Various levels of cross-resistance to Bacillus sphaericus strains in Culex pipiens (Diptera: Culicidae) colonies resistant to B. sphaericus strain 2362.

We studied the cross-resistance to three highly toxic Bacillus sphaericus strains, IAB-59 (serotype H6), IAB-881 (serotype H3), and IAB-872 (serotype H48), of four colonies of the Culex pipiens complex resistant to B. sphaericus 2362 and 1593, both of which are serotype H5a5b strains. Two field-selected highly resistant colonies originating from India (KOCHI, 17,000-fold resistance) and France (SPHAE, 23,000-fold resistance) and a highly resistant laboratory-selected colony from California (GeoR, 36,000-fold resistance) showed strong cross-resistance to strains IAB-881 and IAB-872 but significantly weaker cross-resistance to IAB-59 (3- to 43-fold resistance). In contrast, a laboratory-selected California colony with low-level resistance (JRMM-R, 5-fold resistance) displayed similar levels of resistance (5- to 10-fold) to all of the B. sphaericus strains tested. Thus, among the mosquitocidal strains of B. sphaericus we identified a strain, IAB-59, which was toxic to several Culex colonies that were highly resistant to commercial strains 2362 and 1593. Our analysis also indicated that strain IAB-59 may possess other larvicidal factors. These results could have important implications for the development of resistance management strategies for area-wide mosquito control programs based on the use of B. sphaericus preparations.

Effects of nutritional factors and soil addition on growth, longevity and fecundity of the tadpole shrimp Triops newberryi (Notostraca: Triopsidae), a potential biological control agent of immature mosquitoes.

The notostracan tadpole shrimp (TPS) Triops newberryi Packard has potential to be used as a biocontrol agent of immature mosquitoes. Eggs, nymphal or adult shrimps are considered to be the stages for field introduction. To yield good growth of the shrimp and high production of shrimp eggs under artificial conditions, nutritional requirements of TPS for growth, survival and fecundity need to be elucidated. In the laboratory, we evaluated various nutritional and edaphic regimens, such as soil alone, mosquito larvae or rabbit pellets alone and various combinations of these three components for culturing. These factors influenced the growth, longevity and egg production profoundly. It was shown that the simulated natural conditions, i.e. full combination of all three factors, yielded the largest TPS with longest survival and highest egg production, followed by the combinations of any two components. Any single component, soil, mosquito larvae, or rabbit pellets, did not result in good growth, survival and egg production. By formulating optimal rearing substrates, this species of TPS will yield large numbers of all stages for experimentation and field introductions. Under optimal conditions, they mature in 7-8 days and survive for about one month. Each TPS is capable of producing up to 1,000 eggs during its lifetime. These studies developed nutritional regimens for TPS mass culturing procedures, where the eggs, nymphal and adult TPS can be mass cultured for field introduction and stocking in mosquito developmental sites.

Effects of temperature on development, mortality, mating and blood feeding behavior of Culiseta incidens (Diptera: Culicidae).

Culiseta incidens Thomson is distributed over most of the western USA and Canada northward to Alaska. Because this mosquito is difficult to colonize, its biology has not been well investigated. We colonized this species in 1998 and studied the effects of temperature on various aspects of its life cycle. The time required for egg melanization and the duration of the egg stage were negatively correlated with temperature. The proportion of fertile egg rafts was temperature-independent. An inverse relationship existed between temperature and egg hatch. Molting and stadium duration after hatching were temperature-dependent, with higher temperature accelerating development and molting. Larvae and pupae experienced lower mortality and higher molting success at lower temperatures. Survivorship of adult mosquitoes fed on sugar solution was inversely proportional to temperature, lethal times for 50% mortality (LT50) were greater at the lower temperature than at the higher temperature. Females survived longer than did males at all test temperatures. Because this species is eurygamous, mating only occurred in large cages. Mating success was also affected by temperature. At the test temperatures, 20 degrees C, 25 degrees C and 30 degrees C, mating started from 3-5 days after emergence and reached a peak on days 13-15 after emergence. Maximum mating rates at 20 degrees C and 25 degrees C were higher than at 30 degrees C. Blood feeding, as indicated by cumulative feeding rates, was affected by cage size, mosquito age and temperature. Mosquitoes in large cages exhibited a much higher feeding rate than in small cages. With age, the cumulative blood feeding rate increased, with the highest rate at 25 degrees C, followed by 20 degrees C and 30 degrees C. At all temperatures tested, most of the blood fed females were mated.

Mosquito larval control with Bacillus sphaericus: reduction in adult populations in low-income communities in Nonthaburi Province, Thailand.

During 1999 and 2000 several larvicidal treatments of Bacillus sphaericus strain 2362 water dispersible granular (WDG) formulations were made at 50 to 200 mg/m2 in mosquito developmental sites in low-income communities in Nonthaburi Province, Thailand to determine whether larviciding dense populations would results in a noticeable reduction of adult mosquitoes in small treated areas. In the treated area in 1999 (Soi Jumpa), immature populations were suppressed to extremely low levels for extended periods, especially at the higher dosages. This decline in immature populations was followed by a substantial decline in adult mosquitoes. There was a lag of 7 to 14 days post-larval treatments before maximum decline in adults was noted. Adults that emerged prior to treatments survived for 7-14 days or longer, thus no drastic reduction was noted soon after treatments. Despite a slight resurgence in adult mosquitoes during the middle of the experimental period, adult female mosquitoes (over 98% Cx quinquefasciatus), remained low during the 5-month period of trials. During the last 2 weeks (17 days post last treatment) of the experimental period, female populations reached the pre-treatment level. During the 2000 tests at Wat Pikul reduction in larvae was 87-98% for 7 weeks after first treatment at 200 mg/m2, resulting in a reduction of 24 to 73% (2 and 7 days post-treatment respectively) and 87 to 98 (2-6 weeks) in the adults. In the second and third treatments at 50 mg/m2, larval control and subsequent adult reduction were lower and shorter-lived than at the high dosage, and the fourth treatment at 100 mg/m2 did not yield a high level of reduction in the larvae (18 to 33%), but reduction of adults was still 80%. The final fifth treatment at 200 mg/m2 yielded only 18% control of larvae, suggesting tolerance to B. sphaericus at this site. It was shown that at both treated sites repeated treatments with a larvicide such as B. sphaericus could result in substantial reduction in adult mosquitoes. Vigilance for detection of resistance development should be practiced, as resistance could emerge in certain populations following a few treatments.

Mosquito burden and impact on the poor: measures and costs for personal protection in some communities in Thailand.

To gauge the extent of mosquito problems and their impact on local people in Thailand, a simple questionnaire was designed consisting of 6 questions with multiple choices to be answered in 4 different communities in Thailand in 1998 and 1999. Mosquito biting activity was noted often by respondents. They reported that mosquitoes bit both night and day, and that the insects were abundant both in the dry and the rainy seasons. In all 4 communities, a large proportion of the residents used bed nets, mosquito coils, and aerosol sprays for personal protection; vaporizing mats and repellents were used sparingly. The cost of such measures amounted to dollars 4 to dollars 25 per year per household. For most of the residents, this represented a substantial proportion of their income, and was proportionally greater than the average cost of organized mosquito control in developed countries. This suggests that instituting organized local vector control programs would be cheaper and more effective than the individual use of personal protectants that do not reduce mosquito numbers. An assessment of the available products stocked in neighborhood stores and supermarkets for personal protection was made. A variety of insecticidal aerosols, mosquito coils, liquid sprays, vaporizing mats, and vaporizing liquids was stocked. This ample supply of household insecticides lends support to the preferred methods of protection reported by the respondents. The active ingredients in most of the formulations were synthetic pyrethroids, although a few contained dichlorvos, propoxur, and a few other compounds. Mosquito coils, the most preferred products used by the poor, were evaluated for efficacy, and were found to provide a reduction of 72-96% in landing-biting rates in controlled experiments.

Field evaluation of new water-dispersible granular formulations of Bacillus thuringiensis ssp. israelensis and Bacillus sphaericus against Culex mosquitoes in microcosms.

A variety of formulations of Bacillus thuringiensis var. israelensis de Barjac (B.t.i.) and Bacillus sphaericus Neide (B.s.) have been studied for mosquito control under laboratory and field conditions. High efficacy, specificity, low risk of development of resistance, long shelf-life, and transportability, as well as the safety to nontarget organisms of these 2 microbial agents have been well documented. Some of the currently available formulations of B.t.i. and B.s. have low potency per unit mass. Research and development efforts are focusing on commercializing formulations with high potency and low minimum effective dosage that are suitable for long-distance shipment. To achieve this goal, new water-dispersible granule (WDG) formulations of both microbial agents were prepared and made available by Abbott Laboratories for evaluation. The newly developed WDGs of B.t.i. and B.s. with high potency dispersed readily in water with gentle agitation. These WDGs were evaluated and the minimum effective dosages were determined in microcosms against natural populations of Culex mosquitoes. The minimum effective dosage for B.t.i. WDGs with 4,000 International Toxic Units (ITU)/mg was 0.27-0.53 lb/acre which yielded significant control for up to 7-12 days. The minimum effective dosage for B.s. WDGs with 350-630 ITU/mg was 0.05-0.10 lb/acre, which yielded significant control of immature mosquitoes for up to 14-20 days.

Efficacy of new formulations of the microbial larvicide Bacillus sphaericus against polluted water mosquitoes in Thailand.

Two new water dispersible granular (WDG) formulations of the microbial control agent Bacillus sphaericus (strain 2362) were extensively evaluated in polluted waters against Culex quinquefasciatus in Thailand. The studies were carried out in stagnant as well as flowing waters during August 1997 to July 1998. The trial period covered both rainy and dry seasons. The two WDG formulations of B. sphaericus tested were low potency (350 ITU/mg) and high potency (630 ITU/mg) products. Both formulations were used at various rates to determine initial efficacy and longevity. The high potency formulations provided excellent control (80 to 90%) of immature mosquitoes at the rates of 50 to 100 mg/ m2, while the less potent formulation yielded similar control at the rates of 89 to 250 mg/m2. Longevity of control was anywhere from one week to four weeks or longer depending on the dosage, habitat, and environmental conditions. Two treatments with low dosages of Bacillus thuringiensis ssp. israelensis WDG provided short-term control lasting for one week. Frequent episodes of heavy rains facilitated long-term suppression of immature mosquitoes in klongs by flushing out the larvae. In the absence of rain, the longevity of treatments in flowing waters was shorter than in the presence of rain. Rain, however, had some but not pronounced effect on longevity in the stagnant water habitats. Operation of floodgates controlling water flow from stagnant water habitats into the Chaophrya River had greater influence on the abundance of mosquito immatures in the ponded polluted waters under dwellings. Precipitation, adding large amounts of water, was probably responsible for diluting the control agents and also resulted in lower counts of immatures per unit volume of water. These variables could influence the efficacy (especially longevity) of treatments employing formulations of microbial agents in operational control programs. The WDG formulations are preferred over other formulations, such as granules, because the former contain larger quantities of toxins per unit mass than the latter and are easily applied as aqueous sprays. Potent formulations with greater content of active ingredients are less costly to ship and transport to distant areas.

Activity and biological effects of neem products against arthropods of medical and veterinary importance.

Botanical insecticides are relatively safe and degradable, and are readily available sources of biopesticides. The most prominent phytochemical pesticides in recent years are those derived from neem trees, which have been studied extensively in the fields of entomology and phytochemistry, and have uses for medicinal and cosmetic purposes. The neem products have been obtained from several species of neem trees in the family Meliaceae. Six species in this family have been the subject of botanical pesticide research. They are Azadirachta indica A. Juss, Azadirachta excelsa Jack, Azadirachta siamens Valeton, Melia azedarach L., Melia toosendan Sieb. and Zucc., and Melia volkensii Gürke. The Meliaceae, especially A. indica (Indian neem tree), contains at least 35 biologically active principles. Azadirachtin is the predominant insecticidal active ingredient in the seed, leaves, and other parts of the neem tree. Azadirachtin and other compounds in neem products exhibit various modes of action against insects such as antifeedancy, growth regulation, fecundity suppression and sterilization, oviposition repellency or attractancy, changes in biological fitness, and blocking development of vector-borne pathogens. Some of these bioactivity parameters of neem products have been investigated at least in some species of insects of medical and veterinary importance, such as mosquitoes, flies, triatomines, cockroaches, fleas, lice, and others. Here we review, synthesize, and analyze published information on the activity, modes of action, and other biological effects of neem products against arthropods of medical and veterinary importance. The amount of information on the activity, use, and application of neem products for the control of disease vectors and human and animal pests is limited. Additional research is needed to determine the potential usefulness of neem products in vector control programs.

Mosquito control and bacterial flora in water enriched with organic matter and treated with Bacillus thuringiensis subsp. israelensis and Bacillus sphaericus formulations.

Three tests were conducted during July 17 to October 30, 1998 to study the impact of two mosquitocidal microbial agents on mosquito larvae and their contribution to bacterial flora in aquatic microcosms. Formulations of Bacillus thuringiensis subsp. israelensis (Bti) and Bacillus sphaericus strain 2362 (Bsph) were applied at various rates to outdoor tubs enriched with rabbit pellets and filled with irrigation water from a reservoir. Mosquito larvae were effectively controlled by all treatments; the magnitude of initial and persistent control depended on materials and dosages applied. Bacterial flora were assessed in the irrigation water as well as water in the enriched tubs before and after treatment with the microbial agents. The irrigation water contained 800-1000 total bacterial cells/ml. The populations of total bacteria and spore formers peaked on day 3 after enriching and filling the tubs, then declined progressively to the low levels at the end of the tests. After treatment, the numbers of Bti and Bsph spores in treated tubs prevailed at a dosage-dependent manner, their populations peaked at three hours after treatment, and declined progressively thereafter. The contribution of Bti and Bsph spores to the total bacterial flora was negligible but significant to the counts of spore-forming bacteria. The gram-negative bacteria made up more than 80% of the total bacterial flora during the test periods; and, of these, gram-negative rods constituted the greatest proportion, gradually increasing from the time of flooding to the end of the tests. Gram-negative cocci also occurred in relatively great proportion, but showed a reverse trend as compared with the gram-negative rods, declining gradually from pretreatment to the end of the tests. Gram-positive rods (spore formers), including Bti and Bsph, occurred in low numbers in all the tests but increased slightly in treated tubs due to the addition of Bti and Bsph spores. Gram-positive cocci occurred occasionally in some water samples.

Bacteria and mosquito abundance in microcosms enriched with organic matter and treated with a Bacillus thuringiensis subsp. israelensis formulation.

Bacteria and mosquito abundance were studied in outdoor tubs unenriched and enriched with 0.04% rabbit pellets during the winter, 1999. The irrigation water used to fill the tubs contained a total bacterial count of 1.15-1.35 x 10(3) cells/ml. Adding rabbit pellets for enrichment yielded a total bacterial count of 5.50-7.63 x 10(5) cells/g. Bacterial densities in unenriched water were significantly lower than in enriched tubs on every sampling day. When bacterial densities in both enriched and unenriched regimens reached peak populations on day 3 post-flooding, their numbers in enriched water were 25-fold higher than the unenriched water. Under cool weather conditions, mosquito oviposition activity was low and larval development was very slow. Egg raft counts and larval densities in enriched water were nevertheless higher than those in unenriched water. After reaching peak populations on day 3 post-flooding, the natural decline in bacterial densities in the top portion of enriched water without mosquito larvae was lower compared with that in water with larvae. In water with larval present, the decline of bacterial levels in top water was greater than in bottom water on day 7 post-flooding. VectoBac G, a granular formulation of Bacillus thuringiensis subsp. israelensis caused a reduction in larval numbers of 80, 93, 73% at the rate of 5.5 lb/ac and a reduction of 94, 93, 86% at the rate of 10.6 lb/ac on days, 1, 3, 7 posttreatment, respectively. After treatment, the reductions of bacterial densities in untreated tubs were greater than treated tubs. These results indicate that mosquito larvae play an important role in the decline of bacterial populations by their feeding activity.

Effects of neem products containing azadirachtin on blood feeding, fecundity, and survivorship of Culex tarsalis and Culex quinquefasciatus (Diptera: Culicidae).

When late 3rd or early 4th-instar larvae of Culex tarsalis Coquillett and Culex quinquefasciatus Say mosquitoes were treated with sublethal dosages of neem insecticide until pupation, the blood-feeding activity of the resulting adults was essentially the same as that of untreated controls. In contrast, blood-feeding activity was suppressed when newly emerged adults were fed continuously on 10 parts per million (ppm) or 50 ppm azadirachtin (AZ) in 10% sucrose solution for seven days. Fecundity was also reduced by the various neem treatments. When late 3rd or early 4th-instar larvae were treated with 0.010 ppm AZ to pupation, the resultant females had a lower rate of oviposition than did the untreated controls after a full blood meal. When late instar larvae were treated at 0.005 ppm and 0.010 ppm AZ, the resultant females produced smaller egg rafts after a full blood meal, as compared to the controls, but egg viability was not affected. In newly emerged adults feeding continuously on 10 ppm and 50 ppm AZ in 10% sucrose for seven days (before blood feeding), the oviposition rate, size of egg raft, and hatching rate of the eggs after a full blood meal were all reduced. When newly blood-fed adults were fed continuously on 10 ppm and 50 ppm AZ in 10% sucrose for five days, their oviposition rate was lower than controls in most cases, but the egg raft size and viability of eggs were not affected. In freshly blood-fed females topically treated with AZ with 1 or 5 micrograms/female, the oviposition rate and size of egg rafts were generally reduced. The females receiving topical treatment laid eggs and their hatching was not affected. The longevity of adult females feeding continuously on 10 ppm and 50 ppm AZ in 10% sucrose solution after emergence was reduced, whereas, the longevity of males was only affected at the higher concentration.

Field evaluation of controlled release pellet formulation of methoprene against chironomid midges in man-made lakes.

The duration of the efficacy of 4% methoprene pellets against emergence of chironomid midges, Chironomus stigmaterus Say, Goeldichironomus amazonicus (Fittkau) and Tanypus imperialis Sublette, in man-made lakes supplied with tertiary effluent water in Palm Desert, California, was dose dependent. For C. stigmaterus and T. imperialis, the rate of 4.5 kg/ha of the controlled release pellet formulation of 4% methoprene yielded > 90% control for 3 wk and 75% control in the 4th week, the rate of 3.4 kg/ha provided > 90% control for 2 wk, and the rate of 2.25 kg/ha provided control for 1 wk. Goeldichironomus amazonicus was under 90% control for 2 wk at the rate of 4.5 kg/ha and 1 wk at the rates of 3.4 kg/ha and 2.25 kg/ha.

Ovicidal activity of neem products (azadirachtin) against Culex tarsalis and Culex quinquefasciatus (Diptera: Culicidae).

Bioactive compounds contained in the seed kernel and other parts of the neem tree (Azadirachta indica A. Juss) have been found to show insecticidal activities and other effects in many species of insects. These activities include antifeedancy, growth regulation, fecundity suppression, male sterility, oviposition repellency, changes in biological fitness such as loss of flying ability, immunodepression, enzyme inhibition, splitting of biological rhythms, and so forth. We investigated the ovicidal effects of various formulations of azadrirachtin (AZ) against the mosquitoes Culex tarsalis Coquillett and Culex quinquefasciatus Say. The formulations tested were wettable powder Azad WP10, emulsifiable concentrate Azad EC4.5, and technically pure AZ. The ovicidal activity of the test neem products was influenced by concentration of AZ, age of the egg rafts, and age of the neem preparations. Other factors such as formulation and mosquito species were also involved in the degree of ovicidal activity. When the egg rafts were deposited directly in fresh neem suspension and left there for 4 h before transfer to untreated water, 1 ppm of AZ produced almost 100% mortality in eggs. When egg rafts aged for 0, 4, 8, 12, and 24 h were exposed to 10 ppm neem suspensions for 36 h, the ovicidal activity was only attained in the egg rafts deposited directly (0 h old) in the neem suspension, not in those with ages of 4-24 h. On aging, depending on the formulations and mosquito species, the neem suspensions at 1 ppm completely lost ovicidal activity within 7-20 days. The egg rafts of Cx. quinquefasciatus were more susceptible to the test neem products than those of Cx. tarsalis. The formulated neem products were more persistent and effective than the technical AZ. The wettable powder (WP) formulation was slightly more persistent and effective than the emulsifiable concentrate (EC). The ovicidal activity of the neem products against mosquitoes from the current research clearly demonstrated the potential of neem products as possible ovicides against Culex mosquitoes.

Antifeedancy of neem products containing Azadirachtin against Culex tarsalis and Culex quinquefasciatus (Diptera: Culicidae).

In order to develop and utilize new botanical insecticides originating from neem (Azadirachta indica A. Juss) in mosquito control programs as a potential larvicide, the antifeedant activity of three formulations of neem against the larvae of Culex tarsalis Coquillett and Culex quinquefasciatus Say was investigated under laboratory conditions. A significant antifeedancy was indicated at 5 ppm and 10 ppm azadirachtin (AZ) for all formulations and both species. Within the test concentration range of AZ (1-10 ppm), 5 ppm was the minimum effective concentration for antifeedancy in most cases. Some differences in larval susceptibility in terms of antifeedancy to the test formulations were noted between the two species. The Cx. tarsalis larvae were more susceptible than Cx. quinquefasciatus larvae to Azad EC 4.5 at 1 ppm during the first 15-min exposure period and at 5 and 10 ppm during the whole test period. The Cx. quinquefasciatus larvae were more susceptible than Cx. tarsalis larvae to Azatin WP 4.5 at 1, 5, and 10 ppm during the first 15-min exposure period or to Neemix EC 4.5 at 1 ppm during the first 45-min exposure period. The formulation-related differences in antifeedant activity appeared when the concentration increased from 1 to 10 ppm. In Cx. tarsalis, the Azad EC 4.5 and Neemix EC 4.5 were more effective than Azatin WP 4.5 at 5 ppm during the first 15-min exposure period and at 10 ppm during the whole test period. In Cx. quinquefasciatus, the Azatin WP 4.5 and Neemix EC 4.5 were more effective than Azad EC 4.5 at 1 ppm during the first 30-min exposure period, and at 5 ppm during the first 60-min exposure period, and at 10 ppm during the whole test period.

Organic enrichment of breeding sources for sustained productivity of mosquitoes (Diptera: Culicidae).

Alfalfa pellets and chicken lay mash were used to obtain polluted water for continuous and sustained oviposition by Culex mosquitoes. Water in fiberglass tubs enriched with 0.1 and 0.25 percent (w/v) of either material received heavy oviposition by gravid mosquitoes. The higher level of enrichment resulted in greater oviposition than lower concentrations which in turn received significantly more oviposition than the controls with no enrichment. Oviposition continued in the enriched tubs for more than 43 days where Culex quinquefasciatus and Culex stigmatosoma constituted a major proportion of the eggs laid while Culex tarsalis, as expected, oviposited at much lower rates than the other two species. Supplemental enrichment of mosquito breeding sources with organic material can thus provide uniform and sustained availability of some mosquito larvae for the evaluation of persistent control agents and other related studies.

Selection-dependent trends of autogeny and blood feeding in an autogenous strain of Culex tarsalis (Diptera:Culicidae).

During successive en masse selection of autogeny in Culex tarsalis from parental to 20th generation, the autogenous oviposition capacity changed as follows: mean number of egg rafts/female and mean number of eggs/female increased significantly, from 0.11 to 0.56 and from 6.2 to 20.2, respectively; the mean size of the egg rafts decreased significantly, from 56.2 to 36 eggs/raft; autogeny rates increased significantly, from 18.6 to 65.2%; mean number of autogenous follicles/female decreased significantly, from 57.4 to 34; the hatching rate of autogenous egg rafts reduced significantly, from 88.3 to 75.4%; and the feeding rates of the females on a blood meal source during the 7th day postemergence increased significantly, from 62.8 to 71.1%. The autogeny rate determined by ovarian dissection was higher than that indicated by actual oviposition. As the age of females increased during the observed 10-day oviposition period, the mean number of egg rafts/female, the mean number of eggs/female, and the mean number of eggs/raft decreased successively, and scattered "light color" egg rafts became more common. Successive en masse selection enhanced autogeny and strengthened the bloodfeeding tendency in this facultative autogenous species.

Biological fitness of Culex quinquefasciatus (Diptera:Culicidae) susceptible and resistant to Bacillus sphaericus.

Biological fitness of laboratory and field-collected strains of southern house mosquito, Culex quinquefasciatus Say, susceptible and resistant (37- and 31-fold upon selection) to the microbial agent, Bacillus sphaericus, were compared in the absence of B. sphaericus. The resistant strains showed significantly lower fecundity and fertility, but they had significantly higher survival rates than the susceptible strains. The preadult stages from females of resistant strains developed at slightly faster rates than those of the susceptible strains, which could result in a shorter generation time. However, lower fecundity was likely to lead to overall lower population growth rates than in the susceptible strains. Data provided evidence that the resistant strains exhibited fitness disadvantages in the absence of B. sphaericus. We suggest that once resistance to B. sphaericus is detected in the field, its use should be discontinued until the mosquito population becomes susceptible again because of the decline in number of resistant individuals. A strategy of resistance management by rotation of insecticides is discussed.

Nutritional reserves, body weight, and starvation tolerance of autogenous and anautogenous strains of Culex tarsalis (Diptera:Culicidae).

The nutritional reserves and body weight of autogenous and anautogenous strains of Culex tarsalis Coquillett were determined for 4th instars, pupae, females, and males. Starvation tolerance and survivorship of adult females and males also were studied. The autogenous individuals contained significantly greater amounts of total lipids (except the total lipids in 4th instars), total carbohydrates, glycogen, and total proteins, which resulted in a heavier mean body weight of autogenous than anautogenous mosquitoes (except the fresh weight in pupae). Results of the body fluid coefficient determination were inconclusive. Nutritional reserves in the immature stages, especially the pupal stage, were significantly greater (except the total proteins in 4th instars and female adults) than those in the adult stage. There were no significant differences in median longevity between autogenous and anautogenous females, and between autogenous and anautogenous males, provided with distilled water alone after emergence. The median longevity of females and males was significantly different in both autogenous and anautogenous strains. Our findings strongly indicate a relationship between autogenous reproduction and differences in nutritional reserves of autogenous and anautogenous strains.

Field trials with Bacillus sphaericus formulations against polluted water mosquitoes in a suburban area of Bangkok, Thailand.

Two newly developed Bacillus sphaericus larvicidal formulations, VectoLex CG (corncob granules) and VectoLex WDG (water dispersible granules), were tested against Culex quinquefasciatus larvae in 4 highly polluted breeding sites in Thailand. VectoLex CG, applied at rates of 0.5-2 g/m2, gave satisfactory to complete control of late-instar larvae and pupae for up to 4 wk after treatment. The VectoLex WDG, which had higher potency and was applied at rates of 0.1-0.5 g/m2, gave satisfactory control for 1-4 wk after treatment. Among the factors influencing longevity of control were dosage of a given formulation, precipitation, and flooding of the treated sites; the latter had the greatest impact. Presence of larvivorous fish did not seem to influence larval populations because there were heavy populations of mosquito larvae present in the test sites in the presence of moderate numbers of fish before the application of B. sphaericus treatments.

Physiological aspects of autogeny in Culex tarsalis (Diptera: Culicidae): influences of sugar-feeding, mating, body weight, and wing length.

Sugar feeding and mating promoted the expression of autogeny in Culex tarsalis. In sugar-fed cohorts fed 5, 10, and 20% sucrose solution, the mean number of egg rafts/female, mean number of eggs/raft, mean number of eggs/female, mean number of autogenous follicles/female, and the autogeny rate were all significantly greater than those in sugar-deprived cohorts. There were no significant differences in these parameters among the sugar-fed groups of 5, 10, and 20%. Similarly, magnitudes of the parameters mentioned above in female-male cohorts were significantly higher than those in female-alone cohorts. There was no difference in the hatching rate of the autogenous egg rafts in the cohorts fed on different sucrose solutions and the female-male cohorts, while it was 0 in male-deprived cohorts. There were positive correlations between mean emergence weight and mean number of autogenous eggs/female (r2 = 0.69), mean emergence weight and mean number of autogenous follicles/female (r2 = 0.75), mean wing length and mean number of autogenous eggs/female (r2 = 0.64), and mean wing length and mean number of autogenous follicles/female (r2 = 0.63). The regression analysis of the data indicated that females with emergence weights < 1.92 mg could not oviposit autogenously, and those with emergence weights < 1.89 mg could not develop the ovaries autogenously. Females with wing lengths < 2.89 mm could not oviposit autogenously and those with the wing length of < 2.83 mm could not develop the ovaries autogenously. The coefficient of correlation between emergence weight and autogenous follicles/female was greater than that between emerging weight and autogenous eggs/female. The coefficients of correlation between emergence weight and autogeny were greater than that between wing length and autogeny. Therefore, autogenous follicles/female is a more accurate parameter to describe autogeny expression level than autogenous eggs/female because oviposition is affected by other factors, and emergence weight is a more important factor determining autogeny expression than wing length.