Pintomyia evansi (Diptera: Psychodidae) larvae susceptibility to hydrated lime under laboratory conditions.

This study evaluated the effect of Calcium hydroxide (Ca(OH)2 Mg(OH)2) on third stages Pi. evansi larvae mortality under experimental laboratory conditions. Three treatments containing a mixture of phlebotomine natural breeding soil (substrate) and Calcium hydroxide at different concentrations were used: Treatment 1 (T1), 1 kg of substrate mixed with 56.2 g of lime; Treatment 2 (T2), 1 kg of substrate mixed with 62.5 g of lime; and Treatment 3 (T3), 1 kg of substrate mixed with 70 g of lime. in addition, a sample of substrate without lime was used as a control for each treatment. The mortality in T1 was 1% at 24 h and 12% at 48 h, reaching a maximum of 56% at 72 h of exposure. For T2, mortality was progressive, starting with 12% at 12 h, 36% at 24 h, 52% at 48 h, and 100% at 72 h; while T3 showed mortality percentages of 94% and 100% between 12 and 24 h of exposure. Therefore, T3 was the most effective to according to the Kaplan-Meier survival analysis. This study showed that treatments over 62 g of Calcium hydroxide per 1 kg of substrate offer a starting point for immature stage control under laboratory conditions. With these results, we propose to evaluate the cost-effectiveness and feasibility of the application, of the latter concentration, under field conditions in urban environments for its application in vector control programs.

[Circulación de Leishmania infantum y Trypanosoma cruzi en perros domésticos de áreas urbanas de Sincelejo, región Caribe de Colombia].

Introducción. La enfermedad de Chagas y la leishmaniasis tradicionalmente se han considerado zoonosis endémicas de áreas rurales del país. Sin embargo, la aparición de casos de estas enfermedades en áreas urbanas sugiere nuevos ciclos de circulación de estos parásitos. Por esta razón, se ha propuesto a los perros como centinelas de estos agentes zoonóticos, dado su rol como huéspedes accidentales o reservorios. Objetivo. Evaluar la circulación silenciosa de Leishmania spp. y Trypanosoma cruzi en perros de zonas urbanas de la ciudad de Sincelejo, Sucre. Materiales y métodos. Se analizaron 100 muestras de sangre de perros para amplificar la región ITS1 de Leishmania spp. Las muestras positivas se utilizaron para amplificar la región conservada del minicírculo del ADN del cinetoplasto de Leishmania infantum y para el análisis de polimorfismos de longitud de fragmentos de restricción con la endonucleasa HaeIII. Por otra parte, se amplificó un fragmento del ADN satelital de T. cruzi. Además, se evaluó la presencia de infecciones por Ehrlichia canis y Anaplasma platys, como potencialmente modificadoras de las manifestaciones clínicas. Resultados. De los 100 perros estudiados, se detectó: Leishmania spp. en 32, T. cruzi en 12, ambos parásitos en 7 y L. infantum en 18. Se encontraron infecciones por anaplasmatáceos en 18, y coinfecciones por bacterias y parásitos en 8 de los perros. En general, 47 de los animales estaban infectados por, al menos, un agente etiológico. Conclusión. Se demuestra la circulación de L. infantum y T. cruzi en zonas urbanas de Sincelejo, así como coinfecciones de estos parásitos junto con parásitos de la familia Anaplasmataceae. El presente estudio demuestra la conveniencia del uso de perros en la vigilancia epidemiológica de estos agentes zoonóticos.

Description of Lutzomyia velezi, a new species of phlebotomine sand fly (Diptera: Psychodidae) from the Department of Antioquia, Colombia.

The phlebotomine sand fly Lutzomyia velezi sp.nov. was described and illustrated from male specimens collected by light trap in the Reserva Natural Cañon del Río Claro in the Central Cordillera of the Colombian Andes. The new species belongs to the series sanguinaria of the subgenus Helcocyrtomyia, which is represented in Colombia by Lutzomyia cirrita, Lutzomyia hartmanni, Lutzomyia sanguinaria, Lutzomyia scorzai, Lutzomyia sp. of Pichindé and Lutzomyia tortura. The new species can be differentiated from others of the subgenus by the combination of the following characteristics: long antennal ascoids, reaching level of the papilla, coxite with a single basal seta and fifth palpomere longer than or equal to the sum of the lengths of the third and fourth palpomeres.

Molecular identification of the parasites causing cutaneous leishmaniasis on the Caribbean coast of Colombia.

All clinical manifestations of leishmaniasis exist in Colombia, the cutaneous form being the most frequent in the department of Sucre, where the Leishmania species associated with cutaneous leishmaniasis (CL) is unknown. This study was carried out to determine which Leishmania species was responsible for CL in Sucre, based on amplification and sequencing of the Cyt b gene. Isolates of Leishmania were obtained after CL diagnosis of eight patients who received attention in several health care centers of the study area. The nucleotide sequences obtained from patients were compared to Leishmania reference strains and six of the isolates identified as Leishmania (Viannia) braziliensis, the remaining two being identified as Leishmania (Viannia) panamensis and Leishmania (Viannia) guyanensis. This represents the first report of the presence of L. (V.) guyanensis on the Caribbean coast of Colombia.

Development of Lutzomyia evansi immature stages in peridomiciliary environment in a leishmaniasis urban focus in the Colombian Caribbean.

In the Caribbean region of Colombia, Lutzomyia evansi is recognized as the vector for Leishmania infantum and Leishmania braziliensis. Identifying breeding sites and surveying abundance of immature phlebotomine sand flies in urban foci of leishmaniasis are useful tool to design new vector control strategies. The objective of this study was to describe the natural breeding sites of Lu. evansi in peridomiciliary vegetation in a peri-urban area of the Colombian Caribbean region. Between 2013 and 2015, 466 microhabitats were sampled, collecting 621 kg of soil samples. The explored microhabitats were bases and tree holes, fallen trees, animal caves, leaf litter, domestic animal shelters, and the inside of dwellings. The immature phlebotomines were recovered by direct search under the stereoscope and incubation of soil samples. In total, 103 microhabitats, associated with 17 arboreal species, were identified as natural breeding sites. Of 422 immature sandflies detected, 98.6% were found in soils at the base of the trees. Eight species of the genus Lutzomyia were identified, of which Lu. evansi (52.6%) was the most abundant, followed by Lu. rangeliana, Lu. cayennensis cayennensis, Lu. atroclavata, Lu. micropyga, Lu. trinidadensis, Lu. dubitans and Lu. gomezi. The arboreal species Cordia alba was the most used by phlebotomines for the development of their immature stages. From 63 natural breeding sites identified 268 immatures were recovered including 176 Lu. evansi. The accumulated precipitation showed correlation (R2 = 0.643, p = 0.013) with the abundance of developmental stages, which increased in September and October. The natural breeding sites of Lu. evansi exhibited a local pattern of occurrence dependent on rainfall. The physicochemical analysis of the soil samples showed that the natural sites for C. alba were categorized as fertile loam soils. This is the first systematic study that estimates the temporal variation of immature sand flies in peridomiciliary vegetation in a peri-urban focus of leishmaniasis in Colombia.

DNA barcoding and fauna of phlebotomine sand flies (Diptera: Psychodidae: Phlebotominae) from Los Tuxtlas, Veracruz, Mexico.

Mexico has great diversity of phlebotomine sand flies related to cases of leishmaniasis, yet few studies have dressed the molecular taxonomy of these sand fly species. The use of the cytochrome oxidase subunit 1 (COI) gene, as a DNA Barcode has facilitated the molecular identification of sand flies species worldwide. We use the DNA barcode as a useful tool for the identification of phlebotomine sand flies of the natural reserve Los Tuxtlas from Veracruz, México. A fragment of 536 bp of the COI gene was obtained from 36 individuals belonging to eight species of five genera (Dampfomyia, Lutzomyia, Psathyromyia, Psychodopygus and Brumptomyia) with coverage between 92-100%, and found similarities ranging from 93-98% with other New World phlebotomine sand flies. The NJ dendogram grouped sand flies into eight clusters according to identified species, supported by bootstrap of 97%-100%. In conclusion, all phlebotomine sand flies were correctly identified and agree with the morphological identification, also could separate genetics the isomorphic females of the genus Brumptomyia.

Molecular evidence confirms the taxonomic separation of Lutzomyia tihuiliensis from Lutzomyia pia (Diptera: Psychodidae) and the usefulness of pleural pigmentation patterns in species identification.

The phlebotomine sand flies Lutzomyia pia (Fairchild & Hertig 1961) and Lutzomyia tihuiliensis Le Pont, Torrez-Espejo & Dujardin 1997 (Diptera: Psychodidae) belong to the pia series of the Lu. verrucarum species group, which includes several species that bite humans in Andean foci of leishmaniasis. The females of these two species exhibit isometry and isomorphism in anatomical structures of the head and terminalia commonly used in taxonomic identification of sand flies. They can only be differentiated based on subtle differences in the pigmentation of the pleura. In Lu. tihuiliensis, this is restricted to the basal portions of the katepimeron and katepisternum, whereas in Lu. pia both structures are totally pigmented. Taking into account the subtle morphological differences between these species, the objective of the current study was to evaluate the specific taxonomic status of Lu. tihuiliensis with respect to Lu. pia. A 475-bp portion of the mitochondrial genome was sequenced, composed of the 3' end of the cytochrome b gene, intergenic spacer 1, the transfer RNA gene for serine, intergenic spacer 2, and the 3' end of the gene NAD dehydrogenase 1. Genetic analysis confirms that Lu. tihuiliensis and Lu. pia constitute two distinct species and this is supported by four strong lines of evidence, i.e., the paired genetic distances, size differences and amino acid composition of the cytochrome b protein, presence and absence of intergenic spacer one and divergence observed in the sequence of the transfer RNA gene for serine. It also confirms the validity of the pleural pigmentation pattern as a species diagnostic character and the importance of performing a detailed examination of this character during morphological determination of phlebotomine sand flies in the series pia.

First record of the sand fly Warileya (Hertigia) hertigi from Antioquia, Colombia

Introduction: The genus Warileya is one of the least-known taxa of sandflies, comprising only nine species, i.e., W. (Warileya) phlebotomanica, W. (Hertigia) hertigi, W. (W.) rotundipennis, W. (W.) nigrosacculus, W. (W.) yungasi, W. (W.) fourgassiensis, W. (W.) lumbrerasi, W. (W.) euniceae and W. (W.) leponti. Objective: To document the presence of a species of the genus Warileya in Antioquia, Colombia. Materials and methods: Sandflies were collected in a cavern of the Cañón del Río Claro Natural Reserve, of the municipality of San Francisco, Antioquia department, Colombia. Phlebotomine sampling was carried out using a CDC light trap during three consecutive nights in May of 2008. Taxonomical determination was based on a revision of the type material of the species and through the use of standard keys for American sandflies. Results: Five male and two female sandflies were taxonomically identified as W. (H.) hertigi. In both sexes, the absence of setal scars in the anepisternum, proepimeron and clypeus; the presence of two transverse rows of setal scars in the tergites; and the short length of the vena gamma were notable. Conclusion: The finding of W. (H.) hertigi increases the number of sandfly species found in Antioquia department to 64. In total, 164 sandfly species have been recorded in Colombia.

Genetic variability of Aedes aegypti in the department of Sucre, Colombia, by analysis of the nucleotide sequence of the mitochondrial ND4 gene

Introduction. Aedes aegypti is the most important mosquito species in America for the transmission of viruses of dengue, Zika, Chikungunya and yellow fever. Ecological factors as well as chemical controls can affect the genetic composition of Ae. aegypti populations, which is why its genetic characterization is necessary. Objective. To determine the genetic variability of Ae. aegypti populations in four municipalities of Sucre department, Colombia. Materials and methods. Larvae of Ae. aegypti, collected in the municipalities of Sincelejo, Sampués, Corozal and Guaranda, Sucre department, were reared under laboratory conditions to adult stage. A segment of the mitochondrial ND4 gene which codes for the subunit 4 of the enzyme NADH-dehydrogenase was used as genetic marker. The genetic analysis included the estimation of parameters of nucleotide and haplotype diversity, genetic structure and gene flow. Results. One hundred and eight partial sequences of 357 nucleotides and four nucleotide haplotypes of the ND4 gene of Ae. aegypti were obtained. A significantly high genetic differentiation was found between the Sampués and Guaranda populations (FST=0.59467), Sincelejo and Sampués (FST=0.25637), and Corozal and Guaranda (FST=0.22237). A high gene flow (Nm=infinite) was observed among the populations of Sincelejo and Corozal. Conclusion. There are genetic differences between the Ae. aegypti populations from the municipalities of Sucre department. The presence of a new haplotype of the mitochondrial ND4 gene of Ae. aegypti in Colombia was recorded, detected in the municipality of Sincelejo.

[Analysis of the primary and secondary structure of the mitochondrial serine transfer RNA in seven species of Lutzomyia]. / Análisis de la estructura primaria y secundaria del ARN de transferencia mitocondrial para serina en siete especies de Lutzomyia.

INTRODUCTION: Lutzomyia sand flies are involved in the transmission of the parasite Leishmania spp. in America. The taxonomy of these vectors is traditionally based on morphological features of the adult stage, particularly the paired structures of the head and genitalia. Although these characters are useful to distinguish most species of Lutzomyia, morphological identification may be complicated by the similarities within subgenera and species group. OBJECTIVE: To evaluate the utility of mitochondrial serine transfer RNA tRNA Ser for taxonomic identification of Lutzomyia. MATERIALS AND METHODS: Seven sand fly species, each representing one of the 27 taxonomic subdivisions in genus Lutzomyia, were analyzed including L. trinidadensis (Oswaldoi group), L. (Psychodopygus) panamensis, L.(Micropygomyia) cayennensis cayennensis, L. dubitans (Migonei group), L. (Lutzomyia) gomezi, L. rangeliana (ungrouped) and L. evansi (Verrucarum group). The mitochondrial tRNA Ser gene, flanked by the cytochrome b and NAD dehydrogenase subunit one genes, was extracted, amplified and sequenced from each specimen. Secondary structure of the tRNA Ser was predicted by comparisons with previously described homologous structures from other dipteran species. RESULTS: The tRNA Ser gene ranged in size from 66 base pairs in L. gomezi to 69 base pairs in L. trinidadensis. Fourteen polymorphic sites, including four insertion-deletion events, were observed in the aligned 70 nucleotide positions. The majority of the substitutions were located in the dihydrouridine, ribothymidine-pseudouridine-cytosine and variable loops, as well as in the basal extreme of the anticodon arm. CONCLUSION: Changes of primary sequence of the tRNASer provided useful molecular characters for taxonomic identification of the sand fly species under consideration.

[Estimation of time detection limit for human cytochrome b in females of Lutzomyia evansi].

INTRODUCTION: Molecular biology techniques have allowed a better knowledge of sources of blood meals in vector insects. However, the usefulness of these techniques depends on both the quantity of ingested blood and the digestion process in the insect. OBJECTIVE: To identify the time limit for detection of the human cytochrome b (Cyt b) gene in experimentally fed females of Lutzomyia evansi. MATERIALS AND METHODS: Eight groups of L. evansi females were fed on human blood and sacrificed at intervals of 24 hours post-ingestion. Total DNA was extracted from each female and a segment of 358 bp of Cyt b was amplified. In order to eliminate false positives, amplification products were subjected to a restriction fragment length polymorphism (RFLP) analysis. RESULTS: The human Cyt b gene segment was detected in 86% (49/57) of the females of L. evansi, from 0 to 168 hours after blood ingestion. In 7% (4/57) of the individuals we amplified insect DNA, while in the remaining 7%, the band of interest was not amplified. We did not find any statistical differences between groups of females sacrificed at different times post-blood meal regarding the amplification of the human Cyt b gene segment or the number of samples amplified. CONCLUSION: The human Cyt b gene segment was detectable in L. evansi females up to 168 hours after blood ingestion.

DNA barcode for identification of immature stages of sand flies (Diptera: Psychodidae) collected from natural breeding sites.

Although phlebotomine sand flies breeding sites have been identified and recorded by several studies, the microhabitats exploited by these insects remain little-known and hard to find. In this context, the difficulty of finding immature stages, and the limited number of taxonomic studies to identify immature stages of phlebotomine sand flies, are considered the major obstacles when attempting a complete inventory of Lutzomyia species. The objective of this study is to validate Cytochrome Oxidase I (Barcode region) as a marker for the identification of immature stages of Lutzomyia species recovered from natural breeding sites in Colombia. Among 142 collected sand flies, 18 immature individuals that did not complete their life cycle were identified to species level through sequencing of the COI gene. Values of K2P genetic distance between 0.002-0.031 allowed the identification of larvae at species level. The bootstrap support values (96%) in the Neighbor-Joining dendrogram were consistent for the majority of the established MOTUS of Lutzomyia atroclavata, Lutzomyia micropyga, Lutzomyia serrana, Lutzomyia cayennensis, Lutzomyia rangeliana, Lutzomyia shannoni and some species of the genus Brumptomyia. The COI gene is validated as a marker for the identification of immature stages of the genus Lutzomyia.

Circulación de Leishmania infantum y Trypanosoma cruzi en perros domésticos de áreas urbanas de Sincelejo, región Caribe de Colombia / Silent circulation of Leishmania infantum and Trypanosoma cruzi among urban dogs from Sincelejo city, Caribbean region of Colombia

Introducción. La enfermedad de Chagas y la leishmaniasis tradicionalmente se han considerado zoonosis endémicas de áreas rurales del país. Sin embargo, la aparición de casos de estas enfermedades en áreas urbanas sugiere nuevos ciclos de circulación de estos parásitos. Por esta razón, se ha propuesto a los perros como centinelas de estos agentes zoonóticos, dado su rol como huéspedes accidentales o reservorios. Objetivo. Evaluar la circulación silenciosa de Leishmania spp. y Trypanosoma cruzi en perros de zonas urbanas de la ciudad de Sincelejo, Sucre. Materiales y métodos. Se analizaron 100 muestras de sangre de perros para amplificar la región ITS1 de Leishmania spp. Las muestras positivas se utilizaron para amplificar la región conservada del minicírculo del ADN del cinetoplasto de Leishmania infantum y para el análisis de polimorfismos de longitud de fragmentos de restricción con la endonucleasa HaelII. Por otra parte, se amplificó un fragmento del ADN satelital de T. cruzi. Además, se evaluó la presencia de infecciones por Ehrlichia canis y Anaplasma platys, como potencialmente modificadoras de las manifestaciones clínicas. Resultados. De los 100 perros estudiados, se detectó: Leishmania spp. en 32, T. cruzi en 12, ambos parásitos en 7 y L. infantum en 18. Se encontraron infecciones por anaplasmatáceos en 18, y coinfecciones por bacterias y parásitos en 8 de los perros. En general, 47 de los animales estaban infectados por, al menos, un agente etiológico. Conclusión. Se demuestra la circulación de L. infantum y T. cruzi en zonas urbanas de Sincelejo, así como coinfecciones de estos parásitos junto con parásitos de la familia Anaplasmataceae. El presente estudio demuestra la conveniencia del uso de perros en la vigilancia epidemiológica de estos agentes zoonóticos.

Introduction: Leishmania infantum and Trypanosoma cruzi are considered endemic zoonotic agents in rural areas of the country; however, there is a high risk of urbanization due to anthropogenic processes. For this reason, dogs have been proposed as sentinels of these zoonoses given their role as patients, hosts and/or reservoirs. Objective: To assess the silent circulation of Leishmania spp. and T. cruzi parasites in canines from urban areas of Sincelejo, Sucre. Materials and methods: One hundred canine blood samples were used to amplify the ITS1 region of Leishmania spp. Positive samples were used to amplify the conserved region of the kinetoplast DNA minicircle of L. infantum and for restriction fragment length polymorphism analysis with HaelII endonuclease. In addition, a satellite DNA of T. cruzi was amplified. Also, the presence of Ehrlichia canis and Anaplasma platys was evaluated as infections that can influence clinical symptoms and health of animals. Results: Leishmania spp. was detected in 32% (32/100) and T. cruzi in 12% (12/100) of the animals, and 7% (7/100) of the samples were positive for both parasites. Also, L. infantum and infections with Anaplasmataceae family parasites were both detected in 18 % (18/100) of the samples. In the same way, co-infections with bacteria and parasites were found in 8 % (8/100) of the animals. Overall, 47 % (47/100) of the animals were infected with at least one agent. Conclusion: The circulation of L. infantum and T. cruzi, as well as co-infections of pathogens of the Anaplasmataceae family, is demonstrated in urban areas of Sincelejo. The present study demonstrates the convenience of canines as epidemiological surveillance sentinels of these zoonotic agents.

[Redescription of the female of Lutzomyia vattierae (Diptera: Psychodidae, Phlebotominae) from the serranía de La Macarena, Central Colombia]. / Redescripción de la hembra de Lutzomyia vattierae (Diptera: Psychodidae, Phlebotominae) de la serranía de La Macarena, Colombia.

INTRODUCTION: Lutzomyia species of the subgenus Sciopemyia show restricted distribution and are rarely represented in sand fly captures, with the exception of L. sordellii, which is found from Costa Rica to Argentina. To date, only two of these species. L. sordellii and L. preclara have been reported in Colombia. OBJECTIVE: The female of L. vattierae was redescribed and the record of this Sciopemyia species established for in the Colombian national park, serranía de La Macarena. MATERIALS AND METHODS: The study was carried out in the Serrania de La Macarena, in western province of Meta. The serrania is a small mountain range situated between the Orinoco and Amazon River basins and geographically separated from the Andes. Sand flies were collected during two consecutive nights with a CDC light trap placed from 18:00 to 06:00 hours in a primary forest. RESULTS: Four females belonging to the subgenus Sciopemyia were identified as L. vattierae, which differs from L. preclara by the presence of papillae on the third flagellomere and from L. sordellii by the form and length of the spermathecae and individual sperm ducts. CONCLUSION: The number of known species of Sciopemyia in Colombia is now three and include L. sordellii, L. preclara, and L. vattierae.

[First finding of Lutzomyia tihuiliensis (Diptera: Psychodidae) in the Valle de Aburrá, Colombia]. / Primer hallazgo de Lutzomyia tihuiliensis (Diptera: Psychodidae) en el valle de Aburrá, Colombia.

INTRODUCTION: Three of the seven species that comprise the pia series of the Lutzomyia verrucarum group have been recorded in Colombia, including L. pia, L. limafalcaoae and L. emberai. OBJECTIVE: The aim of this paper is to report the occurrence of an anthropophilic morphospecies of the pia series in the country. MATERIALS AND METHODS: Sand flies were collected with a mouth aspirator on protected human bait in a secondary forest in the municipality of Envigado, department of Antioquia. The entomological survey was performed from 18:00 to 22:00 hours in June and December, 2004. RESULTS: Captured specimens were identified as L. tihuiliensis, which can be distinguished easily from other species of the pia series by its basally pigmented pleura, the length of the labro-epipharynx, (3)350 microm, and the length of the second palpomere, (3)170 microm. In addition to the previous characters, the sand flies collected exhibit a longer common sperm duct than the individual ducts with the ratio of the lengths of the common/individual ducts (3)2. CONCLUSION: The finding of L. tihuiliensis raises to 21 the number of species of the Lutzomyia verrucarum group recorded to date in Colombia, including two endemic species of the pia series. From a biogeographical point of view, the presence of four species of the pia series in Colombia is of great interest for the study of the origin of the taxon.

FAMILY PSYCHODIDAE.

A catalogue is presented of the species of haematophagous and non-haematophagous psychodids recorded in Colombia. The list comprises 199 species distributed among five subfamilies and 16 genera, as follows: Subfamily Bruchomyiinae, genus Nemopalpus Macquart, 1838 (4 species); subfamily Phlebotominae, genera Brumptomyia França & Parrot, 1921 (8 species), Lutzomyia França, 1924 (153 species) and Warileya Hertig, 1948 (2 species); subfamily Psychodinae, genera Arisemus Satchell, 1955 (3 species), Australopericoma Vaillant, 1975 (1 species), Balbagathis Quate, 1996 (1 species), Clogmia Enderlein, 1937 (1 species), Didicrum Enderlein, 1937 (1 species), Feuerborniella Vaillant, 1971 (1 species), Lepidiella Enderlein, 1937 (1 species), Maruina Müller, 1895 (4 species), Paramormia Enderlein, 1935 (1 species), Parasetomima Duckhouse, 1968 (1 species) and Psychoda Latreille, 1796 (7 species); subfamily Sycoracinae, genus Sycorax Haliday, 1839 (5 species); and subfamily Trichomyiinae, genus Trichomyia Haliday, 1839 (5 species).

Comparison of 16S and COX1 genes mitochondrial regions and their usefulness for genetic analysis of ticks (Acari: Ixodidae).

INTRODUCTION: In recent decades the analysis of mitochondrial genes has been used for population and phylogenetic studies of ticks allowing many advances in their systematics. Mitochondrial ribosomal 16S (16S) subunit is one of the most frequently used among those genes available for tick analysis, whereas cytochrome oxidase gene 1 (COX1) has recently been used and proposed as an alternative to the traditional 16S gene marker.  OBJECTIVE: To evaluate the usefulness of 16S and COX1 in genetic studies of ticks by analyzing sequences of three species commonly found in the Caribbean region of Colombia.  RESULTS: The analysis of both genes sequences allowed us to identify the three species with high levels of confidence and interspecific genetic divergence (19-22%), although only COX1 allowed us to detect intraspecific genetic variability (up to ~0.8%). A substitution saturation analysis indicated that the 16S gene was not saturated with transitions while the COX1 gene showed saturation distances starting at ~17%.  CONCLUSION: Our results indicated that the 16S gene seems to have better features for interspecific phylogenetic analyses because of its high level of genetic divergence and low saturation pattern, while the COX1 gene appears to be more useful for intraspecific genetic variability studies. However, as our study was conducted at a local scale, future studies at different biogeographical scales would help to establish its usefulness in wider and more complex scenarios.

A NEW RECORD OF Leishmania (Viannia) guyanensis (Trypanosomatidae) FROM THE PACIFIC COAST OF COLOMBIA / Un nuevo registro de Leishmania (Viannia) guyanensis (Trypanosomatidae) de la costa pacífica de Colombia

ABSTRACT The aim of this study was the identification of Leishmania species that causes cutaneous leishmaniasis in a patient from Buenaventura, Valle del Cauca, on the Pacific coast of Colombia. Clinical samples were obtained from a 29 years-old male who presented a distinct ulcer with raised borders on his neck. Samples were taken for direct microscopic examination, parasite culture, and molecular identification of the infecting Leishmania species by sequencing of the cytochrome b gene. Direct examination was positive for amastigotes of Leishmania but the culture was negative. The infecting parasite species was identified as L. (V.) guyanensis by means of the nucleotide sequence of a 509 bp fragment of the cytochrome b gene. We report the presence of L. (V.) guyanensis in rural areas of Buenaventura in Valle del Cauca, and the expansion of the geographical distribution of this species in the Pacific region of Colombia.

RESUMEN El objetivo de este estudio fue identificar la especie de Leishmania causante de la leishmaniasis cutánea en un paciente de Buenaventura, Valle del Cauca, en la costa Pacífica de Colombia. Se obtuvieron muestras clínicas de un varón de 29 años de edad que presentó una úlcera distintiva con bordes levantados en el cuello. Se tomaron muestras para examen microscópico directo, cultivo de parásitos e identificación molecular de la especie infectante de Leishmania mediante secuenciación del gen del citocromo b. El examen directo fue positivo para amastigotes de Leishmania pero el cultivo fue negativo. La especie parasitaria infectante se identificó como L. (V.) guyanensis por medio de la secuencia de nucleótidos de un fragmento de 509 pb del gen citocromo b. Con este reporte notificamos la presencia de L. (V.) guyanensis en zona rural del municipio de Buenaventura en el Valle del Cauca y la expansión de la distribución geográfica de esta especie en la región Pacífica de Colombia.

[First report of the F1534C mutation associated with cross-resistance to DDT and pyrethroids in Aedes aegypti from Colombia].

INTRODUCTION: The main strategy for the control of Aedes aegypti, vector of dengue, chikungunya and Zika viruses, is based on the use of insecticides to reduce its populations. However, their use has led to insect resistance to these chemicals. Objective: To determine the presence of the F1534C mutation associated with cross-resistance to DDT and pyrethroids in A. aegypti in Sincelejo, Colombia. Materials and methods: We studied nine specimens of A. aegypti that showed resistance to lambdacyhalothrin in bioassays developed by the Secretaría de Salud de Sucre. We used a semi-nested PCR as previously described by Harris, et al., to amplify exon 31 of the para gene of the voltage-dependent sodium channel of A. aegypti. We sequenced, edited, and analyzed PCR products with the MEGA 5 software. Results: We detected the wild and mutant alleles of exon 31 in all of the nine mosquitoes tested, and observed the substitution of thymine for guanine in the nucleotide sequence of the mutant allele, producing a change to UGC in the UUC codon, which led to the replacement of phenylalanine by cysteine in residue 1534 of the protein. Conclusion: The nine mosquitoes analyzed presented a heterozygote genotype for the F1534C mutation, whose phenotypic effect is knockdown resistance (kdr) to DDT and pyrethroids.

Taxonomy and distribution of the series pia of the Lutzomyia verrucarum group (Diptera: Psychodidae), with a description of Lutzomyia emberai n. sp.

A new species of phlebotomine sand fly, Lutzomyia emberai n. sp, is described and illustrated from specimens collected in a dwelling of the Emberá Indian community, situated in the foothills of the Serranía del Baudól on the Colombian Pacific coast. The morphological characteristics of L. emberai n. sp. suggest that it belongs to the series pia of the group verrucarum, easily differentiated from the other members of this group by diagnostic characters on the palps, labro-pharynx, thorax, and spermathecal ducts. The discovery of this new phlebotomine raises to seven the number of species in the series pia, including Lutzomyia pia (Fairchild & Hertig, 1961); Lutzomyia reclusa Fernandez & Rogers, 1991; Lutzomyia suapiensis Le Pont, Torrez-Espejo & Dujardin, 1997; Lutzomyia tihuiliensis Le Pont, Torrez-Espejo & Dujardin, 1997; Lutzomyia tocaniensis Le Pont, Torrez-Espejo & Dujardin, 1997; Lutzomyia limafalcaoae (Wolff & Galati, 2002); and Lutzomyia emberai Bejarano, Duque & Velez, 2004, n. sp. The taxonomy, distribution, and medical importance of this series are reviewed.