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The genus Phlebovirus (order Bunyavirales, family Phenuiviridae) comprises 57 viruses that are grouped into nine species-complexes. Sandfly-transmitted phleboviruses are found in Europe, Africa and the Americas and are responsible for febrile illness and infections of the nervous system in humans. The aim of this study was to assess the genetic diversity of sandfly-transmitted phleboviruses in connected and isolated forest habitats throughout the Panama Canal area in Central Panama. In total, we collected 13 807 sandflies comprising eight phlebotomine species. We detected several strains pertaining to five previously unknown viruses showing maximum pairwise identities of 45-78 % to the RNA-dependent RNA polymerase genes of phleboviruses. Entire coding regions were directly sequenced from infected sandflies as virus isolation in cell culture was not successful. The viruses were tentatively named La Gloria virus (LAGV), Mona Grita virus (MOGV), Peña Blanca virus (PEBV), Tico virus (TICV) and Tres Almendras virus (TRAV). Inferred phylogenies and p-distance-based analyses revealed that PEBV groups with the Bujaru phlebovirus species-complex, TRAV with the Candiru phlebovirus species-complex and MOGV belongs to the proposed Icoarci phlebovirus species-complex, whereas LAGV and TICV seem to be distant members of the Bujaru phlebovirus species-complex. No specific vector or habitat association was found for any of the five viruses. Relative abundance of sandflies was similar over habitat types. Our study shows that blood-feeding insects originating from remote and biodiverse habitats harbour multiple previously unknown phleboviruses. These viruses should be included in future surveillance studies to assess their geographic distribution and to elucidate if these viruses cause symptoms of disease in animals or humans.

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Foram objetos deste estudo a riqueza, a abundância relativa e a frequência mensal das espécies de flebotomíneos (Diptera, Psychodidae) em uma área urbana do município de Codó, estado doMaranhão, que registra elevada prevalência de leishmaniose visceral. Os insetos foram capturados com armadilhas luminosas do tipo CDC, das 18 h às 6 h, duas vezes por mês, de junho de 2012 a maio de 2013. Foram coletados 5.722 espécimes de 11 espécies, sendo 10 pertencentes ao gênero Lutzomyia e 1 ao gênero Brumptomyia. A abundância foi maior no peridomicílio (4.009 exemplares:66 por cento machos e 34 por cento fêmeas) do que no intradomicílio (1.713 exemplares: 55,9 por cento machos e 44,1 por cento fêmeas). A espécie mais abundante foi L. longipalpis (97,7 por cento) e juntamente com L. evandroi, L.sordelli e L. termitophila foram as mais constantes ao longo do ano. A frequência dos flebotomíneos mostrou-se maior no período seco (57,9 por cento), elevando-se com o aumento da temperatura média(p=0,0156; r=0,1007) e reduzindo-se com o aumento da umidade relativa do ar (p=0,0001;r=-0,1740). Sugerem-se ações de controle vetorial durante a estação seca quando os flebotomíneossão mais abundantes.

The richness, relative abundance and the monthly frequency of species of sandflies (Diptera,Psychodidae) in an urban area of the municipality of Codo, Maranhão, a state that records a highprevalence of visceral leishmaniasis, was studied. The insects were captured with CDC light traps,from 18 h to 6 h, twice per month from June 2012 to May 2013 and 5,722 specimens of 11 species, 10belonging to the genus Lutzomyia and one to the genus Brumptomyia were collected. The abundancewas higher outdoors (4,009 samples: 66 percent males and 34 percent females) than in households (1,713 samples:55.9 percent males and 44.1 percent females). The most abundant species was L. longipalpis (97.7 percent), whichtogether with L. evandroi, L.sordellii and L. termitophila were more constant throughout the year. Thefrequency of sandflies was higher in the dry season (57.9 percent), rising with increasing mean temperature(p=0.0156, r=0.1007) and showed a decrease with increasing relative humidity (p=0.0001, r=-0.1740).Vector control during the dry season, when sandflies are most abundant, is suggested.

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La leishmaniasis es una enfermedad parasitaria causada por protozoarios intracelulares del género Leishmania. Es transmitida al humano mediante la picadura de flebótomos. Las leish- maniasis está clasificada en diferentes variantes clínicas: cutánea localizada o difusa, mucocutá- nea y visceral. Es una parasitosis común en Latinoamérica y sobre todo en los países en vías de desarrollo, donde el estilo de vida de la pobla- ción aumenta el riesgo de enfermarse, al vivir en zonas de concentración parasitaria natural, con poco acceso a los servicios de salud. Por esa razón, esta afección constituye un serio proble- ma de salud, cuyo conocimiento es de gran importancia para los médicos que se desempe- ñan en estas áreas dentro de países endémicos. Se presenta caso clínico de niño de siete meses de edad con úlcera no pruriginosa de forma circular de 5mm de diámetro en pómulo dere- cho, de cinco meses de evolución. El paciente es residente permanente de La Puerta, Meren- dón, San Pedro Sula, un área montañosa con abundante vegetación. Detectado en el Institu- to Hondureño de Seguridad Social Regional del Norte en el cual el estudio histopatológico evidenció macrófagos con amastigotes de leishmania. Se administró tratamiento con glucantime por veinte días, citándole a la mitad del tratamiento y al concluirlo obteniendo resultados cicatriciales favorables, sin recidivas. Este es uno de muchos casos de leishmaniasis cutánea localizada en San Pedro Sula, con lo que se confirma el incremento en la propagación de este parásito protozoario en este municipio...(AU)

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Se realizó una investigación en la localidad de Sevilla, ubicada en el municipio costero de Guamá, en Santiago de Cuba, para identificar las especies de insectos causantes de molestias en esta población, así como sus principales fuentes de cría y los horarios de mayor actividad hematófaga de los adultos, con vistas a dirigir eficientemente las estrategias de control de vectores. A tales efectos, se llevaron a cabo encuestas entomológicas larvarias y capturas de insectos adultos sobre cebo humano. Fue identificado el Leptoconops bequaerti (Kieffer) como la especie de mayor actividad hematófaga en el horario diurno, con mayor incidencia entre las 09.30 y 11.30 horas. Este trabajo constituye un nuevo informe para esta provincia, particularmente la mencionada localidad, y un elemento muy útil desde el punto de vista entomológico e higiénico-epidemiológico, pues estos jejenes son importantes transmisores de encefalitis virales y de otros agentes infecciosos, como las filarias y diversos protozoos(AU)

An investigation was carried out in Sevilla town, located in the coastal municipality of Guamá, Santiago de Cuba, to identify the species of insects affecting this population, as well as their main breeding sources and the periods of the adults' greater hematophagus activity, for directing efficiently the strategies to control vectors. To achieve this, larvae entomological interviews and captures of mature insects on human bait were carried out. Leptoconops becquaerti (Kieffer) was identified as the species of greater hematophagus activity during daily hours, with more incidence between 09.30 and 11.30 hours. It was concluded that this new report of the province, and particularly of the mentioned site, constitutes a very useful element from the entomological, hygienic-epidemiological points of view, because gnats are important viral encephalitis and other infectious agents transmitters, as phylarias and diverse protozoos(AU)

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As leishmanioses estão entre as doenças infecciosas parasitárias de maior incidência no mundo. A transmissão para o vertebrado ocorre pela picada de insetos dípteros pertencentes à família Psychodidae e subfamília Phlebotominae, encontrados em toda a região tropical, especialmente nas regiões quentes e úmidas do planeta. No Brasil, o principal gênero é Lutzomyia, com ampla distribuição no Brasil, desde regiões do sul até o norte do país. A taxa de flebotomíneos naturalmente infectados em áreas endêmicas e identificação correta da espécie de Leishmania em uma determinada espécie de flebotomíneo são de grande importância na epidemiologia das leishmanioses. Das 476 espécies de flebotomíneos encontradas nas Américas, aproximadamente 40 estão envolvidas na transmissão das leishmanioses. O processo de armazenamento de insetos para o diagnóstico da infecção natural por Leishmania a partir do conteúdo estomacal de flebotomíneos é de grande significado ecológico e epidemiológico. Para desenvolvimento das pesquisas entomológicas, diversas armadilhas têm sido desenvolvidas e produzidas comercialmente, como a armadilha do tipo "CDC" (Centers on Diseases Control and Prevention), a qual vem sendo bastante utilizada pela sua eficiência. A utilização de métodos moleculares constitui-se em importante ferramenta a ser sugerida ao entomologista. Neste trabalho de revisão literária, ressaltamos alguns aspectos importantes da pesquisa entomológica nas leishmanioses como métodos de prevenção para esta zoonose.

The transmission to the vertebrate occurs by biting flies and insects belonging to the Psychodidae subfamily Phlebotominae family, found throughout the tropics, especially in hot and humid regions of the planet. In Brazil, the main genre is Lutzomyia, with wide distribution in Brazil, from the southern regions to the north. The rate of naturally infected sand flies in endemic areas and correct identification of Leishmania species in a particular sand fly species are important in the epidemiology of leishmaniasis. Of the 476 species of sandflies found in the Americas, approximately 40 are involved in the transmission of leishmaniasis. The process of storage of insects for the diagnosis of natural Leishmania infection from the stomach contents of sand flies is of great ecological and epidemiological significance. For the development of entomological research, several traps have been developed and commercially produced, as the trap of the "CDC" (Centers on Disease Control and Prevention), which has been largely used for their efficiency. The use of molecular methods has become an important tool to be suggested to the entomologist. In this work of literature review, we highlight some important aspects of entomological research in leishmaniasis as methods for preventing this zoonosis.

La leishmaniasis se encuentra entre las enfermedades infecciosas parasitarias con mayor incidencia en el mundo. La transmisión a los vertebrados se produce por la mordedura de insectos pertenecientes a la familia Phlebotominae e subfamília Psychodidae, que se encuentra en las zonas tropicales, sobre todo en las regiones cálidas y húmedas del planeta. En Brasil, el género principal es Lutzomyia, con amplia distribución en Brasil, a partir de las regiones del sur al norte. La tasa de flebótomos infectados de forma natural en las zonas endémicas y la correcta identificación de las especies de Leishmania en un flebótomo en particular son importantes en la epidemiología de las leishmaniasis . De las 476 especies de flebótomos encontradas en las Américas, aproximadamente 40 están involucrados en la transmisión de la leishmaniasis. El proceso de almacenamiento de los insectos para el diagnóstico de la infección natural de Leishmania a partir de los contenidos estomacales de flebótomos es de gran importancia ecológica y epidemiológica. Para el desarrollo de la investigación entomológica, varias trampas se han desarrollado y producido comercialmente, como la trampa de la "CDC " (Centros de Control y Prevención de Enfermedades), que se ha utilizado en gran medida por su eficacia. El uso de métodos moleculares se ha convertido en una herramienta importante para ser sugerido al entomólogo. En este trabajo de revisión bibliográfica, se destacan algunos aspectos importantes de la investigación entomológica en la leishmaniasis como métodos para la prevención de esta zoonosis.

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Granada virus (GRV), a new phlebovirus within the Naples serocomplex, has been recently described in phlebotomine sandflies from Spain. The presence of anti-GRV immunoglobulin G (IgG) antibodies was investigated by indirect fluorescence assay (IFA) and neutralization test (NT) in 920 serum samples from the Granada population. By IFA, an overall GRV seroprevalence of 15.8% (N = 145) was observed, significantly increasing up to 65 years. NT was positive in 18% of anti-GRV IFA-positive samples. IgG antibodies against Toscana virus (TOSV), a hyperendemic phlebovirus within Granada province, were detected in 40% of anti-GRV-positive cases. Anti-GRV IgM antibodies were detected in 36 (6.6%) of 547 acute-phase serum samples from individuals with febrile illness, exanthema, and/or acute respiratory infection. All positives were anti-TOSV IgM-negative. GRV may infect humans, with most cases being asymptomatic. The codetection of anti-GRV and anti-TOSV IgG antibodies could be attributable to cross-reactivity or exposure to the same transmission vector.

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A new member of the phlebovirus genus, tentatively named Granada virus, was detected in sandflies collected in Spain. By showing the presence of specific neutralizing antibodies in human serum collected in Granada, we show that Granada virus infects humans. The analysis of the complete genome of Granada virus revealed that this agent is likely to be a natural reassortant of the recently described Massilia virus (donor of the long and short segments) with a yet unidentified phlebovirus (donor of the medium segment).

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The developmental cycles of five Brazilian populations of the Lutzomyia longipalpis Lutz & Neiva species complex (Diptera: Psychodidae) were compared under laboratory conditions. Three of the populations were derived from insects collected in allopatric sites at Natal (Rio Grande do Norte State), Jacobina (Bahia State) and Lapinha Cave (Minas Gerais State). The other two originated from Sobral (Ceará State), where the males of two sympatric species can be distinguished by the presence of one (1S) or two (2S) pairs of abdominal spots. The results of the present study clearly show that all three populations whose males produce C16 pheromones and use pulse-type copulation songs (Jacobina, Lapinha Cave and Sobral 1S) are more easily adapted to the colonization conditions used in our laboratory, producing larger egg batches, with higher survival and an overall faster developmental cycle. This contrasts with populations producing C20 male pheromones and using burst-type copulation songs (Natal and Sobral 2S) that produce smaller egg batches, have higher oviposition mortality and a slower rate of development under identical laboratory conditions. In conclusion, these phenological differences are a further indication of the differentiation of the siblings within the Lu. longipalpis species complex.

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En la actualidad la emergencia y re-emergencia de nuevos arbovirus es de gran importancia en salud pública. El género Phlebovirus (Bunyaviridae) es parte importante de este grupo de virus constituido en la actualidad por 68 serotipos. La infección causada por Phlebovirus origina una clínica comúnmente compuesta por fiebre, dolor de cabeza y malestar general; rara vez se presentan meningitis o meningoencefalitis, conocida por sus síntomas como la “fiebre del flebótomo”. Estos virus son transmitidos principalmente por dípteros de los géneros Phlebotomus y Lutzomyia de la familia Psychodidae. La dinámica poblacional de los flebovirus indica que su presencia está íntimamente asociada con los sitios donde se encuentra el vector, distribuido en zonas de Asia, África y Europa entre 20° y 45° de latitud, así como en las Américas asociado principalmente a zonas boscosas. Las hembras de estos géneros son hematófagas y capaces de transmitir el virus transovarialmente, por tanto el ciclo del mismo no se encuentra limitado a la transmisión hospedador-reservoriovector, aunado a esto, las larvas de cuarto instar en diapausa mantienen su capacidad infectiva. Actualmente, el diagnóstico se puede realizar por diversas técnicas serológicas y moleculares para algunos virus del grupo, pero dado que éstas no se realizan con frecuencia, los casos suelen pasar desapercibidos.

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Introducción: Los insectos del género Lutzomyia son vectores biológicos de bacterias, protozoarios y virus que atacan tanto a los seres humanos como a los animales. Tienen importancia sobre todo por cuanto las hembras, que son hematófagas, transmiten las diversas formas de leishmaniasis. En Colombia diferentes estudios han comunicado la distribución geográfica de estos dípteros. Con el presente trabajo se informan las especies en distintas localidades y se incluyen varios lugares sin registros conocidos antes. Materiales y métodos: Los ejemplares de este estudio se capturaron cuando pretendían picar a los colectores y con trampas donde el cebo era la luz o un animal. Varios especimenes se atraparon en sitios de reposo. A los flebótomos así obtenidos se les trató con diversos sistemas para poder disecarlos y montarlos entre lámina y laminilla a fin de hacer las identificaciones hasta especie. Resultados: A partir del estudio de 716 representantes, 124 machos y 592 hembras del género Lutzomyia, en 46 sitios de 29 municipios, 11 departamentos, fue posible identificar 33 especies. Discusión: Se amplía en forma considerable la distribución geográfica de los miembros del género Lutzomyia en varios sitios de Colombia. Se menciona la importancia de algunas especies antropofílicas en su responsabilidad como transmisores de enfermedades, sobre todo de las distintas leishmaniasis y se destaca la necesidad de estudios sistemáticos y taxonómicos a fin de fijar sus papeles en la diseminación de agentes patógenos que afectan a los seres humanos.

Introduction: Phlebotomine sand flies are biological vectors of bacteria, protozoa and viruses attacking both human beings and animals. Sand flies are important since females have haematophagous habits and transmit various types of leishmaniases. In Colombia different studies have given the geographic distribution of these insects. This paper reports the finding of species in several localities and includes some places with no previous records. Material and methods: Specimens were captured while attempting to bite on collectors and with traps (light bait and animal bait). Other specimens were trapped in resting places. Sand flies were processed with different techniques regularly used for dissecting and mounting them for species identification. Results: From the study of 716 specimens, 124 males and 592 females, of the genus Lutzomyia in 46 places from 29 municipalities, in 11 departments (= states, provinces) it was possible to identify 33 species. Discussion: The geographic distribution of the genus Lutzomyia is being greatly extended to new Colombian localities. The importance of some anthropophilic species as vectors of diseases, mainly leishmaniases is discussed. The necessity of systematic and taxonomic studies to know the role of these insects in the transmission of pathologies affecting humans is emphasized.

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Durante 1987, nas regiões sul e central do Estado de Roraima, foram realizadas coletas de flebotomineos nas bases de árvores e em armadilhas CDC colocadas a 1,5 e 10m de altura do solo. Foram obtidos 2812 espécimes, de 45 espécies: 35 na região sul e 27 na região cental; 18 espécies foram comuns as duas áreas e 21 estão sendo aqui assinaladas pela primeira vez no Estado. A presente publicação eleva a 54 o número de espécies conhecidad no Estado.

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Six isolates of a new phlebotomus fever serogroup virus, designated Arboledas virus, were obtained from sand flies (Lutzomyia spp.) collected in northeastern Colombia. One of the isolates was made from a pool of male sand flies. By immunofluorescence, Arboledas virus is related to Caimito and Pacui viruses; by neutralization test, it is distinct. Arboledas virus neutralizing antibodies were found in the sera of opossums (Didelphis marsupialis) and humans living in the study area. D. marsupialis inoculated with the virus developed a viremia of four days' duration, and sand flies (Lutzomyia gomezi) feeding on a viremic opossum were readily infected. Transovarial transmission of Arboledas virus was also demonstrated in experimentally infected Lu. gomezi. Results of the above laboratory studies suggest that Arboledas virus is maintained in nature by two mechanisms: vertical (transovarial) transmission in the insect vector, and an alternating marsupial-sand fly cycle. The implications of this complex maintenance cycle for other phleboviruses are discussed.

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Eight new members of the phlebotomus fever arbovirus serogroup (family Bunyaviridae; genus Phlebovirus) from the Amazon region of Brazil are described. One serotype was recovered from a febrile patient, three from small wild animals and four from sand flies. A small serum survey carried out with the human isolate, Alenquer virus, suggests that it rarely infects man. Complement-fixation and plaque reduction neutralization tests were done, comparing the eight new viruses with other members of the phlebotomus fever serogroup. A close antigenic relationship was demonstrated between one of the new agents (Belterra) and Rift Valley fever virus. This finding is of considerable interest and deserves further investigation. Addition of these eight new viruses to the genus Phlebovirus brings to 14 the number of serotypes known to occur in the Amazon region and to 36 the total number reported worldwide. More detailed clinical and epidemiological studies should be conducted in Amazonia in order to define the public health impact caused by phleboviruses.

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