New findings on the geographic distribution of the verrucarum group (Diptera: Psychodidae) in Colombia

The incrimination of sand flies belonging to verrucarum species group in the leishmaniasis transmission underscores the need for a detailed information on the geographical distribution of these species. The current listing adds 34 new records that extend significantly the knowledge of the geographical distribution of the verrucarum group in Colombia. The most important new records pertain to Lutzomyia spinicrassa in the tropical dry forest of the Atlantic coast, Lutzomyia ovallesi in the Orinoco and Amazon River watersheds regions of Colombia, and the sympatric occurrence of Lutzomyia spinicrassa and Lutzomyia quasitownsendi in the eastern chain of the Andes mountains. Additionally, the municipal and altitudinal distributions are compiled for 19 new species recorded for Colombia. The series townsendi of the verrucarum group generally restricted to premontane and lower montane zones based on distribution data on Lutzomyia longiflocosa, L. quasitownsendi, Lutzomyia sauroida, L. spinicrassa, Lutzomyia torvida, Lutzomyia townsendi and Lutzomyia youngi. The series verrucarum is distributed from the tropical lowland to the montane zones, and includes species with wide geographical range (Lutzomyia evansi, Lutzomyia columbiana, L. ovallesi, Lutzomyia nuneztovari, Lutzomyia nevesi), and species of highly endemic distribution (Lutzomyia andina, Lutzomyia disiuncta, Lutzomyia moralesi, Lutzomyia antioquiensis). Members of the series pia (Lutzomyia pia, Lutzomyia limafalcaoae) and the series serrana (Lutzomyia serrana) occur from the tropical lowlands to the lower montane zones. The altitudinal divergences may be intrinsically tied to speciation process, especially as it relates to the climatic and geologic events that have affected the flora and fauna of the Andean region.     

Karyotypic comparison of five species of Lutzomyia (diptera: psychodidae) of the series townsendi and the verrucarum group in Colombia

Cytogenetic characteristics were established for five native species of phlebotomine sand flies (Lutzomyia, series townsendi, verrucarum group): Lutzomyia longiflocosa, Lutzomyia townsendi, Lutzomyia spinicrassa, Lutzomyia torvida and Lutzomyia youngi. Karyotypes and chromosomal morphometry were compared. Using the squash technique, brain tissues from late 4th instar larvae provided the necessary mitotic chromosomes. Chromosomal measurements were made on the following chromosomal characteristics: short arm, long arm, arm ratio, total length, relative length, centromeric index and relative length average of chromosomes. Chromosomes were classified according to their morphometry and position of the centromere. The taxonomic distance was calculated, and the relationships among the species displayed in a phenogram. All five species possessed four pairs of chromosomes as diploid number (2N = 8). None of the karyotypes indicated presence of heteromorphic chromosomes. Statistical analysis of the morphometric data showed highly significant differences among the chromosomes pairs of the five species. However, the total length of the genome was very similar, with the exception of L. youngi. In conclusion, these closely related species were distinguishable at cytological level.     

Leishmaniasis vector potential of Lutzomyia spp. in Colombian coffee plantations

Potential vectors of Leishmania braziliensis Vianna were assessed at four study sites in the mountainous Valle del Cauca, western Colombia, from March to June 1989. In an active focus of transmission at 1450 m altitude, a coffee plantation at Versalles, there were high densities of antropophilic phlebotomines: Lutzomyia columbiana (Ritorcelli & Van Ty) and Lu.townsendi (Ortiz), both in the verrucarum species group, and of Lu.pia (Fairchild & Hertig). At a comparable altitude in a forest reserve at Yotoco where leishmaniasis is unknown, Lu.pia was the prevalent species and Lu.townsendi was absent. In two localities at 1150 m altitude, there were plentiful Lu.lichyi (Floch & Abonnenc) plus both species in the verrucarum group, but Lu.pia was absent. One of these localities, a coffee plantation at Villa Hermosa where a leishmaniasis outbreak occurred in 1986, was compared with a leishmaniasis-free, partly wooded nature reserve at Mateguadua. No natural infections of Leishmania were found in a total of 1896 wild-caught female phlebotomines belonging to at least seven species. It remains unclear why Leishmaniasis transmission is associated with coffee plantations in this part of Colombia. Laboratory-bred Lu.lichyi females were invariably autogenous, and blood-seeking females of this species were always parous. Parity rates in wild-caught females of other species were 55% Lu.pia, 24% Lu.columbiana and 14% Lu.townsendi. Female Lutzomyia infected artificially with Le.braziliensis promastigotes developed peripylarian infections. Higher proportions of Lu.townsendi (96%) and Lu.columbiana (78%) became infected but these species developed lower rates of stomodaeal infections (P less than 0.1) than Lu.lichyi (37%) or Lu.pia (44%).(ABSTRACT TRUNCATED AT 250 WORDS)     

Phlebotomine sandflies associated with a focus of cutaneous leishmaniasis in Valle del Cauca, Colombia

Abstract. A survey was made of the phlebotomine sandfly fauna of La Guaira, a village with coffee plantations near Cali, Colombia, from which cases of American cutaneous leishmaniasis had been reported due to Leishmania (Viannia) panamensis and Le. ( V. ) braziliensis. Among six species of sandfly collected on human bait, Lutzomyia youngi was most important in terms of biting nuisance. Lu. columbiana, Lu. lichyi and Lu. scorzai as well as Lu. youngi adults occurred throughout the year. Sandfly man-biting activity occurred throughout the night and was highest within 2 h of sunset. Despite its abundance in nocturnal samples, Lu. youngi was rarely taken in diurnal resting site collections. In contrast, Lu. lichyi was collected on tree-trunks during the day in large numbers and was the only species biting in daylight. The implications of these and other findings for leishmaniasis control measures in La Guaira are considered.     

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.     

Chorion patterns on eggs of Lutzomyia sandflies from the Peruvian Andes

Abstract. Scanning electron microscopy (SEM) was used to depict and describe the eggs of five species of phlebotomine sandfly from the Andean region of Peru: Lutzomyia caballeroi, Lu.noguchii, Lu.peruensis, Lu.tejadai and Lu.verrucarum. Two new types of chorionic sculpturing of sandfly eggs are reported: these were named disperse (Lu.tejadai) and verrucose (Lu.noguchii). The aeropylar area of the eggs is also described for the first time for neotropical sandflies. These character states appear to vary for ecological rather than phylogenetic reasons and could be used for species identification.     

The re-emergence of American visceral leishmaniasis in an old focus in Venezuela. II. Vectors and parasites.

As part of an epidemiological study in an old focus of American Visceral Leishmaniasis (AVL) in Venezuela (Guayabita, Aragua State), a longitudinal entomological survey (January 1993-June 1994) was carried out. A total of 3,239 males and 6,043 females belonging to 11 phlebotomine sandfly species were collected. The two recognised vectors of AVL in the New World, Lutzomyia evansi and Lu. longipalpis were found to be sympatric. Lutzomyia evansi was the dominant species (86.4%), almost ten fold times more abundant than Lu. longipalpis (10.6%). The two species alternated seasonally: Lu evansi peaked at the end of the rainy season while Lu. longipalpis, almost virtually absent during such period, increased in the dry season. This species seems more greatly influenced by the temperature. Seven of 4,559 Lutzomyia evansi (0.15%) and one of 353 Lu. longipalpis (0.28%) were found positive for suprapyloric promastigotes. Using the polymerase chain reaction (PCR) with universal primers, all isolates were identified as Leishmania spp. Two cultures from Lu. evansi, IEVA/VE/93/UCNA-2 and IEVA/VE/93/UCNA-3, were established. k-DNA restriction analysis showed high homologies between these isolates and Leishmania chagasi. High hybridization signal with L. chagasi specific kDNA confirmed these results. These findings suggest that Lu. evansi may play a role as vector of visceral leishmaniasis in this area. The identity of the parasite carried by Lu. longipalpis needs to be confirmed.     

Association of Lutzomyia columbiana (Diptera: Psychodidae) with a leishmaniasis focus in Colombia due to species of the Leishmania mexicana complex.

In Colombia, Leishmania mexicana has a scattered geographical distribution and no sand fly vectors have been associated with its transmission. During the present study, the anthropophilic sand fly Lutzomyia columbiana was found to be the only species collected using diverse methods, in a small focus of Le. mexicana in the municipality of Samaniego, SW Colombia. Ecological data indicate that this sand fly species is present in both peri and intradomestic habitats, where it readily bites man. Further evidence comes from experimental infections of wild-caught Lu. columbiana with Le. mexicana after feeding on infected hamsters. Based on these results, it is suggested that this sand fly is the most likely vector in the study area, suggesting the existence of a previously unknown sand fly-parasite association.     

The relationship between CDC light-trap and human-bait catches of endophagic sandflies (Diptera: Psychodidae) in the Peruvian Andes

Abstract. A study was carried out in the Peruvian Andes to test the suitability of CDC light traps for monitoring changes in the human-landing rate of endophagic phlebotomine sandflies, following house-spraying with pyrethroid insecticide. On four pairs of consecutive nights, sandflies were caught inside eight sprayed and eight unsprayed houses, either by human bait or by CDC light traps. The sandflies collected were Lutzomyia verrucarum (97%) and Lu. peruensis (3%), both probable vectors of Leishmania peruviana , and the species composition was unaffected by house-spraying. A non-linear relationship was detected between light-trap and human-bait catches, but the relationship did not diverge significantly from linearity within the range of sandfly abundance found in most houses in the endemic area (i.e. between 3 and 200 sandflies/house-night), and did not differ significantly between sprayed and unsprayed houses. However, light trap catches had a significantly lower proportion of blood-fed females in sprayed than in unsprayed houses, probably due to an insecticidal effect on post-blood-feeding behaviour. The proportion of Lu. verrucarum was significantly higher in light trap than in human bait catches, indicating that Lu. peruensis is either more anthropophilic or less phototropic than Lu. verrucarum.     

Presence of American cutaneous Leishmaniasis vectors surrounding the city of Medellín,Colombia

The presence of Lutzomyia (Helcocyrtomyia) hartmanni, as a vector of Leishmania colombiensis and L. columbiana (Verrucarum group), recently incriminated in the transmission of leishmaniasis, and L. pia (Verrucarum group) are reported for the first time in a periurban area of Medellín city. There is thus a risk of leishmaniasis transmission in this town.     

Systematics of the Lutzomyia species of the verrucarum Theodor group, 1965 (Diptera: Psychodiadae)

The verrucarum group of phlebotomine sand flies includes vectors of Leishmania spp. and Bartonella bacilliformis, and from the perspective of public health is considered as one of the most important groups of neotropical phlebotomine sand flies. Due to marked morphological similarity among species constituting this group, the identification based on conventional taxonomic characters can be difficult. Consequently, the verrucarum group has been the focus of numerous taxonomic comparisons which have included the following methods: chaetotaxy, morphometry, larval spiracular system, chorionic structure, morphology of the genital atrium, cytogenetics, morphological phylogenetics, isoenzymes, random amplified polymorphic DNA, cuticular hydrocarbons, DNA probes, and nuclear and mitochondrial nucleotide sequences. Based on morphological characters of the male terminalia, the verrucarum group has been divided in four series, i.e., verrucarum, serrana, townsendi and pia. Since the revision of the group made by Young and Duncan in 1994, ten new species, principally of Andean origin, have been assigned to 3 of the series verrucarum (L. maranonensis, L. cajamarcensis, L. antioquiensis, L. falcaorum), serrana (L. robusta, L. guilvardae) and pia (L. suapiensis, L. tihuiliensis, L. tocaniensis, L. limafalcaoae). The total number of verrucarum group members is now 40. Explanations for this diversity of species include the isolation of ancestral populations in refugia of humid forest during the quaternary period, the Andean cordilleras as geographical barrier, and the appearance of the Isthmus of Panama. Biology systematics and evolution of the verrucarum group is reviewed with emphasis on the 19 species extant in Colombia.     

First record of Lutzomyia evansi (Nuñez-Tovar 1924) in Mexico (Diptera: Psychodidae, Phlebotominae)

The phlebotomine sand fly Lutzomyia evansi is recorded in Mexico for the first time. This species is a suspected vector of Leishmania infantum in other parts of its geographical range and was captured in a focus of American visceral leishmaniasis where the principal vector, Lu. longipalpis sensu lato, was also found. The relative public health importance of the two species in the study area (Chiapas state, Southern Mexico) is discussed.     

Xenodiagnosis using Lutzomyia youngi in Venezuelan cases of cutaneous leishmaniasis due to Leishmania braziliensis

Eight patients infected with Leishmania braziliensis were used for xenodiagnosis with Lutzomyia youngi, before and after specific antileishmanial treatment with "glucantime" and "gabbromycin". All of them infected sandflies fed on the borders of the skin lesions before the treatment, suggesting that infected persons might act as reservoirs of infection for an indoor-biting sandfly species. The negative results obtained by xenodiagnosis carried out after specific treatment of the same individuals indicated cure of the patients, and a reduction of risk for further intradomiciliary transmission.     

Phlebotomine sand flies (Diptera: Psychodidae) associated with the appearance of urban Leishmaniasis in the city of Sincelejo,Colombia

Although once associated only with rural areas, the American leishmaniasis vectors now appear to be associated also with urban and suburban areas of the Neotropics. Following the appearance of the first autochthonous visceral and cutaneous leishmaniasis cases in the urban area of the city of Sincelejo, Colombia, a preliminary entomological survey of the sand fly species composition was performed using Shannon and CDC light traps. A total of 486 sand flies representing six Lutzomyia species were collected. L. evansi, L. panamensis and L. gomezi, known vectors of Leishmania spp. were the predominant sand fly species around dwellings. The finding of these species in relation to the appearance of the first cases of leishmaniasis in the city mentioned is discussed.     

Effect of a second bloodmeal on the oesophagus colonization by Leishmania mexicana complex in Lutzomyia evansi (Diptera: Psychodidae)

Migration and colonization of the oesophagus by Leishmania mexicana parasites were enhanced after digestion of a second bloodmeal intake in Lutzomyia evansi. This event has epidemiological significance since it affects the infection susceptibility of this sand fly species, which is a proven vector of L. chagasi in Colombian and Venezuelan visceral leishmaniasis foci. Also, it may explain the host seeking behaviour displayed by some partially bloodfed flies found inside houses.     

An association between phlebotomine sandflies and aphids in the Peruvian Andes

Abstract. As the composition of natural sugars in the diet of adult sandflies (Diptera: Psychodidae: Phlebotominae) may affect the development of Leishmania (Kinetoplastida: Trypanosomatidae) in sandfly guts, and so play an important role in the epidemiology of leishmaniasis, there is increasing interest in the sources of sugars for wild sandflies. Advanced chromatography techniques have provided indirect evidence that wild sandflies feed on honeydew, a substance released by aphids (Hemiptera: Aphididae) when feeding. Our objective was to determine whether sandfly density can be influenced directly by the local density of aphids. Aphid density was determined by counting absolute numbers of aphids on alfalfa stems in Purisima Valley, Peru, where sandflies transmit Leishmania peruviana causing Andean cutaneous leishmaniasis (uta). Sandfly relative abundance was measured using sticky trap sampling repeatedly in alfalfa fields. Lutzomyia verrucarum accounted for 92% of the total sandflies collected. As there was a female bias in sandflies collected close to houses, only the numbers of male sandflies were used in analysis. Most of the adult aphids found feeding on alfalfa were either Therioaphis trifolii forma maculata (97%) or Acyrthosiphon pisum (3%). By regression analysis, a significant relationship was found between the density of Lu. verrucarum males and the density of adults of both aphid species. This is the first ecological study to support the hypothesis that aphid honeydew may be a source of sugar for sandflies.     

Posterior spiracles of fourth instar larvae of four species of phlebotomine sand flies (Diptera: Psychodidae) under scanning electron microscopy

In the present study, posterior spiracles of laboratory-reared fourth instar larvae of Lutzomyia longipalpis, L. migonei, L. lenti, and L. whitmani (Diptera: Psychodidae) of the State of Ceará, Brazil, were examined under scanning electron microscopy. The number of papillae of spiracles examined varied according to the species examined, but no intraspecific differences were found. The importance of this structure to sand fly larva identification and phylogeny is commented.     

Comparison of three methods for age-grading of female Neotropical phlebotomine sandflies

Abstract. Changes in appearance of the accessory glands (AG), ovaries (OV) and the absence or presence of crease marks at the edges of the genital atrium armature (GA) were compared as methods to age-grade female laboratory-bred (Lutzomyia migonei) and wild-caught (Lu.youngi, Lu.spinicrassa, Lu.ovallesi) Neotropical sandflies.
Dissections at different physiological stages revealed that all parous and high proportions of nulliparous (4/38 unmated, 12/12 mated) females showed similar residual AG secretions. Hence AG appearance (nullipars with AG translucent, versus yellowish opaque granulated AG previously regarded as signs of parousness) is unreliable for age-grading.
Readily recognizable changes in both GA (98% creased) and the OV (100% yellowish with loose ovarioles) indicated that these organs could be employed to discriminate between nulliparous and parous females. Despite finding one misdiagnosed female (a nullipar having creased GA indicative of parity), the GA method of age-grading is recommended because females may be examined satisfactorily at any stage of the gonotrophic cycle. Moreover, the GA characteristics of pickled or dry preserved sandflies as well as fresh slide-mounted specimens can be determined easily.
Among sixty-five wild-caught Lutzomyia females dissected (mostly Lu.spinicrassa and Lu.youngi) , the proportions of specimens yielding preparations suitable for examination were 97% for GA, 89% for OV, but only 58% for AG, which reinforces GA as the preferred character for age-grading.     

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.     

External morphology of sensory structures of fourth instar larvae of neotropical species of phlebotomine sand flies (Diptera: Psychodidae) under scanning electron microscopy.

In the present study, some morphological structures of antennae, maxillary palps and caudal setae of fourth instar larvae of laboratory-reared phlebotomine sand flies (Lutzomyia longipalpis, L. migonei, L. evandroi, L. lenti, L. sericea, L. whitmani and L. intermedia) of the State of Ceará, Brazil, were examined under scanning electron microscopy. The antennal structures exhibited considerable variation in the morphology and position. A prominent digitiform distal segment has been observed only on the antenna of species of the subgenus Nyssomyia. The taxonomic relevance of this and other antennal structure is discussed. The papiliform structures found in the maxillae and the porous structures of the caudal setae of all species examined may have chemosensory function. Further studies with transmission electron microscopy are needed to better understand the physiological function of these external structures.