Laboratory and field evaluation of insect repellents as oviposition deterrents against the mosquito Aedes albopictus

Three experimental approaches were used to evaluate the oviposition deterrency of three insect repellents, AI3-35765, AI3-37220 (piperidine compounds), and the standard N,N-diethyl-3-methylbenzamide (deet) to the mosquito Aedes albopictus Skuse (Diptera: Culicidae). Against laboratory-reared Ae. albopictus gravid females, the EC50 values of AI3-37220, AI3-35765 and deet were 0.004%, 0.008% and 0.011% in laboratory cages and 0.004%, 0.01% and 0.009% in an outdoor screened cage. For a natural population of Ae. albopictus tested in the field, the EC50 values were determined as 0.004%, 0.008% and 0.001%, respectively. Ageing concentrations of 0.1% of each repellent provided >50% effective oviposition deterrency against the laboratory population of Ae. albopictus for 13 days in laboratory cages, for 15 days in the outdoor cage, and for 21 days against field population of Ae. albopictus in Florida. These topical skin repellents are effective oviposition deterrents for Ae. albopictus when employed at relatively low application rates.     

Effect of Novaluron (Rimon 10 EC) on the mosquitoes Anopheles albimanus, Anopheles pseudopunctipennis, Aedes aegypti, Aedes albopictus and Culex quinquefasciatus from Chiapas, Mexico

Dengue fever is a serious problem in Mexico and vector control has not been effective enough at preventing outbreaks. Malaria is largely under control, but it is important that new control measures continue to be developed. Novaluron, a novel host-specific insect growth regulator and chitin synthesis inhibitor, has proved to be effective against agricultural pests, but its efficacy against larval mosquito vectors under field conditions remains unknown. In accordance with the World Health Organization Pesticide Evaluation Scheme, phase I, II and III studies were conducted to evaluate the efficacy and residual effect of Novaluron (Rimon 10 EC, Makhteshim, Beer-Sheva, Israel) on the malaria vectors Anopheles albimanus Wiedemann (Diptera: Culicidae) and Anopheles pseudopunctipennis Theobald, the dengue vectors Aedes aegypti (L) and Aedes albopictus Skuse and the nuisance mosquito Culex quinquefasciatus Say. Laboratory susceptibility tests yielded diagnostic concentrations for all five target species. Field trials to identify the optimum field dosage of Novaluron against Anopheles mosquitoes were carried out under semi-natural conditions in artificial plots and in vessels with wild mosquitoes. Efficacy was measured by monitoring mortality of larvae and pupae and the percentage of inhibition of emergence from floating cages. Dosages of Novaluron for field tests were based on pupal LC(99) (lethal concentration 99%) of An. pseudopunctipennis (0.166 mg/L) in plots and average pupal LC(99) of Ae. aegypti and Ae. albopictus (0.55 mg/L). At all dosages tested, Novaluron significantly reduced larval populations of An. albimanus, Culex coronator Dyar & Knab, Ae. albopictus and Cx. quinquefasciatus by approximately 90%, inhibited adult emergence of An. albimanus and An. pseudopunctipennis by approximately 97% for almost 4 months in experimental plots, and inhibited adult emergence of Ae. aegypti and Ae. albopictus by approximately 97% for up to 14 weeks. Recommended dosages of Novaluron for non-container breeding and container breeding mosquitoes are 0.166 mg/L and 0.55 mg/L, respectively. Overall, the residual effect was more sustained than that of temephos. The lowest dosage of Novaluron had less of an impact on non-target organisms than did temephos. Small-scale field trials in natural breeding sites treated with Novaluron at 0.6 L/ha eliminated adult emergence of An. albimanus and Cx. coronator for 8 weeks. For phase III studies, Novaluron was tested at the local and village levels, applying the optimum field rate to all natural breeding habitats within 1 km of a pair of neighbouring villages. Village-scale trials of Novaluron at 0.6 L/ha reduced An. albimanus larval populations for at least 8 weeks and, more importantly, sharply reduced the densities of adult host-seeking mosquitoes approaching houses. We conclude that Novaluron is effective and environmentally safer than temephos.     

Efficacy of three insect repellents against the malaria vector Anopheles arabiensis

Three commercial repellents marketed in South Africa: Bio-Skincare (BSC, oils of coconut, jojoba, rapeseed and vitamin E), Mosiguard towelletes with 0.574 g quwenling (p-menthane-3,8-diol, PMD) and the standard deet (15% diethyl-3-methylbenzamide, Tabard lotion), were compared against a laboratory colony of the mosquito Anopheles arabiensis Patton (Diptera: Culicidae), the predominant malaria vector in South Africa. Human forearms were treated with 1.2 g BSC, 0.8 g PMD towelette or 0.5 g deet and exposed to 200 hungry An. arabiensis females for 1 min, at intervals of 1-6 h post-treatment. Tests were conducted by three adult male volunteers (aged 30-45 years, crossover controlled test design for 3 consecutive days), using their left arm for treatment and right arm for untreated control. Biting rates averaged 39-52 bites/min on untreated arms. All three repellents provided complete protection against An. arabiensis for up to 3-4 h post-application; deet and PMD gave 90-100% protection up to 5-6h, but BSC declined to only 52% protection 6h post-treatment. These results are interpreted to show that all three repellent products give satisfactory levels of personal protection against An. arabiensis for 4-5 h, justifying further evaluation in the field.     

Aedes albopictus (Diptera: Culicidae) in Rome: experimental study of relevant control strategy parameters

Since 1997, Aedes albopictus has colonised and then rapidly invaded the city of Rome (Italy) and its peripheral areas. Presently, the control of this mosquito in Italy relies mainly on larvicidal treatment of street storm sewer catch basins with the organophosphate temephos. We have therefore obtained baseline data on the susceptibility to temephos of the Roman Ae. albopictus population by laboratory bioassays on F1 fourth-instar larvae following standard WHO protocols. Estimated lethal concentrations were 0.014 mg/l (LC50) and 0.022 mg/l (LC90) indicating a lack of resistance to this compound. The persistence of temephos in sewer catch basins was evaluated by follow-up of catch basins treated with a dose of 1.5 mg of active ingredient. Mosquito larvae were recovered in 10% and 50% of the treated basins at 9 and 18 days posttreatment, respectively. In order to understand the relative contribution of this larval habitat to adult populations, we conducted a survey in the Zoo of Rome to estimate the larval density of mosquitoes breeding in sewer catch basins. A complete census of a 16.5 ha area mapped 243 catch basins, but only 25 (10.3%) contained water; of the latter 8 (32.0%) hosted mosquito larvae. All positive catch basins contained larvae of Culex pipiens, which were associated with Culiseta longiareolata and/or Ae. albopictus in 6 and 3 cases, respectively. A longitudinal survey of one catch basin over 4 months showed that the mean larval density of Ae. albopictus was markedly lower than that of Cx pipiens and Cs. Iongiareolata, ranging between 0 and 1.3 larvae/dip as compared to 0-33.2 and 0-22.7 larvae/dip, respectively. However, adult densities of Ae. albopictus in this area estimated during the same period with 20 ovitraps showed consistently high values (proportion of positive ovitraps around 100%). These preliminary observations suggest that whenever alternative larval biotopes other than sewer catch basins are widely available, they might be more productive and/or preferred substrates to catch basins for Ae. albopictus breeding.     

Laboratory evaluation of Mesocyclops annulatus (Wierzejski, 1892) (Copepoda: Cyclopidea) as a predator of container-breeding mosquitoes in Argentina

In laboratory bioassays we tested the predatory capacity of the copepod Mesocyclops annulatus on Aedes aegypti and Culex pipiens larvae. A single adult female of M. annulatus caused 51.6% and 52.3% mortality of 50 first instar larvae of Ae. aegypti and Cx. pipiens respectively, in a 72 h test period. When alternative food was added to the containers, mortality rates declined to 16% and 10.3% for Ae. aegypti and Cx. pipiens respectively. When 50 first instar larvae of each of the two mosquito species tested were placed together with a single adult female of M. annulatus, mortality rates were 75.5% for Ae. aegypti larvae and 23.5% for Cx. pipiens larvae in a three day test period. Different density of adult females of M. annulatus ranged from 5 to 25 females produced mortality rates of Ae. aegypti first instar larvae from 50% to 100% respectively. When a single adult female of M. annulatus was exposed to an increasing number of first-instar Ae. aegypti larvae ranging from 10 to 100, 100% mortality was recorded from 1 to 25 larvae, then mortality declined to 30% with 100 larvae. The average larvae killed per 24 h period by a single copepod were 29.     

Thanaka (Limonia acidissima) and deet (di-methyl benzamide) mixture as a mosquito repellent for use by Karen women

The prevention and treatment of drug-resistant malaria is becoming increasingly difficult. On the Thai–Myanmar border multi-drug resistant strains of falciparum malaria are increasing and, because the malaria vector Anopheles bite outdoors during early evening, insecticide house-spraying or impregnated bednets provide only limited protection. Therefore, the protective efficacy of repellent formulations containing di-methyl benzamide (deet) and permethrin against local vectors was estimated, when applied to the skin, and their acceptability amongst pregnant Karen women who are at relatively high risk from malaria was assessed. Human landing catches of mosquitoes showed that almost complete protection was achieved using different formulations of 20% deet and 0.5% permethrin for up to 6 h. All-night collections from human subjects indicated that this repellent combination reduced exposure to malaria parasites by at least 65 and 85% for those transmitted by Anopheles minimus and An. maculatus , respectively, the two principal vectors in this area. Pregnant women in the camps preferred repellents which were mixed with 'thanaka', a root paste made from pulp of the wood apple tree, Limonia acidissima , used locally as a cosmetic. Apart from a temporary warming sensation where repellent thanaka was applied to the skin, the repellents were well tolerated. An intervention trial is currently in progress to determine whether deet mixed with thanaka can protect pregnant women against malaria in this part of the world. Bioassays using a laboratory strain of Aedes aegypti demonstrated that thanaka is itself slightly repellent at high dosages and the mixture with deet provides protection for over 10 h. This treatment would therefore also provide some personal protection against dengue, which is increasing locally, transmitted by Ae. aegypti and Ae. albopictus biting during the daytime.     

Horizontal transfer of the insect growth regulator pyriproxyfen to larval microcosms by gravid Aedes albopictus and Ochlerotatus triseriatus mosquitoes in the laboratory

The insect growth regulator (IGR) pyriproxyfen is highly active against mosquitoes (Diptera: Culicidae). Through continuous emersion of large larvae (instars 3-4) the concentration causing 50% inhibition of adult emergence (EI50) was determined as 0.200 p.p.b. for Aedes albopictus (Skuse) and 3.5 to 7 times less for Ochlerotatus triseriatus (Say): IE50 0.0288 p.p.b. As a possible method of application to larval microcosms of these species that oviposit in water containers and phytotelmata, the horizontal transfer of pyriproxyfen to larval microcosms by adult mosquitoes was evaluated under laboratory conditions. Gravid females were forced to walk on surfaces treated with pyriproxyfen (tarsal contact exposure) and then allowed to oviposit in larval microcosms. Using replicate bioassay cages, each with an oviposition container, and a factorial experimental design, we assessed Ae. albopictus for the effects of (i) pyriproxyfen concentration (0.2, 0.3 and 0.4 mg/cm2) contacted by gravid females, and (ii) the number of treated gravid females added to bioassay cages (one, three or five females/cage), on the mortality of larvae in oviposition containers. Only 0.2 mg/cm2 treatment rate was tested on Oc. triseriatus. A significant (P < 0.05) curvilinear response in inhibition of emergence (IE) was achieved on both species. Densities of one or three treated Oc. triseriatus females/cage yielded IE rates of only 21-27%, whereas five treated females/cage resulted in 70% inhibition. With Ae. albopictus, densities of three or five treated females/cage yielded 48-67% and 59-73% IE, respectively, whereas one treated female/cage gave only 4-30% inhibition. Use of IGR-treated oviposition containers to achieve horizontal transfer of pyriproxyfen to mosquito oviposition sites could be a field management technique based on mosquito biology and behaviour. In binary choice tests with Ae. albopictus, horizontal transfer of pyriproxyfen from a container with a treated ovistrip (0.3 or 0.4 mg/cm2) to an untreated microcosm resulted in 14-38% inhibition. In larval bioassays, pyriproxyfen activity declined markedly within 10 days. Forcibly exposing gravid female mosquitoes to pyriproxyfen-treated paper surface did not affect their fecundity. However, from the 1st to 2nd gonotrophic cycles the egg hatch rate declined by 30% (P < 0.05). Some variation of results could be due to interactions between females at the oviposition site, possibly causing disproportionate transfer of pyriproxyfen to larval microcosms. Comparative studies of the oviposition behaviour of each mosquito are warranted and would potentially provide information needed to improve the technique.     

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.     

An experimental field study of delayed density dependence in natural populations of Aedes albopictus

Aedes albopictus, a species known to transmit dengue and chikungunya viruses, is primarily a container-inhabiting mosquito. The potential for pathogen transmission by Ae. albopictus has increased our need to understand its ecology and population dynamics. Two parameters that we know little about are the impact of direct density-dependence and delayed density-dependence in the larval stage. The present study uses a manipulative experimental design, under field conditions, to understand the impact of delayed density dependence in a natural population of Ae. albopictus in Raleigh, North Carolina. Twenty liter buckets, divided in half prior to experimentation, placed in the field accumulated rainwater and detritus, providing oviposition and larval production sites for natural populations of Ae. albopictus. Two treatments, a larvae present and larvae absent treatment, were produced in each bucket. After five weeks all larvae were removed from both treatments and the buckets were covered with fine mesh cloth. Equal numbers of first instars were added to both treatments in every bucket. Pupae were collected daily and adults were frozen as they emerged. We found a significant impact of delayed density-dependence on larval survival, development time and adult body size in containers with high larval densities. Our results indicate that delayed density-dependence will have negative impacts on the mosquito population when larval densities are high enough to deplete accessible nutrients faster than the rate of natural food accumulation.     

Aedes albopictus, vector of chikungunya and dengue viruses in Reunion Island: biology and control

Chikungunya virus (CHIKV) and dengue virus (DENV) are mosquito-borne viruses transmitted by the Aedes genus. Dengue is considered as the most important arbovirus disease throughout the World. Chikungunya, known from epidemics in continental Africa and Asia, has up to now been poorly studied. It has been recently responsible for the severe 2004-2007 epidemic reported in the Indian Ocean (IO), which has caused several serious health and economic problems. This unprecedented epidemic of the IO has shown severe health troubles with morbidity and death associated, which had never been observed before. The two major vectors of those arboviruses in the IO area are Aedes aegypti and Aedes albopictus. The latest is considered as the main vector in most of the islands of the area, especially in Reunion Island. Ae. albopictus showed strong ecological plasticity. Small disposable containers were the principal urban breeding sites, and preferred natural developmental sites were bamboo stumps and rock holes in peri-urban and gully areas. The virus has been isolated from field collected Ae. albopictus females, and in two out of 500 pools of larvae, demonstrating vertical transmission. Experimental works showed that both Ae. albopictus and Ae. aegypti from west IO islands are efficient vectors of dengue and chikungunya viruses. Since 2006 and all along the epidemic of CHIKV, measures for the control of larvae (temephos then Bacillus thuringiensis) and adults (fenitrothion, then deltamethrine) of Ae. albopictus where applied along with individual and collective actions (by the use of repellents, and removal of breeding sites around houses) in Reunion Island. In order to prevent such epidemics, a preventive plan for arboviruses upsurge is ongoing processed. This plan would allow a quicker response to the threat and adapt it according to the virus and its specific vector.     

Parasitism of Ascogregarina taiwanensis and Ascogregarina culicis (Apicomplexa: Lecudinidae) in larvae of Aedes albopictus and Aedes aegypti (Diptera: Culicidae) from Manaus, Amazon region, Brazil

The aim of the study was to determine the existence of Ascogregarina spp. in larvae of Aedes albopictus and Aedes aegypti collected in urban and suburban areas of Manaus, Amazon region, Brazil. Between May 2004 and July 2005, the mid-gut of 3rd and 4th instar larvae, collected in tire traps in six neighborhoods of Manaus, was examined for the presence of trophozoites of Ascogregarina. Coexistence of Ae. albopictus larvae infected by A. taiwanensis, and Ae. aegypti larvae by A. culicis, was detected in traps in the field. The percentage of Ae. albopictus larvae infected by A. taiwanensis ranged from 21% to 93.5% and of Ae. aegypti larvae infected by A. culicis from 22% to 95%. The mean infection intensity was similar in both species of Aedes. In traps located in Mauazinho, the replacement of Ae. aegypti by Ae. albopictus larvae was observed. In Manaus, Ae. albopictus larvae were parasitized by A. taiwanensis, and Ae. aegypti larvae by A. culicis. Infection rates were high when the species of Aedes were found separately.     

Better to Be in Bad Company than to Be Alone? Aedes Vectors Respond Differently to Breeding Site Quality in the Presence of Others

This study focuses on two competing species, Aedes aegypti and Aedes albopictus (Diptera: Culicidae), both invasive mosquitoes of the New World. Context-specific competition between immature forms inside containers seems to be an important determinant of the coexistence or displacement of each species in different regions of the world. Here, competition experiments developed at low density (one, two or three larvae) and receiving four different resource food concentration, were designed to test whether Ae. albopictus and Ae. aegypti respond differently to competition, and whether competition can be attributed to a simple division of resources. Three phenotypic traits - larval development, adult survival under starvation and wing length - were used as indicators of performance. Larvae of neither species were limited by resource concentration when they were alone, unlike when they developed with competitors. The presence of conspecifics affected Ae. aegypti and Ae. albopictus, inducing slower development, reduced survival and wing length. The response to resource limitation was different when developing with heterospecifics: Ae. aegypti developing with one heterospecific showed faster development, producing smaller adults with shorter lives, while in the presence of two competitors, development increased and adults lived longer. Aedes albopictus demonstrated a better performance when developing with heterospecifics, with no loss in their development period and improved adult survival. Overall, our results suggest that response to competition can not simply be attributed to the division of resources, and that larvae of both species presented large phenotypic plasticity in their response to the presence or absence of heterospecifics and conspecifics.     

Repellency of deet and SS220 applied to skin involves olfactory sensing by two species of ticks

Responses of host-seeking nymphs of the blacklegged tick, Ixodes scapularis Say and lone star tick, Amblyomma americanum (Linnaeus) (Acari: Ixodidae) to the repellents N,N-diethyl-3-methylbenzamide (deet) and (1S, 2'S)-2-methylpiperidinyl-3-cyclohexene-1-carboxamide (SS220) were studied using fingertip laboratory bioassays. Ethanol solutions of both compounds applied to the skin strongly repelled both species of ticks at 0.8 and 1.6 micromole of compound/cm2 skin. The ticks were also repelled when two layers of organdie cloth covered the portion of a finger treated with either deet or SS220. Gas chromatographic analyses of the outer layer of cloth that had covered skin treated with 1.6 micromole compound/cm2 skin revealed only 0.1 nmole SS220/cm2 cloth and 2.8 nmole deet/cm2 cloth. However, in bioassays in which a single layer of cloth was treated with a dose of deet or SS220 equivalent to the amount found in the outer layer of cloth, ticks were not repelled. Results unequivocally demonstrated that these ticks responded to the repellents in the vapour phase when repellent treated skin was covered with cloth to obviate tactile contact with them, and made it clear that the ticks detect the repellents by olfactory sensing. Heretofore, the mode of action of deet and SS220 was unclear.     

Larval development of Aedes aegypti and Aedes albopictus in peri-urban brackish water and its implications for transmission of arboviral diseases

Aedes aegypti (Linnaeus) and Aedes albopictus Skuse mosquitoes transmit serious human arboviral diseases including yellow fever, dengue and chikungunya in many tropical and sub-tropical countries. Females of the two species have adapted to undergo preimaginal development in natural or artificial collections of freshwater near human habitations and feed on human blood. While there is an effective vaccine against yellow fever, the control of dengue and chikungunya is mainly dependent on reducing freshwater preimaginal development habitats of the two vectors. We show here that Ae. aegypti and Ae. albopictus lay eggs and their larvae survive to emerge as adults in brackish water (water with <0.5 ppt or parts per thousand, 0.5-30 ppt and >30 ppt salt are termed fresh, brackish and saline respectively). Brackish water with salinity of 2 to 15 ppt in discarded plastic and glass containers, abandoned fishing boats and unused wells in coastal peri-urban environment were found to contain Ae. aegypti and Ae. albopictus larvae. Relatively high incidence of dengue in Jaffna city, Sri Lanka was observed in the vicinity of brackish water habitats containing Ae. aegypti larvae. These observations raise the possibility that brackish water-adapted Ae. aegypti and Ae. albopictus may play a hitherto unrecognized role in transmitting dengue, chikungunya and yellow fever in coastal urban areas. National and international health authorities therefore need to take the findings into consideration and extend their vector control efforts, which are presently focused on urban freshwater habitats, to include brackish water larval development habitats.     

Bean alpha-amylase inhibitors in transgenic peas inhibit development of pea weevil larvae

This glasshouse study used an improved larval measurement procedure to evaluate the impact of transgenic pea, Pisum sativum L., seeds expressing a-amylase inhibitor (AI)-1 or -2 proteins on pea weevil, Bruchus pisorum L. Seeds of transgenic 'Laura' and 'Greenfeast' peas expressing alpha-(AI)-1 reduced pea weevil survival by 93-98%. Larval mortality occurred at an early instar. Conversely, in nontransgenic cultivars, approximately 98-99% of the pea weevils emerged as adults. By measuring the head capsule size, we determined that larvae died at the first to early third instar in alpha-(AI)-1 transgenic peas, indicating that this inhibitor is highly effective in controlling this insect. By contrast, transgenic Laura and 'Dundale' expressing alpha-(AI)-2 did not affect pea weevil survival, but they did delay larval development. After 77 d of development, the head capsule size indicated that the larvae were still at the third instar stage in transgenic alpha-(AI)-2 peas, whereas adult bruchids had developed in the nontransgenic peas.     

几种驱避化合物对白纹伊蚊寄主搜寻能力的影响

研究了白纹伊蚊Aedes albopictus在含不同浓度的驱避化合物的空间内停留不同时间后,对其寄主搜寻能力的影响。结果显示：在密闭空间中, 分别以0.013～0.500 μg/cm³浓度的柠檬醛、丁香酚、芳樟醇、香叶醇、茴香醛处理白纹伊蚊24～96 h后,试蚊均出现不同程度的定向寄主抑制,其中以茴香醛及香叶醇抑制效果最佳,但香茅醛各处理组试蚊均未受到影响。白纹伊蚊搜寻定向寄主能力受到抑制的个体,在实验室正常饲养后,可逐渐恢复;但茴香醛0.250 μg/cm³处理试虫96 h后,一直未能恢复正常的吸血行为。研究结果提示,茴香醛与香叶醇作为优良的空间驱避剂,在白纹伊蚊防控技术领域可发挥重要作用。     

Larval growth and biosynthesis of reserves in mosquitoes

Developmental instars of four species of mosquitoes have been analyzed for growth and synthesis of biomass with respect to their caloric content of protein, lipids, and carbohydrates for each instar of Aedes aegypti and Culex pipiens of the subfamily Culicinae, and Anopheles albimanus, and An. gambiae of the subfamily Anophelinae. The diameter of the thorax grows during the intermolt, reflecting continuous increase in biomass because it correlates significantly with the larval synthesis of total protein, lipids, and carbohydrates. For Ae. aegypti the fourth instar was sexed to disclose the sex-specific synthetic potential. In Ae. aegypti the protein increased in linear proportion with larval body size, whereas lipid synthesis followed a significant, exponential regression, which was clearly steeper in male larvae and most pronounced in the last instar. When normalized for size, the size-specific protein and lipid contents showed minimal levels of 0.25 and 0.1, respectively, regardless of standard or crowded rearing conditions. The rate of lipid synthesis in Ae. aegypti was determined by incubating fourth instar larvae with (14)C-acetate and estimating the lipids. The rate was highest in the early larvae and decreased towards the end shortly before pupation; in male larvae incorporation was twice the rate of female larvae. Cx. pipiens reached the largest body sizes of all species tested, with protein and lipids increasing linearly with size. Their minimal levels of size-specific caloric contents were around 0.35 for protein and 0.25 for lipids. Anopheles also showed a linear relationship between larval size and caloric protein and lipid contents. Their minimal threshold levels in size-specific contents were 0.35 for protein and 0.2 for total lipids, similar to Culex, but slightly higher than in Aedes. Starvation of Ae. aegypti larvae and subsequent feeding partially improved their lipid contents, but never to the levels of non-starving, optimal controls. Conversely, well-fed final instars exposed to complete starvation showed a tremendous reduction of the protein and lipids contents in the surviving imagines, accompanied by 73% mortality. These results demonstrate the biosynthetic plasticity and the significance of the phagoperiod in Ae. aegypti during the final fourth instar for growth. The characteristic differences between these two subfamilies in their larval physiology are discussed in relation to ecological factors as encountered in the field under natural conditions, and in relation to our earlier findings on the reproductive physiology.     

Organochlorines and organophosphates susceptibility of Aedes albopictus Skuse larvae from agricultural and non-agricultural localities in Peninsular Malaysia

Aedes albopictus larvae obtained from different types of agricultural and non-agricultural localities in Peninsular Malaysia were subjected to several larvicides at World Health Organization (WHO) recommended dosages. Upon 24 h of WHO larval bioassay using two organochlorines and six organophosphates, high resistance against dichlorodiphenyltrichloroethane (DDT), temephos, chlorpyrifos and bromophos were demonstrated among all larval populations. Aedes albopictus larvae from both paddy growing areas (92.33% mortality) and rubber estates (97.00% mortality) were moderately resistant to dieldrin while only Ae. albopictus larvae from dengue prone residential areas (89.00% mortality) showed high resistance against dieldrin. All Ae. albopictus larval populations also developed either incipient or high resistance to both malathion (33.67%–95.33% mortality) and fenitrothion (73.00%–92.67% mortality). Only Ae. albopictus larvae from fogging-free residential areas that were tolerant to fenthion (97.33% mortality), whereas Ae. albopictus larvae from dengue prone residential areas were highly resistant to the same organophosphate (88.33% mortality). Cross resistance between intraclass and interclass larvicides of organochlorines and organophosphates were also exhibited in this study. The present study provided baseline data on various susceptibility levels of Ae. albopictus larval populations from different types of agricultural and non-agricultural localities against organochlorines and organophosphates at WHO recommended dosages. Nevertheless, further susceptibility investigations are suggested using revised doses of larvicides established from the local reference strain of Ae. albopictus to prevent the underestimation or overestimation of insecticide resistance level among Ae. albopictus field strains of larvae.     

Dosage-mortality and time-mortality studies of a granulosis virus in a laboratory strain of the codling moth,Laspeyresia pomonella

Different doses of a granulosis virus were administered to first- and fifth-instar larvae of the codling moth Laspeyresia pomonella. Virus was very pathogenic for both larval instars. The LD₅₀ values for first- and fifth-instar larvae were 5 and 49 capsules/larva, respectively. However, fifth-instar larvae were much more variable in their response to virus than first-instar larvae. Using probit methods it was calculated that 1 capsule could cause death in about 25% of both larval instars but 1578 capsules were required to cause 70% mortality of fifth-instar larvae as compared to 12 capsules for first-instar larvae. This is the first report of a decided difference in variability of response to virus by two larval instars of the same species. A bimodal response by both larval instars was observed in time-mortality studies. Apparently, about 20% of the larvae were very resistant to virus infections.     

Distributions of Competing Container Mosquitoes Depend on Detritus Types, Nutrient Ratios, and Food Availability

Coexistence of competitors may result if resources are sufficiently abundant to render competition unimportant, or if species differ in resource requirements. Detritus type has been shown to affect interspecific competitive outcomes between Aedes albopictus (Skuse) and Aedes aegypti (L.) larvae under controlled conditions. We assessed the relationships among spatial distributions of detritus types, nutrients, and aquatic larvae of these species in nature. We collected mosquitoes, water, and detritus from artificial containers across 24 Florida cemeteries that varied in relative abundances of Ae. aegypti and Ae. albopictus.We measured nutrient content of fine particulate organic matter in water samples as total N, P, and C and ratios of these nutrients. We quantified food availability via a bioassay, raising individual Aedes larvae in the laboratory in standard volumes of field-collected, particulate-containing water from each cemetery. Quantities of detritus types collected in standard containers were significant predictors of nutrients and nutrient ratios. Nutrient abundances were significant predictors of relative abundance of Ae. aegypti, and of larval survival and development by both species in the bioassay. Survival and development of larvae reared in particulate-containing water from sites decreased with decreasing relative abundance of Ae. aegypti. These data suggest that N, P, and C availabilities are determined by detritus inputs to containers and that these nutrients in turn determine the feeding environment encountered by larvae, the intensity of interspecific competition among larvae, and subsequent relative abundances of species at sites. Detritus inputs, nutrients, and food availability thus seem to contribute to distributions of Ae. aegypti and Ae. albopictus in cemetery containers throughout Florida.