Cost of mating and insemination capacity of a genetically modified mosquito Aedes aegypti OX513A compared to its wild type counterpart

The idea of implementing genetics-based insect control strategies modelled on the traditional SIT is becoming increasingly popular. In this paper we compare a genetically modified line of Aedes aegypti carrying a tetracycline repressible, lethal positive feedback system (OX513A) with its wild type counterpart with respect to their insemination capacities and the cost of courtship and mating. Genetically modified males inseminated just over half as many females as the wild type males during their lifetime. Providing days of rest from mating had no significant effect on the total number of females inseminated by males of either line, but it did increase their longevity. Producing sperm had a low cost in terms of energy investment; the cost of transferring this sperm to a receptive female was much higher. Continued mating attempts with refractory females suggest that males could not identify refractory females before investing substantial energy in courtship. Although over a lifetime OX513A males inseminated fewer females, the number of females inseminated over the first three days, was similar between males of the two lines, suggesting that the identified cost of RIDL may have little impact on the outcome of SIT-based control programmes with frequent releases of the genetically modified males.     

Predictable modification of body size and competitive ability following a host shift by a seed beetle

Interfertile populations of the seed beetle Callosobruchus maculatus differ genetically in several behavioral, morphological, and life-history traits, including traits that affect the intensity of larval competition within seeds. Previous studies have suggested that this variation depends on differences in host size. I performed a selection experiment in which replicate beetle lines were either maintained on a small, ancestral host (mung bean) or switched to a larger, novel host (cowpea). After 40 generations, I estimated survival, development time, and adult mass on each host, both in the presence and absence of larval competition. The shift to cowpea substantially reduced body size; irrespective of rearing host, adults from the cowpea lines were more than 10% lighter than those from the mung bean lines. Switching to cowpea also improved survival and reduced development time on this host, but without decreasing performance on the ancestral host. The most striking effect of the shift to a larger host was a reduction in larval competitiveness. When two even-aged larvae co-existed within a seed, the probability that both survived to adult emergence was > or = 65% if larvae were from the cowpea lines but < or = 12% if they were from the mung bean lines. The adverse effects of competition on development time and adult mass were also less severe in the cowpea lines than in the mung bean lines. By rapidly evolving smaller size and reduced competitiveness, the cowpea lines converged toward populations chronically associated with cowpea. These results suggest that evolutionary trajectories can be predictable, and that host-specific selection can play a major role in the diversification of insect life histories. Because host shifts by small, endophagous insects are comparable to the colonization of new habitats, adaptive responses may often include traits (such as larval competitiveness) that are not directly related to host use.     

Studies on Aedes albopictus larval mass-rearing optimization

To set up a sterile male technique program to control Aedes albopictus (Skuse) in areas in northern Italy, a pilot mass-rearing facility is under development. For this purpose, experiments were carried out to find the optimal larval density for the optimization of the rearing parameters, i.e., to obtain the fastest larval development, the highest larval and pupal survival rate, and large-sized pupae. Several different larval densities, from 40 to 2874 larvae per liter, were tested. For densities from 40 to 600 larvae per liter significant size differences were found among pupae obtained under different larval densities. The larvae raised at the lowest density tended to be smaller and to develop most slowly, i.e., longer pupation time. Also, increasing water volume and depth seemed to negatively affect the pupation success. Compared with the other larval densities tested, the larvae reared at a density of 2874 larvae per liter developed slightly faster and showed higher survival rates, indicating this density as appropriate for the development of a mass rearing, at least using the current larval diet.     

Open Field Release of Genetically Engineered Sterile Male Aedes aegypti in Malaysia

Background

Dengue is the most important mosquito-borne viral disease. In the absence of specific drugs or vaccines, control focuses on suppressing the principal mosquito vector, Aedes aegypti, yet current methods have not proven adequate to control the disease. New methods are therefore urgently needed, for example genetics-based sterile-male-release methods. However, this requires that lab-reared, modified mosquitoes be able to survive and disperse adequately in the field.

Methodology/Principal Findings

Adult male mosquitoes were released into an uninhabited forested area of Pahang, Malaysia. Their survival and dispersal was assessed by use of a network of traps. Two strains were used, an engineered ‘genetically sterile’ (OX513A) and a wild-type laboratory strain, to give both absolute and relative data about the performance of the modified mosquitoes. The two strains had similar maximum dispersal distances (220 m), but mean distance travelled of the OX513A strain was lower (52 vs. 100 m). Life expectancy was similar (2.0 vs. 2.2 days). Recapture rates were high for both strains, possibly because of the uninhabited nature of the site.

Conclusions/Significance

After extensive contained studies and regulatory scrutiny, a field release of engineered mosquitoes was safely and successfully conducted in Malaysia. The engineered strain showed similar field longevity to an unmodified counterpart, though in this setting dispersal was reduced relative to the unmodified strain. These data are encouraging for the future testing and implementation of genetic control strategies and will help guide future field use of this and other engineered strains.     

THE RESPONSE TO SELECTION FOR FAST LARVAL DEVELOPMENT IN DROSOPHILA MELANOGASTER AND ITS EFFECT ON ADULT WEIGHT: AN EXAMPLE OF A FITNESS TRADE-OFF

A selection experiment using Drosophila melanogaster revealed a strong trade-off between adult weight and larval development time (LDT), supporting the view that antagonistic pleiotropy for these two fitness traits determines mean adult size. Two experimental lines of flies were selected for a shorter LDT (measured from egg laying to pupation). After 15 generations LDT was reduced by an average of 7.9%. The response appeared to be controlled primarily by autosomal loci. A correlated response to the selection was a reduction in adult dry weight: individuals from the selected populations were on average 15.1% lighter than the controls. The lighter females of the selected lines showed a 35% drop in fecundity, but no change in longevity. Thus, there is no direct relationship between LDT and adult longevity. The genetic correlation between weight and LDT, as measured from their joint response to selection, was 0.86. Although there was weak evidence for dominance in LDT, there was none for weight, making it unlikely that selection acting on this antagonistic pleiotropy could lead to a stable polymorphism. In all lines, sex differences in weight violated expectations based on intrasex genetic correlations: Females, being larger than males, ought to require a longer LDT, whereas there was a slight trend in the opposite direction. Because the sexual dimorphism in size was not significantly altered by selection, it appears that the controlling loci are either invariant or have very limited pleiotropic effect on developmental time. It is suggested that they probably control some intrinsic, energy-intensive developmental process in males.     

Food availability and parasite infection influence the correlated responses of life history traits to selection for age at pupation in the mosquito Aedes aegypti

We selected six lines of mosquito Aedes aegypti for earlier or later pupation and measured the correlated responses of several life history traits: adult size, two fecundity measures and pre-adult survival. We further examined the influence of two environmental parameters – larval food availability and infection by the microsporidian parasite Edhazardia aedis– on the correlated responses. Pre-adult survival did not respond to selection for age at pupation in any environment. For all of the other traits, the environment influenced the correlated response, though the contribution of the different environmental aspects differed among traits. While the correlated response of adult size was influenced only by larval food availability, the likelihood that a female laid eggs was influenced by parasite infection, and the correlated response of the number of eggs was influenced by the interaction of the two environmental parameters. Generally, a deteriorating environment moved the correlated response from one favouring later pupation to one favouring earlier pupation. Larval food availability and parasite infection also influenced the association between the mean wing length and fecundity of the selected lines. At high food availability, there was a positive relationship between adult size and fecundity, while infected mosquitoes reared at low food availability showed the opposite trend. We discuss these results in light of the coevolutionary potential of the host–parasite interaction.     

The effects of selection for larval behavior on adult life-history features in Drosophila melanogaster

Selection for late-life fecundity and longevity in adult Drosophila melanogaster is well known to modify numerous characteristics of life history and physiology. We report experiments here in which selection applied to behavior affects features in an identical fashion. Selection for feeding rate of larval D. melanogaster modifies caloric intake, as measured by the uptake and incorporation of labeled glucose. Selection for slow larval feeding produced lines of D. melanogaster in which larvae synthesized significantly less lipid prior to pupation and eclosed to have low early-life fecundity and a long life as adults. They also had greater lifetime fecundity, but lower viability of egg to hatched adult. Alternatively, fast-feeding larvae incorporated more lipid before pupation and eclosed with high early-fecundity that declined rapidly throughout their short adult life. Slow-feeding populations also had a significantly enhanced expression of the stress-resistance genes CuZn-SOD, CATALASE, and HSP70. Selection on larval feeding behavior reproduced the antagonistic evolutionary trade-off found under selection for adult life span and mimicked the physiological response in life span as seen in many species when dietary restriction is imposed on adults. Thus, nutrient acquisition during development appears to share a common evolutionary and genetic basis with the allocation processes that determine adult life-history traits and the related phenotypic dietary restriction phenomena.     

Age and size thresholds for pupation and developmental polymorphism in the browntail moth, Euproctis chrysorrhoea (Lepidoptera: Lymantriidae), under conditions that either emulate diapause or prevent it

Size and age thresholds for pupation are important life history traits of insects. They are the ultimate consequences of the underlying physiological mechanism that optimize resource allocation. Such thresholds may have a plastic response under time-varying environmental conditions, developmental polymorphism (i.e., plasticity in the number of instars before pupation) being a common strategy adopted by insects to overcome this challenging situation. In this study, we systematically explore the variables related with both age and size thresholds for pupation and developmental polymorphism in the browntail moth, Euproctis chrysorrhoea (Lepidoptera: Lymantriidae), by rearing a group of caterpillars under conditions that either emulate larval diapause or prevent it. As an innovative approach, we evaluated the importance of predictor variables by means of generalized linear modeling in a multi-model inference framework. Our results show that (i) rearing conditions affect fitness, (ii) rearing conditions, size of hatchlings, size and age at maturity and sex are related to the number of instars before pupation, and (iii) there are both age and size thresholds for pupation which differ between sexes and between larvae reared under different conditions. Results are discussed in the context of lepidopteran plasticity in life history traits and its relationship with optimal molting strategies.     

Maternal effects and their consequences for offspring fitness in the Yellow Dung Fly

1. Maternal adult diet and body size influence the fecundity of a female and possibly the quality and the performance of her offspring via egg size or egg quality. In laboratory experiments, negative effects in the offspring generation have often been obscured by optimal rearing conditions.
2. To estimate these effects in the Yellow Dung Fly, Scathophaga stercoraria , how maternal body size and adult nutritional status affected her fecundity, longevity and egg size were first investigated.
3. Second, it was investigated how female age and adult nutritional experience, mediated through the effects of egg size or egg quality, influenced the performance of offspring at different larval densities.
4. Maternal size was less important than maternal adult feeding in increasing reproductive output. Without food restriction, large females had larger clutch sizes and higher oviposition rates, whereas under food restriction this advantage was reversed in favour of small females.
5. Offspring from mothers reared under nutritional stress experienced reduced fitness in terms of egg mortality and survival to adult emergence. If the offspring from low-quality eggs survived, the transmitted maternal food deficiency only affected adult male body size under stressful larval environments.
6. Smaller egg sizes due to maternal age only slightly affected the performance of the offspring under all larval conditions.     

Selection for adult desiccation resistance in Drosophila melanogaster: fitness components, larval resistance and stress correlations

In previous experiments we found that Drosophila melanogaster lines selected for increased adult desiccation resistance had increased resistance to other environmental stresses at the adult stage including starvation, intense ⁶⁰Co-γ radiation and a toxic ethanol level. In further studies on these lines, we now show that selection did not alter resistance to desiccation and ethanol at the larval stage. As well as having a lower early fecundity, selected lines showed increased adult male longevity and increased viability at high larval densities compared with control lines. There were no changes in development time or mating success. The increased male longevity is consistent with the reduced metabolic rate of the selected lines.
A genetic correlation between resistance to different stresses was confirmed by an analysis of isofemale lines derived from a population founded by flies from a stress-resistant line and an unselected line. The results are consistent with the existence of genes segregating in natural populations conferring increased general stress resistance.     

Evolutionary responses of Drosophila melanogaster life history to differences in larval density

The study examined the effects of evolution at two different larval densities on pre-adult and adult fitness traits. Five replicate selection lines each were cultured at either 50 or 150 larvae per vial, avoiding selection on development time, age at breeding or for adaptation to adult density, one or more of which factors has been a confounding variable in previous studies. Low density selection lines evolved extended development times at both growth densities. The extended development times were associated with greater adult body size at the lower growth density only, and particularly in females. The lines did not differ significantly in larval competitive ability at either growth density. At neither growth density did the early adult fertility of females or the lifespan of either sex differ between the lines from the two selection regimes, but at the lower growth density the late fertility of low density line females was significantly enhanced. The results suggest that larval density does have important effects on the expression and resolution of life history trade-offs in Drosophila melanogaster, but that these may be somewhat different from those reported in previous studies.     

六种昆虫生长调节剂对葱蝇生长发育和繁殖力的影响

用灭蝇胺、氟铃脲等6种昆虫生长调节剂和对比药剂辛硫磷LC_40-50浓度处理葱蝇1龄和3龄幼虫,观察对其生长发育和繁殖的影响;并且研究了葱蝇成虫取食药剂对其繁殖力的控制效应。结果表明：6种药剂中除虫酰肼外,其余都对葱蝇幼虫发育历期、化蛹率、成虫产卵前期、成虫寿命、产卵量有很大影响,对其生长发育和繁殖表现出显著不利性。其中灭蝇胺和氟铃脲处理1龄幼虫后,存活幼虫的历期和成虫产卵前期各延长3天,化蛹率降低26.7%～30.0％,成虫寿命缩短22.3～24.3天,产卵量降低35.5％～49.9％;处理3龄幼虫主要导致产生畸形蛹,且正常羽化的成虫寿命降低18.0～20.67天,产卵量降低45.7%～57.8％。成虫连续取食用牛奶稀释的２ 000倍的药液,能导致产卵前期延长、产卵量减少、卵孵化率降低,幼虫死亡率极高,达95%以上。结果提示,利用成虫补充营养的特性,将灭蝇胺和氟铃脲等混入牛奶或糖水中诱集成虫取食,可有效降低成虫的繁殖力,有助于实现对葱蝇的无公害治理。     

No impact of transgenic nptII-leafy Pinus radiata (Pinales: Pinaceae) on Pseudocoremia suavis (Lepidoptera: Geometridae) or its endoparasitoid Meteorus pulchricornis (Hymenoptera: Braconidae)

To investigate the biosafety to insects of transgenic Pinus radiata D. Don containing the antibiotic resistance marker gene nptII and the reproductive control gene leafy, bioassays were conducted with an endemic lepidopteran pest of New Zealand plantation pine forests and a hymenopteran endoparasitoid. Larvae of the common forest looper, Pseudocoremia suavis (Butler), were fed from hatching on P. radiata needles from either one of two nptII-leafy transgenic clones, or an isogenic unmodified control line. For both unparasitized P. suavis and those parasitized by Meteorus pulchricornis (Wesmael), consuming transgenic versus control pine had no impact on larval growth rate or mass at any age, larval duration, survival, pupation or successful emergence as an adult. Total larval duration was 1 d (3%) longer in larvae fed nptII-2 than nptII-1, but this difference was considered trivial and neither differed from the control. In unparasitized P. suavis larvae, pine type consumed did not affect rate of pupation or adult emergence, pupal mass, or pupal duration. Pine type had no effect on the duration or survival of M. pulchricornis larval or pupal stages, mass of cocoons, stage at which they died, adult emergence, or fecundity. Parasitism by M. pulchricornis reduced P. suavis larval growth rate, increased the duration of the third larval stadium, and resulted in the death of all host larvae before pupation. The lack of impact of an exclusive diet of nptII-leafy transgenic pines on the life history of P. suavis and M. pulchricornis suggests that transgenic plantation pines expressing nptII are unlikely to affect insect populations in the field.     

Effect of larval rearing temperature on adult female morphology of Anopheles (Nyssorhynchus) albimanus (Diptera: Culicidae)

Summary

Time to pupation, percent survival to pupation, and percent adult emergence of Anopheles albimanus Wiedemann decreased at higher larval rearing temperature. Mosquitoes reared at 30°C experienced higher mortality during the pupal stage than did mosquitoes reared at 22°C. Analysis of variance revealed that wing length and costal wing spot patterns of adult female A. albimanus were affected by larval rearing temperature. Female A. albimanus reared at 22°C had longer wings, and larger basal pale + prehumeral pale, prehumeral dark, and humeral pale costal wing spots than did female siblings reared at 30°C. Female A. albimanus reared at 30°C had larger subcostal pale spots than did female siblings reared at 22°C. Analyses of 2x2 contingency tables indicated that sex ratio was independent of larval rearing temperature, whereas survival to the adult stage and coalescence of wing spots were not independent of rearing temperature. The need to examine stability of morphological characters under differing environmental conditions is discussed.     

Temperature dependent larval resource allocation shaping adult body size in Drosophila melanogaster

Geographical variation in Drosophila melanogaster body size is a long-standing problem of life-history evolution. Adaptation to a cold climate invariably produces large individuals, whereas evolution in tropical regions result in small individuals. The proximate mechanism was suggested to involve thermal evolution of resource processing by the developing larvae. In this study an attempt is made to merge proximate explanations, featuring temperature sensitivity of larval resource processing, and ultimate approaches focusing on adult and pre-adult life-history traits. To address the issue of temperature dependent resource allocation to adult size vs. larval survival, feeding was stopped at several stages during the larval development. Under these conditions of food deprivation, two temperate and two tropical populations reared at high and low temperatures produced different adult body sizes coinciding with different probabilities to reach the adult stage. In all cases a phenotypic trade-off between larval survival and adult size was observed. However, the underlying pattern of larval resource allocation differed between the geographical populations. In the temperate populations larval age but not weight predicted survival. Temperate larvae did not invest accumulated resources in survival, instead they preserved larval biomass to benefit adult weight. In other words, larvae from temperate populations failed to re-allocate accumulated resources to facilitate their survival. A low percentage of the larvae survived to adulthood but produced relatively large flies. Conversely, in tropical populations larval weight but not age determined the probability to reach adulthood. Tropical larvae did not invest in adult size, but facilitated their own survival. Most larvae succeeded in pupating but then produced small adults. The underlying physiological mechanism seemed to be an evolved difference in the accessibility of glycogen reserves as a result of thermal adaptation. At low rearing temperatures and in the temperate populations, glycogen levels tended to correlate positively with adult size but negatively with pupation probability. The data presented here offer an explanation of geographical variation in body size by showing that thermal evolution of resource allocation, specifically the ability to access glycogen storage, is the proximate mechanism responsible for the life-history trade-off between larval survival and adult size.     

Variations in the life-history parameters of Hemipyrellia ligurriens (Diptera: Calliphoridae) in response to larval competition for food

Abstract. 1. Larval rearing densities of Hemipyrellia ligurriens (Wiedemann) (Diptera: Calliphoridae) in standardized carrion were manipulated in order to investigate changes in life-history parameters in response to larval competition for food.
2. Competition was of the typical scramble type. Survivorship remained high at densities up to 32 larvae g liver^-1 but decreased rapidly as larval density increased further.
3. Emergent adults were undersized with reduced fecundity and longevity. Variations in adult body size apparently reduced the effects of competition on larval mortality.
4. Females of dry weight corresponding to only 10.4% of the potential maximum emerged at the highest rearing densities of 128 larvae g liver-1. However, these females had a nearly four-fold increase in reproductive investment (per unit weight) when compared to the largest individuals.
5. The duration of larval development declined when competition was intense (i.e. at high larval densities).
6. The short adult life of H.ligurriens, combined with the unpredictability of larval habitat availability, may reduce the value of long-range dispersal so that females 'do better' by maintaining reproductive investment despite a concomitant decline in dispersal ability.     

Effects of Hydrilla verticillata (L.f.) Royle (Hydrocharitaceae) growing conditions and nutrient content on the performance of a leaf-mining fly,Hydrellia purcelli Deeming (Diptera: Ephydridae)

Host plant quality for insects used in weed biological control influences their performance and hence their ability to suppress target host populations. Determining the specific response of these insects to the quality of their host is important because phytophagous insects have variable tolerances of the different constituents of host plant quality, most notably dietary nitrogen (N), but also other physical and chemical components. The invasive aquatic weed Hydrilla verticillata (L.f.) Royle (Hydrocharitaceae) was cultivated under varying nutrient conditions to determine the influence of plant quality on immature survival, development, larval mining, reproductive output and adult longevity of a leaf-mining fly Hydrellia purcelli Deeming (Diptera: Ephydridae). Additionally, field-collected H. verticillata was included in the investigations to assess the potential performance of H. purcelli in the field. Variation in plant tissue N and phosphorus (P) concentrations had no effects on larval survival, female fecundity, or adult longevity, but high levels of N and P were associated with reduced immature development times and higher body mass of females. Overall, plant quality factors not measured in this study appeared to have a greater impact on the performance of the fly, rather than dietary N and P. The results provided insights into optimal mass-rearing conditions for H. purcelli and the potential performance of the fly in the field in South Africa. Furthermore, the results demonstrate the importance of considering other aspects of plant quality for insect agents, in addition to dietary N and P, when developing mass-rearing protocols or predicting their potential impact in the field.     

Dietary wheat germ oil and age influences fatty acid compositions in adult oriental fruit flies

The oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) is a serious pest of fruit crops in Asia, several Pacific Islands and sometimes the western United States, particularly California. Sterile insect technique programs have been developed for management of several tephritid fruit fly pests. These programs are based on continuous production of adult fruit flies. The high expense of mass-rearing oriental fruit flies drives research to improve the cost effectiveness of rearing programs. One recent improvement for mass rearing oriental fruit flies involves adding wheat germ oil (WGO) to the larval culture medium, which improved several parameters of biological performance. The performance enhancing influence of WGO is due to the presence of polyunsaturated fatty acids (PUFAs), some of which are nutritionally essential for many insect species. We considered the issue of whether WGO supplementation of the larval culture medium influences the fatty acid make up of adult tissues. We report that WGO supplementation led to substantial increases in adult tissue C18 PUFAs. Unlike the outcomes of unrelated nutritional studies on moths, the PUFA components of WGO did not improve adult fruit fly performance. Taken with recent publications reporting that WGO in larval diets influences gene expression, we conclude that dietary WGO improves biological performance of adults through changes in tissue C18 PUFAs and gene expression.     

A semi-synthetic diet for rearing Dipha aphidivora (Lepidoptera: Pyralidae), a promising predator of woolly aphid in sugarcane

Dipha aphidivora (Meyrick), a lepidopteran predator of sugarcane woolly aphid (SWA) Ceratovacuna lanigera Zehntner was successfully reared for three successive generations, for the first time on a freeze-dried beef liver-based larval semi-synthetic diet. We compared biological parameters viz., larval survival, adult weight, pre-and oviposition period, fertility, fecundity and female longevity of D. aphidivora reared on the semi-synthetic diet with the predators reared on SWA. Development time of larvae reared from first instar to pupation was 20.6 days on the semi-synthetic diet and 12 days on SWA, while survival of the larvae to adults was 61.8 and 91.8% on larval semi-synthetic diet and SWA, respectively. Fecundity recorded from semi-synthetic diet (41 eggs/female) was significantly less than those produced on SWA (58 eggs/female). However, fertility and longevity of the predators reared on SWA and semi-synthetic diet did not differ significantly. The study revealed the possibility of rearing D. aphidivora larvae using synthetic diet.     

Evolution of increased adult longevity in Drosophila melanogaster populations selected for adaptation to larval crowding

In holometabolous animals such as Drosophila melanogaster, larval crowding can affect a wide range of larval and adult traits. Adults emerging from high larval density cultures have smaller body size and increased mean life span compared to flies emerging from low larval density cultures. Therefore, adaptation to larval crowding could potentially affect adult longevity as a correlated response. We addressed this issue by studying a set of large, outbred populations of D. melanogaster, experimentally evolved for adaptation to larval crowding for 83 generations. We assayed longevity of adult flies from both selected (MCUs) and control populations (MBs) after growing them at different larval densities. We found that MCUs have evolved increased mean longevity compared to MBs at all larval densities. The interaction between selection regime and larval density was not significant, indicating that the density dependence of mean longevity had not evolved in the MCU populations. The increase in longevity in MCUs can be partially attributed to their lower rates of ageing. It is also noteworthy that reaction norm of dry body weight, a trait probably under direct selection in our populations, has indeed evolved in MCU populations. To the best of our knowledge, this is the first report of the evolution of adult longevity as a correlated response of adaptation to larval crowding.