Molecular detection and identification of Bartonella in the cat flea Ctenocephalides felis felis collected from companion animals in a border area in northeastern Argentina

Molecular methods were used to detect and identify Bartonella species in the cat fleas Ctenocephalides felis felis from Puerto Iguazú, a border area in northeastern Argentina. The fleas were collected from 12 household animals, 9 dogs (Canis lupus familiaris) and 3 cats (Felis silvestris catus) during July 2016. Out of 15C. f. felis analyzed for PCR, only one flea collected from a cat was positive (6.66%) in screened for Bartonella spp. based on the gltA gene. Bartonella clarridgeiae was identified in the genetic analyses, this specimen clustered monophyletically with others B. clarridgeiae isolated from different geographical origins (1.0 PP), even, all shared the same haplotype. The results obtained provide evidence of the presence of B. clarridgeiae in cat fleas from Argentina suggesting the probable presence of related flea-borne diseases in the region and the role of cat fleas in the transmission of Bartonella among mammals including humans.Centro de Estudios Parasitológicos y de Vectore

Lutzomyia longipalpis (Diptera: Psychodidae) Argentina-Bolivia border: new report and genetic diversity

American visceral leishmaniasis (AVL) has two main scenarios of transmission as follows: scattered cases in rural areas andurban outbreaks. Urban AVL is in active dispersion from the northeastern border of Argentina-Paraguay-Brazil to the South.The presence of Lutzomyia longipalpis was initially reported in urban environments in the northwestern border of the country.The presence of Lu. longipalpis, environmental variables associated with its distribution, and its genetic diversity were assessedin Salvador Mazza, Argentina, on the border with Bolivia. The genetic analysis showed high haplotype diversity, low nucleotidediversity, and low nucleotide polymorphism index. We discuss the hypothesis of an expanding urban population with introgressivehybridisation of older haplogroups found in their path in natural forest or rural environments, acquiring a new adaptability tourban environments, and the possibility of changes in vector capacity.Fil: Quintana, María Gabriela. Ministerio de Salud. Instituto Nacional de Medicina Tropical; Argentina. Red de Investigación de la Leishmaniasis en la Argentina; ArgentinaFil: Pech May, Angélica del Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Red de Investigación de la Leishmaniasis en la Argentina; Argentina. Ministerio de Salud. Instituto Nacional de Medicina Tropical; ArgentinaFil: Fuenzalida, Ana Denise. Ministerio de Salud. Instituto Nacional de Medicina Tropical; Argentina. Red de Investigación de la Leishmaniasis en la Argentina; Argentina. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo. Instituto Superior de Entomología; ArgentinaFil: Direni Mancini, José Manuel. Red de Investigación de la Leishmaniasis en la Argentina; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo. Instituto Superior de Entomología; ArgentinaFil: Barroso, Paola Andrea. Ministerio de Salud. Instituto Nacional de Medicina Tropical; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; ArgentinaFil: Yadon, Zaida Estela. Ministerio de Salud. Instituto Nacional de Medicina Tropical; ArgentinaFil: Zaideneberg, Mario. Ministerio de Salud y Acción Social; ArgentinaFil: Salomón, Oscar Daniel. Ministerio de Salud. Instituto Nacional de Medicina Tropical; Argentina. Red de Investigación de la Leishmaniasis en la Argentina; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentin

Development and evaluation of a duplex TaqMan qPCR assay for detection and quantification of Trypanosoma cruzi infection in domestic and sylvatic reservoir hosts

Background: A question of epidemiological relevance in Chagas disease studies is to understand Trypanosoma cruzi transmission cycles and trace the origins of (re)emerging cases in areas under vector or disease surveillance. Conventional parasitological methods lack sensitivity whereas molecular approaches can fill in this gap, provided that an adequate sample can be collected and processed and a nucleic acid amplification method can be developed and standardized. We developed a duplex qPCR assay for accurate detection and quantification of T. cruzi satellite DNA (satDNA) sequence in samples from domestic and sylvatic mammalian reservoirs. The method incorporates amplification of the gene encoding for the interphotoreceptor retinoid-binding protein (IRBP), highly conserved among mammalian species, as endogenous internal amplification control (eIAC), allowing distinction of false negative PCR findings due to inadequate sample conditions, DNA degradation and/or PCR interfering substances. Results: The novel TaqMan probe and corresponding primers employed in this study improved the analytical sensitivity of the assay to 0.01 par.eq/ml, greater than that attained by previous assays for Tc I and Tc IV strains. The assay was tested in 152 specimens, 35 from 15 different wild reservoir species and 117 from 7 domestic reservoir species, captured in endemic regions of Argentina, Colombia and Mexico and thus potentially infected with different parasite discrete typing units. The eIACs amplified in all samples from domestic reservoirs from Argentina and Mexico, such as Canis familiaris, Felis catus, Sus scrofa, Ovis aries, Equus caballus, Bos taurus and Capra hircus with quantification cycles (Cq´s) between 23 and 25. Additionally, the eIACs amplified from samples obtained from wild mammals, such as small rodents Akodon toba, Galea leucoblephara, Rattus rattus, the opossums Didelphis virginiana, D. marsupialis and Marmosa murina, the bats Tadarida brasiliensis, Promops nasutus and Desmodus rotundus, as well as in Conepatus chinga, Lagostomus maximus, Leopardus geoffroyi, Lepus europaeus, Mazama gouazoubira and Lycalopex gymnocercus, rendering Cq´s between 24 and 33. Conclusions: This duplex qPCR assay provides an accurate laboratory tool for screening and quantification of T. cruzi infection in a vast repertoire of domestic and wild mammalian reservoir species, contributing to improve molecular epidemiology studies of T. cruzi transmission cycles.Fil: Wehrendt, Diana Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Gómez Bravo, Andrea. Fundación Mundo Sano; ArgentinaFil: Ramirez Gomez, Juan Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Cura, Carolina Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Pech May, Angélica del Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; ArgentinaFil: Ramsey, Janine M.. Instituto Nacional de Salud Pública; MéxicoFil: Abril, Marcelo. Fundación Mundo Sano; ArgentinaFil: Guhl, Felipe. Universidad de los Andes; ColombiaFil: Schijman, Alejandro Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentin

Primera detección de Rickettsia asembonensis en Ctenocephalides felis felis en Argentina : Un estudio epidemiológico en pulgas de animales de compañía y sinantrópicos en el trifinio del Noreste argentino

Las rickettsiosis son enfermedades zoonóticas causadas por bacterias del género Rickettsia y transmitidas a humanos por medio de artrópodos vectores. En este sentido, las pulgas son importantes en salud pública debido a su rol como parásitos y como vectores de bacterias patógenas.Trabajo publicado en Cagliada, Maria del Pilar Lilia y Galosi, Cecilia Mónica (comps.). I Congreso de Microbiología Veterinaria. Libro de resúmenes. La Plata: Facultad de Ciencias Veterinarias, 2021.Facultad de Ciencias Veterinaria

Genética poblacional y filogeografía de Lutzomyia longipalpis (Diptera: Psychodidae), vector de Leishmania infantum

El complejo Lutzomyia longipalpis tiene una distribución amplia, aunque discontinua, por toda la región neotropical entre México y el norte de Argentina y Uruguay. En América, este flebotomino es el principal vector de Leishmania infantum, parásito responsable de la leishmaniasis visceral (LV). Estudios genéticos proporcionan evidencia de al menos cuatro especies hermanas en el complejo de Lu. longipalpis, sin embargo, no hay consenso actual sobre el número de haplogrupos, o sobre su tiempo de divergencia. A pesar de su expansión hacia el sur y la reciente colonización de ambientes urbanos, son escasos los estudios genéticos de Lu. longipalpis en Argentina. Por tal motivo, en la presente tesisse analizó la diversidad genética y la estructura de Lu. longipalpis en Argentina, y estos datos se integraron con datos tomados de la bibliografía para re-evaluar la filogeografía del complejo Lu. longipalpis utilizando marcadores mitocondriales a escala de América Latina. La diversidad genética fue estimada a partir de muestreos provenientes de seis localidades en Argentina, utilizando un fragmento de los genes ND4 y cyt b. Se encontraron altos valores de diversidad genética en Tartagal, Santo Tomé y San Ignacio, así como una altadiferenciación genética entre poblaciones usando ambos marcadores. Los análisis genéticos espaciales de Geneland revelan la existencia de dos grupos genéticos en Argentina. Los análisis filogeográficos con ambos genes concatenados identificaron tres haplogrupos en Argentina, mientras que cuando se incluyeron secuencias de genes ND4 y cyt b de diversossitios geográficos de la región neotropical (disponibles en GenBank), se encontró una mayor divergencia entre haplogrupos que la reportada en otros estudios. Con el gen ND4 se diferenciaron al menos ocho haplogrupos a lo largo de la región neotropical, cada uno separado por múltiples pasos mutacionales, mientras que con el gen cyt b (para el que hay menos secuencias disponibles en GenBank), se diferenciaron cinco haplogrupos que tuvieron menos pasos mutacionales entre ellos. La divergencia del complejo de Lu. longipalpis con respecto a su ancestro común más reciente (MRCA) se estimó en 0.70 Ma (intervalo de credibilidad de HPD del 95% = 0,48 - 0,99 Ma) y 0.45 Ma (intervalo decredibilidad de HPD del 95% = 0,22 - 0,83 Ma) para el gen ND4 y cyt b, respectivamente. A partir de los resultados de esta tesis doctoral surge la necesidad de analizar la competencia vectorial de acuerdo con el haplogrupo, entre otros parámetros intrínsecos de la zoonosis, yconsiderarlos en el diseño y vigilancia de las estrategias de control de vectores y de transmisión de la LV en la región.The Lutzomyia longipalpis complex has a wide but discontinuous distribution throughout the Neotropical realm between Mexico and northern Argentina and Uruguay. In the Americas, this phlebotomine sandfly is the main vector of Leishmania infantum, the parasite responsible for Visceral Leishmaniasis (VL). Genetic studies provide evidence for at least four sibling species in the Lu. longipalpis complex, although there is no current consensus on the number of haplogroups, or on their divergence. Despite its southern expansion and recent colonization of urban environments, there have been few genetic analyses of Lu. longipalpis in Argentina. In this PhD thesis, the genetic diversity and structure of Lu. longipalpis from key sites in Argentina were analyzed, and the data were used to reevaluate the phylogeography of the Lu. longipalpis complex at a Latin American scale using mitochondrial markers. Genetic diversity was estimated from six sites in Argentina, using a fragment of the ND4 and the cyt b genes. Highest genetic diversity was found in Tartagal, Santo Tomé and San Ignacio.There was high genetic differentiation of Lu. longipalpis in Argentina using both markers. Genetic and spatial Geneland analyses reveal the existence of two primary genetic clusters in Argentina. Phylogeographic analyses using the ND4 and the cyt b fragments available in GenBank from diverse geographic sites, indicate greater divergence than previously reported. At least eight haplogroups each separated by multiple mutational steps using the ND4, are differentiated across the Neotropical realm. While, with the cyt b gene, five haplogroups are differentiated (fewer sequences available in GenBank), although they had fewer mutational steps between them. With both genes, three haplogroups were identified in Argentina. Divergence of the Lu. longipalpis complex from its most recent common ancestor (MRCA) was estimated to have occurred 0.70 MYA (95% HPD interval = 0.48 - 0.99 MYA) and 0.45 MYA (95% HPD interval = 0.22 - 0.83 MYA) for the ND4 and cyt b genes, respectively. This PhD study provides new evidence for Lu. longipalpis complex, these findings suggest the need to analyze vector competence, among other parameters intrinsic to a zoonosis, based on vector haplogroup, and to incorporate this evidence in the design and surveillance of vector and transmission control strategies in the region.Fil: Pech May, Angélica del Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Ministerio de Salud. Instituto Nacional de Medicina Tropical; Argentin

Phlebotomine sand flies (Diptera: Psychodidae) of Chiapas collected near the Guatemala border, with additions to the fauna of Mexico and a new subgenus name

Collections from four localities, two of the High Plateau and two of the Eastern Mountains Municipality of Chiapas, near the border with Guatemala, included 26 species with four new species records for Mexico: Lutzomyia (Helcocyrtomyia) hartmanni (Fairchild & Hertig, 1957), Dampfomyia (Coromyia) disneyi (Williams, 1987), Psychodopygus bispinosus (Fairchild & Hertig, 1951), and Psychodopygus corossoniensis (LePont & Pajot, 1978). These records represent an updated total of 50 species in Mexico, 48 of which are extant species and the remaining two fossils. The name Xiphopsathyromyia n. n. is proposed in substitution of Xiphomyia Artemiev, 1991, a homonym of Xiphomyia Townsend, 1917, a genus of Tachinidae (Diptera).Fil: Ibañez Bernal, Sergio. Instituto de Ecología; MéxicoFil: Muñoz, José. Instituto Nacional de Salud Pública; MéxicoFil: Rebollar Téllez, Eduardo A.. Universidad Autónoma de Nuevo León; MéxicoFil: Pech May, Angélica del Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Ministerio de Salud. Instituto Nacional de Medicina Tropical; Argentina. Instituto Nacional de Salud Pública; MéxicoFil: Marina, Carlos F.. Instituto Nacional de Salud Pública; Méxic

Haplotipos del fragmento barcode del gen COI de Migonemyia migonei

Nuevos haplotipos generados a partir del análisis de diversidad en Migonemyia migonei, a partir de la base de datos (secuencias) armada con datos propios y de terceros disponibles en bases de datos online.Fil: Moya, Sofía Lorián. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Administracion Nacional de Laboratorios E Institutos de Salud "dr. Carlos G. Malbran". Instituto Nacional de Medicina Tropical.; ArgentinaFil: Pech May, Angélica del Rosario. Facultad de Medicina Veterinaria y Zootecnia; México. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Quintana, María Gabriela. Administracion Nacional de Laboratorios E Institutos de Salud "dr. Carlos G. Malbran". Instituto Nacional de Medicina Tropical.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Salomón, Oscar Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Administracion Nacional de Laboratorios E Institutos de Salud "dr. Carlos G. Malbran". Instituto Nacional de Medicina Tropical.; Argentin

Cryptic Diversity in Sympatric Migonemyia migonei (Diptera: Psychodidae), Eventual Meaning for Leishmaniasis Transmission

Migonemyia migonei (Franҫa, 1920) (Diptera: Psychodidae) belongs to the subfamily Phlebotominae, of epidemiological importance due to its role as a vector in leishmaniasis transmission cycles and its broad geographic distribution in South America. Few morphometric and genetic studies have demonstrated the existence of variability among geographically distant populations in Brazil. The aim of the study was to estimate the genetic distance within the morphospecies Mg. migonei through the analysis of cytochrome C oxidase subunit I (COI) sequences of specimens captured in Argentina and those available in online databases. The COI sequences from specimens collected in different localities of Argentina and sequences available in online databases were utilized. Genetic distances were analyzed and a median-joining haplotype network was constructed. Finally, phylogenetic reconstruction was performed according to Bayesian inference. The analyses led to the identification of at least two haplogroups: haplogroup I with sequences of specimens from Colombia, Brazil and Argentina, and haplogroup II with sequences of specimens from Argentina. Interestingly, specimens from Argentina whose haplotypes corresponded to both haplogroups, were collected in sympatry. The results suggest that Mg. migonei could be a species complex with at least two distinct members. This hypothesis could explain the known characteristics of adaptability and vector permissiveness of the species, as the putative cryptic species of the complex could differ in traits of epidemiological importance.Fil: Moya, Sofía Lorián. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud "Dr. C. G. Malbrán"; ArgentinaFil: Pech May, Angélica del Rosario. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud "Dr. C. G. Malbrán"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Quintana, María Gabriela. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud "Dr. C. G. Malbrán"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Tucumán; ArgentinaFil: Salomón, Oscar Daniel. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud "Dr. C. G. Malbrán"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Phylogenetic relationships of closely-related phlebotomine sand flies (diptera: psychodidae) of nyssomyia genus and Lutzomyia subgenus

BACKGROUND The Nyssomyia genus and Lutzomyia subgenus include medical important species that are Latin American leishmaniases vectors. Little is known about the phylogenetic relationships of closely-related species in each of these taxonomic groups that are morphologically indistinguishable or differentiated by very subtle details.OBJECTIVES We inferred the phylogenetic relationships of closely-related species within both the Nyssomyia genus and the Lutzomyia subgenus using a cytochrome c oxidase subunit I (COI) fragment.METHODS The sampling was carried out from 11 Argentinean localities. For genetic analyses, we used GenBank sequences in addition to our sequences from Argentina. Kimura 2-parameter (K2P) genetic distance and nucleotide divergence (Da) was calculated between closely-related species of Nyssomyia genus, Lutzomyia subgenus and between clades of Lutzomyia longipalpis complex.FINDINGS The K2P and Da values within species of Nyssomyia genus and Lutzomyia subgenus were lower than the divergence detected between clades of Lu. longipalpis complex. The haplotype network analyses within Lutzomyia subgenus showed shared haplotypes between species, contrary to Nyssomyia genus with none haplotype shared. Bayesian inference within Nyssomyia genus presented structuring by species.MAIN CONCLUSIONS This study evidences the phylogenetic proximity among closely-related species within Nyssomyia genus and Lutzomyia subgenus. The COI sequences of Nyssomyia neivai derived from the present study are the first available in GenBank.Fil: Pech May, Angélica del Rosario. Administracion Nacional de Laboratorios E Institutos de Salud "dr. Carlos G. Malbran". Instituto Nacional de Medicina Tropical.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Moya, Sofía Lorián. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Administracion Nacional de Laboratorios E Institutos de Salud "dr. Carlos G. Malbran". Instituto Nacional de Medicina Tropical.; ArgentinaFil: Quintana, María Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Administracion Nacional de Laboratorios E Institutos de Salud "dr. Carlos G. Malbran". Instituto Nacional de Medicina Tropical.; ArgentinaFil: Manteca Acosta, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Administracion Nacional de Laboratorios E Institutos de Salud "dr. Carlos G. Malbran". Instituto Nacional de Medicina Tropical.; ArgentinaFil: Salomón, Oscar Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Administracion Nacional de Laboratorios E Institutos de Salud "dr. Carlos G. Malbran". Instituto Nacional de Medicina Tropical.; Argentin

Inferring distributional shifts of epidemiologically important North and Central American sandflies from Pleistocene to future scenarios

Nine sandfly species (Diptera: Psychodidae) are suspected or proven vectors of Leishmania spp. in the North and Central America region. The ecological niches for these nine species were modelled in three time periods and the overlaps for all time periods of the geographic predictions (G space), and of ecological dimensions using pairwise comparisons of equivalent niches (E space), were calculated. Two Nearctic, six Neotropical and one species in both bioregions occupied a reduced number of distribution areas. The ecological niche projections for most sandfly species other than Lutzomyia shannoni and Lutzomyia ovallesi have not expanded significantly since the Pleistocene. Only three species increase significantly to 2050, whereas all others remain stable. Lutzomyia longipalpis shared a similar ecological niche with more species than any other, although both L. longipalpis and Lutzomyia olmeca olmeca had conserved distributions over time. Climate change, at both regional and local levels, will play a significant role in the temporal and spatial distributions of sandfly species.Fil: Moo Llanes, David A.. Universidad Nacional Autónoma de México; MéxicoFil: Pech May, Angélica del Rosario. Ministerio de Salud. Instituto Nacional de Medicina Tropical; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario; ArgentinaFil: Ibarra Cerdeña, C. N.. Cinvestav Unidad Merida; México. Universidad Nacional Autónoma de México; MéxicoFil: Rebollar Téllez, E. A.. Universidad Autónoma de Nuevo León; MéxicoFil: Ramsey, Janine. Instituto Nacional de Salud Pública/crisp; Méxic