Lutzomyia longipalpis naturally infected by Leishmania (L.) chagasi in Várzea Grande, Mato Grosso State, Brazil, an area of intense transmission of visceral leishmaniasis

The American visceral leishmaniasis (AVL) is caused by parasites belonging to the genus Leishmania (Trypanosomatidae) and is transmitted to humans through the bite of certain species of infected phlebotomine sand flies. In this study, we investigated the natural infection ratio of Lutzomyia longipalpis, the main vector species of AVL in Brazil, in Várzea Grande, Mato Grosso State. Between July 2004 and June 2006, phlebotomine sand flies were captured in peridomestic areas using CDC light-traps. Four hundred and twenty (420) specimens of Lu. longipalpis were captured. 42 pools, containing 10 specimens of Lu. longipalpis each, were used for genomic DNA extraction and PCR (polymerase chain reaction) amplification. Leishmania spp. DNA was detected in three out of the 42 pools tested, resulting in a minimal infection ratio of 0.71%. Restriction fragment length polymorphism (RFLP) analysis indicated that Leishmania (L.) chagasi was the infective agent in the positive pools.     

Leishmaniasis in Brazil. XXI. Visceral leishmaniasis in the Amazon Region and further observations on the role of Lutzomyia longipalpis (Lutz & Neiva, 1912) as the vector

Further evidence is presented incriminating the sandfly Lutzomyia longipalpis as the vector of Leishmania chagasi, the causative agent of American visceral leishmaniasis, in the Amazon Region of Brazil. During an outbreak of the disease in Santarém, Pará State, this insect was shown to be the only species of sandfly consistently present in and around the patient's homes, where it often occurred in very large numbers. Of 491 specimens dissected, 35 (7.14%) proved to be infected, and isolates of L. chagasi were made from 16 of 27 of these sandflies following the inoculation of the promastigotes into hamsters. Finally, the parasite was transmitted to four other hamsters which had been subjected to the bites of large numbers of wild-caught Lu. longipalpis. Isolates of Leishmania from Lu. longipalpis captures in Santarém, and in another focus of visceral leishmaniasis on the Island of Marajó, Pará, have been shown to be biologically and biochemically indistinguishable from the parasite infecting man, dogs and foxes in Pará, and from stocks obtained from man elsewhere in Brazil (Bahia and Ceará States).     

Copulation songs in three siblings of Lutzomyia longipalpis (Diptera: Psychodidae)

We present the results of recording male courtship songs of the sandfly Lutzomyia longipalpis. The striking differences in the songs from 3 Brazilian populations of this sandfly with 3 distinct male pheromones support the 3 sibling species previously proposed based on this characteristic.     

Antibodies against Lutzomyia longipalpis saliva in the fox Cerdocyon thous and the sylvatic cycle of Leishmania chagasi

Sera of 11 wild Cerdocyon thous foxes from an endemic area for American visceral leishmaniasis were tested for the presence of antibodies against salivary gland homogenates (SGH) of Lutzomyia longipalpis. All foxes had higher levels of anti-Lu. longipalpis SGH antibodies than foxes from non-endemic areas, suggesting contact between foxes and the vector of visceral leishmaniasis. Sera of humans and dogs living in the same area were also tested for reactivity against Lu. longipalpis SGHs and had a lower proportion of reactivity than foxes. Antibodies against Leishmania chagasi were not detected in any of the foxes, but three foxes showed the presence of parasites in the bone marrow by direct examination, PCR or by infecting the vector. Both humans and dogs had higher levels of anti-Le. chagasi IgG antibodies than C. thous. The finding of an antibody response against saliva of Lu. longipalpis among C. thous together with the broad distribution of the vector in resting areas of infected foxes suggests that the natural foci of transmission of Le. chagasi exists independently of the transmission among dogs and humans.     

Genetic structure and divergence in populations of Lutzomyia cruciata,a phlebotomine sand fly (Diptera: Psychodidae) vector of Leishmania mexicana in southeastern Mexico

The low dispersal capacity of sand flies could lead to population isolation due to geographic barriers, climate variation, or to population fragmentation associated with specific local habitats due to landscape modification. The phlebotomine sand fly Lutzomyia cruciata has a wide distribution throughout Mexico and is a vector of Leishmania mexicana in the southeast. The aim of this study was to evaluate the genetic diversity, structure, and divergence within and among populations of Lu. cruciata in the state of Chiapas, and to infer the intra-specific phylogeny using the 3′ end of the mitochondrial cytochrome b gene. We analyzed 62 sequences from four Lu. cruciata populations and found 26 haplotypes, high genetic differentiation and restricted gene flow among populations (Fst = 0.416, Nm = 0.701, p < 0.001). The highest diversity values were recorded in populations from Loma Bonita and Guadalupe Miramar. Three lineages (100% bootstrap and 7% overall divergence) were identified using a maximum likelihood phylogenetic analysis which showed high genetic divergence (17.2–22.7%). A minimum spanning haplotype network also supported separation into three lineages. Genetic structure and divergence within and among Lu. cruciata populations are hence affected by geographic heterogeneity and evolutionary background. Data obtained in the present study suggest that Lu. cruciata in the state of Chiapas consists of at least three lineages. Such findings may have implications for vector capacity and hence for vector control strategies.     

Natural infection of Lutzomyia longipalpis by Leishmania sp. in Teresina, Piauí State, Brazil

In Brazil, control of the vector Lutzomyia longipalpis is one of the main strategies used to limit the expansion of American visceral leishmaniasis. However, studies on the ecology of this sand fly are rare, especially regarding its natural infection with species of Leishmania. A study of the natural infection of Lu. longipalpis by Leishmania sp. was carried out in the Bela Vista neighborhood in the city of Teresina, Piauí State, Brazil, an important area of American visceral leishmaniasis transmission. From February 2004 to January 2005, sand flies were captured with CDC light traps. Approximately 10 female sand flies in each capture were dissected and examined for the presence of evolutionary forms of Leishmania sp. Two sand fly species were identified: 1,832 were Lu. longipalpis and six Lu. whitmani. Twenty female sand flies (1.1%), all Lu. longipalpis, were infected with procyclic and nectomonad forms of Leishmania sp., found mostly in the hindgut. Higher proportions of infected sand flies were found four months after the rainy season, suggesting that environmental factors may predict not only vector abundance (as already known) but also their level of infection.     

Leishmaniasis in Brazil. XIX: visceral leishmaniasis in the Amazon Region, and the presence of Lutzomyia longipalpis on the Island of Marajó, Pará State

Sporadic cases of visceral leishmaniasis in Amazonian Brazil appear limited to Pará State, in the lower Amazon valley and principally near the Atlantic coast. The fox Cerdocyon thous (L.) has been incriminated as a natural host of the causative parasite, Leishmania donovani chagasi, but past doubts have existed over the identification of the most likely vector as Lutzomyia (Lutzomyia) longipalpis (Lutz & Neiva, 1912). Investigations on two of five recent cases of visceral leishmaniasis of man in the Districts of Cachoeira do Arari and Salvaterra, on the eastern part of the Island of Marajó, Pará showed undoubted Lu. longipalpis to be abundant in one house and in numerous chicken-houses. This is the first record of Lu. longipalpis on Marajó Island, and the finding supports previous implication of this sandfly in the epidemiology of visceral leishmaniasis in other parts of Pará. Morphological differences have been noted between this insect from Marajó and other specimens from more highly endemic regions in the States of Ceará and Minas Gerais, Brazil.     

Leishmaniasis in Bolivia. I. Lutzomyia longipalpis (Lutz & Neiva, 1912) as the vector of visceral leishmaniasis in Los Yungas

A relatively high leishmanial infection rate was found in the phlebotomine sandfly Lutzomyia longipalpis collected from three villages of the Los Yungas region (Department of La Paz, Bolivia). 2,578 female sandflies were dissected. In three houses surveyed in Santa Barbara promastigote infection rates of Lu. longipalpis were 4.2, 2.2 and 3.2% respectively. Anatomical localization of the infection in the insect, and biochemical characterization of the strains indicate that the parasite belongs to the Leishmania donovani complex. The geographical area and the biotopes of Lu. longipalpis are discussed in relation to the vector-parasite relationship.     

Leishmaniasis in Brazil. XXII: Characterization of Leishmania from man, dogs and the sandfly Lutzomyia longipalpis (Lutz & Neiva, 1912) isolated during an outbreak of visceral leishmaniasis in Santarém, Pará State

During epidemiological studies on an outbreak of visceral leishmaniasis in Santarém, Pará State, north Brazil, isolates of Leishmania from two children, three dogs and six naturally infected specimens of the sandfly Lutzomyia longipalpis were compared, biochemically, by starch-gel enzyme electrophoresis. They have proved to be indistinguishable from each other, and from a reference strain of Leishmania chagasi Cunha & Chagas, 1937 from a case of human visceral leishmaniasis from Bahia State, north-east Brazil, on their enzyme profiles for ASAT, ALAT, PGM, GPI, MDH and MPI. Lu. longipalpis is the principal, and possibly the only vector to man in the Amazon Region of Brazil.     

Phlebotomine sandfly species from an American visceral leishmaniasis area in the Northern Rainforest region of Pernambuco State, Brazil

The aim of this study was to identify the fauna of phlebotomine sandflies in an area with incidence of American visceral leishmaniasis (AVL) in S?o Vicente Férrer, a municipality (county) located in the northern Zona da Mata in Pernambuco State, Brazil. Sandfly captures were conducted monthly for four nights/month from December 2002 to November 2003, by means of manual captures (with a Shannon trap) and by CDC light traps. A total of 23,156 specimens of phlebotomines sandflies were collected in a remnant of the Atlantic rainforest, houses, and animal shelters. Lutzomyia complexa was the most prevalent species, with a frequency of 62.5% (14,445/23,156), followed by Lutzomyia migonei, with 33.2% (7,677/23,156). No Lutzomyia longipalpis (the main vector of Leishmania chagasi in Brazil and the Americas) was found in the target area. Some specimens were dissected, but no Leishmania parasites were found.     

First record of finding Lutzomyia longipalpis (Lutz & Neiva, 1912) in the urban area of Brazil

This is a report of the first finding of visceral leishmaniasis' vector Lutzomyia longipalpis in the urban area of Campo Grande, State of Mato Grosso do Sul. The importance of this finding regarding the transmission of the disease in this area is discussed.     

Seasonal and nocturnal domiciliary human landing/biting behaviour of Lutzomyia (Lutzomyia) evansi and Lutzomyia (Psychodopygus) panamensis (Diptera; Psychodidae) in a periurban area of a city on the Caribbean coast of eastern Venezuela (Barcelona; Anzoátegui State)

In recent years, in addition to American cutaneous leishmaniasis (ACL), a significant number of cases of American visceral leishmaniasis (AVL) have been reported in periurban areas of Barcelona city (Anzoátegui State, Venezuela). We studied the bionomics of Lutzomyia (Lutzomyia) evansi and Lutzomyia (Psychodopygus) panamensis, possible vectors of AVL and ACL, respectively, in El Rincón, a periurban village of that city. To evaluate the seasonal domiciliary landing/biting activity of sandflies on human bait, a house was chosen in El Rincón. Landing catches were carried out between 18:00 and 06:00, once a month for a year. The results show the presence of 2 species, Lu. (Lu.) evansi (89.9%) and Lu. (Psy.) panamensis (10.1%). Lu. evansi was most abundant in the months of October and July, associated with the bimodal cycle of annual rainfall in the area. Maximum landing/biting activity of Lu. evansi was observed at 24:00 and 03:00. These findings suggest that at this time of the year and at these hours there is heightened risk of the transmission of AVL. Lu. panamensis monthly abundance also shows a direct association with rainfall and maximum landing/biting activity was observed between 02:00 and 03:00. The lower domiciliary abundance of Lu. panamensis suggests its greater importance in the extradomiciliary transmission of ACL.     

Molecular analysis of natural infection of Lutzomyia longipalpis in an endemic area for visceral leishmaniasis in Brazil

The main purpose of this study was to investigate natural infection by Leishmania chagasi in female sand flies in a visceral leishmaniasis (VL) focus on S?o Luís Island, Maranh?o State, Brazil. Molecular analysis by polymerase chain reaction (PCR) was applied to determine the rate of natural infection of Lutzomyia longipalpis by L. chagasi in areas of old and recent human settlement on S?o Luís Island. Based on a sample of 800 female specimens captured from March to August 2005, the natural infection rate was 1.25% in an area of old settlement and 0.25% in two recently settled areas. Infection of L. longipalpis was detected in both areas, regardless of the number of reported human VL cases, indicating that other factors modulating infection in the wild need to be investigated. The results confirm PCR as a specific technique and an important tool for epidemiological surveillance.     

Correlated morphologic and genetic diversity among Lutzomyia longipalpis (Diptera: Psychodidae) collections in Venezuela

Diversity among Lutzomyia longipalpis populations in Venezuela was characterized using 2 methods: larval mouthpart morphology-morphometry and isoenzyme electrophoresis. Analysis of the results suggested the presence of 2 morpho-genotypes. The mentum, maxillary comb, mandibular ventral teeth, and adenylate kinase and hexokinase enzyme-encoding loci suggested that a population from the northwestern Coriano System (Curarigua) is a distinct lineage within the L. longipalpis complex. Three widely separated populations from the Llanos (savanna), Andes, and northcentral Coastal Cordillera showed no significant substructure. These studies provide morphologic markers that are congruent with genetic data and suggest that the morphologic markers may be used to characterize and differentiate populations within this species complex.     

Sandflies (Diptera, psychodidae) in a secondary forest area in the Paco do Lumiar city, Maranhao, Brazil: a leishmaniasis transmission area

This paper analyzes the wealth of species, relative abundance, seasonal fluctuation, and nocturnal activity of sandflies. The field survey was conducted in a "capoeira" (secondary forest) area in the county of Pa?o do Lumiar, Maranh?o, where cutaneous and transmission of visceral leishmaniasis frequently occurs. Sandflies were captured by CDC-type light traps from 6:00 PM to 6:00 AM, once a month, from March 1997 to February 1998. A total of 489 specimens were collected (251 males and 238 females), distributed among 10 species: Lutzomyia antunesi (45.19%), Lutzomyia whitmani (29.4%), Lutzomyia longipalpis (7.56%), Lutzomyia sordelli (6.34%), Lutzomyia flaviscutellata (4.5%), Brumptomyia avellari (4.09%), Lutzomyia evandroi (1.85%), Lutzomyia umbratilis (0.61%), Lutzomyia corossoniensis (0.41%), and Lutzomyia trispinosa (0.41%). The sandflies were present year round, with higher abundance during the rainy season. They were present in all intervals studied, with the highest frequency between 12:00 PM and 1:00 AM (31%).     

Evolution,epidemiology, and population genetics of black flies (Diptera: Simuliidae)

More than 2000 species of black flies feed on vertebrate blood; 1.5% of all species are vectors of pathogens that cause human diseases. Of nine simuliid-borne animal diseases, only two, mansonellosis and onchocerciasis, afflict humans. Onchocerciasis is a debilitating disease infecting an estimated 40 million people in Africa, Latin America, and Yemen, whereas mansonellosis is a mild disease in the Neotropics. Cytogenetic studies of natural populations of more than 500 species of black flies have revealed that the classic morphospecies of taxonomists is typically a complex of two or more reproductively isolated entities, or sibling (cryptic) species. Most vectors of human pathogens are sibling species, each ecologically unique in traits such as breeding habitats, dispersal capabilities, and degree of vector competence. We review the evolution of black flies, the cytogenetics that have revealed about 260 cytologically distinct entities, the molecular studies that continue to expose additional hidden biodiversity, and a case study of the epidemiology of the Simulium damnosum complex, the largest species complex of blood-feeding arthropods on Earth and the premier group of black flies responsible for human onchocerciasis.     

The Sero-Prevalence of Antibodies to <Emphasis Type="Italic">Trypanosoma cruzi</Emphasis> in Latin American Refugees and Immigrants to Canada

Background: Chagas’ disease is caused by infection with the protozoan agent Trypanosoma cruzi. An estimated sixteen to eighteen million people are infected in Latin America. Outside of endemic regions, Chagas’ disease may be transmitted through the transfusion of infected blood components, congenital infection and organ transplantation. We sought to determine the sero-prevalence of antibodies to T. cruzi in a community sample of Latin American refugees and immigrants to Canada. Methods: This was a sero-prevalence study in Latin American refugees and immigrants living in Canada. Eligible subjects were born in South America, Central America or in Mexico. Participants were recruited from a variety of community settings, as well as from medical clinics. Serum was tested by enzyme-linked immunoassay for antibodies to T. cruzi. Results: A total of 102 participants were enrolled. One sample tested positive for antibodies for T. cruzi. The seroprevalence in our sample was 1.0% (95% CI: 0.2%- 5.3%). Interpretation: We found a low sero-prevalence of Chagas’ disease in a community sample of Latin American immigrants and refugees. Physicians who treat Latin American immigrants should consider the risk profile and clinical status of the individual in their decision to screen for Chagas’ disease.     

Phylogeography of the neotropical sand fly Lutzomyia longipalpis inferred from mitochondrial DNA sequences.

Sand flies in the Lutzomyia longipalpis species complex include the primary vector of Leishmania chagasi, the etiologic agent of visceral leishmaniasis in the Neotropics. Twelve L. longipalpis populations from South and Central America were compared using the cytochrome c oxidase I (COI) gene from the mitochondrial genome. The haplotype profiles for each population revealed that the majority of sequence variation was inter-population (98%) rather than intra-population, suggesting that sequence polymorphisms at the COI locus should provide excellent characters for the study of phylogenetic relationships among populations. Phylogenetic reconstruction using distance (neighbor-joining) and maximum parsimony analysis revealed the existence of four clades among the L. longipalpis populations studied: (1) Laran, (2) Brazilian, (3) cis-Andean and (4) trans-Andean. We suggest that these clades represent species. A biogeographical interpretation of the molecular phylogeny suggests that the process of speciation in the L. longipalpis complex began in the Pliocene, from a sub-Andean-Amazonian gene pool resulting from the Andean orogeny (formation of the East Andean Cordillera). The four clades probably diverged as a result of vicariance events that occurred throughout the late Pliocene and Pleistocene. We propose and discuss several historical scenarios, based on the biogeography and historical geology of Central and South America.     

Leishmaniasis in Brazil. VI. Observations on the seasonal variations of Lutzomyia flaviscutellata in different types of forest and its relationship to enzootic rodent leishmaniasis (Leishmania mexicana amazonensis)

The seasonal fluctuation of Lu. flaviscutellata is shown to be closely related to rainfall, the population of this species declining during the rainy season, between May and March, and increasing again during the dry season, reaching a peak towards December and January. A comparison of the numbers of Lu. flaviscutellata caught in different types of forest showed that this species is more common in the wetter forests of the “igapo” than in the drier “terra firme” forests, although similar seasonal fluctuations are noted in both types of forest. The close correlation between the numbers of Lu. flaviscutellata and the incidence of initial infections of Leishmania mexicana amazonensis in Oryzomys capito indicates that the major factor controlling the incidence of leishmaniasis is the number of sandflies.     

Leishmaniasis in Brazil: XX. Prevalence of “enzootic rodent leishmaniasis” (Leishmania mexicana amazonensis), and apparent absence of “pian bois” (Le. braziliensis guyanensis), in plantations of introduced tree species and in other non-climax forests in eastern Amazônia

In Amazonian Brazil most human leishmaniasis is due to Leishmania braziliensis s.l. and is acquired during the clearing of primary climax forest. One of the largest deforestation projects has taken place on the JARI property where plantations of exotic tree species are grown for paper pulp. The ability of the regional leishmaniasis enzootics to invade plantations was investigated. CDC light-trap catches indicated the phletobomine vectors of Le. b. guyanensis (causing “pian bois” in man) to be very scarce in JARI plantations compared to native-forest controls. It is concluded (drawing on other observations) that the vectors of “pian bois” are unlikely to thrive in any secondary forest. In contrast, catches from mammal traps and rodent-baited (Disney) traps demonstrated the presence in JARI plantations of infected Proechimys guyannensis and large populations of Lutzomyia flaviscutellata, respectively the major rodent reservoir and sandfly vector of Le. mexicana amazonensis. Alone amongst the local vectors of human cutaneous leishmaniasis, Lu. flaviscutellata is adapted to non-climax forests (primary or secondary, natural or man-made; synopsis given). It is predicted that the public health importance of Le. m. amazonensis is unlikely to diminish following the development of Amazônia. This is worrying because ca. 30% of Le. m. amazonensis infections in man cause highly-disfiguring, incurable “diffuse cutaneous leishmaniasis”.