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PubMed2006

Role of kdr and esterase-mediated metabolism in pyrethroid-resistant populations of Haematobia irritans irritans (Diptera: Muscidae) in Brazil.

The horn fly, Haematobia irritans irritans (L.) (Diptera: Muscidae), has become a problem for Brazilian cattle producers even though its introduction into Brazil is relatively recent. Failure to control this cattle pest is becoming a concern, and horn fly populations from several ranches from the state of Mato Grosso do Sul were surveyed for pyrethroid resistance. Susceptibility bioassays revealed that cypermethrin resistance was widespread and reached high levels in horn fly populations throughout the state, with resistance factors (RFs) ranging from 50.4 to 704.8. Synergist bioassays failed to detect a major role for esterases as a pyrethroid resistance mechanism in these populations, except for the highly pyrethroid-resistant Estrela do Oeste population (RF = 704.8). The kdr sodium channel gene mutation was not detected in eight of the 13 populations, but < 7% of individuals from four populations and 50% of the flies from Estrela do Oeste exhibited this mutation. Neither the superkdr sodium channel gene mutation nor a resistance-associated gene mutation in the HialphaE7 carboxylesterase were found in any of the fly populations. Although target site insensitivity (kdr) and esterase-mediated metabolism occur in horn fly populations from Mato Grosso do Sul state, it seems that they are not the major mechanism causing pyrethroid resistance in most of these populations.

Animals↗
PubMed2006

Bloodmeal hosts of Anopheles species (Diptera: Culicidae) in a malaria-endemic area of the Brazilian Amazon.

Hosts of blood-fed anophelines (Diptera: Culicidae) were determined in three riverine villages, 1.5-7.0 km apart, along the Matapí River, Amapá state, Brazil, by enzyme-linked immunosorbent assay midgut analysis for IgG of common vertebrates. Anopheles marajoara Galvão & Damsceno and Anopheles darlingi Root had higher human blood indices (HBI) than Anopheles nuneztovari Gabaldón, Anopheles triannulatus (Neiva and Pinto), and Anopheles intermedius (Chagas), which were relatively zoophilic. HBIs of An. darlingi varied significantly among villages, attributable to a low proportion of human-fed mosquitoes in Santo Antônio. Significantly higher incidence of An. marajoara and An. nuneztovari fed on pig blood at two villages, associated with a low number of pigs in Santo Antônio. The incidences of bovine blood varied significantly among villages for all three of the most common anopheline species. The incidence of mixed meals ranged from 7.1 to 27.6% among common species, and, for An. marajoara, varied significantly among villages. This study demonstrates differences in host selection patterns among villages only a few kilometers apart, which may be influenced by host availability and have important epidemiological consequences.

Animals↗
PubMed2006

The potential of Aedes triseriatus (Diptera: Culicidae) as an enzootic vector of West Nile virus.

The susceptibility of Aedes triseriatus (Say) (Diptera: Culicidae) to low levels of West Nile virus (family Flaviviridae, genus Flavivirus, WNV) was determined and compared with that of Culex pipiens L. to assess the likelihood of its participation in an enzootic cycle involving mammals. Ae. triseriatus and Cx. pipiens were exposed to WNV by feeding on baby chickens with WNV serum titers ranging from 10(4.1 +/- 0.1) to 10(8.6 +/- 0.1) plaque-forming units (PFU)/ml and from 10(4.1 +/- 0.1) to 10(7.0) PFU/ml, respectively. Infection rates and 95% confidence intervals (CIs) of 8% (4, 14) and 25% (15, 38) occurred in Ae. triseriatus and Cx. pipiens after feeding on chickens with WNV titers of 10(4.1 +/- 0.1) PFU/ml and increased to 65% (49, 79) and 100% (72, 100) in Ae. triseriatus and Cx. pipiens after feeding on chickens with titers of 10(7.1 +/- 0.1) PFU/ml. The mean infection rate of Ae. triseriatus ranged from 97% (84, 100) to 100% (79, 100) after feeding on chickens with WNV titers of > or = 10(8.2) PFU/ml. The infectious dose (ID)50 values for Ae. triseriatus and Cx. pipiens were 10(6.5) (6.4, 6.7) and 10(4.9) (4.6, 5.1) PFU/ml, respectively. The combined estimated transmission rate of Ae. triseriatus at 14 and 18 d after feeding on chickens with a mean WNV titer of 10(8.6 +/- 0.1) PFU/ml was 55%. Although Ae. triseriatus is significantly less susceptible to WNV than Cx. pipiens, the susceptibility of Ae. triseriatus to WNV titers < 10(5.0) PFU/ml and its ability to transmit WNV suggest that Ae. triseriatus has the potential to be an enzootic vector among mammalian populations.

Aedes↗
PubMed2006

Spatial analysis of Aedes albopictus (Diptera: Culicidae) oviposition in suburban neighborhoods of a Piedmont community in North Carolina.

Temporal and spatial distribution of egg-laying by Aedes albopictus (Skuse) (Diptera: Culicidae) was investigated in suburban neighborhoods in Raleigh, NC, by using oviposition traps (ovitraps) at fixed sampling stations during the 2002 and 2003 mosquito seasons. Variations in the phenology of oviposition between the two mosquito seasons resulted from differences in the patterns and amounts of rainfall early in the season. Aerial images of each study neighborhood were digitized, and the proportions of specific types of land cover within buffer zones encompassing ovitraps were estimated. Retrospective analyses showed that in some neighborhoods, oviposition intensity was significantly associated with specific types of land cover. However, in general, it seemed that gravid Ae. albopictus searched throughout the landscape for water-filled containers in which to lay eggs. Peridomestic surveys were carried out concurrently with ovitrap collections to estimate production of Ae. albopictus pupae in discarded water-filled containers and the abundance of females in vegetation that made up the resting habitat. Results of linear regression analyses indicated that the mean standing crop of pupae (total and per container) per residence was not a significant predictor of mean egg densities in ovitraps. However, the mean standing crop of adult females was a significant but weak predictor variable, because the magnitude and sign of regression coefficients varied between neighborhoods. Linear spatial regression analyses revealed that oviposition intensity was not spatially dependent on pupal standing crop or the numbers of pupae-positive containers distributed peridomestically. However, a weak spatial dependence on the standing crop of adult females was found in some neighborhoods. Based on spherical variogram models, kriging was carried out to predict the spatial patterns of oviposition in suburban neighborhoods. Focal areas of high and low oviposition intensity were evident in most neighborhoods; however, the spatial patterns of oviposition changed between mosquito seasons. Kriging predictions were evaluated, using cross-validation, by systematically removing each data point from our data set and predicting the removed point by using the remaining points. The root mean square (standardized) error values of best fitting variogram models approximated 1, and plots of standardized PRESS residuals showed no distinct pattern for most neighborhoods, indicating that predictions of the spatial distribution of oviposition intensity were valid. Spherical variogram models are a satisfactory method for describing the spatial distribution of Ae. albopictus oviposition, and kriging can be a useful technique for predicting oviposition intensity at locations that have not been sampled.

Aedes↗
PubMed2000

Coadaptation of male aedeagal filaments and female spermathecal ducts of the old world phlebotomine sand flies (Diptera: Psychodidae).

The morphology of insect genitalia is often highly species-specific, and its variation has been suggested as an important impetus for evolution. Structural variation of the male genitalia and the female spermathecae in Phlebotomine sand flies is unique among the blood sucking Diptera. We describe the fine structures involved in mating for Phlebotomus papatasi (Scopoli). Relationships among the length of the spermathecal duct and aedeagal filament were studied in 26 species of Old World phelebotomine sand flies comprising 12 subgenera. In most taxa the aedeagal filaments were very long allowing direct insemination into the spermathecae, indicating that intrasexual competition among males occurs as sperm precedence. There was a positive correlation between the lengths of the spermathecal ducts and aedeagal filaments, indicating that an evolutionary "arms race" occurs between males and females over the control of fertilization. The pattern of genitalia variation in phlebotomine sand flies also indicated that differences in the lengths of the spermathecal ducts and aedeagal filaments are distributed unevenly among closely related species. Genital differences do not seem to occur in populations of the Phlebotomus argentipes Annandale & Brunetti complex, whereas differences showed strongly in closely related Phlebotomus papatasi and P. bergeroti Parrot, and in sympatric Phlebotomus martini Parrot and P. celiae Minter.

Animals↗
PubMed2000

Sensilla on the terminalia of Lutzomyia spp. (Diptera: Psychodidae) sand flies.

Sensilla on the male terminalia of four different species of sand flies (Diptera: Psychodidae) are described. Two species belonged to the same species complex: Lutzomyia longipalpis (Lutz & Neiva) and L. cruzi (Mangabeira). Other Brazilian species complex was represented by L. lenti (Mangabeira) and L. carmelinoi (Ryan, Fraiha, Lainson & Shaw). Approximately 5-15 sensilla were found on the initial part of the styles of all species. Structurally, the sensory organs were similar to small coeloconica sensilla. Their number varied both individually and interspecifically. These sensilla may play an important role in the copulation behavior of these species of sand flies.

Animals↗
PubMed2002

Sand flies (Diptera: Phlebotominae) in Sanliurfa, Turkey: relationship of Phlebotomus sergenti with the epidemic of anthroponotic cutaneous leishmaniasis.

Sand fly (Diptera: Phlebotominae) fauna were surveyed in various districts of Sanliurfa in southeast Turkey for 3 yr immediately after an epidemic of cutaneous leishmaniasis (Leishmania tropica). Sticky papers and CDC light traps collected a total of 10,937 sand flies, of which 10,919 (4,158 females and 6,761 males) were identified as Phlebotomus and 18 (11 females and seven males) as Sergentomyia (S. theodori Parrot; S. adleri Theodor). Eight Phlebotomus spp. were identified: P. sergenti Parrot (72.3%), P. papatasi (Scopoli) (27.2%), P. brevis Theodor & Mesghali (0.20%), P. neglectus Leger & Pesson (0.13%), P. perfiliewi Parrot (0.05%), P. mascitti Grassi, P. halepensisTheodor, and P. alexandri Sinton (0.01%). Phlebotomus mascitti and P. neglectus, along with both Sergentomyia sp., have not been previously described from the study area. Similar results were obtained when both trapping methods were applied in the same houses, indicating that local P. sergenti and P. papatasi populations were equally attracted to the light. P. sergenti was consistently abundant, agreeing with the general view that this species is the vector of leishmaniasis in the region. There was no apparent decrease in the relative abundance of this vector versus the other species, suggesting that factor (s) other than a change in the dynamics of sand fly populations precipitated the decline of the human leishmaniasis epidemic in Sanliurfa.

Animals↗
PubMed2002

Post-bloodmeal diuretic shedding of hepatitis B virus by mosquitoes (Diptera: Culicidae).

Persistence and diuretic shedding of hepatitis B virus (HBV) by mosquitoes (Diptera: Culicidae) was studied by using infectious blood feedings, intrathoracic inoculations, and detection of virus by polymerase chain reaction (PCR) and Southern hybridization. Results showed that both Anopheles stephensi Liston and Ochlerotatus triseriatus (Say) shed HBV during diuresis for up to 72 h after feeding on an HBV-positive serum drawn from a human donor. HBV did not persist in the bodies of either An. stephensi or Oc. triseriatus past 72 h by infectious feeding or intrathoracic inoculation of HBV suspension. Viral dissemination did not occur by infectious feeding in An. stephensi or Oc. triseriatus, or by intrathoracic inoculation in An. stephensi, Oc. triseriatus, or Culex quinquefasciatus Say. These results suggest that HBV could be transmitted to humans by a stercorarian route, especially if mosquitoes that fed on an HBV-positive human are interrupted during feeding and move to another person to resume feeding.

Animals↗
PubMed2002

Genetic differentiation and diagnostic loci among Anopheles (Nyssorhynchus) rangeli, An. (Nys.) nuneztovari, and An. (Nys.) dunhami (Diptera: Culicidae) in the Brazilian Amazon.

Genetic variability and divergence were estimated for populations of Anopheles rangeli Gabaldón, Cova-Garcia & Lopes, Anopheles nuneztovari Gabald6n cytotype A, and Anophels dunhami Causey from the Brazilian Amazon using isozyme electrophoresis. These species are included in the Oswaldoi subgroup, subgenus Nyssorhynchus (Diptera: Culicidae). Thirteen enzymes yielded a total of 22 loci, of which 14 were monomorphic in the three species. Three diagnostic loci (Gpi-1, Hk-1, and Me) and a strong differentiation in the Mdh locus were found between An. rangeli and An. nuneztovari. Five diagnostic loci (Mdh, Gpi-l, Hk-1, Gpd, and Me) separated An. rangeli from An. dunhami, whereas one diagnostic locus (Gpd) separated An. nuneztovari from An. dunhami. Moderate differentiation was observed in the Est-5 and Pgm loci between An. rangeli and An. nuneztovari, and between An. nuneztovari and An. dunhami. Anopheles dunhami had the highest values for three indices of genetic variability, whereas An. rangeli showed the lowest values for mean number of alleles per locus and mean heterozygosity. Nei's genetic distance was highest between An. rangeli and An. dunhami (0.280) and lowest between An. nuneztovari and An. dunhami (0.072). Between An. rangeli and An. nuneztovari the genetic distance was 0.237. Anopheles dunhami and An. nuneztovari are sister species very closely related and may have a recent evolutionary origin. Anopheles rangeli probably diverged before the other two species separated. This is the first record of An. dunhami in Coari (Amazonas, Brazil) and only the third collection site of this species since its description in 1945.

Animals↗
PubMed2002

Sampling for sand flies (Diptera: Psychodidae) among prairie dog colonies on ranches with histories of vesicular stomatitis in new Mexico and Colorado.

The possible presence of sand flies (Diptera: Psychodidae) among colonies of Gunnison's prairie dogs, Cynomys gunnisoni Baird, was investigated on or near ranches with histories of vesicular stomatitis (VS) in domestic livestock in the mid-Rio Grande River Valley, NM, and west-slope region of Colorado. Sampling was conducted at six locations, three in each region, using three methods: standard down-draft suction/light traps, up-draft suction/light traps (both supplemented with CO2), and burrow-emergence traps. Two male and four female sand flies, Lutzomyia apache, Young and Perkins were recovered from prairie dog colony and sand-sagebrush association habitats on a cattle ranch in Socorro County, NM. Sand flies were captured only in up-draft suction/light traps. This record is of interest for the following reasons: (1) sand flies have not been reported from the Rio Grande River Valley of New Mexico, (2) sand flies are biological vectors of VS viruses in other areas, (3) the mid-Rio Grande River Valley has been a focus of VS virus activity in domestic livestock during recent outbreaks, and (4) the source of vesicular stomatitis viruses that infect livestock in the western United States is unknown, but may involve introductions of virus from Mexico or an endemic cycle maintained by vector insects.

Animals↗
PubMed2003

Influence of sugar availability and indoor microclimate on survival of Anopheles gambiae (Diptera: Culicidae) under semifield conditions in western Kenya.

The influence of indoor microclimate on survival of female Anopheles gambiae sensu stricto Giles (Diptera: culicidae) mosquitoes fed on different nutrition sources was evaluated in a semifield experimental hut exposed to ambient climate in western Kenya. Cages of mosquitoes (n approximately 50 per cage) were placed in nine positions within the hut combining three different sides and three different heights. At each height and side, mosquitoes were offered either human blood (once every 2 d), glucose (6% wt:vol) or a combination of the two diets over three experiments so that each cage position received one diet source. The effect of diet on survival was significant with mean survival times of 14 d for mosquitoes fed blood alone, 29 d for sugar alone and 33 d for blood plus sugar. Sugar availability decreased the odds of mortality approximately 85% compared with the blood group. Micro heterogeneities of temperature but not relative humidity also influenced survival although to a much lesser extent. The side but not height within the hut at which mosquitoes were placed, influenced survival but could not be explained by either temperature or relative humidity differences. The potential influence of seemingly minor heterogeneities of indoor microclimate upon vector longevity and vectorial capacity may merit further investigation. Also, the availability of sugar was shown to be a potentially crucial determinant of vectorial capacity. Compared with blood alone, the availability of sugar served to increase survival potential of vectors beyond ages at which they are old enough to transmit malaria.

Animals↗
PubMed2003

Biting rates and developmental substrates for biting midges (Diptera: Ceratopogonidae) in Iquitos, Peru.

Biting midges (Diptera: Ceratopogonidae) were collected at 16 periurban and rural sites around Iquitos, Peru, between 17 October 1996 and 26 May 1997. Culicoides paraensis (Goeldi), the principal vector of Oropouche virus, was the most commonly collected species (9,086 flies) with Culicoides insinuatus Wirth & Blanton second (7,229 flies). Although both species were collected at all sampling sites (linear (distance surveyed approximately 25 km), C. paraensis dominated at northern collection sites (> 90%), whereas C. insinuatus prevailed at southern collection sites (> 60%). C. paraensis were collected from human sentinels at a constant rate throughout daylight hours, at similar rates during wet and dry months, and regardless of rainfall. Larval developmental substrates for C. paraensis included decaying platano (Musa x paradisiaca L. [Musaceae]) stems, stumps, flowers, fruits, and debris beneath platano trees as well as from soil beneath a fruiting mamay (Syzygium malaccense Merr. & Perry [Myrtaceae] ) tree and organic-rich mud along a lake shoreline. C. insinuatus adults likewise emerged from decaying platano and organic-rich mud along a lake shoreline, but also from debris accumulated in the axils of aguaje (Mauritia flexuosa L. [Palmae]) fronds and decaying citrus fruit. Despite high numbers of biting adults near putative substrates, adults of neither species emerged from other decomposing plant material, soil, phytotelmata, or artificial containers. Because both species of biting midges emerged in high numbers from all parts of platano (ubiquitous in Iquitos), it will be challenging to control them through sanitation.

Animals↗
PubMed2004

Spatial and temporal patterns of phlebotomine sand flies (Diptera: Psychodidae) in a cutaneous leishmaniasis focus in northern Argentina.

Phlebotomine sand flies (Diptera: Psychodidae) were captured in an area of Argentina endemic for American cutaneous leishmaniasis (ACL). A total of 44,944 flies were collected during a 130-wk interepidemic period from 1990 through 1993. These sand flies included Lutzomyia neivai (Pinto) (97.8%), Lutzomyia migonei (Franca) (1.2%), Lutzomyia cortelezzii (Brèthes) (0.8%), Lutzomyia shannoni (Dyar) (0.1%), and Lutzomyia punctigeniculata (Floch and Abonnenc) (0.1%). Lutzomyia neivai was more abundant in secondary forests and peridomestic environments associated with human cases than in primary forest or xeric thorn scrub areas. Time series analyses of species densities suggested a bimodal or trimodal annual pattern related to rainfall peaks, a 5-wk reproductive cycle, and peridomestic local populations that were located adjacent to secondary forests. In general, sand fly abundance was correlated with the rainfall of the previous year. Lutzomyia neivai spatial distributions were consistent with ACL incidence patterns during the study and in the recent outbreaks in Argentina. However, Lu. migonei also may be involved in peridomestic transmission. Our results suggest that there is a need for improved, long-term surveillance of sand flies and ACL cases, as well as development of effective intervention strategies.

Animals↗
PubMed2004

Description and bionomics of Anopheles (Cellia) ovengensis (Diptera: Culicidae), a new malaria vector species of the Anopheles nili group from south Cameroon.

Mosquito species of the Anopheles nili group (Diptera: Culicidae) transmit malaria to humans along rivers in Africa. To date, the An. nili group includes the species Anopheles nili s.s. and its pale-winged variant known as the "Congo form," Anopheles somalicus and Anopheles carnevalei. Larval and adult mosquito collections in the forest region of Campo, in southern Cameroon, uncovered an additional morphological variant provisionally called "Oveng form" that was subsequently found to be genetically distinct from the other members of the An. nili group. In this study, we provide further biological data that characterizes this new taxon and justifies elevation to specific rank. We propose calling this new species Anopheles ovengensis, after its geographical origin. We present a morphological description of the adult female and fourth instars and original data on the biology, ecology, and role as a human malaria vector of this new species in its type location. We provide dichotomous keys for identification of adult females and fourth instars that can be used at least in tropical areas of west and central Africa.

Animals↗
PubMed2004

Analysis of reproductive isolation between sibling species anopheles Albitarsis sensu stricto and Anopheles deaneorum, two malaria vectors belonging to the Albitarsis complex (Diptera: Culicidae).

Complexes of sibling species are common among mosquitoes, and their existence within vector species can have important epidemiological consequences. Anopheles albitarsis sensu stricto and Anopheles deaneorum Rosa-Freitas are two putative vectors of malaria parasites belonging to the Albitarsis species complex (Diptera: Culicidae). Using an induced mating technique, we studied the reproductive isolation between these two closely related species and their reciprocal hybrids. Evidence for hybrid male sterility consistent with Haldane's rule was found. The results indicate that male hybrids show very low insemination rates, probably due to abnormalities in their reproductive organs. In addition, the data show that hybrid males carrying an X chromosome derived from An. deaneorum perform significantly worse than hybrid males carrying an An. albitarsis s.s. X chromosome.

Animals↗
PubMed2004

Temporal and geographic patterns of Aedes aegypti (Diptera: Culicidae) production in Iquitos, Peru.

Large-scale longitudinal cohort studies are necessary to characterize temporal and geographic variation in Aedes aegypti (L.) (Diptera: Culicidae) production patterns and to develop targeted dengue control strategies that will reduce disease. We carried out pupal/demographic surveys in a circuit of approximately 6,000 houses, 10 separate times, between January 1999 and August 2002 in the Amazonian city of Iquitos, Peru. We quantified the number of containers positive for Ae. aegypti larvae and/or pupae, containers holding pupae, and the absolute number of pupae by 4-mo sampling circuits and spatially by geographic area by using a geographic information system developed for the city. A total of 289,941 water-holding containers were characterized, of which 7.3% were positive for Ae. aegypti. Temporal and geographic variations were detected for all variables examined, and the relative importance of different container types for production of Ae. aegypti was calculated. Ae. aegypti larvae and pupae were detected in 64 types of containers. Consistent production patterns were observed for the lid status (lids: 32% wet containers, 2% pupal production), container location (outdoor: 43% wet containers, 85% pupal production), and method by which the container was filled with water (rain filled: 15% wet containers, 88.3% pupal production); these patterns were consistent temporally and geographically. We describe a new container category (nontraditional) that includes transient puddles, which were rare but capable of producing large numbers of pupae. Because of high variable pupal counts, four container categories (large tank, medium storage, miscellaneous, and nontraditional) should be targeted in addition to outdoor rain-filled containers that are not covered by a lid. The utility of targeted Ae. aegypti control is discussed, as well as the ability to achieve control objectives based on published but untested threshold values.

Aedes↗
PubMed2006

Field evaluation of deet, Repel Care, and three plant based essential oil repellents against mosquitoes, black flies (Diptera: Simuliidae) and land leeches (Arhynchobdellida: Haemadipsidae) in Thailand.

Diethyl methyl benzamide, or deet, a commercial plant-based repellent (Repel Care), and essential ils from 3 species of plants (finger root rhizomes, guava leaves, and turmeric rhizomes), steam distillated and formulated as insect repellents, were evaluated in the field on human volunteers against hematophagous mosquitoes, black flies, and land leeches in Thailand. Field trials were conducted against wild mosquitoes in Bang Bua Thong District, Nonthaburi Province, and in the Thap Lan National Park Headquarters, Nadee District, Pranchinburi Province; anthroophilic black flies (Diptera: Simuliidae) at the Forestry Fire Control Station in Doi Inthanon National Park, Chomthong district, Chiang Mai Province; and land leeches (Arhynchobdellida: Haemadipsidae) in the Khao Yai National Park, Pak Chong District, Nakhon Ratchasima Province. The 3 experimental plant-based essential oil formulations as well as Repel Care and deet provided complete protection from mosquito landing and biting for up to 9 h (duration of the experiment). Similar results were obtained with the 5 products against black flies, providing 100% protection for 9 h but 96-82% protection after 10 and 11 h posttreatment. The 5 repellent products also provided 100% protection against land leeches for at least 8 h. Thi is the 1st report of repellency of plant-based repellents against black flies and land leeches in Thailand. The identification and availability of inexpensive sources of plant-based oils, i.e., finger root rhizomes, guava leaves, and turmeric rhizomes providing long-lasting repellency against blood-sucking organisms are promising leads into commercial production of relatively safe and effective repellents.

Aedes↗
PubMed1996

[Identification and characterization of allergens of Chironomus flaviplumus adults (Chironomidae, Diptera) in mice].

Non-biting midges (Chironomidae, Diptera) are one of the largest insect families, which are distributed worldwidely and are found in nearly all types of inland waters. They are known to be aggressive inhalant allergens which cause allergenic diseases. In this study, the crude antigens of Chironomus flaviplumus adults which are most widely distributed in Korea were extracted, and their allergens were analyzed with the sera from experimentally sensitized mice. The mice were immunized with 1 microg or 10 microg of the crude antigens, respectively, and the specific serum IgE levels were measured by both ELISA and passive cutaneous anaphylaxis (PCA) techniques. The highest levels of both total IgE and chironomid-specific IgE were found in the mouse sera obtained after 9 weeks of the first injection with 1 mu g crude antigen. The crude antigen was separated into 16-18 protein bands on gel by SDS-PAGE. The crude extract was assessed by SDS-PAGE/immunoblot analysis. One IgE-binding band (65 kDa) was detected by developing with colorimetric substrate, and 4 IgE-binding bands (65, 52, 35 IgE-binding and 25 kDa) by developing with CSPD chemiluminescent substrate. The SDS-PAGE gel of the crude extract of chironomid adults was equally cut into 30 pieces and each of them was eluted to isolate proteins by molecular weight, and the allergenicity of each eluate was assessed by applying P-K test on rats. Proteins of 65, 35 and 15 kDa showed the highest P-K titer (x 512) which was 16 times higher than that of the crude extract (x 32). The P-K titer of 52 kDa protein was also 4 times higher (x 128) than that of the crude extract, whereas the 25 kDa protein poorly responded, which seemed not antigenic. In conclusion, the present result in mice demonstrated that adults of Chironomus flaviplumus, a predominent species in Korea, cause allergenic diseases and the main allergens are 65, 52, 35 and 15 kDa proteins, of which 65 kDa protein seems to be a main allergen.

Allergens↗