Bioinsecticide and leaf litter combination increases oviposition and reduces adult recruitment to create an effective ovitrap for Culex mosquitoes

Mosquito egg traps, aquatic habitats baited with oviposition attractant and insecticide, are important tools for surveillance and control efforts in integrated vector management programs. The bioinsecticide Bacillus thuringiensis subsp. israelensis (Bti) is increasingly used as an environmentally friendly alternative to chemical insecticides and the combination of Bti with a simple oviposition attractant like leaf litter to create an effective egg trap seems appealing. However, previous research suggests that Bti may itself alter oviposition, and that leaf litter may dramatically reduce Bti toxicity. Here we present results from field experiment designed to link the effects of litter and Bti on mosquito oviposition habitat selection and post‐colonization survival to production of adult mosquitoes. Tripling litter increased Culex spp. oviposition nearly nine‐fold, while Bti had no effect on oviposition. Neither factor altered egg survival, thus larval abundance reflected the effects of litter on oviposition. Both Bti and litter reduced larval survival by ～60%. We found no evidence that increased litter reduced Bti toxicity. Adult production was dependent upon both litter and Bti. In the absence of Bti, effects of litter on oviposition translated into three‐fold more adults. However, in the presence of Bti, initial increases in oviposition were erased by the combined negative effects of Bti and litter on post‐colonization survival. Thus, our study provides field evidence that combined litter and Bti application creates an effective ovitrap. This combined treatment had the highest oviposition and the lowest survival, and thus removed the greatest number of mosquitoes from the landscape.     

EFFECTS OF ELEVATED ATMOSPHERIC CO(2) ON WATER CHEMISTRY AND MOSQUITO (DIPTERA: CULICIDAE) GROWTH UNDER COMPETITIVE CONDITIONS IN CONTAINER HABITATS

We investigated the direct and indirect effects of elevated atmospheric CO(2) on freshwater container habitats and their larval mosquito occupants. We predicted that a doubling of atmospheric CO(2) would (1) alter the chemical properties of water in this system, (2) slow degradation of leaf litter, and (3) decrease larval growth of Aedes albopictus (Skuse) mosquitoes raised on that litter under competitive conditions. Effects of elevated CO(2) on water quality parameters were not detected, but the presence of leaf litter significantly reduced pH and dissolved oxygen relative to water-filled containers without litter. Degradation rates of oak leaf litter from plants grown under elevated CO(2) atmospheres did not differ from breakdown rates of litter from ambient CO(2) conditions. Litter from plants grown in an elevated CO(2) atmospheres did not influence mosquito population growth, but mosquito production decreased significantly with increasing larval density. Differences among mosquito density treatments influenced survivorship most strongly among male Ae. albopictus and time to emergence most strongly among females, suggesting fundamental sex-determined differences in response to competition. Results of this and other studies indicate that direct and indirect effects of doubled atmospheric CO(2) are minimal in artificial containers with freshwater.     

Influence of molting on efficacy of two functionally different larvicides: Bti and temephos

The sensitivity of the second, third (L3), and fourth (L4) instars of Culex pipiens L. and Ochlerotatus sticticus (Meigen) to two larvicides, temephos and Bacillus thuringiensis variety israelensis (Bti) was tested with and without molting occurring in the experiments. In the first experiment for both tested mosquito species, LC50 values increased from the second to the fourth instar when tested against temephos, whereas for Bti the opposite trend was observed. The highest LC50 value was found for fourth instars of Cx. pipens (0.00405 mg/liter) against temephos and for second instars of Oc. sticticus (0.267 mg/liter) against Bti. The determined LC50 values for the second and third instars of both species decreased with an increased number of molted larvae in the experiments with temephos. For the experiments with Bti molting did not have any significant influence on LC50 values, except a small increase in toxicity during the L3/L4 molt of Oc. sticticus. These findings could help assess and define larviciding, as well as influence the quantity of larvicides needed for an efficient treatment.     

The naturally derived insecticide spinosad is highly toxic to Aedes and Anopheles mosquito larvae

Spinosad is a naturally derived biorational insecticide with an environmentally favourable toxicity profile, so we investigated its potency against mosquito larvae (Diptera: Culicidae). By laboratory bioassays of a suspension concentrate formulation of spinosad (Tracer), the 24 h lethal concentration (LC50) against Aedes aegypti (L.) third and fourth instars was estimated at 0.025 p.p.m. following logit regression. The concentration-mortality response of third- and fourth-instar Anopheles albimanus Weidemann did not conform to a logit model. The LC50 value of spinosad in Anopheles albimanus was 0.024 p.p.m. by quadratic linear regression. A field trial in southern Mexico demonstrated that spinosad 1 p.p.m. compared with the standard temephos (Abate) 1% granules 100 g/m3 water prevented Ae. aegypti breeding in plastic containers of water for 8 weeks; at 10 p.p.m. spinosad prevented breeding for > 22 weeks. In another field trial, spinosad at 5 p.p.m. and temephos both completely eliminated reproduction of Ae. aegypti for 13 weeks. In contrast, the bacterial insecticide Bacillus thuringiensis var. israelensis (Bti, Vectobac) AS) performed poorly with just 2 weeks of complete inhibition of Ae. aegypti breeding. Spinosad also effectively prevented breeding of Culex mosquitoes and chironomids in both trials to a degree similar to that of temephos. We conclude that spinosad merits evaluation as a replacement for organophosphate or Bti treatment of domestic water tanks in Mesoamerica. We also predict that spinosad is likely to be an effective larvicide for treatment of mosquito breeding sites.     

The efficacy of a chitin synthesis inhibitor against field populations of organophosphate-resistant Aedes aegypti in Brazil

The mosquito Aedes aegypti is the main focus of dengue control campaigns. Because of widespread resistance against conventional chemical insecticides, chitin synthesis inhibitors (CSIs) are considered control alternatives. We evaluated the resistance status of four Brazilian Ae. aegypti populations to both the organophosphate temephos and the pyrethroid deltamethrin, which are used in Brazil to control larvae and adults, respectively. All vector populations exhibited high levels of temephos resistance and varying rates of alterations in their susceptibility to pyrethroids. The effect of the CSI novaluron on these populations was also investigated. Novaluron was effective against all populations under laboratory conditions. Field-simulated assays with partial water replacement were conducted to evaluate novaluron persistence. Bioassays were continued until an adult emergence inhibition of at least 70% was attained. We found a residual effect of eight weeks under indoor conditions and novaluron persisted for five-six weeks in assays conducted in an external area. Our data show that novaluron is effective against the Ae. aegypti populations tested, regardless of their resistance to conventional chemical insecticides.     

Cross-induction of detoxification genes by environmental xenobiotics and insecticides in the mosquito Aedes aegypti: impact on larval tolerance to chemical insecticides

The effect of exposure of Aedes aegypti larvae to sub-lethal doses of the pyrethroid insecticide permethrin, the organophosphate temephos, the herbicide atrazine, the polycyclic aromatic hydrocarbon fluoranthene and the heavy metal copper on their subsequent tolerance to insecticides, detoxification enzyme activities and expression of detoxification genes was investigated. Bioassays revealed a moderate increase in larval tolerance to permethrin following exposure to fluoranthene and copper while larval tolerance to temephos increased moderately after exposure to atrazine, copper and permethrin. Cytochrome P450 monooxygenases activities were induced in larvae exposed to permethrin, fluoranthene and copper while glutathione S-transferase activities were induced after exposure to fluoranthene and repressed after exposure to copper. Microarray screening of the expression patterns of all detoxification genes following exposure to each xenobiotic with the Aedes Detox Chip identified multiple genes induced by xenobiotics and insecticides. Further expression studies using real-time quantitative PCR confirmed the induction of multiple CYP genes and one carboxylesterase gene by insecticides and xenobiotics. Overall, this study reveals the potential of xenobiotics found in polluted mosquito breeding sites to affect their tolerance to insecticides, possibly through the cross-induction of particular detoxification genes. Molecular mechanisms involved and impact on mosquito control strategies are discussed.     

Evaluation of various control agents against mosquito larvae in rice paddies in Taiwan.

A field test was conducted in rice paddies adjacent to Wufeng, Taichung County in Central Taiwan to evaluate the efficacy of control agents against mosquito larvae. The agents included Bacillus thuringienesis israelensis (Bti), two Lagenidium giganteum products (Lg product A and T), and temephos. The major mosquito species found in the rice paddies were Culex tritaeniorhynchus/vishnui and Anopheles sinensis. Compared to controls, a 7-day treatment with Bti or Lg products A and T caused overall reductions in the number of immatures (larvae and pupae) of Cx. tritaeniorhynchus/vishnui of 77.5%, 49.7%, and 21.9%, respectively, whereas temephos caused an increase of 66.9%. The overall reductions in An. sinensis were 85.4%, 8.6%, 44.6%, and 92.1%, respectively. There was no significant reduction in the number of mosquito larvae following 42 days of treatment with these agents. In summary, 1-week treatments with both biological control agents produced moderate overall reductions in mosquito larvae in rice paddies. The insecticide temephos, on the other hand, was very effective at suppressing the larvae of An. sinensis but significantly increased the number of Cx. tritaeniorhynchus/vishnui larvae in temephos-treated plots.     

Comparative histopathology of some Diptera and Crustacea of aquatic alpine ecosystems, after treatment with Bacillus thuringiensis var. israelensis

Species of Diptera (larval Culicidae, Chironomidae and Simuliidae) and Crustacea (cladoceran adults) that are representative of the fauna of aquatic alpine ecosystems are routinely treated with Bacillus thuringiensis var. israelensis (Bti) for mosquito control. The toxicity of Bti on these organisms was investigated. Bioassays indicated that Bti used at the concentration for operational field application is deleterious to all dipteran species, but not to Cladocera. Histopathological observations reveal that, in all cases, the midgut epithelium is affected by Bti treatment. However, the vulnerability of epithelial cells to the microbial insecticide is different from one cell-type to another, clear cells being more vulnerable than dark cells. Because of the concentration of clear cells within the anterior midgut of dipteran larvae, this region appears to be the first to show symptoms of intoxication through enhancing a natural process of cell turnover. The relative importance of such a vulnerable region in the midgut of dipteran larvae may account for the general efficacy of Bti to these species. Its harmlessness to Cladocera may be correlated to the relative scarcity of clear cells and their patchy distribution along the whole midgut together with important cellular renewal capacity of the epithelium.     

INCREASE IN LARVAL GUT PROTEOLYTIC ACTIVITIES AND Bti RESISTANCE IN THE DENGUE FEVER MOSQUITO

The bioinsecticide Bacillus thuringiensis var. israelensis (Bti) is increasingly used worldwide for mosquito control. Although no established resistance to Bti has been described in the field so far, a resistant Aedes aegypti strain (LiTOX strain) was selected in the laboratory using field‐collected leaf litter containing Bti toxins. This selected strain exhibits a moderate resistance level to Bti, but a high resistance level to individual Cry toxins. As Bti contains four different toxins, generalist resistance mechanisms affecting mosquito tolerance to different toxins were expected in the resistant strain. In the present work, we show that the resistant strain exhibits an increase of various gut proteolytic activities including trypsins, leucine‐aminopeptidases, and carboxypeptidase A activities. These elevated proteolytic activities resulted in a faster activation of Cry4Aa protoxins while Cry4Ba or Cry11Aa were not affected. These results suggest that changes in proteolytic activities may contribute to Bti resistance in mosquitoes together with other mechanisms.     

Comparison of the insecticide susceptibilities of laboratory strains of Aedes aegypti and Aedes albopictus

A susceptible strain of Aedes albopictus derived from the Gainesville strain (Florida, USA) was established in our laboratory. The larvicidal efficacies of the neurotoxic insecticides temephos, permethrin and the pure cis and trans-permethrin isomers and the microbial insecticide Bacillus thuringiensis israelensis (Bti) against Ae. albopictus were estimated and compared to a susceptible strain of Aedes aegypti. The larvicidal effect of insect growth regulator pyriproxyfen was also evaluated in both mosquito strains. The median lethal concentration/median emergency inhibition values for Ae. aegypti and Ae. albopictus, respectively, were: temephos, 3.058 and 6.632 ppb, permethrin, 3.143 and 4.933 ppb, cis-permethrin, 4.457 and 10.068 ppb, trans-permethrin, 1.510 and 3.883 ppb, Bti, 0.655 and 0.880 ppb and pyriproxyfen, 0.00774 and 0.01642 ppb. Ae. albopictus was more tolerant than Ae. aegypti to all six larvicides evaluated. The order of susceptibility for Ae. aegypti was pyriproxyfen > Bti > trans-permethrin > temephos > permethrin > cis-permethrin and for Ae. albopictus was pyriproxyfen > Bti > trans-permethrin > permethrin > temephos > cis-permethrin. Because both species can be found together in common urban, suburban and rural breeding sites, the results of this work provide baseline data on the susceptibility of Ae. albopictus to insecticides commonly used for controlling Ae. aegypti in the field.     

Effect of Leaf Type and Pesticide Exposure on Abundance of Bacterial Taxa in Mosquito Larval Habitats

Lentic freshwater systems including those inhabited by aquatic stages of mosquitoes derive most of their carbon inputs from terrestrial organic matter mainly leaf litter. The leaf litter is colonized by microbial communities that provide the resource base for mosquito larvae. While the microbial biomass associated with different leaf species in container aquatic habitats is well documented, the taxonomic composition of these microbes and their response to common environmental stressors is poorly understood. We used indoor aquatic microcosms to determine the abundances of major taxonomic groups of bacteria in leaf litters from seven plant species and their responses to low concentrations of four pesticides with different modes of action on the target organisms; permethrin, malathion, atrazine and glyphosate. We tested the hypotheses that leaf species support different quantities of major taxonomic groups of bacteria and that exposure to pesticides at environmentally relevant concentrations alters bacterial abundance and community structure in mosquito larval habitats. We found support for both hypotheses suggesting that leaf litter identity and chemical contamination may alter the quality and quantity of mosquito food base (microbial communities) in larval habitats. The effect of pesticides on microbial communities varied significantly among leaf types, suggesting that the impact of pesticides on natural microbial communities may be highly complex and difficult to predict. Collectively, these findings demonstrate the potential for detritus composition within mosquito larval habitats and exposure to pesticides to influence the quality of mosquito larval habitats.     

Pulse-exposure effects of selected insecticides to juvenile Australian crimson-spotted rainbowfish (Melanotaenia duboulayi)

Laboratory toxicity studies were conducted in southeastern Queensland, Australia, to determine the acute lethal effects of a 1-h pulse exposure of selected insecticides to adult and juvenile (<72 h old) crimson-spotted rainbowfish, Melanotaenia duboulayi (Castlenau). In addition, to its ecological significance, this native fish is a predator of mosquitoes. Two organophosphate (OP) compounds (temephos and pirimiphos-methyl), an entomophathogenic bacterium (Bacillus thuringiensis variety israelensis [Bti]) and two insect growth regulators (IGRs) (s-methoprene and pyriproxyfen) were evaluated. Although none of the five insecticides were acutely toxic to adult M. duboulayi under the test conditions, temephos and pirimiphos-methyl were found to be toxic to juveniles, with 24 h pulse exposure LC50 values of 27 and 15 microg/liter (ppb), respectively. Of the two OPs, pirimiphos-methyl was the most toxic, with a lethal dose ratio (pulse exposure LC50 temephos/ pulse exposure LC50 pirimiphos-methyl) of 1.8 (95% CL 0.5-6.4). These pulse exposure LC50 values represented 40 and 4.5% of the estimated environmental concentrations (EEC) for a 15 cm deep water body, respectively. Bti and the two IGRs had no acute toxic effects at up to 10 and 12.5 times the EEC, respectively. Accordingly, in the interests of environmental conservation and integrated pest management (IPM), preference should be given to the latter three insecticides for control of mosquito larvae in juvenile M. duboulayi habitat.     

Involvement of cytochrome P450 monooxygenases in the response of mosquito larvae to dietary plant xenobiotics

The response of mosquito larvae to plant toxins found in their breeding sites was investigated by using Aedes aegypti larvae and toxic arborescent leaf litter as experimental models. The relation between larval tolerance to toxic leaf litter and cytochrome P450 monooxygenases (P450s) was examined at the toxicological, biochemical and molecular levels. Larvae pre-exposed to toxic leaf litter show a higher tolerance to those xenobiotics together with a strong increase in P450 activity levels. This enzymatic response is both time- and dose-dependent. The use of degenerate primers from various P450 genes (CYPs) allowed us to isolate 16 new CYP genes belonging to CYP4, CYP6 and CYP9 families. Expression studies revealed a 2.3-fold over-expression of 1 CYP gene (CYP6AL1) after larval pre-exposure to toxic leaf litter, this gene being expressed at a high level in late larval and pupal stages and in fat bodies and midgut. The CYP6AL1 protein has a high level of identity with other insect's CYPs involved in xenobiotic detoxification. The role of CYP genes in tolerance to natural xenobiotics and the importance of such adaptive responses in the capacity of mosquitoes to colonize new habitats and to develop insecticide resistance mechanisms are discussed.     

Comparative sensitivity of larval mosquitoes to vegetable polyphenols versus conventional insecticides

The larvicidal effects of polyphenols of natural crude decomposed alder leaf litter and commercially available tannic acid were experimentally compared with those of two common conventional insecticides (Bacillus thuringiensis ssp. israelensis: microbial insecticide; temephos: organophosphate insecticide). Comparative standard bioassays using third instar larval Aedes aegypti, A. albopictus, Culex pipiens and Coquillettidia richiardii as references indicated that Aedes and Culex taxa are far more sensitive to alder leaf litter than to tannic acid and conventional insecticides. C. richiardii is far more resistant to conventional insecticides than Aedes and Culex taxa, but its sensitivity to tannic acid is close to that of those taxa. Dietary vegetable polyphenols are thus proposed as new, practical, alternative chemicals for mosquito control when conventional insecticides are difficult and costly to be used (e.g., in the management of Aedes and Culex populations in man-made breeding sites and Coquillettidia control strategy).     

Effects of Bacillus thuringiensis var. israelensis on Target and Nontarget Organisms: a Review of Laboratory and Field Experiments

Since the discovery of Bacillus thuringiensis var. israelensis (Bti) in 1976, extensive literature has proved its efficacy to control mosquitoes and black flies, of which many species are known as important vectors of diseases or simply as pests of humans and animals. Since 1978, Bti has been used in many countries on all continents and numerous studies have been made on target mosquitoes and black flies, as well as nontarget organisms (NTO). This review analyses the results of 75 studies on these organisms covering approximately 125 families, 300 genera and 400 species. Different factors such as species, instar, feeding behaviour and environmental parameters (larval density, water temperature, suspended matter etc.) may drastically affect the efficacy of the Bti products. This is addressed in detail by reviewing the main factors affecting mosquitoes as well as black flies. The results of a wide range of laboratory and field experiments using different target and nontarget species, various preparations and formulations of Bti and different biotic or abiotic factors are present in the literature, making the data difficult to compare on a common basis. Our analysis shows that, under different application conditions, the effects of Bti on target and nontarget organisms may be hard to predict. Although Bti has been proclaimed to be relatively highly specific, some studies show that some NTO are affected either by single or repeated Bti treatments. Present use against black flies seems ecologically acceptable. High frequencies of application and/or overdosages against mosquitoes may result in some persistence of the toxin crystals and ultimately this may have adverse effects on the food web. A long-term study (published in 1998) in mosquito habitats has shown that intensive Bti treatments over three years did in fact produce an impact on the food web in wetlands. This raises questions, for the first time, on Bti environmental specificity. The importance of this impact is discussed and the alternatives for practical pest control are considered. Some modifications of Bti use against mosquitoes, guided by research, is probably the best of these alternatives.     

Differential toxicity of leaf litter to dipteran larvae of mosquito developmental sites

The relative toxicity of leaf litter to nematocerous dipteran larvae characteristic of mosquito developmental sites was investigated. Culicidae, Chironomidae, and Simuliidae taxa originating from alpine hydrosystems were tested together with two laboratory nonindigenous culicid taxa. Bioassays indicate that ingestion of 10-month-old decaying leaves from Alnus glutinosa, Populus nigra, and Quercus robur by larvae is more deleterious for Aedes aegypti, A. albopictus, Culex pipiens, Simulium variegatum, and Chironomus annularius than for A. rusticus. Histopathological observations reveal that the midgut epithelium is the main target organ of the toxic effect of dietary leaf litter, which appears to be stronger than that of previously reported tannic acid. There is a general response of the nematocerous larval midgut epithelium to dietary tannins-phenolic compounds: clear cells of the anterior midgut showing symptoms of intoxication before dark cells of the posterior midgut.     

Salt-marsh Crustaceans, Gammarus duebeni and Palaemonetes varians as Predators of Mosquito Larvae and Their Reaction to Bacillus thuringiensis subsp. israelensis

The predatory potential of two onmnivorous crustaceans, Gammarus duebeni and Palaemonetes varians, has been examined to investigate their effect as mosquito larval predators. Mature gammarids ate 4-8 Aedes detritus larvae/24 h and palaemonids, 22-30/1 h, often killing larvae when not hungry. The crustaceans were exposed to Bacillus thuringiensis subsp. israelensis (Bti) and to mosquito larvae killed by Bti with no adverse effects, endorsing the safety of this microbial pesticide. Crustacean faecal pellets, collected post-feeding and placed in fresh marsh water, were toxic to mosquito larvae the following day. After placing the crustaceans in fresh salt-marsh water for 6 days, the fresh faecal pellets failed to kill mosquitoes apart from pellets obtained from crustaceans originally fed on the highest concentration of Bti, where there was a 50% kill after 3 days of incubation. Mosquito larvae on salt-marshes are not easy to control with ecologically undesirable toxic chemicals. Encouraging the breeding of predators such as crustaceans, or even their release, could prove to be a useful method of mosquito control to supplement the periodic inundative use of the ecologically acceptable Bti.     

Seasonal availability of resources and habitat degradation for the western tree-hole mosquito, <Emphasis Type="Italic">Aedes sierrensis</Emphasis>

The nutrient base of aquatic tree-hole communities is derived from leaf litter, benthic detritus, and water flowing down the tree trunk (stemflow water). Previous studies in eastern North America with the mosquito, Aedes triseriatus, have identified leaf litter as a major and stemflow water as a minor source of mosquito nutrition, but did not consider the role of the benthic detritus or how the aggregate or relative contribution of these sources of mosquito nutrition changed during the year. We use the leaf litter, benthic detritus, and stemflow water from tree holes in western Oregon (USA) to determine how these substrates affect mass at metamorphosis, biomass yield, and fitness (cohort replacement rate; R ₀) of the mosquito, Aedes sierrensis, through both natural and simulated winters, the normal growing season for larvae in tree holes. We found that fresh leaf litter constitutes the major determinant of mosquito fitness by a factor of >15:1 over any other substrate taken directly from tree holes in nature. The other substrates, including the benthic detritus, individually make only a meager contribution to mosquito fitness but, when added to the leaf litter, can sustain yield and improve fitness at high, limiting larval densities. Nutritional quality of tree-hole substrates declines by >90% from early (fall) to late (spring) in the larval growing season. At both times of year, the coarse or fine detritus provide minor resources, and stemflow water provides no detectable contribution to mosquito nutrition. The resources in the litter are not transported during the year to the benthic detritus; rather, these resources are either exploited by mosquitoes when they first become available, or they deteriorate and become progressively more unavailable to them. Growth and development of A. sierrensis feeding on dried and reconstituted tree-hole contents during a 6-month simulated winter in the laboratory showed: (1) the same relative contributions of leaf litter, benthic detritus, and stemflow water to mosquito nutrition, (2) that the winter deterioration of substrate quality is a direct consequence of microbial decomposition, and (3) that pre-emptive competition from pre-existing A. sierrensis greatly increases substrate deterioration. We conclude that the progressive winter deterioration of larval resources in combination with the dry summers of western North America are the most likely environmental factors that limit species diversity in tree holes and that have selected for early recruitment (autumnal hatching) of A. sierrensis and for its univoltine life cycle from Mexico to Canada. Received: 28 December 1999 / Accepted: 10 April 2000     

Influence of Stream Profile and Abiotic Factors on the Performance and Residual Dosages of Two Commercial Formulations of Bacillus thuringiensis subsp. israelensis During a 2-Year Experiment

Recently, results of a 3-year study showed the importance of abiotic factors such as water temperature, stream discharge and the hyporheic zone on the behaviour, performance and loss of the residual dosages (amount of the injected dosage left at each station) of two commercial liquid formulations of Bacillus thuringiensis subsp. israelensis ( Bti ), Teknar HP-D and Vectobac 1200L. Experiments were performed in the same stream and under environmental conditions that allowed comparisons of results between formulations. To improve our understanding of the influence of abiotic factors on the behaviour and the performance of these formulations, a 2-year experiment using the same products was conducted in another river characterized by a much different profile, higher discharges and colder temperatures. A field procedure based on a system of gutters located on the bank of the river already used in the previous 3-year experiment was utilized. Black fly larval mortalities were recorded along the river at different distances (stations) to evaluate the behaviour and the performance of the products while water samples taken at the stations were tested in the laboratory against mosquito neonate larvae to evaluate the residual dosages of Bti at each station. Results showed that higher residual dosages of the Vectobac 1200L were recovered compared to Teknar HP-D along the river. Although higher dosages were recovered, higher mortality was observed only for the Vectobac 1200 L in high discharge conditions. As seen before, the hyporheic zone (interstitial water between the streambed and groundwater) produced a major loss of the dosages in the first meters of the river. But because of the river profile, the hyporheic zone had a lesser effect on the loss of the dosages further in the river resulting in very long carries for both Teknar HP-D and Vectobac 1200L.     

Combination of Mesocyclops thermocyclopoides and Bacillus thuringiensis var. israelensis: a better approach for the control of Aedes aegypti larvae in water containers.

The efficacy of a local Thai-strain of the copepod, Mesocyclops thermocyclopoides and the larvicide, Bacillus thuringiensis var. israelensis (Bti), used jointly and singly, was studied against Aedes aegypti in water containers. In a laboratory test, copepods alone produced mortality of 98-100% in 1st instar larvae of Ae. aegypti at copepod:larvae ratios ranging from 1:1 to 1:4. In an outdoor field simulated experiment that ran for 16 wk, after a single inoculation, the treatment of copepods and Bti combined yielded the better, more sustainable results than the agents used individually. Numbers of mosquito larvae per sample in the combined treatment were zero during the first 8 wk; larval numbers then increased but were maintained at a very low level for the next 4 wk after which the larval numbers increased moderately but still remained below numbers in the control. Bti alone kept the larvae at the zero level for the first 4 wk after which their numbers increased slightly and were at low levels up to 12 wk. Copepods alone maintained larval numbers at a low level as compared with those of the control. During the course of the experiment the larval numbers in the control were greater than 20 per sample. Statistically significant differences were noted among treatment means (F = 23.083, df = 3/60, P<0.01) over the total period of the study. The number of copepods in the joint treatment was significantly higher than in the copepod alone treatment for the first 8 wk (t = -4.97, df = 14, P<0.01). The density of copepods, however, for the whole 16-wk period was not significantly different in these two treatments (t = -1.51, df = 30, P>0.1).