Experimental formulations of Bacillus sphaericus and B. thuringiensis israelensis against Culex quinquefasciatus and Anopheles gambiae (Diptera: Culicidae) in Burkina Faso

Efficacies of locally produced, sustained release granular formulations of Bacillus sphaericus (BS) strain 2362 and B. thuringiensis israelensis (BTI) were compared with commercial liquid concentrates of the same bacteria in cesspits and rain-filled puddles in Ouagadougou, Burkina Faso. Duration of control was dependent on the product, the transient nature of some sites, and the target mosquito larvae. BS granules applied at the rate of 3.0 g/m2 (30 kg/ha) reduced Culex quinquefasciatus Say 99% for at least 28 d in cesspits, whereas the same dosage of 2 BTI granules and commercial liquid formulations of BS and BTI gave 95% control for 8-14 d. The levels of control obtained with the 2 liquid products were not different. Accordingly, the reported inferiority of BTI to BS in polluted water was attributed to low dosages of BTI. Because products were compared at equal application rates, recycling seemed to play a minor role compared with product formulation. BTI was more broad spectrumed than BS also killing Cx decens Theobald, Cx cinereus Granpré & Charmoy, and psychodid larvae. Depending on the method of estimation, granular and liquid BS gave 60-97% control of Anopheles gambiae s.l. Giles for 10 d in rain puddles. The transient nature of the rain puddles was probably more important for the duration of control than formulation type. Cesspits and puddles, respectively, were the most important larval habitats for these 2 species in Ouagadougou during the rainy season, and these trials showed that An. gambiae was not controlled easily with larvicides alone. However, biological larvicides may play a central role in the control of Cx. quinquefasciatus provided that most breeding sites are treated.     

Impact of inorganic pollutants perchlorate and hexavalent chromium on efficacy of Bacillus sphaericus and Bacillus thuringiensis subsp. israelensis against Culex quinquefasciatus (Diptera: Culicidae)

The effects of two widespread environmental pollutants, perchlorate and hexavalent chromium, were assessed on the efficacy of Bacillus thuringiensis subsp. israelensis (Bti) and Bacillus sphaericus (Bsph) against fourth instars of Culex quinquefasciatus Say (Diptera: Culicidae) in 24-h laboratory bioassays. Although 250 mg/liter perchlorate, a level somewhat higher than would be considered ecologically relevant, did not affect the control provided by either larvicide, presence of 1.04 mg/liter hexavalent chromium, an ecologically relevant concentration, increased the efficacy of both Bti and Bsph by 21 and 80%, respectively. In the presence of hexavalent chromium, improved suppression could be expected from Bacillus applications at the current label rates. However, because hexavalent chromium has been shown to affect many taxa, we propose that the potential exists for increased susceptibility of nontarget organisms to Bacillus products in polluted habitats.     

Laboratory selection for resistance to Bacillus sphaericus in Culex quinquefasciatus (Diptera: Culicidae) from California, USA

A previously untreated field population of Culex quinquefasciatus Say, collected near Bakersfield, CA, was subjected to intensive laboratory selection with the bacterial insecticide Bacillus sphaericus Neide (strain 2362) at a level producing 95% mortality. Resistance rapidly appeared and resistance levels increased such that fourth instars of generation 12 were able to survive a concentration of B. sphaericus that was 7,000 times higher than the median lethal concentration (LC50) of the susceptible reference colony. Similar resistance levels were detected in first instars. Cross-resistance in the selected colony was detected toward B. sphaericus strains 1593 and 2297, but little or no cross-resistance was observed toward B. sphaericus strains IAB59 or ISPC5 (= WHO 2173). Cross-resistance also was not detected toward the bacterial insecticide Bacillus thuringiensis subsp. israelensis, toward a recombinant strain expressing both B. thuringiensis subsp. israelensis and B. sphaericus (strain 1593) toxins, toward individual or multiple toxins from B. thuringiensis subsp. israelensis, or toward conventional synthetic insecticides. Genetic analysis revealed that B. sphaericus resistance was inherited as a recessive trait and controlled by a single major locus. These data are discussed in relation to cases of field resistance toward this biopesticide in the Cx. pipiens (L.) complex.     

Inheritance and mechanism of resistance to Bacillus sphaericus in Culex quinquefasciatus (Diptera: Culicidae) from China and Brazil

Investigations on the inheritance and mechanism of resistance to Bacillus sphaericus Neide in Culex quinquefasciatus Say colonies, selected with strains C3-41 (RLCq1/C3-41) and 2362 (CqRL1/2362), were performed in China and Brazil, respectively. The progeny of reciprocal F1 crosses (susceptible female x resistant male and vice versa) from both resistant colonies responded alike in bioassays, indicating recessive inheritance. Data on larvae susceptibility from the backcross offspring between F1 and their respective susceptible and resistant parental colonies are consistent with a monofactorial and autosomal mode of inheritance. In vitro binding assays between 125I binary (Bin2) toxin and the brush border membrane fractions (BBMF) from CqRL1/2362 and RLCq1/C3-41 larvae showed that resistance, in both colonies, is caused by a failure in the binding step of the B. sphaericus Bin2 toxin to its specific midgut receptor. The specific and saturable binding of Bin2 toxin to BBMF from F1 larvae (CqRL1/2362 X susceptible counterpart) confirms the recessive inheritance of the resistance gene. Further studies are needed to advance understanding of B. sphaericus resistance.     

Mosquitocidal toxin from Bacillus sphaericus induces stronger delayed effects than binary toxin on Culex quinquefasciatus (Diptera: Culicidae)

We studied the toxicity and delayed effects of a mosquitocidal toxin (Mtx1) and a binary toxin (Bin) produced in Escherchia coli E-TH21 and Bacillus thuringiensis B-CW1, respectively, on Culex quinquefasciatus (Diptera: Culicidae). Bioassay results showed that both E-TH21 powder and B-CW1 sporulated culture were highly toxic against susceptible Cx. quinquefasciatus, with LC50 values of 0.65 and 1.70 mg/liter against third and fourth instars at 48 h, respectively. After initial 48-h exposure of larvae to different concentrations of Mtx1 and Bin, significant continued mortality could be observed in larval, pupal, and emergence stages of Cx. quinquefasciatus. Importantly, the Mtx1 could induce higher cumulative larval and preadult mortalities than Bin toxin on the target mosquito. This finding is important for understanding the mode of action of Mtx1 and Bin toxins and for developing a new bioassay procedure for evaluation of toxicity of Bacillus sphaericus Neide, some strains of which produce Mtx1 and Bin, in the laboratory and field.     

Susceptibility profile of Culex quinquefasciatus (Diptera: Culicidae) to Bacillus sphaericus on selection with rotation and mixture of B. sphaericus and B. thuringiensis israelensis

Several strains of Bacillus sphaericus (Neide) (Bsph), a spore-forming bacterium that produces potent mosquitocidal toxins, have been widely used against mosquito larvae globally. In some locations in the field and in the laboratory, moderate to high levels of resistance in Culex pipiens complex mosquito larvae to Bsph have been reported. Practical strategies for dealing with the resistance problem and more importantly to delay or possibly prevent development of resistance to Bsph in the first place are urgently needed. From published reports, we deduced that Bsph-resistant mosquito larvae are completely susceptible to Bacillus thuringiensis ssp. isreaelensis (Bti) and that there is no appearance of cross-resistance to Bti. On the basis of this information, we hypothesized that Bti offers a good potential for the management of resistance to Bsph, especially when it comes to delaying or preventing emergence of resistance. We conducted laboratory studies to see whether rotation of Bsph and Bti or mixture of both delays or prevents resistance development in Bsph-susceptible California Cx. quinquefasciatus. Development of resistance using Bsph alone increased after F15 and fluctuated slightly on further selections. Rotation of Bsph and Bti resulted in much higher level and rapid emergence of resistance to Bsph. However, selection with mixtures of Bti and Bsph for 36 generations showed no emergence of resistance. On the basis of this information, we believe mixtures have a good potential in delaying or preventing Bsph resistance.     

Laboratory and simulated field evaluation of a new recombinant of Bacillus thuringiensis ssp. israelensis and Bacillus sphaericus against Culex mosquito larvae (Diptera: Culicidae)

In the laboratory, three microbial mosquito larvicidal products consisting of Bacillus thuringiensis ssp. israelensis de Barjac (Bti), Bacillus sphaericus (Neide) (Bsph) (strain 2362), and the University of California Riverside (UCR) recombinant (producing toxins of both Bacillus sphaericus and Bacillus thuringiensis ssp. israelensis) were bioassayed against larvae of Culex quinequefasciatus Say (susceptible and resistant to Bsph 2362), and Aedes aegypti (L.). Bti proved highly effective against Cx. Quinequefasciatus susceptible and resistant strains, with LC50 values of 0.009 and 0.011 ppm and LC90 values of 0.057 and 0.026 ppm for Bsph-susceptible and -resistant strains, respectively. Bti was also highly active against Ae. eagypti with LC50 and LC90 values of 0.014 and 0.055 ppm, respectively. The UCR recombinant was equally active against both Bsph-susceptible and -resistant strains of Cx. Quinquefasciatus; LC50 values were 0.005 and 0.009 and LC90 values were 0.030 and 0.043 ppm, respectively. Bti and the UCR recombinant essentially showed similar activity against Bsph-susceptible and -resistant strains. UCR recombinant showed high toxicity against Ae. eagypti with LC50 and LC90 values of 0.023 and 0.064 ppm, respectively. Bsph was highly active against susceptible strain of Cx. quinequefasciatus with LC50 and LC9o values of 0.006 and 0.024 ppm, respectively. Bsph exhibited little toxicity against Ae. eagypti larvae and also no toxicity to Bsph resistance. In the field, we evaluated four experimental corn grit formulations of Bti (VBC 60021), Bsph (VBC 60022), UCR recombinants VBC 60023 (7.89%), and VBC 60024 (1.87%) in simulated field (microcosms) against Bsph-susceptible Culex mosquitoes. Bti and low-concentrate UCR recombinant showed similar initial activity as well as persistence. Both materials provided high-to-moderate level of control for 2-7 d posttreatment at low treatment rates. At low dosages, residual activity of Bti and UCR recombinant lasted for <7 d. Bsph and high-concentrate UCR recombinant (VBC 60023), however, were more effective against natural populations of Cullex and achieved longer control (7-21 d) than the other two materials.     

Overproduced esterases in Culex pipiens quinquefasciatus (Diptera: Culicidae) from Vietnam

The electrophoretic polymorphism of loci encoding for 10 enzymes was studied in Culex p. quinquefasciatus Say from six localities of Vietnam. The analysis of 11 "neutral genes" showed that differentiation among samples was low, but significant (Fst = 0.06), and significantly related to geographic distance between sample sites. These results are similar to those observed in other countries (Europe and west Africa). A single type of overproduced esterases (A2-B2) was observed, and its frequency was high (60-100%) in all samples. This situation is in sharp contrast with that observed in other countries of South East Asia (China, South Korea and Japan), where two or more types of overproduced esterases have been reported. A map summarizing the geographic distribution of Asian Cr. p. quinquefasciatus with overproduced esterases is provided.     

Synergistic activity of Cyt1A from Bacillus thuringiensis subsp. israelensis with Bacillus sphaericus B101 H5a5b against Bacillus sphaericus B101 H5a5b-resistant strains of Anopheles stephensi Liston (Diptera: Culicidae)

Integrated vector control is an effective and essential part of any successful vector control program. Increasing insecticide resistance requires strategies to prolong the use of highly effective vector control compounds. Synergistic activity between current effective pesticides is a powerful tool and is one such strategy. In the current study, Cyt1A from Bacillus thurigiensis subsp. israelensis and Bacillus sphaericus B101H5aH5b Bs were evaluated for the synergistic activity against B. sphaericus-resistant strains of Anopheles stephensi. The combinations of Cyt1A and B. sphaericus were found to act synergistically and were greatly enhanced at different ratios. A ratio of 1:4 of Cyt1A and Bs was 13,200-fold more toxic at LC₉₀ against the Bs-resistant strains of A. stephensi than was Bs alone and this high level of activity resulted from synergism between Cyt1A toxin and Bs. Our results therefore suggest that Cyt1A may enhance toxicity by facilitating the binding or insertion of the binary toxins to the microvillar membrane.     

Stable, germ-line transformation of Culex quinquefasciatus (Diptera: Culicidae)

A Hermes-based transposable element transformation system incorporating an enhanced green fluorescent protein (EGFP) marker was used to produce two transgenic lines of Culex quinquefasciatus (Say). The transformation frequency was approximately 12% and transformation of Culex was shown to be dependent on the presence of Hermes transposase. Injected Culex embryos were treated with four different heat shock regimes, two of which produced transformed individuals. These individuals were mated with wild-type mosquitoes and produced offspring which expressed the dominant EGFP gene in Mendelian ratios predicted for the stable integration of a gene at a single locus. The two transformed lines displayed distinct patterns of phenotypic expression, the expression of which has remained stable after fifteen generations. In these transgenic lines both the Hermes element and flanking plasmid DNA integrated into the Culex genome, as has been previously seen in Hermes-mediated transgenic strains of Aedes aegypti (L.). The high frequency of Culex transformation together with the dependence on the presence of Hermes transposase suggests that, as for Ae. aegypti, this mode of transposition into the germ-line genome occurs by an alternate mechanisms to the cut and paste type of transposition seen for this element in other insect species and in the somatic nuclei of mosquitoes. This is the first report of the genetic transformation of a species in the genus Culex and demonstrates that this medically important mosquito species can now, along with several other Culicine and Anopheline mosquito species, be genetically manipulated.     

Larvicidal activity of saponin from Achyranthes aspera against Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae)

The acetone, chloroform, ethyl acetate, hexane and methanol leaf extracts of Acalypha indica, Achyranthes aspera, Leucas aspera, Morinda tinctoria and Ocimum sanctum were studied against the early fourth-instar larvae of Aedes aegypti L and Culex quinquefasciatus Say. The larval mortality was observed after 24 h exposure. All extracts showed moderate larvicidal effects; however, the highest larval mortality was found in the ethyl acetate extract of A. aspera. In the present study, bioassay-guided fractionation of A. aspera led to the separation and identification of a saponin as a potential mosquito larvicidal compound, with LC₅₀ value of 18.20 and 27.24 ppm against A. aegypti and C. quinquefasciatus, respectively. ¹H NMR, ¹³C NMR and mass spectral data confirmed the identification of the active compound. This is the first report on the mosquito larvicidal activity of the saponin from the ethyl acetate extract of A. aspera. This study investigates the potential of crude extracts from commonly used medical herbs in India as an environmentally safe measure to control the vector of dengue and lymphatic filariasis.     

Larvicidal effects of various Euro-Asiatic plants against Culex quinquefasciatus Say larvae (Diptera: Culicidae)

Extracts from 56 species of plants in the Euro-Asiatic region were tested for larvicidal activity against the fourth larval instar of the mosquito Culex quinquefasciatus Say (Diptera: Culicidae). All plant extracts showed larvicidal activity after 24 h of exposure to the plant extracts in a maximal dose of 500 ppm. The extracts of the plants Otanthus maritimus and Ammi visnaga displayed the highest larvicidal effect (LD₅₀ 7 and 9 ppm, respectively) followed by Acer pseudoplatanus, Humulus japonicus, Acer platanoides, Satureja hortensis, Ocimum basilicum and Thymus vulgaris (LD₅₀ 23, 25, 28, 28, 32 and 48 ppm respectively). For eight species, the appraisal value of LD₅₀ was between 51 and 100 ppm, another eight species from 101 to 200 ppm, 15 species from 201 to 500 ppm and for 17 species, low mortality showed no lethal dose (LD₅₀ > 500 ppm).     

Persistence of hepatitis B antigen in Culex quinquefasciatus (Diptera: Culicidae)

Groups of wild-caught Culex quinquefaciatus Say, previously tested for the presence of hepatitis B surface antigen (HBsAg), were tested for the presence of hepatitis B e antigen (HBeAg). This antigen was detected at low levels in blood-fed, half-gravid, and gravid groups. A colony of Cx. quinquefasciatus was established in the laboratory and tested for the persistence of HBsAg and HBeAg. Five days after feeding on blood infected with HBsAg and HBeAg, 9 of 20 (45%) pools of Cx. quinquefasciatus were HBeAg-positive and 5 of 20 (25%) pools were HBeAg-positive; low levels of HBsAg and HBeAg were still detectable 28 d after the infective meal in 2 of 20 (10%) and 1 of 20 (5%) pools, respectively. A crude protease extract was prepared from colony mosquitoes, and the effect of this extract on HBsAg and HBeAg present in human serum was tested in vitro. After 20 h, tests for both antigens were still strongly positive. Low levels of HBsAg were detected in ovaries 7 d after infection. Salivary glands were HBsAg- and HBeAg-negative.     

Mosquito larvicidal and pupicidal efficacy of Solanum xanthocarpum (Family: Solanaceae) leaf extract and bacterial insecticide, Bacillus thuringiensis, against Culex quinquefasciatus Say (Diptera: Culicidae)

The bio-efficacy of Solanum xanthocarpum leaf extract and bacterial insecticide, Bacillus thuringiensis, were assessed against the first to fourth instar larvae and pupae of Culex quinquefasciatus, under the laboratory conditions. The medicinal plants were collected from the outskirt Bharathiar University, Coimbatore, Tamil Nadu, India. The shade dried plant materials were extracted by employing the Soxhlet apparatus with ethanol (organic solvent) for 8 h and filtered. The extracts were concentrated at reduced temperature on a rotary evaporator and stored at a temperature of 4°C. Both S. xanthocarpum and B. thuringiensis show varied degree of larvicidal and pupicidal activity against various stages of C. quinquefasciatus. The LC(50) and LC(90) of S. xanthocarpum against the first to fourth instar larvae and pupae were 155.29, 198.32, 271.12, 377.44, and 448.41 ppm and 687.14, 913.10, 1,011.89, 1,058.85, and 1,141.65 ppm, respectively. On the other hand, the LC(50) values of B. thuringiensis against the first to fourth instar larvae and pupae were 133.88, 157.14, 179.44, 206.80, and 240.74 ppm; the LC(90) values were 321.04, 346.89, 388.86, 430.95, and 492.70 ppm, respectively. However, the combined treatment of S. xanthocarpum?+?B. thuringiensis (1:2) material shows highest larvicidal and pupicidal activity of the LC(50) values 126.81, 137.62, 169.14, 238.27, and 316.02 ppm and the LC(90) values 476.36, 613.49, 705.29, 887.85, and 1,041.73 ppm against C. quinquefasciatus in all the tested concentrations than the individuals and clearly established that there is a substantial amount of synergist act. Therefore, the present investigation clearly exhibit that both S. xanthocarpum and B. thuringiensis materials could serve as a potential of highest mortality rate against the mosquito larvae laboratory as well as the field conditions. Since C. quinquefasciatus is a ditch breeder vector mosquito, this is a user and eco-friendly biopesticide for the control of mosquito vector management program.     

Dispersal of Culex quinquefasciatus (Diptera: Culicidae) in a Hawaiian rain forest

Introduced mosquito-borne pathogens avian malaria (Plasmodium relictum Grassi and Feletti) and avian pox virus (Avipoxvirus) have been implicated in the past extinctions and declines of Hawaiian avifauna and remain significant obstacles to the recovery and restoration of endemic Hawaiian birds. Effective management of avian disease will require extensive mosquito control efforts that are guided by the local ecology of the vector Culex quinquefasciatus Say (Diptera: Culicidae). During October and November 1997 and September through November 1998 five mark-release-recapture experiments with laboratory-reared Cx. quinquefasciatus were conducted in a native rain forest on Hawaii Island. Of the overall 66,047 fluorescent dye-marked and released females, 1,192 (1.8%) were recaptured in 43-52 CO2-baited traps operated for 10-12-d trapping periods. Recaptured mosquitoes were trapped in all directions and at distances up to 3 km from the release site. The cumulative mean distance traveled (MDTs) over the trapping period ranged from a high of 1.89 km after 11 d (September 1998) to a low of 0.81 km after 11 d (November 1998). Released mosquitoes moved predominately in a downwind direction and they seemed to use forestry roads as dispersal corridors. Applying an estimated MDT of 1.6 km to a geographical information system-generated map of the Hakalau Forest National Wildlife Refuge clearly demonstrated that the effective refuge area could be reduced 60% by mosquitoes infiltrating into managed refuge lands. These findings should have significant implications for the design of future refuges and development of effective mosquito-borne avian disease control strategies.     

Larvicidal activity of some Euphorbiaceae plant extracts against Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae)

Larvicidal activity of ethyl acetate, butanol, and petroleum ether extracts of five species of Euphorbiaceae plants, Jatropha curcas, Pedilanthus tithymaloides, Phyllanthus amarus, Euphorbia hirta, and Euphorbia tirucalli, were tested against the early fourth instar larvae of Aedes aegypti L. and Culex quinquefasciatus (Say). The larval mortality was observed after 24 h of exposure. All extracts showed low larvicidal effects; however, the highest larval mortality was found in petroleum ether extract. The LC₅₀ value of petroleum ether extracts of J. curcas, P. tithymaloides, P. amarus, E. hirta, and E. tirucalli were 8.79, 55.26, 90.92, 272.36, and 4.25 ppm, respectively, against A. aegypti and 11.34, 76.61, 113.40, 424.94, and 5.52 ppm, respectively, against C quinquefasciatus. Of the various ratios tested, the petroleum ether extracts of J. curcas and E. tirucalli were observed to be more efficient than the other plant extracts. It is, therefore, suggested that E. tirucalli can be applied as an ideal potential larvicide against A. aegypti and C. quinquefasciatus. This is an ideal ecofriendly approach for the control of the dengue vector, A. aegypti, and the lymphatic filariasis vector, C. quinquefasciatus.     

Temporal and geographic genetic variation in Culex pipiens quinquefasciatus (Diptera: Culicidae) from Florida

Culex (Culex) pipiens quinquefasciatus Say field population from Vero Beach, FL, sampled monthly over a period of 8 mo, a colony sample, and six geographic samples were analyzed for genetic variation at 12 enzymes (10 "neutral" gene enzymes with 11 putative loci and two "complex" gene enzymes) by using polyacrylamide gel electrophoresis. The analysis of the 11 putative loci in both temporal and geographic samples showed that the four loci (Gpi, Hk, Mdhp-2, and Pgm) diagnostic of Cx. p. quinquefasciatus in the southern United States are present in similar frequencies in Florida samples. The Cx. p. quinquefasciatus colony sample showed significantly lower genetic variation than the temporal field samples, measured by mean number of alleles per locus (colony 1.2 +/- 0.1 versus field 1.44 +/- 0.03), percentage of polymorphic loci (colony 18.2% versus field 28.4%), mean observed heterozygosity (H(o) = colony 0.027 +/- 0.02 versus field 0.09 +/- 0.01), and mean Hardy-Weinberg expected heterozygosity (H(e) = colony 0.025 +/- 0.02 versus field 0.085 +/- 0.01). Three of the 11 loci (Acoh, Pgd, and Pgm) from the Vero Beach field samples showed bimodal patterns in their frequencies of the most common allele during peak density of the population. The low value of F(st) of 0.058 indicated minimum population substructuring among the temporal samples. Genetic variability values between geographic samples from the Florida panhandle and south Florida were not significant. Gene flow estimates based on F(ST), = 0.05, indicating low levels of gene flow among the geographic samples of Cx. p. quinquefasciatus. The average Nei's and modified Rogers' genetic distances among the six populations were 0.005 +/- 0.001 and 0.077 +/- 0.007, respectively. The cluster analysis did not suggest geographic clustering. The analysis of the "complex" gene enzymes in both temporal and geographic samples of Cx. p. quinquefasciatus from Florida showed the presence of two highly amplified esterases (Estbeta1 and Estalpha2\Estbeta2), indicating resistance to organophosphate insecticides and highly amplified Aldox enzyme (an enzyme that indicates resistance to at least one insecticide and a herbicide). Comparison of our results with previous studies on Cx. p. quinquefasciatus populations in the United States indicates that the genetic characteristics of the Florida populations of Cx. p. quinquefasciatus are very similar to populations from areas where ecological conditions are very different.     

Mosquitocidal properties of nereistoxin against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae)

Emergence of resistance among mosquitoes is a recent problem. Safe and eco-friendly agents from biological origins are the need of the hour. Nereistoxin, a naturally occurring substance, was first isolated from the marine annelid Lumbriconereis heteropoda and stored in the freezer. In the present study, the toxicity of nereistoxin was evaluated against vector mosquitoes. The larvicidal, ovicidal and adulticidal activities of nereistoxin were assayed for their toxicity against three important vector mosquitoes, viz., Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae). The nereistoxin was inversely proportional to the concentration and directly proportional to the mosquitoes. The larvicidal activity after 24 h LC₅₀ value was observed at 0.467, 0.535 and 0.601 ppm for A. stephensi, A. aegypti and C. quinquefasciatus, respectively. The ovicidal activity after 120 h zero percentage of egg hatchability was observed at a concentration of 0.8 ppm for A. stephensi and 1.0 ppm for A. aegypti and C. quinquefasciatus. The results of the adulticidal activity after 24 h LD₅₀ value were observed at 0.022, 0.028 and 0.034 ppm for A. stephensi, A. aegypti and C. quinquefasciatus, respectively. The extracted nereistoxin was characterized and identified by ultraviolet, infrared, nuclear magnetic resonance, mass spectroscopic methods and high pressure liquid chromatography. These results clearly reveal that the nereistoxin served as a potential larvicidal, ovicidal and adulticidal activity against vector mosquitoes.     

Feeding patterns of Culex quinquefasciatus (Diptera: Culicidae) from eastern Santa Catarina state, Brazil

Blood-feeding sources of Culex quinquefasciatus Say (Diptera: Culicidae) collected in the eastern region of the Brazilian state of Santa Catarina were analyzed by precipitin technique. One hundred fifty-four female mosquitoes collected by CDC traps in the Navegantes municipality 13-15 February 2005 reacted to one or more of eight antisera, including chicken, dog, goat, sheep, horse, opossum, human and rodent antisera. One hundred thirty-seven specimens (89%) reacted to only one source, and 17 (11%) specimens reacted to two sources. Among the 137 specimens reacting to only one source, reactions to rodent (50.4%), sheep (5.8%), chicken (5.1%), goat (5.1%), dog (2.2%), horse (3.6%), and human (3.6%) antisera were observed. The analyzed species demonstrated a high degree of opportunistic feeding behavior in relation to host preference. Results are compared with results from similar studies, and the low proportion of reactions to human antisera is discussed.