Laboratory evaluation of insecticide-treated sugar baits for control of phlebotomine sand flies (Diptera: Psychodidae)

The purpose of this study was to evaluate the use of boric acid, imidacloprid, ivermectin, or abamectin incorporated into sugar baits as oral toxicants for adult phlebotomine sand flies. Variable toxicity of insecticide-sugar bait solutions to adult male and female sand flies was demonstrated, based on male female median lethal concentration values of 0.10-0.08, 6.13-9.53, and 9.03-18.11 mg/liter of imidacloprid, ivermectin, and abamectin, respectively. Complete control of sand flies could not be achieved with as high as 40 g/liter of boric acid in sugar bait solution; concentrations >40 g/liter were found repellent to the sand flies. Uranine O (a fluorescent tracer dye that can be used to measure the ingestion of sugar baits by sand flies) did not interact negatively with imidacloprid, ivermectin, or abamectin when it was combined with the insecticides in a sugar bait. Also, incorporation of imidacloprid, ivermectin, or abamectin into sugar baits did not reduce the effect whether adult male and female sand flies fed on these sugar baits. We propose that imidacloprid, ivermectin, or abamectin could be used to control adult sand fly populations with targeted use of insecticide-treated sugar baits.     

Brain cell karyotypes of six new world sand flies (Diptera: Psychodidae)

The brain cell karyotypes of six New World sand flies are described and compared. Lutzomyia trapidoi has three pairs of chromosomes, 2N = 6. The other species, L. gomezi, L. erwindonaldoi, L. carmelinoi, L. walkeri and L. columbiana, have four pairs of chromosomes, 2N = 8. No heteromorphic chromosomes were observed. Cytogenetic similarities among sand fly species are discussed.     

Ivermectin as a rodent feed-through insecticide for control of immature sand flies (Diptera: Psychodidae)

Ivermectin was evaluated as a potential rodent feed-through for the control of immature stages of Phlebotomus papatasi. The survival of sand fly larvae fed feces of Syrian hamsters (Mesocricetus auratus) that had been fed a diet containing 0, 2, 6, 10, 20, 60, or 100 ppm ivermectin was measured. Sand fly larvae fed the feces of ivermectin-treated hamsters had significantly reduced survival, with 100% mortality of larvae fed feces of hamsters fed a diet containing 20, 60, and 100 ppm ivermectin. The results of this study suggest that a control strategy using rodent baits containing ivermectin to control phlebotomine sand flies may be possible. Because rodent reservoirs and sand fly vectors of Leishmania major live in close association in many parts of the Middle East, the control of transmission of the agent of zoonotic cutaneous leishmaniasis also may be possible.     

Diversity and species composition of sand flies (Diptera: Psychodidae) in a Venezuelan urban focus of cutaneous leishmaniasis

The present study examined the spatial and temporal abundance and diversity of phlebotomine sand flies in an area of Venezuela that is an ancient focus of leishmaniasis. The study was conducted in 6 stations in urban localities in Trujillo City, located in northwestern Venezuela (9 degrees 22' 24" N, 70 degrees 26' 08" W), which is located in a mountain range in the Andean ecoregion (altitude = 600-1,010 m). During 1995-99, entomological surveys were conducted after and before the rainy season. Shannon light traps were operated from 1800 to 2000 h in peridomestic site trap locations. Twelve species were captured, and Lutzomyia youngi, L. ovallesi, L. scorzai, L. gomezi, L. lichyi, and L. shannoni occurred at all localities in each year. The abundance of these species showed low variation over time but high variation between localities. The S?rensen similarity index, used to compare diversity between years within each locality, ranged from 0.60 at Carmona to 0.84 at La Hacienda. Sand fly communities exhibited annual variation in species richness and diversity. Variations were affected more by changes in species abundance than by changes in species composition. Lutzomyia ovallesi, L. lichyi, and L. scorzai had the highest coefficient of variation between years (63, 38, and 23%, respectively).     

Standardization of conditions for PCR detection of Leishmania spp. DNA in sand flies (Diptera, Psychodidae)

The correct identification of etiological agents in vector insects is crucial for epidemiological studies. Identification of flagellates in such vectors, usually by dissection of the digestive tract and microscopic observation of the contents as well as attempts at parasite isolation from insects in culture media, have proven operationally inadequate and with poor diagnostic specificity, since female sand flies are also hosts for other flagellates like Trypanosoma and Endotrypanum. Due to the efficiency and specificity of DNA target sequence amplification by polymerase chain reaction (PCR), the latter could be used to investigate the presence of Leishmania in sand flies, although the insects need to be properly stored and the Leishmania DNA extracted using appropriate methodology. This paper describes methodologies to standardize sand fly storage and Leishmania DNA extraction in such specimens as a more practical method in field studies.     

Attraction of sand flies (Diptera: Psychodidae) to light traps in rural areas of Minas Gerais State,Brazil

The objective of this study was to detect phlebotomine sand fly vector species by using light traps, and thus determine the risk of cutaneous leishmaniasis (CL) transmission in rural areas of the Vi?osa microregion, Minas Gerais State, Brazil. Sand flies were captured with Shannon-like and Falc?o light traps hung between 1830 and 2230 h on 22 nights. Four sampling points where CL transmission occurs naturally were chosen. A total of 888 sand flies was collected. The most abundant species was Lutzomyia intermedia. Shannon-like traps caught significantly more L. intermedia, perhaps because of a greater surface area on which the insects can land or because they reflect more light. The positive phototaxis observed in L. intermedia and L. whitmani may pose a greater risk of transmission of Leishmania in houses where an external light source is situated close to a light-colored wall that reflects light and that have adjacent bushes or trees and domestic animal shelters within 50 m.     

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.     

Frequency of Wolbachia infection in laboratory and field sand fly (Diptera: Psychodidae) populations

Using a polymerase chain reaction (PCR) assay with primers designed for the Wolbachia 16S rRNA and outer surface protein (wsp) gene, we screened 11 laboratory colonies and 4 field samples of 6 sand fly species for Wolbachia infection. Infection was only detected in 3 laboratory colonies of Phlebotomus papatasi Scopoli, originally collected in Israel, Egypt, and Saudi Arabia.     

Evaluation of repellent and anti-feeding effect of garlic oil (Allium sativum) against the bite of phlebotomine sandflies Diptera: Psychodidae

The repellent and anti-feeding effect of garlic oil was evaluated in laboratory conditions against the bite of Phlebotomus papatasi females. The effectiveness was evaluated by two different laboratory procedures: (i) topical application of garlic oil on five human volunteers, using the "standard cage test", and (ii) feeding sandflies on artificial membranes treated with the compound. Garlic oil showed a significant protection by topical application on the skin of volunteers, being the protection 97% and 40%, respectively at 1% and 0.005% dilution. Garlic oil showed also an anti-feeding effect when tested on the artificial membrane feeding apparatus; the anti-feeding effect was dose dependent, being 100% at 1%. The results of the present study confirm previous observations on the repellent effect of garlic oil against haematophagous arthropods.     

Circadian activity of Lutzomyia shannoni (Diptera: Psychodidae) during late season population peaks

The phlebotomine sand fly, Lutzomyia shannoni (Dyar), is considered both zoophillic and anthropophillic and may be a potential vector of human and livestock pathogens. While the known range of this species in North America has recently been expanded, many aspects of its natural history, which are fundamental components for assessments of disease risk or development of integrated pest management programs for the vector, remain unclear. One is the time of day when individuals are active and searching for blood meals. Through the use of a rotating trap mechanism, male L. shannoni were found to be active earlier in the evening (1-4 h after sunset), whereas the majority of female activity occurred after midnight.     

Hybridization in Phlebotominae (Diptera: Psychodidae): A mini-review

Taxonomy based only on morphology, although extremely important for the classification of sandflies, has been shown to be insufficient for the delimitation of some taxa. Thus, integrative taxonomy could play a fundamental role in clarifying these and other taxonomic issues, since data from different areas are used to aggregate greater reliability in species classification. Experimental crosses are important taxonomic tools, since the presence of reproductive barriers when associated with divergence between two evolutionary lineages, confirms the specific status of taxa based on the biological species concept. In the subfamily Phlebotominae, experimental crosses were mostly focused on the study of the Lutzomyia longipalpis complex, which helped to identify different evolutionary lineages for that group of vectors. Considering the difficulty of classifying some Phlebotominae species and the importance of hybridization studies for taxonomy, we grouped all the information associated with experimental crosses in sandflies in a mini-review. In view of the results grouped in this review, it is evident that i) experimental crossings are important tools to aggregate studies of integrative taxonomy in the Phlebotominae subfamily; ii) these analyses should be applied in the taxonomic studies of cryptic species; iii) Lu longipalpis populations have pre and/or post-zygotic reproductive barriers; iv) Lu. longipalpis represents more than one species and efforts must be applied to differentiate the taxa of the Lu. longipalpis complex; v) Phlebotomus populations do not present intraspecific reproductive barriers; vi) the absence of reproductive barriers between Ph. sergenti from Israel and Turkey (representing populations of the same evolutionary lineage) does not rule out the possible existence of cryptic species, it being necessary to perform experimental crosses between the different strains indicated by the molecular markers; and finally, vii) different species of Phlebotomus have post-zygotic barriers, confirming the specific status of Ph. duboscqi, Ph. papatasi, and Ph. bergeroti.     

Exclusion of phlebotomine sand flies from inhabited areas by means of vertical mesh barriers

Vector control constitutes an important component of integrated disease control campaigns. Source reduction is not an option for phlebotomine sand fly vectors of leishmaniasis, because larval breeding sites remain either unknown or inaccessible. Thus, all control efforts are directed against the adult sand flies, mostly attempting to limit their contact with humans. We describe experiments using an insecticide-treated vertical barrier to prevent sand flies from reaching inhabited areas of an agricultural settlement. A 400 meter long section of the peripheral fence of Kibbutz Sde Eliyahu, Jordan Valley, Israel was draped with a deltamethrin-impregnated net that is impenetrable to sand flies (polyester net, 450 holes/inch²). Sand flies were captured before and after construction of the barrier using CO₂-baited CDC traps. Sand fly numbers, as monitored around three houses internal to the barrier, exhibited an 84.9% decrease once the barrier was erected (P = 0.003). Concurrently, the neighboring control group of three houses, not protected by the barrier, exhibited a 15.9% increase in sand fly numbers (P = 0.974). These results corroborate previous findings of field tests conducted on a smaller scale in an arid suburban setting. Campaigns for reducing the burden of sand fly bites and curtailing the transmission of leishmaniasis, should consider integrating vertical fine-mesh nets with other sand fly control measures.     

Description of a new species of sand fly Sergentomyia (Parrotomyia) vadhanurensis (Diptera: Psychodidae) from Puducherry Union Territory, India

A new species of phlebotomine sand fly, Sergentomyia (Parrotomyia) vadhanurensis, is described from Puducherry Union Territory, India, with illustrations of adult female and male specimens. This species was collected in tree holes, tree buttresses, termite mounds, cattle sheds, and human dwellings from rural areas.     

Vertical stratification of phlebotomine sandfly fauna (Diptera,Psychodidae) in a primary non-flooded forest of the Central Amazon,Amazonas State,Brazil

Studies on the vertical stratification of phlebotomine sandfly fauna were conducted in a non-flooded primary forest at a Tropical Forest Experimental Station of the National Institute of Amazonian Research from October 1998 to March 1999. CDC light traps were placed at one, 10, and 20 meters above ground. A total of 2,859 sandflies were captured, belonging to the Lutzomyia (99.93%) and Brumtomyia (0.07%) genera, represented by 38 species. In the Lutzomyia gender, the most frequent sub-gender was Nyssomyia (43.4%), followed by Psychodopygus (22.8%). Lutzomyia umbratilis, L. anduzei, L. rorotaensis, L. trichopyga, and L. olmeca nociva predominated at one meter above ground, while L. davisi, L. infraspinosa, L. umbratilis, L. trichopyga, and L. anduzei predominated at 10 meters. L. anduzei, L. tuberculata, L. dendrophyla, and L. dreisbachi were the most abundant species at 20 meters. L. umbratilis, which appeared at all three levels of vertical stratification, has great epidemic significance as a vector of Leishmania (Viannia) guyanensis.     

Study of sand flies (Diptera,Psychodidae, Phlebotominae) in the urban area of Campo Grande,Mato Grosso do Sul State,Brazil, from 1999 to 2000

From February 1999 to February 2000, sand flies were captured weekly with CDC light traps at five sites in the urban area of Campo Grande, Mato Grosso do Sul State, Brazil. Traps were placed in 11 different ecotopes in the environment (ground level, tree canopies, and forest edge) and the peridomicile (chicken coops and banana trees). A total of 1,245 sand flies were captured, belonging to 28 species: 4 species from genus Brumptomyia Fran a & Parrot, 1921 and 24 from genus Lutzomyia Fran a, 1924. The species were: B. avellari, B. brumpti, B. galindoi, B. pintoi, L. aragaoi, L. bourrouli, L. campograndensis, L. cerradincola, L. christenseni, L. claustrei, L. cortelezzii, L. corumbaensis, L. cruzi, L. damascenoi, L. flaviscutellata, L. hermanlenti, L. lenti, L. longipalpis, L. longipennis, L. migonei, L. punctigeniculata, L. quinquefer, L. renei, L. shannoni, L. sordellii, L. teratodes, L. termitophila, and L. whitmani. L. longipalpis and L. cruzi, vectors of visceral leishmaniasis, and L. whitmani, L. flaviscutellata and L. migonei, vectors of cutaneous leishmaniasis, were captured in the urban area. The most frequent species were L. termitophila, L. aragaoi, L. lenti, L. longipennis, and L. longipalpis.     

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.     

Laboratory tests of three repellents against Phlebotomus perniciosus (Diptera:Psychodidae)

Three topical repellents were evaluated against Phlebotomus perniciosus using dose-response techniques. A total of 1800 laboratory reared nulliparous females of P. perniciosus, 2 to 15 days old, and 21 human volunteers were used. The effective dose (ED50 and ED90) for Deet, Indalone and MGK 11 has been determined. Indalone and MGK 11 were more effective than standard Deet. The data obtained indicate that P. perniciosus is highly sensitive to the repellents compared with other blood-sucking arthropods.