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PubMed2001

Phlebotomine sandfly responses to carbon dioxide and human odour in the field.

Responses of Lutzomyia sandflies (Diptera: Psychodidae: Phlebotominae) to carbon dioxide (CO2) and human odour were investigated by field experiments in Parana State, southern Brazil. Catches of two predominant species: Lu. intermedia (Antunes & Coutinho) and Lu. whitmani Lutz & Neiva, were compared between traps baited with a human adult or with CO2 emitted at the human-equivalent rate. When the baits were only 40 cm apart, no difference of attractiveness was detected. When baits were separated by 20 m, however, significantly fewer sandflies (44% Lu. intermedia, 46% Lu. whitmani) were trapped with CO2 compared with human bait. This is the first field evidence that anthropophilic sandflies are attracted by human kairomones in addition to CO2. For both species [Lutzomyia intermedia and Lu. whitmani] [corrected], the proportion of human attractiveness attributable to CO2 was significantly more [corrected] for males than females; for Lu. intermedia males human bait was no more attractive than CO2 alone. Gender differences in sandfly olfactory sensitivity are likely to be associated with behavioural differences on the host, where females feed on blood and males find mates. With traps 20 m apart, both Lutzomyia spp. showed roughly linear increased responses (log-log scale) to 0.08-0.55% CO2 equivalent to 0.5-4 humans. This would explain why host size is generally proportional to attractiveness, as observed for other species of phlebotomine sandflies.

Adult↗
PubMed1994

Phlebotomine sandflies associated with households of human visceral leishmaniasis cases in Baringo District, Kenya.

A 12-month field study on sandflies was conducted from April 1992 to March 1993 at three locations in Baringo District, Rift Valley Province, Kenya. Study sites were selected based on their close proximity to the households of three patients who had had parasitologically confirmed visceral leishmaniasis in 1988, 1990 or 1991. Sandfly populations were highest during the two rainy seasons, April-June and November-December. A significant association was detected between the monthly abundance of sandflies and rainfall in the previous month. A significantly higher proportion of females than males was collected in light traps than on sticky paper traps set in termite mounds or animal burrows: approximately twice as many males as females were collected from the sticky paper traps whereas twice as many females as males were collected in light traps. Over 10% of the female sandflies collected from animal burrows and termite mounds were gravid, indicating that these flies were seeking oviposition sites at these locations. A total of 2231 sandflies was collected and five females were found to have flagellates in their midguts. The flagellates cultured from four Sergentomyia spp. were tentatively identified as Crithidia sp. by cellulose acetate electrophoresis (CAE). The promastigotes from the one Phlebotomus martini female found with an infected midgut were identified as Leishmania donovani by CAE.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2024

Characterization of West Nile virus Koutango lineage from phlebotomine sandflies in Kenya.

The West Nile virus (WNV), primarily transmitted by mosquitoes, is one of the most widespread flaviviruses globally, with past outbreaks occurring in the USA and Europe. Recent studies in parts of Africa, including Kenya, have identified the West Nile virus Koutango lineage (WN-KOUTV) among phlebotomine sandfly populations, however, our understanding of this virus remains limited. This study aimed to characterize WN-KOUTV from phlebotomine sandflies. Sandflies were sampled between 12th -16th March 2021 and 16th -20th March 2023 from six villages each in Baringo and Isiolo Counties, using CDC light traps. Female sandflies were taxonomically identified and pooled based on genus and site of collection. Virus isolation was performed in Vero cells. Viral genomes were determined using next-generation sequencing. Phylogenetic and molecular clock analyses were done to decipher the virus's evolutionary relationships. Comparative analyses of amino acid sequences were performed to determine variations. Protein modeling in Pymol was conducted to elucidate variations in key protein regions. Evolutionary pressure analysis investigated the selection pressures on the virus. In vitro experiments were done to investigate the virus growth kinetics in mammalian Vero E6 and mosquito C6/36 cells. We report the isolation of WN-KOUTV from Salabani in Baringo and Aremet in Isiolo, Kenya. The isolated WN-KOUTVs clustered with previously identified WN-KOUTV strains. Comparative analysis revealed a unique amino acid at NS5 653. The WN-KOUTV lineage as a whole is under purifying selective pressure, with diversifying pressure acting at site NS3 267. The current WN-KOUTV replicated in Vero E6 and C6/36 cells comparable to West Nile virus Lineage 1a, isolated from mosquitoes. Subsequent isolations of WN-KOUTV in phlebotomine sandflies suggest potential vectors, however, vector competence studies would confirm this. Replication in mammalian and insect cell lines suggests there may exist a vector/host relationship. We speculate the close genetic relationship of WN-KOUTV strains from East and West Africa may potentially be enabled by bird migratory routes between the two regions. If proven, this could point to a potential future pandemic pathway for this virus.

Animals↗
PubMedDate not supplied

Visceral leishmaniasis in Teresina, State of Piauí, Brazil: preliminary observations on the detection and transmissibility of canine and sandfly infections.

A Leishmania donovani-complex specific DNA probe was used to confirm the widespread dissemination of amastigotes in apparently normal skin of dogs with canine visceral leishmaniasis. When Lutzomyia longipalpis were fed on abnormal skin of five naturally infected dogs 57 of 163 (35%) flies became infected: four of 65 flies (6%) became infected when fed on apparently normal skin. The bite of a single sandfly that had fed seven days previously on a naturally infected dog transmitted the infection to a young dog from a non-endemic area. Within 22 days a lesion had developed at the site of the infective bite (inner ear): 98 days after infection organisms had not disseminated throughout the skin, bone marrow, spleen or liver and the animal was still serologically negative by indirect immunofluorescence and dot-enzyme-linked immunosorbent assay. When fed Lu. longipalpis were captured from a kennel with a sick dog known to be infected, 33 out of 49 (67%) of flies contained promastigotes. In contrast only two infections were detected among more than 200 sandflies captured in houses. These observations confirm the ease of transmissibility of L. chagasi from dog to sandfly to dog in Teresina. It is likely that canine VL is the major source of human VL by the transmission route dog-sandfly-human. The Lmet2 DNA probe was a useful epidemiological tool for detecting L. chagasi in sandflies.

Animals↗
PubMed2004

[Influence of rearrangement and cleaning of the peridomiciliary area and building disinsectization on sandfly population density in the municipality of Doutor Camargo, Paraná State, Brazil].

Phlebotomine sandfly captures were performed in the domicile, domestic animal shelters, and peridomicile before and after rearrangement, cleaning, and disinsectization. Phlebotomine sandflies were captured with Falcão traps from October 1998 to April 2000 in the Recanto Marista district, Doutor Camargo municipality, Paraná State. The results were compared with those from October 1996 to September 1997. In this survey, 122,616 phlebotomine sandflies were captured using 10 traps, as compared to 157,983 in the previous period, with 6 traps. Lutzomyia neivai predominated in both periods. During the former period an hourly average of 1,641.9 phlebotomine sandflies were captured as compared to 806.7 in the latter. The results with four traps installed in the same locations shows that the hourly average was 1,318.8 in the former period and 156.0 in the latter. The large reduction in the number of phlebotomine sandflies captured in the second period may be due to the impact of alterations in the environment and the disinsectization of the buildings after the first period.

Animals↗
PubMed2006

[Evaluation of sandfly control in Lobato municipality, Paraná State, southern Brazil].

Sandfly control measures were evaluated at the Da Barra Ranch, Lobato municipality, Paraná State, Brazil. The insects were captured with Falcão traps in houses, a cattle corral, and a forest area from July to November 1999 and from February to June 2000. In December 1999 and January 2000 the following measures were taken to decrease the sandfly density in peridomiciliary areas and domiciles: (i) screens were placed on windows of buildings (domiciles, dormitory, cafeteria); (ii) all organic material was cleaned from the peridomiciliary area; and (iii) buildings were sprayed for insects. There were more Nyssomyia whitmani, Pintomyia fischeri, P. monticola, and Brumptomyia brumpti specimens in the former period, while N. neivai, Migonemyia migonei, and N. pessoai predominated in the latter. In the initial period most of the sandflies were captured in the forest and in the latter period in the houses. The sandfly control measures did not reduce the sandfly population on the Da Barra Ranch, but they led to significant proportional changes in the insect fauna composition.

Animals↗
PubMed2003

Distribution of sandflies (Diptera: Phlebotominae) in forest remnants and adjacent matrix habitats in Brazilian Amazonia.

We studied the distribution of sandflies (Diptera: Phlebotominae)--insect vectors of several diseases, including leishmaniasis--at the interface between primary forest and cattle pasture and between primary forest and secondary forest (< 15 yr old) in Southern Brazilian Amazonia. Sandflies were collected by using a combination of light traps and traps having vertebrates as baits. Strong differences in abundance and species richness were found between primary forests and pastures. Very few sandfly species were found in the pastures, and those that were found generally occurred at lower densities when compared to the adjacent forest. At least one species (Lutzomyia lainsoni), however, can become extremely abundant in pastures, possibly depending on the presence of cattle and water bodies. Differences between primary forests and secondary forests were not so strong, although the latter usually had fewer species and lower population abundances. No species were exclusively found in pastures or secondary forests; the species present in these two habitats were a subset of those found in primary forests. The distance to the edge did not affect the abundance, richness and composition of sandfly species in primary forests. The abundance and richness of sandflies, however, was greater in forest edges facing pastures than those facing secondary forests. This pattern could not be explained by an influx of species and individuals from the adjacent pasture, suggesting the existence of in situ differences between the different types of forest edges studied.

Animals↗
PubMed1992

Studies on the epidemiology of leishmaniasis in Kenya: flight range of phlebotomine sandflies in Marigat, Baringo District.

A study on the flight range of phlebotomine sandflies using the mark-release-recapture technique was undertaken in Marigat, Baringo District, Kenya, to determine the distance sandflies are able to travel and assess their importance in disease transmission. Over a period of 17 months, a total of 108,429 sandflies collected from their natural habitats were marked and released from a central point in a valley. Of these, 842 (0.8%) sandflies comprising 11 species were recaptured. Ninety-five percent of the recaptured sandflies were recovered within a radius of 50 m from the release point and 54% within a radius of 10 m. Two Sergentomyia ingrami females were recaptured at a distance of 1000 m from the release point.

Animals↗
PubMedDate not supplied

Bioecologic aspects of Phlebotomine sandflies in relation to human and canine visceral leishmaniasis in northern Tunisia.

In northern Tunisia 29 sandfly collecting places were studied for two years, in five human leishmaniasis foci, seven canine foci and in the nature. Nine species were found among the 4942 specimens collected. P. perniciosus, the well known vector of visceral leishmaniasis and the herpetophile S. minuta parroti were commonest. The sandfly season extended from april to october. P. perniciosus showed a numerical curve with one peak. It occupied various types of shelters in nature and human settlements, but prefer shady and humid shelters. In settlements it is frequently found around dwellings and has been identified in human and canine leishmaniasis foci. The highest sandfly densities and greatest species diversity were noticed in towns, especially in the cracks of the old walls or old inhabited ruins, where rodents as well as the small domestic animals provide a favourable environment for the sandflies. The historical urban ruins are the main biotope of the sandfly vectors. Infantile visceral leishmaniasis occurs during summer when the children sleep in front of the houses or in large, opened rooms exposed to P. perniciosus flies which have bitten infected dogs.

Animals↗
PubMed1995

[Meningitis after travel in the Mediterranean region, caused by the sandfly fever virus].

Sandfly fever virus is the causative agent of sandfly fever (pappataci fever). The virus is endemic in the Mediterranean region and is transmitted by bites of Phlebotomus species. The three sandfly fever virus serotypes, Sicilian (SFS), Naples (SFN) and Toscana (TOS), are of travel-related significance. In addition to a flu-like illness, the sandfly fever virus may result in a serious neurological disease with meningitis and encephalitis. Due to the high number of travelers may occur in nonimmune persons. Since many clinicians are not aware of this disease, sandfly fever is still rarely diagnosed.

Adult↗
PubMed2002

Molecular evolution of the period gene in sandflies.

The molecular evolution of the clock gene period was studied in Phlebotomine sandflies (Diptera: Psychodidae). Comparison of the synonymous and nonsynonymous substitution rates between sandflies and Drosophila revealed a significantly higher evolutionary rate in the latter in three of the four regions analyzed. The differences in rate were higher in the sequences flanking the Thr-Gly repetitive domain, a region that has expanded in Drosophila but remained stable and short in sandflies, a result consistent with the coevolutionary scenario proposed for this region of the gene. An initial phylogenetic analysis including eight neotropical sandfly species and one from the Old World was also carried out. The results showed that only the subgenus Nyssomyia is well supported by distance (neighbor-joining) and maximum parsimony analysis. The grouping of the other species from the subgenus Lutzomyia and Migonei group shows very low bootstrap values and is not entirely consistent with classical morphological systematics of the genus Lutzomyia.

Amino Acid Sequence↗
PubMed2004

Distribution of sandfly species in relation to canine leishmaniasis from the Ebro Valley to Valencia, northeastern Spain.

In Spain, only two of the 12 recorded species of sandflies, Phlebotomus (Larroussius) ariasi Tonnoir and P. (L.) perniciosus Newstead, are proven vectors of Leishmania infantum Nicolle, the causative agent of endemic leishmaniasis. Studies of the distributions of phlebotomine sandflies are important for evaluating the possible effects of climate warming on any northward or altitudinal range shifts of leishmaniasis or the other diseases they transmit. We describe a recent sandfly survey in Spain, starting in the northern Ebro Valley and continuing southeast into the Levante region of the Mediterranean coast. Sandflies (P. ariasi only) were found for the first time in the northern province of Alava, in the upper Ebro Valley, where cases of canine leishmaniasis have been described during the last decade. Throughout the provinces sampled, P. ariasi predominated over P. perniciosus in cooler bioclimatic zones, and this statistically significant pattern was more marked than that with higher altitudes.

Altitude↗
PubMed2005

Nocturnal activity of phlebotomine sandflies (Diptera: Psychodidae) in a cutaneous leishmaniasis focus in Chichaoua, Morocco.

The nocturnal activity of phlebotomine sandflies (Diptera: Psychodidae) was studied "at an epidemic focus" on human cutaneous leishmaniasis due to Leishmania tropica Wright in Chichaoua province, in Morocco. Sandflies were collected using light and sticky-paper traps changed at 2-h intervals, inside and around houses, in August and October 2004. Overall, 633 sandflies, belonging to six species of Phlebotomus and three of Sergentomyia, were collected. Sandfly activity was nocturnal and higher at twilight. Several activity patterns were observed according to the species. Phlebotomus (Paraphlebotomus) sergenti Parrot, 1917, the suspected vector of L. tropica in this focus, was caught during each collection performed from 1900 to 0500 hours, the numbers of species caught peaked at 1900-2100 hours. There were seasonal variations of the nocturnal activity, which could be related to the variations in temperature and relative humidity.

Animals↗
PubMed1996

The effect of immune sera from hamsters immunized with sandfly gut and whole body extract antigens on the fecundity and mortality of Phlebotomus duboscqi (Diptera: Psychodidae).

Phlebotomus duboscqi were fed on hamsters previously immunized with different concentrations of homogenized crude sandfly gut antigen and supernatant obtained from whole body extract. The humoral response in the rodents was quantified at different times post-immunization by the enzyme-linked immunosorbent assay. Sandflies were fed on either immunized or saline control hamsters and the effect of the blood meals on sandfly feeding, survival and fecundity was investigated. The humoral response in immunized hamsters as measured by the presence of P. duboscqi-specific IgG antibodies was significantly greater (P < 0.05) as compared to the controls. This difference was noted in sera collected on 15 and 25 days post-immunization. Sandflies fed on immunized hamsters had a significantly higher mortality (P < 0.05) and decreased egg production (P < 0.05) than those fed on unimmunized control hamsters.

Animals↗
PubMed2006

Detection and identification of Leishmania species in field-captured phlebotomine sandflies based on mini-exon gene PCR.

Leishmaniasis is one of the most diverse and complex of all vector-borne diseases. Because it involves several overlapping species and sandfly vectors, the disease has a complex ecology and epidemiology. Adequate therapy and follow-up depend on parasitological diagnosis, but classical methods present several constraints when identifying species. We describe a polymerase chain reaction (PCR) which uses primers designed from mini-exon repetitive sequences that are specific for subgenus LeishmaniaViannia (PV), as well as sequences with specificity for genus (PG) that can distinguish between Leishmania species from other insect flagellates with minor differences in PCR products. For standardization, these PCR were tested in experimentally infected sandflies, and Leishmania infection in these insects was successfully confirmed. This methodology identified a 3.9% infection rate in field-captured phlebotomine sandflies from an endemic region in Brazil. Natural infection by Leishmania species was identified in three samples of Lutzomyia longipalpis, of which two were Leishmania (L.) chagasi and one Leishmania (L.) amazonensis. Irrespective of specific epidemiological conclusions, the method used in this study was able to identify Leishmania infections both in experimentally infected and field-captured phlebotomine sandflies, and could be a useful tool in epidemiological studies and strategic planning for the control of human leishmaniasis.

Animals↗
PubMed2005

Pool screen PCR for estimating the prevalence of Leishmania infantum infection in sandflies (Diptera: Nematocera, Phlebotomidae).

Prevalence studies of infection in the sandfly vector can be used as an indicator of a change in the intensity of Leishmania transmission. However, these studies are difficult to carry out as prevalence in the vector is usually low and its estimation requires a large number of sandflies to be dissected. Our objective was to establish whether a L. infantum-specific PCR-ELISA applied to pools of female sandflies and a previously described algorithm could be useful tools to study the prevalence of infection by this parasite in natural vector populations. We collected sandflies from six collection points in two stable foci of leishmaniasis in southern (N=3) and north-eastern (N=3) Spain, following standard procedures. A fraction of the collected females was dissected and morphologically identified. Another fraction was used for pool screening. In total, 127 pools of 30 females (3810 specimens) were studied by PCR-ELISA and 1764 specimens were individually dissected. The prevalence of infection determined by dissection does not differ from that determined by pool screen PCR. The results suggest that pool screen PCR can be of practical use in the epidemiological surveillance of leishmaniasis in European countries of the western Mediterranean basin, associated with control interventions or global change.

Animals↗
PubMedDate not supplied

Species-specific detection of Leishmania in sandflies using an enzyme-linked immunosorbent assay.

A direct enzyme-linked immunosorbent assay (ELISA) using species-specific monoclonal antibodies was developed for the detection and identification of Leishmania in sandflies. A titration of mock-infected Phlebotomus papatasi showed that fewer than 2000 L. major promastigotes could be detected. The percentage of infected P. papatasi collected in the field, as determined by dissection, was compared to that revealed by the ELISA. Both methods gave similar results, irrespective of whether the flies were caught by sticky papers or light-traps. The percentage of infected flies determined by either method was also similar in experimentally infected colony reared sandflies. The ELISA can be carried out using multiple species-specific antibodies, and is as accurate as identification of infected sandflies by microscopical examination. The technique should be useful for identifying sandfly species involved in transmitting different species of Leishmania, and for rapid assessment of leishmanial infection rates in endemic regions.

Animals↗
PubMed1999

Filamentous proteophosphoglycan secreted by Leishmania promastigotes forms gel-like three-dimensional networks that obstruct the digestive tract of infected sandfly vectors.

Development of Leishmania parasites in the digestive tract of their sandfly vectors involves several morphological transformations from the intracellular mammalian amastigote via a succession of free and gut wall-attached promastigote stages to the infective metacyclic promastigotes. At the foregut midgut transition of Leishmania-infected sandflies a gel-like plug of unknown origin and composition is formed, which contains high numbers of parasites, that occludes the gut lumen and which may be responsible for the often observed inability of infected sandflies to draw blood. This "blocked fly" phenotype has been linked to efficient transmission of infectious metacyclic promastigotes from the vector to the mammalian host. We show by immunofluorescence and immunoelectron microscopy on two Leishmania/sandfly vector combinations (Leishmania mexicana/Lutzomyia longipalpis and L. major/Phlebotomus papatasi) that the gel-like mass is formed mainly by a parasite-derived mucin-like filamentous proteophosphoglycan (fPPG) whereas the Leishmania polymeric secreted acid phosphatase (SAP) is not a major component of this plug. fPPG forms a dense three-dimensional network of filaments which engulf the promastigote cell bodies in a gel-like mass. We propose that the continuous secretion of fPPG by promastigotes in the sandfly gut, that causes plug formation, is an important factor for the efficient transmission to the mammalian host.

Acid Phosphatase↗