Protection of dogs from bites of phlebotomine sandflies by deltamethrin collars for control of canine leishmaniasis

Abstract. Dog collars made of PVC plastic impregnated with the pyrethroid insecticide deltamethrin at 40 mg/g were investigated for their protective efficacy against phlebotomine sandflies. Collared dogs were kept separately (two untreated control dogs lived together) in outdoor enclosures, each with a kennel, in the Cevennes, southern France. To measure sandfly mortality and anti-feeding effects due to the deltamethrin-impregnated collars worn continuously by the dogs for up to 8 months, each dog was periodically sedated and exposed for 2h to 150–200 laboratory-reared Phlebotomus perniciosus females (plus c. 25 males) inside a net (1.2m square, 1.8 m high) indoors. After dogs were removed from the nets, allowed to recover and returned to their kennels, any dead sandflies were collected from inside the net and counted. Surviving flies were kept overnight, then scored according to whether they were still alive or dead, unfed or blood-fed. From tests 2, 3, 4, 13, 20, 26 and 34 weeks after the dogs began wearing collars, the overall numbers of blood-fed female sandflies recaptured were 75 from two dogs with collars, compared with 1911 from two collarless dogs. Thus, for every 100 flies which fed on collarless dogs, only 4 fed on collared dogs, i.e. the collars protected dogs from 96% of the bites and this activity was maintained for up to 34 weeks. During the same period, the percentage of recaptured female sandflies that had fed on collared dogs was 0–12% compared to 55–95% on collarless dogs. Immediately after dogs were taken out of the nets, 21–60% of flies confined with the collared dogs were found dead, compared to 0–12% with the controls. It is concluded that, at least in the Mediterranean subregion, this insecticidal collar would protect a dog from the majority of sandfly bites and retain a killing effect for a complete sandfly season. Moreover, it seems likely that the use of collars on all dogs in a focus of Leishmania infantum would reduce contact between sandfly vectors and canine reservoir hosts sufficiently to diminish the risk of infection for humans as well as dogs.     

Host preferences of the phlebotomine sandfly Lutzomyia longipalpis in Amazonian Brazil

Experiments were undertaken to determine the relative attractiveness of humans, dogs and chickens to Lutzomyia longipalpis, the principal vector of Leishmania chagasi causing American visceral leishmaniasis. Field experiments in two villages on Marajó Island, Pará State, Brazil, showed that one boy attracted significantly more flies than one dog or chicken, and slightly fewer flies than a group of six chickens. Experiments with laboratory-bred female flies showed that a significantly greater number of flies engorged on a single human than on either a single dog or chicken, and man-biting catches demonstrated the willingness of flies to bite in the field. It appears that Lu.longipalpis has catholic feeding habits, the attractiveness of different hosts being largely a function of their relative sizes. These results are discussed with reference to the epidemiology of visceral leishmaniasis in Brazil.     

Copulatory courtship song in Lutzomyia migonei (Diptera: psychodidae)

Lutzomyia migonei is a vector of leishmaniasis with a wide distribution in South America, which could favour population differentiation and speciation. Cryptic species of the Lutzomyia longipalpis complex, the widely distributed sand fly vector of visceral leishmaniasis in Latin America, have previously been shown to display distinct copulation songs. We found that Lu. migonei males also produce a song during copulation. This "lovesong" presents short trains (6-8 pulses) with an inter-pulse interval around 26 ms and is potentially involved in cryptic female choice and insemination success.     

Susceptibility of laboratory-reared female Lutzomyia longipalpis (Lutz & Neiva, 1912) to infection by different species and strains of Leishmania Ross, 1903

A study was undertaken to compare the susceptibility of laboratory-reared female Lutzomyia longipalpis to infection by different species or strains of New World Leishmania. The sand flies proved to be highly susceptible to infection by a strain of Le. guyanensis, with flagellates developing in all (18/18) of the specimens examined. A lower infection rate of 37% (10/27) was recorded in flies exposed to infection by a strain of Le. amazonensis. Flagellates developed in 13% (6/46) of the sand flies that blood fed on dogs in the early stage of experimental infection with an old laboratory strain of Le. chagasi. In contrast, promastigotes did not develop in sand flies that blood fed on dogs with naturally acquired Le. chagasi. The naturally infected dogs were in an advanced stage of disease. Flagellates developed in 9% (3/32) of the sand flies that blood fed on lesions of hamsters infected with a strain of Le. braziliensis and in 9% (3/34) of those that fed on hamsters with lesions due to a parasite of the mexicana complex (strain MHOM/BR/73/BH121). Sand flies did not develop flagellate infections after blood feeding on hamsters bearing lesions induced by strain MHOM/BR/71/BR49. Factors influencing the susceptibility of Lu. longipalpis to infection by New World species of Leishmania are discussed.     

Detection of Leishmania infantum in naturally infected Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae) and Canis familiaris in Misiones, Argentina: the first report of a PCR-RFLP and sequencing-based confirmation assay

In this study, a genotypification of Leishmania was performed using polimerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing techniques to identify species of Leishmania parasites in phlebotomine sand flies and dogs naturally infected. Between January-February of 2009, CDC light traps were used to collect insect samples from 13 capture sites in the municipality of Posadas, which is located in the province of Misiones of Argentina. Sand flies identified as Lutzomyia longipalpis were grouped into 28 separate pools for molecular biological analysis. Canine samples were taken from lymph node aspirates of two symptomatic stray animals that had been positively diagnosed with canine visceral leishmaniasis. One vector pool of 10 sand flies (1 out of the 28 pools tested) and both of the canine samples tested positively for Leishmania infantum by PCR and RFLP analysis. PCR products were confirmed by sequencing and showed a maximum identity with L. infantum. Given that infection was detected in one out of the 28 pools and that at least one infected insect was infected, it was possible to infer an infection rate at least of 0.47% for Lu. longipalpis among the analyzed samples. These results contribute to incriminate Lu. longipalpis as the vector of L. infantum in the municipality of Posadas, where cases of the disease in humans and dogs have been reported since 2005.     

Further evidence that deltamethrin-impregnated collars protect domestic dogs from sandfly bites

In many foci of zoonotic visceral leishmaniasis (ZVL), domestic dogs are important reservoir hosts of the causative Leishmania parasites transmitted by phlebotomine sandflies (Diptera: Psychodidae). We tested the protective value of impregnated dog collars (20 g plastic containing deltamethrin 800 mg ai) against Phlebotomus papatasi (Scopoli) sandflies in Iran. For each assay, the dog was sedated and caged in a net with 70-100 wild-caught sandflies overnight (23.30-06.30 hours). Dogs wearing the collars were bitten by approximately 80% fewer sandflies than before collars were fitted, i.e. 51% vs. 11% of hungry female flies exposed. Sandfly mortality rates following 20 h exposure to dogs with collars (18%) or without collars (17%) were not significantly different. Effects of collars were tested when dogs had been wearing them for 8 days. A previous trial against the sandfly P. perniciosus Newstead in France, using smaller dogs, showed that effects of such collars were not fully realized until they had been worn for 2 weeks or more; they remained effective for at least 8 months and killed significant proportions of the sandflies exposed. Present results with P. papatasi, confirming that this simple device provides effective protection against sandflies, are considered sufficiently encouraging to justify a community-wide field trial of deltamethrin-impregnated dog collars against ZVL vector sandflies in Iran.     

Density of sand flies (Diptera: psychodidae) in domestic and wild animal shelters in an area of visceral Leishmaniasis in the state of Rio Grande do Norte, Brazil.

The objective of the present study was to determine the association of sand flies with the presence of domestic and wild animals in the peridomiciliary area. The sand flies were collected using direct aspiration and CDC light traps placed in animal shelters. The results suggest that different sand flies species have different behavioral characteristics in an apparent preference for animal baits and that Lutzomyia longipalpis and Lu. evandroi were the most eclectic species regarding their biotope choice. Lu. longipalpis showed a distinct preference for horses and Lu. evandroi for armadillos.     

Reactive oxygen species scavenging by catalase is important for female Lutzomyia longipalpis fecundity and mortality

The phlebotomine sand fly Lutzomyia longipalpis is the most important vector of American visceral leishmaniasis (AVL), the disseminated and most serious form of the disease in Central and South America. In the natural environment, most female L. longipalpis are thought to survive for less than 10 days and will feed on blood only once or twice during their lifetime. Successful transmission of parasites occurs when a Leishmania-infected female sand fly feeds on a new host. Knowledge of factors affecting sand fly longevity that lead to a reduction in lifespan could result in a decrease in parasite transmission. Catalase has been found to play a major role in survival and fecundity in many insect species. It is a strong antioxidant enzyme that breaks down toxic reactive oxygen species (ROS). Ovarian catalase was found to accumulate in the developing sand fly oocyte from 12 to 48 hours after blood feeding. Catalase expression in ovaries as well as oocyte numbers was found to decrease with age. This reduction was not found in flies when fed on the antioxidant ascorbic acid in the sugar meal, a condition that increased mortality and activation of the prophenoloxidase cascade. RNA interference was used to silence catalase gene expression in female Lu. longipalpis. Depletion of catalase led to a significant increase of mortality and a reduction in the number of developing oocytes produced after blood feeding. These results demonstrate the central role that catalase and ROS play in the longevity and fecundity of phlebotomine sand flies.     

Differential application of lambda-cyhalothrin to control the sandfly Lutzomyia longipalpis

Abstract. To study the impact of residual pyrethroid insecticide on the abundance and distribution of peridomestic Lutzomyia longipalpis , the sandfly vector of visceral leishmaniasis in Brazil, lambda-cyhalothrin was applied at 20 mg a.i.m^-2 in the following interventions: (i) spraying of all animal pens in a village (blanket coverage); (ii) treatment of a subset of animal pens, either by spraying, or by installation of insecticide-impregnated 1 m² cotton sheets as 'targets' (focal coverage).
By sampling with CDC light traps, and using a novel analytical approach, we detected a 90% reduction in Lu. longipalpis abundance in sprayed sheds of the focal intervention. However, there was no discernible effect on the abundance of other phlebotomines trapped in sheds, or on the abundance of Lu. longipalpis in untreated dining-huts and houses. This differential impact on Ludongipalpis abundance is explained in terms of the disruption of male pheromone production. Treated targets were approximately half as effective as residual spraying in reducing the abundance of Lu.longipalpis in sheds.
Following blanket intervention, the abundance of Lu.longipalpis in traps fell by only 45% (not significant): catches at untreated dining-huts actually increased, possibly because the blanket coverage diverted Lu. longipalpis away from major aggregation sites at animal pens. It is recommended that care be taken during vector control programmes to ensure that all potential aggregation sites are treated. The possible consequences of leaving some sites untreated include poor control of peridomestic sandfly abundance and an increase in the biting rate on dogs and humans.     

Gene silencing in phlebotomine sand flies: Xanthine dehydrogenase knock down by dsRNA microinjections

Lutzomyia longipalpis are vectors of medically important visceral leishmaniasis in South America. Blood-fed adult females digest large amounts of protein, and xanthine dehydrogenase is thought to be a key enzyme involved in protein catabolism through the production of urate. Large amounts of heme are also released during digestion with potentially damaging consequences, as heme can generate oxygen radicals that damage lipids, proteins and nucleic acids. However, urate is an antioxidant that may prevent such oxidative damage produced by heme. We investigated xanthine dehydrogenase by developing the RNAi technique for sand flies and used this technique to knock down the Lu. longipalpis xanthine dehydrogenase gene to evaluate its role in survival of adult females after blood feeding. The gene sequence of Lu. longipalpis xanthine dehydrogenase is described together with expression in different life cycle stages and RNAi knock down. Semi-quantitative RT-PCR of xanthine dehydrogenase expression showed a significant increase in expression after bloodmeal ingestion. Microinjection of dsRNA via the thorax of 1-day-old adult female sand flies resulted in approximately 40% reduction of xanthine dehydrogenase gene expression in comparison to flies injected with a control dsRNA. A significant reduction of urate in the whole body and excretions of Lu. longipalpis was observed after dsRNA xanthine dehydrogenase microinjection and feeding 96h later on rabbit blood. Sand flies injected with XDH dsRNA also exhibit significantly reduced life span in comparison with the mock-injected group when fed on sucrose or when rabbit blood fed, showing that urate could be indeed an important free radical scavenger in Lu. longipalpis. The demonstration of xanthine dehydrogenase knock down by dsRNA microinjection, low mortality of microinjected insects and the successful bloodfeeding of injected insects demonstrated the utility of RNAi as a tool for functional analysis of genes in phlebotomine sand flies.     

Caspar-like gene depletion reduces Leishmania infection in sand fly host Lutzomyia longipalpis

Female phlebotomine sand flies Lutzomyia longipalpis naturally harbor populations of the medically important Leishmania infantum (syn. Leishmania chagasi) parasite in the gut, but the extent to which the parasite interacts with the immune system of the insect vector is unknown. To investigate the sand fly immune response and its interaction with the Leishmania parasite, we identified a homologue for caspar, a negative regulator of immune deficiency signaling pathway. We found that feeding antibiotics to adult female L. longipalpis resulted in an up-regulation of caspar expression relative to controls. caspar was differentially expressed when females were fed on gram-negative and gram-positive bacterial species. caspar expression was significantly down-regulated in females between 3 and 6 days after a blood feed containing Leishmania mexicana amastigotes. RNA interference was used to deplete caspar expression in female L. longipalpis, which were subsequently fed with Leishmania in a blood meal. Sand fly gut populations of both L. mexicana and L. infantum were significantly reduced in caspar-depleted females. The prevalence of L. infantum infection in the females fell from 85 to 45%. Our results provide the first insight into the operation of immune homeostasis in phlebotomine sand flies during the growth of bacterial and Leishmania populations in the digestive tract. We have demonstrated that the activation of the sand fly immune system, via depletion of a single gene, can lead to the abortion of Leishmania development and the disruption of transmission by the phlebotomine sand fly.     

Evaluation of deltamethrin-impregnated bednets and curtains against phlebotomine sandflies in Valle del Cauca, Colombia

Abstract. The effectiveness of bednets and curtains (nylon mesh 64 per cm²) impregnated with deltamethrin at 26 mg a.i./m² in reducing the biting nuisance caused by three phlebotomine sandfly species: Lutzomyia columbiana, Lu. lichyi and the predominant Lu. youngi (Diptera: Psychodidae), was evaluated at La Guaira, a rural settlement in Valle de Cauca near Cali, Colombia.
Pairs of volunteers collected sandflies under impregnated bednets, in rooms protected by impregnated curtains or in unprotected rooms in a randomized matched design. Collections were made in three houses per night on three consecutive nights, so that each house was sampled under each of the three treatments. This routine was repeated at 2-week intervals for 6 months. There was no significant difference between the overall numbers of sandflies collected in rooms with or without impregnated curtains. Only 0.14 sandflies/man-hour were caught on human bait under impregnated bednets, significantly fewer than the numbers collected on human bait outside the nets in the same room (1.91) or in unprotected rooms (3.29).
In a second set of experiments carried out in La Guaira and the neighbouring community of Jiguales, the effect of deltamethrin impregnation was evaluated by comparing numbers of sandflies collected on human bait under treated and untreated nets. Significantly fewer were collected under the impregnated nets (0.25 v. 0.69/man-hour). Wild-caught female Lu. youngi exposed to treated netting for 2 min in the laboratory all died with 24 h. The impact of deltamethrin-impregnated bednets was considered to be useful against Lu. youngi and other potential vectors of leishmaniasis in such communities.     

Constitutive and blood meal-induced trypsin genes in Lutzomyia longipalpis

Trypsins constitute some of the most abundant midgut digestive proteases expressed by hematophagous insects upon blood feeding. In addition to their role in the digestion of the blood meal, these proteases also have been implicated in the ability of certain pathogens to infect their natural vector. In sand flies, digestive proteases including trypsins were associated with early killing of Leishmania and are believed to play a role in the species-specificity dictating sand fly vectorial capacity. Our group is involved in studies of midgut digestive proteases in the sand fly Lutzomyia longipalpis, the principal vector of visceral leishmaniasis in Brazil. Here we report on the identification of two cDNAs, Lltryp1 and Lltryp2, which code for putative midgut trypsins in L. longipalpis. Analyses of RNA abundance using semi-quantitative RT-PCR show a different pattern of expression between the two genes. Lltryp1 expression remains undetected until blood feeding and reaches a peak at 12 h post-blood meal (PBM), returning to pre-blood meal levels at 72 h PBM. Additionally, Lltryp1 expression is undetected during larval development. Lltryp2, on the other hand, is constitutively expressed as high levels in the non-blood fed female, but is reduced upon blood feeding. At the end of the digestive cycle, Lltryp2 regains its pre-blood meal levels. This cDNA also is present in all developmental stages and in adult males. This pattern of expression is reminiscent of what is seen in mosquitoes and Old World sand flies, but has characteristics that are unique to L. longipalpis.     

Deltamethrin-impregnated bednets reduce human landing rates of sandfly vector Lutzomyia longipalpis in Amazon households

The entomological efficacy of using 25% deltamethrin EC insecticide-treated bednets (ITNs) was evaluated against the sandfly Lutzomyia longipalpis Lutz and Neiva (Diptera: Psychodidae), the principal vector of zoonotic visceral leishmaniasis (ZVL) in Latin America. A crossover field study in Amazon Brazil (Marajó Island, Pará State) demonstrated that, compared with untreated nets, the insecticide increased the barrier effect of nets by 39% (95% confidence interval [CI] 34-44%), reduced human landing rates by 80% (95% CI 62-90%) and increased the 24-h mortality rate from 0% to 98% (95% CI 93-99%) inside ITNs. The presence of an ITN also reduced the human landing rate on unprotected persons outside the net in the same room by 56% (95% CI 52-59%), and increased 24-h mortality to 68% (95% CI 62-73%) compared to 0.4% (0.1-2.0%) in untreated houses. The reduction in human landing rates in ITN rooms was associated with a doubling in the proportion of sandflies alighting on walls compared with that in untreated rooms, which was attributed to insecticide-induced excito-repellency. There was no evidence that sandflies were diverted onto unprotected hosts. Human landing catches inside houses peaked between 19.00 hours and 23.00 hours and declined steadily to zero at 02.00 hours and thereafter. House-to-house questionnaires established that only 34% of households owned at least one net (median two, range 1-8), only 20% of the population slept under a net (33% of 0-5-year-old children), and the majority (73%) of the population slept in hammocks. Combined data pertaining to sleeping times for children and sandfly activity period indicate that > 50% of sandfly bites inside houses, and substantially more outside houses, were taken before a third of children were potentially protected by a net. This study demonstrates the clear entomological efficacy of ITNs against Lu. longipalpis in this endemic region. The effectiveness of ITNs at preventing ZVL infection and disease has still to be evaluated.     

Diversity, distribution and abundance of sandflies (Diptera: Psychodidae) in Parana State, Southern Brazil

Due to the spread of Visceral leishmaniasis in Brazilian territory, this research was undertaken with the objective to identify the sandfly fauna and aspects of the species behavior in forest and anthropic environments in Paraná State, Brazil. The sandfly captures were sporadically carried out in 37 municipalities of the Paraná state, during the period from March 2004 to November 2005. Each municipality was once visited, when Falc?o traps were installed during three consecutive nights in forested fragments and anthropic environments; captures with Shannon traps installed in forests and edge of forests, inspection of walls and electric aspiration in domiciles and forested areas were sporadically undertaken. The data analysis was based on estimates of frequency and abundance of the species, by five native landscapes. A total of 38,662 specimens belonging to 23 species were captured. Nyssomyia neivai (Pinto) (75.6%), Ny. whitmani (Antunes & Coutinho) (10.1%), Migonemyia migonei (Fran?a) (7.8%), Expapillata firmatoi (Barreto et al.) (2.1%) and Pintomyia fischeri (Pinto) (1.6%) together constituted 97.2% of the phlebotomine collected. High frequencies of sand flies were found in forests, henhouses, pig-pens and domiciles. The three most frequent species have been implicated in transmission of tegumentary leishmaniasis in Paraná and in the Brazilian Southeastern and Southern Regions. Lutzomyia longipalpis (Lutz & Neiva), the visceral leishmaniasis vector, was not found, however Lutzomyia gaminariai (Cordero et al.), female of which is similar to that of Lu. longipalpis was captured, raising the need to investigate its behavior, including as regard to the vectorial competence.     

Seasonality of Lutzomyia intermedia and Lutzomyia whitmani (Diptera: Psychodidae: Phlebotominae), occurring sympatrically in area of cutaneous leishmaniasis in the State of Rio de Janeiro,Brazil

Lutzomyia intermedia and Lutzomyia whitmani occur sympatrically in the district of Posse, municipality of Petrópolis, in the State of Rio de Janeiro (114 km from the city of Rio de Janeiro), where sand flies were captured weekly for three consecutive years in both the peridomicile and forest, in two first years from 18:00 to 22:00 h, using a Shannon trap and CDC light traps. The anthropophilic habit was assessed by recording the specimens that bit the human captors. Considering the different capture types and sites, the following species were identified: Brumptomyia brumpti, B. cardosoi, B. guimaraesi, Lutzomyia amarali, L. longipalpis, L. microps, L. costalimai, L. edwardsi, L. firmatoi, L. migonei, L. termitophila, L. tupynambai, L. fischeri, L. pessoai, L. lutziana, L. shannoni, L. barrettoi, L. intermedia, L. whitmani, L. lloydi, and L. minasensis. L. intermedia (38.3%) and L. whitmani (38.6%) predominated. L. intermedia predominated in the peridomicile, where it displayed more intense anthropophily, while L. whitmani displayed greater density in the banana plantations and forest, preferential sites for biting humans. Studies on seasonality showed that while L. intermedia displayed greater density in the hottest months of the year (December, January, and February), L. whitmani was abundant in the coolest months (June, July, and August), although both occurred throughout the year.     

Ootaxonomic investigation of five Lutzomyia species (Diptera, Psychodidae) from Venezuela

The eggshell fine structure of five sand fly species from Venezuela belonging to the genus Lutzomyia (L. migonei, L. ovallesi, L. absonodonta, L. gomezi and L. panamensis) was examined by scanning electron microscopy. The chorionic sculpturing of L. migonei, L. ovallesi, L. absonodonta and L. gomezi was characterized by series of columns arranged in palisade to form sinuous ridges. In inter-ridge areas, the basal layer was covered with fibrous material. The outer chorion of L. panamensis had a pattern known as "mountain- or volcano-like". The morphology of the posterior pole and aeropyle had a common structure in the five species, with some species-specific characters. The eggshell features of the five species are compared with those of other phlebotomine sand flies.     

Vertical stratification and development aspects of phlebotomine sand flies (Diptera: Psychodidae) in an area of Atlantic Forest tree species in a metropolitan region in northeastern Brazil.

In the state of Rio Grande do Norte in northeast Brazil, cases of visceral leishmaniasis (VL) occur mainly in the periurban areas of the city of Natal. Lutzomyia longipalpis Lutz & Neiva 1912 (Diptera: Psychodidae), a vector of Leishmania chagasi (Protozoa: Trypanosomatidae) to humans, is found throughout the state. Flora and fauna influence the distribution of sand fly species, whose horizontal or vertical stratification can be used as a parameter for identifying potential vectors, considering the presence of vertebrate hosts in the area. The purpose of this study was to obtain information about the vertical stratification of phlebotomine sand flies in an endemic area of leishmaniasis in Rio Grande do Norte, and associate it with the presence of other animals in the peridomiciliary environment as well as to analyze, under laboratory conditions, aspects of L. longipalpis reproduction in wild females. The sand flies were captured with light traps hung at different heights in species of Atlantic Forest trees and in a peridomiciliary environment in animal shelters. The traps were placed between 17:30 and 6:00 of the following day, in a peridomiciliary and extradomiciliary area of a forest fragment in both dry and rainy months. In the extradomiciliary environment, the traps were installed at 1, 3 and 5 m above the ground. The biological cycle of L. longipalpis was followed from the eggs of 200 wild females. Specimens of L. lenti, L. walkeri, and L. migonei were captured. The comparison and statistical analysis showed that L. longipalpis is more abundant at a height of 3 m and L. evandroi at 1 m. In the animal shelters (chickens, horses, and armadillos), we captured mainly specimens of L. longipalpis and L. evandroi. The duration of the biological cycle of L. longipalpis was approximately 38 days at a temperature of 28 degrees C.     

Lutzomyia longipalpis and the eco-epidemiology of American visceral leishmaniasis, with particular reference to Brazil: a review

An historical review is given of American visceral leishmaniasis (AVL), with particular reference to the eco-epidemiology of the disease in Brazil. Following the first records of AVL in this country, in 1934, the sandfly Lutzomyia longipalpis (Lutz and Neiva, 1912) was incriminated as the principal vector. It is now generally accepted, however, that there exist a number of cryptic species under the name of Lu. longipalpis s.l. and that variations in the quantity of the vasodilatory peptide maxadilan in the saliva of flies from different populations of Lu. longipalpis s.l., may account for the variable clinical manifestations of AVL seen in different geographic regions. Distribution of AVL has been shown to extend throughout most of South and Central America, with the domestic dog serving as the principal reservoir of infection for man. However, while one hypothesis suggests that the causative parasite is Leishmania infantum, imported from Europe with the Portuguese and Spanish colonists, the demonstration of a high rate of benign, inapparent infection in foxes in Amazonian Brazil raised an opposing suggestion that the parasite is indigenous to the Americas. Recent reports of similar infections in native marsupials, and possibly rodents, tend to support this view, particularly as Lu. longipalpis is primordially a silvatic sandfly. Although effective control measures in foci of the disease will diminish the number of canine and human infections, the presence of such an enzootic in a variety of native animals will render the total eradication of AVL unlikely.