Suppression of Aedes aegypti by predatory Toxorhynchites moctezuma in an island habitat

Larval populations of the mosquito Aedes aegypti were suppressed by predatory Toxorhynchites moctezuma mosquito larvae released systematically in a village on Union Island (Saint Vincent and the Grenadines) during March-December 1988. Eggs and larvae of Tx.moctezuma were transported from Trinidad and introduced into all semi-permanent and permanent water-holding containers in the experimental village at Clifton. The semi-isolated village of Ashton served as control. Base-line Ae.aegypti indices (house, ovitrap, Breteau, cistern/tank, drum/barrel, small containers) were obtained for the two villages over a 4-month period prior to the introduction of the predatory Tx.moctezuma mosquito larvae. After sustained releases of predators for 5 months, all indices of Ae.aegypti were lower in the treated village than in the untreated village during the last 3 months of the year.     

A laboratory investigation of mosquito larval predation by Toxorhynchites moctezuma on Aedes aegypti

1. Functional responses of predatory Toxorhynchites moctezuma (Dyar & Knab) larvae feeding on Aedes aegypti (L.) larvae (Diptera: Culicidae) were found to be type II of Holling (1959) and Rogers (1972). 2. Estimates of searching rate were generally higher for later instar predators. The search rate of second instar predators declined as prey instar increased, but fourth instar Tx. moctezuma had the highest search rate for second instar Ae. aegypti. 3. Prey handling times were higher for early instar predators and late instar prey. 4. When presented with mixtures of two instars of Ae. aegypti, second instar Tx. moctezuma showed frequency independent selectivity for the early instars, whereas fourth instar predators showed frequency independent selectivity for the late instars of Ae. aegypti. There was no evidence of frequency dependent predation. Preferences appeared to be transitive. 5. Extended random predator equations, using parameters derived from the functional responses, did not adequately describe the outcome of predation in the prey mixture experiment, even when the possibility of optimal switching behaviour was accounted for.     

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.     

Biochemical and cytoimmunological evidence for the control of Aedes aegypti larval trypsin with Aea-TMOF

Trypsin and chymotrypsin-like enzymes were detected in the gut of Aedes aegypti in the four larval instar and pupal developmental stages. Although overall the amount of trypsin synthesized in the larval gut was 2-fold higher than chymotrypsin, both enzymes are important in food digestion. Feeding Aea-Trypsin Modulating Oostatic Factor (TMOF) to Ae. aegypti and Culex quinquefasciatus larvae inhibited trypsin biosynthesis in the larval gut, stunted larval growth and development, and caused mortality. Aea-TMOF induced mortality in Ae. aegypti, Cx. quinquefasciatus, Culex nigripalpus, Anopheles quadrimaculatus, and Aedes taeniorhynchus larvae, indicating that many mosquito species have a TMOF-like hormone. The differences in potency of TMOF on different mosquito species suggest that analogues in other species are similar but may differ in amino acid sequence or are transported differently through the gut. Feeding of 29 different Aea-TMOF analogues to mosquito larvae indicated that full biological activity of the hormone is achieved with the tetrapeptide YDPA. Using cytoimmunochemical analysis, intrinsic TMOF was localized to ganglia of the central nervous system in larvae and male and female Ae. aegypti adults. The subesophageal, thoracic, and abdominal ganglia of both larval and adult mosquitoes contained immunoreactive cells. Immunoreactive cells were absent in the corpus cardiacum of newly molted 4th instar larvae but were found in late 4th instar larvae. In both males and females, the intrinsic neurosecretory cells of the corpus cardiacum were filled with densely stained immunoreactive material. These results indicate that TMOF-immunoreactive material is synthesized in sugar-fed male and female adults and larvae by the central nervous system cells.     

Occurrence of Toxorhynchites guadeloupensis (Dyar Knab) in oviposition trap of Aedes aegypti (L.) (Diptera: Culicidae)

Toxorhynchites guadeloupensis (Dyar Knab), a poorly known mosquito species, was observed preying upon Aedes aegypti (L.) larvae, in an oviposition trap placed for routine dengue entomological surveillance, during 2003-2004 in the urban area of Boa Vista, Roraima, Brazil. This is the first report for Tx. guadeloupensis using Ae. aegypti oviposition traps as breeding places. This finding may have important consequences in the epidemiology and local dengue control since Ae. aegypti density is a basic variable in dengue prediction. Whether predation of Ae aegypti by Tx. guadeloupensis in the Amazon is of significance, is a question to be examined. Also, larval predation may be a cause for underestimation of the actual Ae aegypti numbers. Together these hypotheses need to be better investigated as they are directly related to dengue epidemiology, to the success of any outbreak prediction and surveillance program.     

Oviposition habitat preferences of Toxorhynchites moctezuma mosquitoes in four types of tropical forest in Trinidad

1. Oviposition of the mosquito Toxorhynchites moctezuma Dyar & Knab was investigated in four types of tropical forest in Trinidad, West Indies, using surrogate and natural ovitraps. Larvae of Tx. moctezuma are obligate predators that might be useful for the biological control of Aedes aegypti (L). 2. Significantly more oviposition occurred in seasonal-deciduous forest than in either montane or evergreen-seasonal forest. 3. Oviposition in surrogate containers (black-painted polystyrene cups, 90 mm diameter) was compared with that occurring in typical natural containers (nutpots of Lecythis zapucajo Aublet). Surrogate ovipots were relatively insensitive indicators of oviposition activity, and would be an inefficient means of harvesting Tx. moctezuma eggs. 4. Implications for the collection, culture and mass release of Tx. moctezuma are discussed.     

Laboratory experiments on factors affecting oviposition site selection in Toxorhynchites amboinensis (Diptera: Culicidae), with a report on the occurrence of egg cannibalism

Laboratory experiments tested the effects of water quality and the presence of conspecific and heterospecific immatures on oviposition by Toxorhynchites amboinensis (Doleschall). The females showed a highly significant preference for oak leaf infusion water over distilled water. When twenty starved third and fourth instar Tx.amboinensis larvae were present in the water, substantially fewer eggs were counted from pots containing these conspecifics, than from controls in which no larvae were present. Numbers of eggs from pots containing starved second instar larvae did not differ significantly from controls. Observations of larval behaviour while oviposition was occurring suggested that egg numbers were reduced in containers because of egg cannibalism with third and fourth instar larvae, and not because the larvae caused a deterrent effect. Subsequent experiments confirming the occurrence of substantial egg cannibalism by third and especially fourth instar larvae are described. As with larvae, the presence of Tx.amboinensis pupae in the water had little effect on oviposition. If placed in the water 24 h prior to test, pupae very slightly enhanced its attractiveness, but if introduced immediately before test there appeared to be a slight deterrent effect. With heterospecific larvae, twenty fourth instar Ae.aegypti larvae introduced into infusion water 24 h before test rendered the water slightly attractive, while water in which Ae.aegypti larvae had been reared for 48 h proved highly repellent.     

Physiological and morphological aspects of Aedes aegypti developing larvae: effects of the chitin synthesis inhibitor novaluron

Population control of the dengue vector mosquito, Aedes aegypti, is difficult due to many reasons, one being the development of resistance to neurotoxic insecticides employed. The biosynthesis of chitin, a major constituent of insect cuticle, is a novel target for population control. Novaluron is a benzoylphenylurea (BPU) that acts as a chitin synthesis inhibitor, already used against mosquitoes. However, information regarding BPU effects on immature mosquito stages and physiological parameters related with mosquito larval development are scarce. A set of physiological parameters were recorded in control developing larvae and novaluron was administered continuously to Ae. aegypti larvae, since early third instar. Larval instar period duration was recorded from third instar until pupation. Chitin content was measured during third and fourth instars. Fourth instars were processed histochemically at the mesothorax region, stained with hematoxylin and eosin (HE) for assessment of internal tissues, and labeled with WGA-FITC to reveal chitinized structures. In control larvae: i) there is a chitin content increase during both third and fourth instars where late third instars contain more chitin than early fourth instars; ii) thoracic organs and a continuous cuticle, closely associated with the underlying epidermis were observed; iii) chitin was continuously present throughout integument cuticle. Novaluron treatment inhibited adult emergence, induced immature mortality, altered adult sex ratio and caused delay in larval development. Moreover, novaluron: i) significantly affected chitin content during larval development; ii) induced a discontinuous and altered cuticle in some regions while epidermis was often thinner or missing; iii) rendered chitin cuticle presence discontinuous and less evident. In both control and novaluron larvae, chitin was present in the peritrophic matrix. This study showed quantitatively and qualitatively evidences of novaluron effects on Ae. aegypti larval development. To our knowledge, this is the first report describing histological alterations produced by a BPU in immature vector mosquitoes.     

Parasitoid-induced behavioral alterations of Aedes aegypti mosquito larvae infected with mermithid nematodes (Nematoda: Mermithidae).

A wide range of parasites are known to cause behavioral changes in their hosts and parasitized insects are especially amenable to the study of such changes. The majority of studies addressing parasite-induced behavioral alterations have focused on parasites with complex life cycles and the adaptive nature of such changes. Behavioral changes caused by parasitoids, single-host parasites that kill their host upon emergence, have been studied less and the adaptive nature of these changes is likely to be different than those in complex life cycles. I investigated behavioral alterations in Aedes aegypti mosquito larvae infected with parasitoid nematodes (family Mermithidae). I conducted several experiments in which I tested the following hypotheses: 1) Mermithid nematodes induce behavioral changes in mosquito larvae and the changes are density dependent. 2) Different species of mermithid nematodes induce similar changes in mosquito larvae behavior. 3) Behavioral alterations vary with mermithid developmental stage. 4) Mosquito larvae infected with mermithid nematodes behave similarly to uninfected food-deprived mosquito larvae. I found that 4th instar Ae. aegypti infected with Romanomermis culicivorax or Strelkovimermis spiculatus exhibited resting behaviors significantly more often than uninfected controls but that intensity of infection did not affect activity levels. In earlier instars, infected mosquito larvae were more active than uninfected control larvae in some behaviors associated with feeding. There was no significant difference between infected and uninfected food-deprived mosquitoes in nine of the ten behaviors observed. The decrease in activity of late instar Ae. aegypti larvae infected with mermithids may be a parasitoid adaptation that reduces the risk of predation and thus increases host and parasitoid survival. The increase in feeding activity in earlier instars as well as the similarity between uninfected food-deprived and infected Ae. aegypti behavior may indicate that these behaviors are adaptive for the parasitoid, increasing nutritional acquisition for successful parasitoid development.     

Evaluation of some adhesives for collecting Musca domestica and Chrysomya megacephala adults or mosquito larvae in sticky traps

1. Seven types of water-insoluble adhesives were evaluated in sticky traps for collecting adults of Musca domestica L. and Chrysomya megacephala (Fabricius) or mosquito larvae (Aedes aegypti (L.) and Culex quinquefasciatus Say). 2. Adhesive viscosity affected the tackiness of the glues and this determined their trapping efficiency in air or water. 3. From the 'Hyvis' range of adhesives tested, 'Hyvis 200' was most effective for trapping adult flies. 4. With 24 h exposure to fourth instar Ae.aegypti larvae in tapwater, submerged plates coated with 'Hyvis 10', 'Hyvis 30' or 'Hyvis 200' formulations trapped the majority of larvae. In polluted water the highest rates of trapping were 17.3% of Ae.aegypti and 18.7% of Cx quinquefasciatus with 'Hyvis 200'. Floating traps were consistently less productive than submerged traps under laboratory conditions. 5. In a heavily polluted natural breeding-site of Cx quinquefasciatus, floating traps were more productive than submerged sticky traps with four of seven adhesives tested, the most efficient being 'Hyvis 200' (4.2 mosquitoes per hour) and Hyvis:polyethylene 90:10 (4.5/h). Despite the relative inefficiency of aquatic traps, emergent adults, pupae and second to fourth instars of larvae were collected quickly from the habitat.     

Biological control of Culicidae with the copepod Mesocyclops aspericornis and larvivorous fish (Poeciliidae) in a village of French Polynesia

The copepod Mesocyclops aspericornis Daday and the larvivorous fishes Gambusia affinis (B. & G.) and Poecilia reticulata R. & B., were released into mosquito breeding sites in Tuherahera village, Tikehau atoll, French Polynesia, to control larvae of Aedes aegypti (L.), Ae.polynesiensis Marks, Culex annulirostris Skuse and Cx quinquefasciatus Say. Treatments were completed within a week, in January 1990. Fish quickly eliminated mosquito larvae from the open breeding sites (ponds, wells). The impact of copepods in water tanks, drums and covered wells was inconsistent, apparently depending on the availability of microfaunal diet for growth of copepod nauplii. As the biting rate of adult Ae.aegypti seemed to be unaffected by the biological control of larvae, this village-scale experiment was judged to be unsuccessful as a means of vector control.     

Larvicide and oviposition deterrent effects of fruit and leaf extracts from Melia azedarach L. on Aedes aegypti (L.) (Diptera: Culicidae)

Aedes aegypti (L.) (Diptera: Culicidae), the main urban vector of dengue, has developed resistance to various insecticides, making its control increasingly difficult. We explored the effects of Argentine Melia azedarach L. (Meliaceae) fruit and senescent leaf extracts on Ae. aegypti larval development and survival, by rearing cohorts of first instar mosquitoes in water with different extract concentrations. We also analysed oviposition deterrent activity in choice tests with extract-treated ovitraps. The leaf extract showed a strong larvicide activity, with all larvae dying before pupation, and significantly delayed development time. It strongly inhibited oviposition by Ae. aegypti females. The fruit extract showed much weaker effects. This first report of highly effective larvicidal, growth regulating and oviposition deterrent activity of a senescent leaf extract of M. azedarach against Ae. aegypti, suggests that such extract could represent a promising tool in the management of this mosquito pest.     

Role of gregarine parasite Ascogregarina culicis (Apicomplexa: Lecudinidae) in the maintenance of Chikungunya virus in vector mosquito

Ascogregarina culicis and Ascogregarina taiwanensis are common gregarine parasites of Aedes aegypti and Aedes albopictus mosquitoes, respectively. These mosquito species are also known to transmit dengue and Chikungunya viruses. The sporozoites of these parasites invade the midgut epithelial cells and develop intracellularly and extracellularly in the gut to complete their life cycles. The midgut is also the primary site for virus replication in the vector mosquitoes. Therefore, studies were carried out with a view to determine the possible role of these gregarines in the vertical transmission of dengue and Chikungunya viruses from larval to adult stage. Experiments were performed by exposing first instar mosquito larvae to suspensions containing parasite oocysts and viruses. Since Ascogregarina sporozoites invade the midgut of first instar larvae, the vertical transmission was determined by feeding the uninfected first instar larvae on the freshly prepared homogenates from mosquitoes, which were dually infected with viruses and the parasite oocysts. Similarly, the role of protozoan parasites in the vertical transmission of viruses was determined by exposing fresh first instar larvae to the dried pellets of homogenates prepared from the mosquitoes dually infected with viruses and the parasite oocysts. Direct vertical transmission and the vertical transmission of CHIK virus through the oocyst of the parasites were observed in the case of Ae. aegypti mosquitoes. It is suggested that As. culicis may have an important role in the maintenance of CHIK virus during the inter-epidemic period.     

Surveillance of the mosquito Aedes aegypti and its biocontrol with the copepod Mesocyclops aspericornis in Australian wells and gold mines

Abstract. A survey of the dengue vector mosquito Aedes aegypti was undertaken using runnel traps to detect immature stages (larvae and pupae) in flooded disused mine shafts and wells in Charters Towers, Queensland, Northern Australia. The town has a history of dengue fever since 1885 when goldminers were the first recorded victims. During the latest dengue epidemic in 1993, 2% of the population had laboratory-confirmed dengue virus Type 2, despite source reduction of Ae.aegypti breeding-sites at ground level or above. This led to suspicions that dengue vector Ae.aegypti breeding-sites might be below ground level. When surveyed in March 1994, Ae.aegypti immatures were found in 9/10 wells and 1/6 mine shafts. The water in wells and mines had similar characteristics -except that turbidity was higher in the mines, which more often contained predators of mosquito immatures.
The copepod Mesocyclops aspericornis was collected from water in 1/10 wells and 2/6 mine shafts. Laboratory predation trials resulted in 95.5–100% predation by 25 copepods/1 on Ae.aegypti first-instar larvae up to 200 larvae/1. Five wells containing Ae.aegypti in the survey were inoculated with fifty indigenous M.aspericornis , and five wells (one positive and four negative in the survey) were left untreated as controls. Nine months later, in December 1994, Ae.aegypti had been eliminated from all five treated wells but all untreated control wells contained Ae.aegypti , except for one well that contained a natural population of M.aspericornis. The role of wells and mines as winter/ dry season refuges of Ae.aegypti in northern Australia is reviewed, and we recommend the use of M.aspericornis as a cost-effective, environmentally acceptable and persistent agent for the sustainable control of Ae.aegypti , especially in inaccessible breeding sites.     

Susceptibility of ten species of mosquito larvae to the parasitic nematode Romanomermis iyengari and its development

Ten species of mosquitoes (Diptera: Culicidae) from five genera were exposed to preparasites of the tropical mermithid nematode species Romanomermis iyengari (Welch) (Nematoda: Mermithidae), a strain isolated in 1978 from Pondicherry. By exposing mosquito larvae during the second instar, nematode infection was invariably lethal, the rate being highest in Culex sitiens Wiedemann (95%) followed by Cx. quinquefasciatus Say (90%), Aedes aegypti (L.) (79%), Anopheles subpictus Grassi (64%), Ae. albopictus (Skuse) and Armigeres subalbatus Coquillett (62%), Cx. tritaeniorhynchus Giles (57%), Mansonia annulifera (Theobald) (46%), An. stephensi Liston (40%) and An. culicifacies Giles (36%). When fourth-instar larvae were exposed, the infection was highest in Ar. subalbatus (66%), followed by An. stephensi (52%), Cx. quinquefasciatus (47%), Ae. aegypti and An. subpictus (42%), Ae. albopictus (30%), An. culicifacies (29%), Cx. sitiens (24%), Cx. tritaeniorhynchus (19%) and Ma. annulifera (8%), with 2-45% of infected culicines surviving to adulthood. The parasitic phase of the nematode lasted 5-7 days in all the host species, yielding 1.1-3.2 parasites per II instar and 1.1-2.5 parasites per IV instar. The overall output of parasites per 100 mosquito larvae (infected + uninfected) was highest for Ae. aegypti when mosquitoes were exposed during II instar (2.53 parasites/larva) and for Ar. subalbatus when mosquitoes were exposed during IV instar (1.65/larva), and lowest for Ma. annulifera exposed during IV instar (0.09/larva). For routine laboratory culture of R. iyengari it is convenient to employ Cx. quinquefasciatus as the host yielding 90-190 parasites/100 larva.     

Chemical composition and anti-mosquito potential of rhizome extract and volatile oil derived from Curcuma aromatica against Aedes aegypti (Diptera: Culicidae).

Crude rhizome extracts and volatile oils of Curcuma aromatica were evaluated for chemical composition and anti-mosquito potential, including larvicidal, adulticidal, and repellent activities against the Aedes aegypti mosquito. Chemical identification achieved by GC/MS analysis revealed that xanthorrhizol, 1H-3a, 7-methanoazulene and curcumene at 35.08 and 13.65%, 21.81 and 30.02%, and 13.75 and 25.71%, were the main constituents in hexane extracts and volatile oils, respectively. Volatile oil of Cu. aromatica possessed a significantly higher larvicidal activity against the 4th instar larvae of Ae. aegypti than that of hexane extracts, with LC50 values of 36.30 and 57.15 ppm, respectively. In testing for adulticidal activity, on the other hand, hexane-extracted Cu. aromatica (LC50: 1.60 microg/mg female) was found to be slightly more effective against female Ae. aegypti than volatile oil (LC50: 2.86 microg/mg female). However, the repellency of these two products against Ae. aegypti adult females differed significantly. The hexane-extracted Cu. aromatica, with a median complete protection time of 1 h (range = 1-1.5 h) when applied at a concentration of 25%, appeared to have significantly higher repellency than that of distillate oil (0.5 h, range = 0-0.5 h). The different results obtained from both products of Cu. aromatica were probably due to variety in quantity and type of active ingredients as well as the biological and physiological characteristics that differed between both developmental stages of mosquitoes, larvae, and adults.     

The first report of Aedes (Stegomyia) albopictus in Haiti

Aedes albopictus was found in six of the 10 departments of Haiti and in 14 of the 35 communes surveyed. The survey found the larvae of Ae. albopictus in 13 different types of containers. Used tires and tins were by far the most common breeding sites used by this mosquito species. At the breeding sites, Ae. albopictus was associated with other mosquito species, such as Aedes aegypti, Culex nigripalpus and Aedes mediovittatus. The highest proportion of association was with Ae. aegypti. This study represents the first report of Ae. albopictus in Haiti.     

Evaluations of larvicidal activity of medicinal plant extracts to Aedes aegypti （Diptera： Culicidae） and other effects on a non target fish

A preliminary study was conducted to investigate the effects of the extracts of 112 medicinal plant species, collected from the southern part of Thailand, on Aedes aegypti. Studies on larvicidal properties of plant extracts against the fourth instar larvae revealed that extracts of 14 species showed evidence of larvicidal activity. Eight out of the 14 plant species showed 100% mosquito larvae mortality. The LC50 values were less than 100μg/mL （4.1μg/ mL-89.4μg/mL）. Six plant species were comparatively more effective against the fourth instar larvae at very low concentrations. These extracts demonstrated no or very low toxicity to guppy fish （Poecilia reticulata）, which was selected to represent most common non-target organism found in habitats ofAe. aegypti, at concentrations active to mosquito larvae. Three medicinal plants with promising larvicidal activity, having LC50 and LC50 values being 4.1 and 16.4 μg/mL for Mammea siamensis, 20.2 and 34.7 μg/mL forAnethum graveolens and 67.4 and 110.3μg/mL forAnnona muricata, respectively, were used to study the impact of the extracts on the life cycle ofAe. aegypti. These plants affected pupal and adult mortality and also affected the reproductive potential of surviving adults by reducing the number of eggs laid and the percentage of egg hatchability. When each larval stage was treated with successive extracts at the LC50 value, the first instar larvae were found to be very susceptible to A. muricata and the second instar larvae were found to be susceptible to A. graveolens, while the third and fourth instar larvae were found to be susceptible to M. siamensis. These extracts delayed larval development and inhibited adult emergence and had no adverse effects on P. reticulata at LC50 and LC50 values, except for the M. siamensis extract at its LC50 value.     

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.     

Mosquito larval consumption of toxic arborescent leaf-litter,and its biocontrol potential

Previously we described the mosquito larvicidal properties of decomposed leaf-litter from deciduous trees, especially the alder Alnus glutinosa (L) Gaertn., due to toxic polyphenols and other secondary compounds. To further examine the biocontrol potential of toxic leaf-litter for mosquito control, feeding rates of third-instar mosquito larvae were assessed for examples of three genera: Anopheles stephensi Liston, Aedes aegypti (L) and Culex pipiens L. (Diptera: Culicidae). When immersed in a suspension of non-toxic leaf-litter particles (approximately 0.4 mm), pre-starved larvae of all three species ingested sufficient material in 30 min to fill the anterior gut lumen (thorax plus two to three abdominal segments). Gut filling peaked after 1-2 h ingestion time, filling the intestine up to six to seven abdominal segments for Ae. aegypti, but maxima of five abdominal segments for Cx. pipiens and An. stephensi. Using three methods to quantify consumption of three materials by third-instar larvae of Ae. aegypti, the average amount of leaf-litter (non-toxic 0.4 mm particles) ingested during 3 h was determined as approximately 20 microg/larva (by dry weight and by lignin spectrophotometric assay). Consumption of humine (approximately 100 microm particles extracted from leaf-litter) during 3 h was approximately 80 microg/larva for Ae. aegypti, but only approximately 30 microg/larva for Cx. pipiens and 15 microg/larva for An. stephensi, with good concordance of determinations by dry weight and by radiometric assay. Cellulose consumption by Ae. aegypti was intermediate: approximately 40 microg/larva determined by radiometric assay. Apparent differences between the amounts of these materials ingested by Ae. aegypti larvae (humine four-fold, cellulose two-fold more than leaf-litter) may be attributed to contrasts in palatability (perhaps related to particle size or form), rather than technical discrepancies, because there was good concordance between results of both methods used to determine the amounts of humine and leaf-litter ingested. Bioassays of toxic leaf-litter (decomposed 10 months) with 4-h exposure period (ingestion time) ranked the order of sensitivity: Ae. aegypti (LC50 < 0.03 g/L) > An. stephensi (LC50 = 0.35 g/L) > Cx. pipiens (LC20 > 0.4 g/L). When immersed in the high concentration of 0.5 g/L toxic leaf-litter (0.4 mm particles), as little as 15-30 min ingestion time (exposure period) was sufficient to kill the majority of larvae of all three species, as soon as the gut lumen was filled for only the first few abdominal segments. Possibilities for mosquito larval control with toxic leaf-litter products and the need for standardized ingestion bioassays of larvicidal particles are discussed.