. Según los estudios consultados, los tres métodos de diagnóstico evaluados presentabanuna adecuada eficacia diagnóstica para detectar la tuberculosis multirresistente y la extremadamente resistente.

Abstract Introduction: Multi-drug resistant (MDR-TB) and extensively drug-resistant (XDR-TB) tuberculoses are a global public health problem. Their timely detection might reduce the burden of the disease and the economic impact on health systems worldwide. Objective: To conduct a literature review of the diagnostic accuracy of three molecular tests to detect multi-drug resistant and extensively drug-resistant tuberculoses. Materials and methods: A systematic literature review following Cochrane methodology was carried out to study the diagnostic accuracy of three molecular tests to detect MDR-TB and XDR-TB in previous studies among immunocompetent population. Articles indexed in Medline and Embase were reviewed starting in 2007. Diagnostic accuracy was reported by sensitivity, specificity, and positive and negative predictive values of each test. Results: In total, 8, 12 and 13 studies were included to assess the diagnostic accuracy of GeneXpert MTB/RIF(r), GenoType MTBDRplus (r) and GenoType MTBDRsl (r), respectively. The specificity of GeneXpert MTB/RIF(r) ranged between 91 and 100%, and its sensitivity between 33.3 and 100%. The sensitivity of GenoType(r) MTBDRplus (r) ranged between 88 and 100%. The sensitivity and specificity of GenoType MTBDRsl (r) to evaluate drug resistance ranged between 56 and 100% and 21 and 100%, respectively. Conclusion: The three diagnostic tests evaluated have shown an adequate diagnostic accuracy to detect MDR and XDR tuberculoses.

Coexistence of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in Peninsular Florida Two Decades After Competitive Displacements.

The spread of Aedes albopictus (Skuse) eastward in the mid-1980s from its initial establishment in Houston, TX, was associated with rapid declines and local disappearances of Aedes aegypti (L.) in Gulf Coast states and Florida where annual larval surveillance during the early 1990s described temporal and spatial patterns of competitive displacements in cemeteries and tire shops. Approximately 20 yr later in 2013-2014, we re-visited former collection sites and sampled aquatic immatures of these two species from tire shops in 10 cities on State Route 441 and from 9 cemeteries from Lakeland to Miami in southwest Florida. In the recent samples Ae. aegypti was recovered from three central Florida cities where it had not been detected in 1994, but its northern limit on Rte. 441, Apopka, did not change. Other evidence, such as trends at a few cemeteries, suggested a moderate resurgence of this species since 1994. Cage experiments that exposed female progeny of Ae. aegypti from recent Florida collection sites to interspecific mating by Ae. albopictus males showed that females from coexistence sites had evolved resistance to cross-mating, but Ae. aegypti from sites with no Ae. albopictus were relatively susceptible to satyrization. Habitat classifications of collection sites were reduced by principal component (PC) analysis to four variables that accounted for > 99% of variances; PCs with strong positive loadings for tree cover and ground vegetation were associated with collection sites yielding only Ae. albopictus Within the coexistence range of the two species, the numbers of Ae. aegypti among total Aedes collected were strongly correlated in stepwise logistic regression models with two habitat-derived PCs, distance from the coast, and annual rainfall and mean maximum temperatures at the nearest weather station. Subtle increases in the range of Ae. aegypti since its previous displacements are interpreted in the context of the evolution of resistance to mating interference, realized versus fundamental niches of the two species, and persisting competition with Ae. albopictus.

Análisis de impacto presupuestal del uso de anticuerpos bloqueadores de acetilcolina receptores, comparado con la prueba de Tensilon, de estímulo repetitivo, electromiografía con electrodo de fibra única e I CE test para pacientes con miastenia gravis / Budget impact analysis of the use of acetylcholine receptor blocking antibodies, compared to the Tensilon test, repetitive stimulus, electromyography with single fiber electrode and IEC test for patients with myasthenia gravis

Tecnologías evaluadas: -Tecnologías actuales: electromiografía con electrodo de fibra única e ICE test;\r\n-Tecnología nueva: anticuerpos bloqueadores de acetilcolina receptores, prueba de Tensilon, prueba de estímulo repetitivo. Población: Esta prueba se puede aplicar a todas las edades y a todos los sexos, ya que la aparición de la enfermedad puede presentarse en toda la población. Perspectiva: Tercer pagador - Sistema General de Seguridad en Salud (SGSSS) colombiano. Horizonte temporal: El horizonte temporal de este AIP en el caso base corresponde a un año. Adicionalmente se reportan las estimaciones del impacto presupuestal para los años 2 y 3, bajo el supuesto de la inclusión en el POS en el año 1. Costos incluidos: Solo se tuvieron en cuenta los costos de las pruebas: -Electromiografía con electrodo de fibra única: $71.262,1; -Ice test: $26.223,3; Prueba completa de Tensilon: $24.000; -Prueba de estímulo repetitivo: $46.219,1; -Test de anticuerpos contra receptor de acetilcolina por RIA (ACRA): $45.416. Fuente de costos: Para todas las pruebas diagnósticas se utilizó el promedio ponderado estimado desde los registros de uso de servicios de 2014 SISPRO (módulo de prestación de servicios, mediante conexión OBDS), teniendo como corte de búsqueda la fecha del desarrollo de este impacto (20/10/2015). Todos los costos de las pruebas son ponderados por el número de unidades utilizadas que reporta la misma base de datos. Además, todas\r\nlas tecnologías son costeadas desde bases de aseguradores, para confirmación de precios. Resultados: Actualmente, el mercado se encuentra dominado por la electromiografía con electrodo de fibra única, la cual se encuentra dentro del plan de beneficios, pero por opinión de los realizadores, una vez que la prueba de acetilcolina receptores y de Lambert entre al plan de beneficios, se aumentará su participación, lo cual repercutirá en un ahorro al sistema, dado que dichas pruebas son menos costosas.(AU)

Análisis de impacto presupuestal del uso de inmunoglobulina humana normal para los pacientes con deficiencia inmune primaria en Colombia / Análisis de impacto presupuestal del uso de inmunoglobulina humana normal para los pacientes con deficiencia inmune primaria en Colombia

Tecnologías Evaluadas: Inmunoglobulina humana normal. Población: Pacientes con inmunodeficiencia primaria. Perspectiva: La perspectiva del presente AIP corresponde al tercero pagador, que en este caso es el Sistema General de Seguridad Social en Salud (SGSSS) en Colombia. Horizonte Temporal: El horizonte temporal de este AIP en el caso base corresponde a un año. Adicionalmente se reportan las estimaciones del impacto presupuestal para los años 2 y 3, bajo el supuesto de la inclusión en el POS en el año 1. Costos incluidos: El costo anual de la inmunoglobulina humana normal es de $41.883.552,73 pesos colombianos. Fuentes de costos: Dado que se estimó el costo de medicamentos, se utilizó la información de sistema deI nformación de Precios de Medicamentos y Dispositivos Médicos para el año 2014. Escenarios: Se plantearon los siguientes escenarios: adopción del 100% a su ingreso al plan de beneficios y un segundo escenario donde la adopción para el año uno es del 50%, hasta alcanzar el 100% al tercer año comparadora. Resultados: Debido a que no existe tecnología comparador dentro del análisis, se evidencia para el primer escenario donde la adopción es del 100%, se estima un impacto presupuestal por $185.125.303.066,60 pesos colombianos y para el escenario en el que comienza con el 50% y creciendo se evidencia un impacto de $92.562.651.533,3 pesos colombianos.(AU)

Satyrization without evidence of successful insemination from interspecific mating between invasive mosquitoes.

Previous research has documented low frequencies of interspecific mating in nature between the invasive vectors Aedes aegypti and Aedes albopictus. It is also known that heterospecific male accessory gland substances transferred during mating sterilize A. aegypti but not A. albopictus females, leading to satyrization, a form of reproductive interference. This paper demonstrates that satyrization of A. aegypti by A. albopictus may occur without evidence of successful insemination. Our results show that A. aegypti females, previously exposed to A. albopictus males, are rendered refractory to subsequent conspecific mating even though their spermathecae contain no heterospecific sperm. Additional experiments demonstrating transfer of labelled semen from A. albopictus males to A. aegypti females and low production of viable eggs of females housed with conspecific males, following exposure to A. albopictus males, confirm higher incidences of satyrization than expected, based on heterospecific insemination rates. We conclude that frequencies of satyrization based on detection of interspecific sperm in spermathecae may underestimate the impact of this form of reproductive interference.

Detection of insemination status in live Aedes aegypti females.

Using the technique described in this report, the presence or absence of sperm in spermathecae of female Aedes aegypti is detectable without dissection. Spermathecae of a lightly anesthetized female can be visualized by phase contrast microscopy through the distended abdomen, after the intersegmental membranes are stretched by ventral placement of a glass cover slip. Most females recovered after the procedure were capable of subsequent reproductive activities. Albeit tedious, this technique preserves the female alive for subsequent experiments or observations. Its extension to other mosquito species, or other Diptera and insects, will depend on spermathecal and sperm visibility through the distended abdomen.

Evolution of resistance to satyrization through reproductive character displacement in populations of invasive dengue vectors.

Recently, the highly invasive Asian tiger mosquito, Aedes albopictus, rapidly displaced resident populations of the yellow fever mosquito, Aedes aegypti in the southeastern United States and in Bermuda. Although multiple mechanisms of competitive displacement have been hypothesized, recent evidence of cross-insemination between these species in nature and the sterilizing effects of male accessory gland products asymmetrically favoring A. albopictus in interspecific matings support a role for satyrization (a form of reproductive interference) to explain the rapid displacements. Because of the drastic reproductive loss of A. aegypti females satyrized by A. albopictus males, we predicted selection for prezygotic isolation in populations of A. aegypti sympatric with A. albopictus. Exposures in cages demonstrated that female A. aegypti from populations in Florida sympatric with A. albopictus for the past 20 y were significantly less likely than nearby allopatric populations to mate with heterospecific males. Cross-inseminations of A. albopictus females by A. aegypti males were significantly less common, supporting the one-way direction of displacements observed in nature. Our results indicate rapid sexual selection leading to reproductive character displacement and the potential for satyr-resistant A. aegypti to recover from competitive displacements. These results have implications for increased risks of dengue transmission where these vector species meet worldwide.

Description of Lutzomyia (Helcocyrtomyia) tolimensis, a new species of phlebotomine sandfly (Diptera: Psychodidae) from Colombia

A description is presented of Lutzomyia tolimensis sp. nov., a new species of the subgenus Helcocyrtomyia, series sanguinaria. It was collected in dwellings, peridomestic environment and in nearby forest patches located in the foothills of the Andean Central Cordillera, where in 2004-2006 occurred the largest epidemic ever recorded of leishmaniasis in Colombia. The male of this species is differentiated from other members of the series sanguinaria based on the following combination of characters: (i) base of coxite with 0-3 subequal setae, (ii) spines of gonostyle organized in positions 2.1.2, (iii) spines inserted on distal half of gonostyle and (iv) relationship of alar indices. The female is recognized principally by the following characters: (i) palpomere V longer than III, (ii) length of labro-epipharynx and (iii) relationship of the alar indices.

Description of Lutzomyia (Helcocyrtomyia) tolimensis, a new species of phlebotomine sandfly (Diptera: Psychodidae) from Colombia.

A description is presented of Lutzomyia tolimensis sp. nov., a new species of the subgenus Helcocyrtomyia, series sanguinaria. It was collected in dwellings, peridomestic environment and in nearby forest patches located in the foothills of the Andean Central Cordillera, where in 2004-2006 occurred the largest epidemic ever recorded of leishmaniasis in Colombia. The male of this species is differentiated from other members of the series sanguinaria based on the following combination of characters: (i) base of coxite with 0-3 subequal setae, (ii) spines of gonostyle organized in positions 2.1.2, (iii) spines inserted on distal half of gonostyle and (iv) relationship of alar indices. The female is recognized principally by the following characters: (i) palpomere V longer than III, (ii) length of labro-epipharynx and (iii) relationship of the alar indices.

Phlebotomine vector ecology in the domestic transmission of American cutaneous leishmaniasis in Chaparral, Colombia.

Phlebotomine vector ecology was studied in the largest recorded outbreak of American cutaneous leishmaniasis in Colombia in 2004. In two rural townships that had experienced contrasting patterns of case incidence, this study evaluated phlebotomine species composition, seasonal abundance, nocturnal activity, blood source, prevalence of Leishmania infection, and species identification. CDC miniature light traps were used to trap the phlebotomines. Traps were set indoors, peridomestically, and in woodlands. Natural infection was determined in pools by polymerase chain reaction-Southern blot, and blood sources and species identification were determined by sequencing. Large differences were observed in population abundance between the two townships evaluated. Lutzomyia longiflocosa was the most abundant species (83.1%). Abundance was higher during months with lower precipitation. Nocturnal activity was associated with human domestic activity. Blood sources identified were mainly human (85%). A high prevalence of infection was found in L. longiflocosa indoors (2.7%) and the peridomestic setting (2.5%). L. longiflocosa was responsible for domestic transmission in Chaparral.

Description of Culicoides lisicarruni (Diptera: Ceratopogonidae), a new species from Cundinamarca, Colombia

A new species of Culicoides of the subgenus Diphaomyia Vargas from high altitudes of the Andes in Colombia is described and photographied. The species is compared with its similar congener Culicoides marinkellei Wirth & Lee. Data on the collecting site and notes on the species daily activity are also provided.

Breeding sites of Culicoides pachymerus Lutz in the Magdalena River basin, Colombia.

The breeding sites of Culicoides pachymerus are described for the first time in western Boyacá Province, Colombia, where this species is a public health problem. In addition to being a nuisance due to its enormous density and its high biting rates, C. pachymerus cause dermatological problems in the human population. Analysis of microhabitats by the sugar flotation technique and the use of emergence traps allowed us to recover 155 larvae of Culicoides spp and 65 adults of C. pachymerus from peridomiciliary muddy substrates formed by springs of water and constant rainwater accumulation. These important findings could aid in the design of integrated control measures against this pest.

Breeding sites of Culicoides pachymerus Lutz in the Magdalena River basin, Colombia

The breeding sites of Culicoides pachymerus are described for the first time in western Boyacá Province, Colombia, where this species is a public health problem. In addition to being a nuisance due to its enormous density and its high biting rates, C. pachymerus cause dermatological problems in the human population. Analysis of microhabitats by the sugar flotation technique and the use of emergence traps allowed us to recover 155 larvae of Culicoides spp and 65 adults of C. pachymerus from peridomiciliary muddy substrates formed by springs of water and constant rainwater accumulation. These important findings could aid in the design of integrated control meas-ures against this pest.