The effect of larval and adult nutrition on successful autogenous egg production by a mosquito

Females of most mosquito species require a blood meal to provision eggs and can be medical problems because of this dependency. Autogenous mosquitoes do not require blood to mature an initial egg batch and, instead, acquire nutrients for egg provisioning as larvae. We studied the importance of larval and adult nourishment for Ochlerotatus atropalpus which is obligatory autogenous for its first egg cycle but may ingest blood for subsequent cycles. Larval nourishment strongly influenced autogenous egg production: female larvae that were nutritionally stressed emerged as smaller adults, produced fewer eggs and emerged with less protein, lipid and glycogen stores. Female Oc. atropalpus are 100% autogenous, regardless of larval diet quality or whether females are fed sugar or water at emergence. Upon completion of the first egg batch, only females emerging from a poor larval diet ingested blood and produced a second egg batch.     

Inhibitory effects of oostatic hormone on ovarian maturation and ecdysteroid production in diptera

Extracts from post-vitellogenic ovaries of Musca domestica, Aedes atropalpus and Drosphila melanogaster were examined for the ability to inhibit the vitellogenic phase of growth in newly emerged A. atropalpus and D. melanogaster adult females. All extracts were inhibitory in A. atropalpus. D. melanogaster females were less sensitive than A. atropalpus females, only showing inhibition at a two-fold greater concentration of M. domestica extract. The titre of oostatic hormone in A. atropalpus ovaries correlated with the decline in ecdysteroid synthesis by these ovaries. A. atropalpus ovaries containing post-vitellogenic follicles were shown to inhibit ovarian ecdysteroid synthesis in vitro by vitellogenic ovaries of both A. atropalpus and D. melanogaster. D. melanogaster ovaries were not capable of eliciting such an inhibition in vitro, at least with the numbers of ovaries utilized in our experiments.     

INDUCTION OF OVARIAN DEVELOPMENT IN AUTOGENOUS AEDES ATROPALPUS BY JUVENILE HORMONE AND 20-HYDROXYECDYSONE

Aedes atropalpus is an autogenous mosquito characterized by a first gonadotropic cycle which results in approximately 200 mature oocytes without a bloodmeal. Ovarian development is completely inhibited if these animals are decapitated or ligated between the thorax and abdomen shortly after adult emergence. Injection of 4.8 ng of 20-hydro- xyecdysone into decapitated females 12 h after eclosion restores ovarian development in all females so treated. However, the same amount of 20-hydroxyecdysone injected into isolated abdomens obtained shortly after adult emergence had no discernible effect on vitellogenesis. In contrast, all abdomens which received 0.5 ng of topically applied JH I followed by the injection of 4.8 ng 20-hydroxyecdysone produced mature oocytes. Isolated abdomens were also capable of oocyte maturation when treated with excess amounts of JH alone; JH I was the most effective followed by JH II and then JH III.

Polyacrylamide gel electrophoretic analysis of vitellin extracted from the ovaries of hormonally-treated animals did not reveal any qualitative differences compared to intact normal controls. However, less yolk protein was present in the former. This was verified by counting the number and measuring the size of ovarian follicles in individual females.     

Genetic determinants of host ranges of Bacillus sphaericus mosquito larvicidal toxins.

The 51.4-kDa-41.9-kDa binary toxin produced by different strains of Bacillus sphaericus shows differential activity toward Culex quinquefasciatus, Aedes atropalpus, and Aedes aegypti mosquito larvae. The patterns of larvicidal activity toward all three mosquito species and growth retardation in A. aegypti have been shown to be due to the 41.9-kDa protein. By using mutant toxins expressed in Escherichia coli, insecticidal activity and growth retardation correlated with amino acids centered around position 100 of the 41.9-kDa protein. In its response to these toxins, A. atropalpus resembled C. quinquefasciatus rather than its congener, A. aegypti.     

Insulin signaling is necessary for vitellogenesis in Drosophila melanogaster independent of the roles of juvenile hormone and ecdysteroids: female sterility of the chico1 insulin signaling mutation is autonomous to the ovary

It has been suggested that insulin signaling mutations of Drosophila melanogaster are sterile and long-lived because of juvenile hormone (JH) and ecdysteroid deficiency. However, female sterility of an insulin/IGF-like signaling mutant (chico(1)) of D. melanogaster is not mediated by downstream systemic signaling in terms of major alterations in JH or ecdysteroid levels. chico(1) is a null mutation in the insulin substrate protein (CHICO) gene of D. melanogaster. Homozygous chico(1) females are sterile and their oocytes do not mature beyond the last previtellogenic stage. Homozygous chico(1) females exhibit approximately wild-type rates of JH biosynthesis, ovarian release of ecdysteroids and haemolymph ecdysteroid levels, suggesting that these two major hormone systems play no role in producing the sterility. Previtellogenic wild-type ovaries transplanted into homozygous chico(1) females underwent vitellogenesis, showing that systemic factors present in mutant females are sufficient to support normal vitellogenesis. chico(1) ovaries transplanted into wild-type females did not undergo vitellogenesis indicating that CHICO is necessary in the ovary for vitellogenic maturation. The ovary transplant experiments corroborate the endocrine results and demonstrate that insulin/insulin-like signaling (IIS) is necessary for vitellogenesis even when sufficient levels of JH, ecdysteroids or other factors are present.     

Juvenile hormone and ovarian maturation in the diptera: A review of recent results

The roles of the JHs and ecdysteroids in ovarian maturation of adult Aedes aegypti, A. atropalpus, Musca domestica and Drosophila melanogaster have been characterized by comparing the effects of surgical (allatectomy, ovariectomy, decapitation, abdominal ligation), genetic, and nutritional (sugar feeding) manipulations. The results show that in all species JHs, or their mimics, and 20-hydroxyecdysone act in combination to stimulate ovarian maturation and vitellogenin (Vg) synthesis and that high doses of exogenous JHs, or their mimics, stimulate ovarian ecdysteroid synthesis, at least in A. aegypti, A. atropalpus, and D. melanogaster. Thus the gonadotropic regulatory mechanisms that exist in various dipteran species are more similar than originally suspected. In M. domestica, and possibly D. melanogaster, 20-hydroxyecdysone is present in the hemolymph at vitellogenic levels in newly emerged females and may persist in ovariectomized adults. If true for D. melanogaster, this could explain why topical application of JH, or its mimics, to ovariectomized, isolated abdomens is effective in stimulating Vg synthesis in the absence of the ovaries.     

OVARIAN DEVELOPMENT AND OOSORPTION IN AUTOGENOUS FEMALES OF THE BLOWFLY,LUCILIA CUPRINA (WIED.)

Nearly 90% of nulliparous females of a laboratory selected autogenous strain of the sheep blowly, Lucilia cuprina, developed mature oocytes when fed only sucrose and water. The mean number of oocytes matured by those females which reached maturity was about 100. In the few females which did not reach maturity, ovarian development ceased early in vitellogenesis. Females had approximately 250 ovarioles, and when given ad libitum access to sheep liver, matured virtually all their primary oocytes. Flies given limited amounts of protein-rich material matured more oocytes than individuals given only sucrose and water; the apparent efficiency of conversion of ingested protein to ovary protein was of the order of 50%.

In females fed only sucrose and water or limited amounts of protein-rich material, the reduction in the number of oocytes resulted from oosorption initiated early in vitellogenesis. Oosorption was initiated earlier in females fed only sucrose and water, which were destined to resorb of the order of 150 oocytes, than in females which resorbed about 65 oocytes after having consumed a limited amount of protein-rich material.

Mating increased slightly both the proportion of females expressing autogeny and the number of oocytes matured.     

Effects of a methanolic extract of the plant Haplophyllum tuberculatum and of teflubenzuron on female reproduction in the migratory locust, Locusta migratoria (Orthoptera: Oedipodinae)

The effects of a methanolic extract of the plant Haplophyllum tuberculatum (ME-Ht) and of teflubenzuron (TFB) were compared on several reproductive variables and ecdysteroid titers in the females of Locusta migratoria. The test products were administered orally to newly emerged females at doses of 1500 (ME-Ht) and 10μg/female (TFB). The methanolic extract and TFB had comparable effects on several of the variables examined. Both significantly delayed the first oviposition and reduced fecundity and fertility. ME-Ht and TFB also displayed similar effects on ovarian growth, vitellogenesis and ecdysteroid titers. Both treatments induced a drop in hemolymph protein levels as well as a reduction in vitellogenin uptake by oocytes. This delay in oogenesis was accompanied by a resorption of terminal oocytes. However, whereas TFB completely blocked egg hatch, ME-Ht only had a modest inhibitory effect on this variable. Hemolymph and ovarian ecdysteroid titers, as measured by radioimmunoassay, were similar and low in both control and treated females, except for a peak observed only in control females at the end of vitellogenesis. We discuss the functional significance of the observed effects in the context of the putative modes of action of the methanolic plant extract and TFB.     

Ecdysteroid titers in mated and unmated Drosophila melanogaster females

Radioimmunoassay was used to determine ecdysteroid titers in mated or unmated Drosophila melanogaster females. Whole-body ecdysteroid titers increase after mating and this response is more pronounced after 12-24 hours than it is immediately after mating. In one experiment, females were mated to transgenic males deficient in accessory gland proteins to test whether these peptides mediate the observed increase in female whole-body ecdysteroid titers. Females mated to such transgenic males do not show a pronounced increase in whole-body ecdysteroid titers. The effect of mating on female hemolymph ecdysteroid titers was also investigated. Hemolymph ecdysteroid titers decrease after mating. The ecdysteroid titer change in the hemolymph may result from yolk protein uptake of ecdysteroids into developing vitellogenic oocytes as a consequence of male accessory gland protein stimulation of female oocyte maturation and yolk protein synthesis following mating.     

Seasonal life cycle and autogeny in the mosquitoAedes togoi in northern Kyushu,Japan, with experimental analysis of the effects of temperature,photoperiod and food on life-history traits

We studied seasonal changes in the larval population structure, adult size, and autogeny (egg production without a bloodmeal) of the mosquitoAedes togoi on the seacoast of northern Kyushu, Japan. The effects of temperature, photoperiod and food conditions on larval development, adult size and autogeny were studied in the laboratory.Aedes togoi overwintered in both the egg and larval stages and was multivoltine. Adult size was greater in spring and autumn and smaller in summer. Autogeny occurred in spring and autumn but not in summer. Autogenous females were larger than anautogenous females, and larger autogenous females produced more eggs. Laboratory experiments showed that autogeny was promoted under low temperature and short-day conditions which corresponded with spring/autumn conditions in the field. Experimental food limitation in the larval stage greatly reduced adult size and autogenous egg production. In the field, large inter-pool variations in adult size and autogeny rate probably resulted from variations in nutrient and crowding conditions. Seasonal autogeny ofAedes togoi was discussed in terms of life-history strategy under fluctuating environmental conditions where relative advantages of autogenous and anautogenous reproduction alternate seasonally.     

Sensilla basiconica (grooved pegs) on the antennae of female mosquitoes: electrophysiology and morphology

The sensilla basiconica or grooved pegs of mosquitoes consist of two morphologically and functionally distinct classes. Short grooved pegs house sensory cells that are excited by lactic acid whereas lactic acid-excited cells are absent in long grooved pegs. This correlation was found in four species of mosquito examined:Aedes aegypti, Aedes atropalpus, Aedes epactius, andCulex pipiens. InAedes epactius, both short and long grooved pegs house receptors that are sensitive to butyric acid.     

Ecdysteroid levels throughout the life cycle of the desert locust, Schistocerca gregaria

Moulting hormone levels for all stages of the life cycle of the desert locust, Schistocerca gregaria, have been determined using gas chromatography with electron capture detection of the trimethylsilylated hormones. During larval development, the major hormone detected is 20-hydroxyecdysone with smaller quantities of ecdysone present. In mature adult females the major ecdysteroid observed is a polar conjugate of ecdysone, with smaller quantities of conjugated 20-hydroxyecdysone also present. During embryonic development the pattern changes from a high proportion of conjugated ecdysone in the early stages to give more free hormone and a higher proportion of 20-hydroxyecdysone in later stages. The highest titre of 20-hydroxyecdysone found in this insect is during the 5th larval instar. Maximal levels of ecdysteroid per insect are found in mature females just before oviposition, while the highest level of ecdysteroid per g of tissue is found in the eggs.     

Autogeny in populations of blood-sucking mosquitoes (Culicidae) on the southern Yamal

After three-year studies a degree of autogeny in populations of Aedes communis. A. pullatus and A. hexodontus was defined. In various seasons a share of autogenous females in A. communis was stable enough amounting to 30-37%. At the same time considerable variations in the degree of autogeny in some micropopulations of this species were observed. Variations in the ratio between autogenous and non-autogenous individuals reflect a dissimilar effect of varied combinations of ecological factors upon larvae of mosquitoes.     

The influence of mating on autogenous egg development in the mosquito, Aedes taeniorhynchus

In several strains of Aedes taeniorhynchus, the presence of males increased the levels of autogeny. To stimulate autogenous egg production, the males had to mate with the females. Ageing further enhanced the rates of autogeny in mated females but not in virgins. Males did not influence the size of the autogenous egg batch. Even after the sixth day following emergence some females still possessed the capacity to respond to the male stimulus. With regard to ovarian maturation in A. taeniorhynchus, there appeared to be 3 types of females: (1) those that did not require a mating stimulus to be autogenous, (2) those that needed this stimulus and (3) those that remained anautogenous whether mated or not.     

Reproductive Status of Females in the Eusocial Wasp Polistes ferreri Saussure (Hymenoptera: Vespidae)

In the subfamily Polistinae, caste dimorphism is not pronounced and differences among females are primarily physiological and behavioral. We investigated factors that indicate the reproductive status in females of Polistes ferreri Saussure. We analyzed females from nine colonies and evaluated morphometric parameters, ovarian development, occurrence of insemination, relative age, and cuticular chemical profile. The colony females showed three kinds of ovarian development: type A, filamentous ovarioles; type B, ovarioles containing partially developed oocytes; and type C, long and well-developed ovarioles containing two or more mature oocytes. The stepwise discriminant analysis of the cuticular chemical profile showed that it was possible to distinguish the three groups of females: workers 1, workers 2, and queens. However, the stepwise discriminant analysis of the morphological differences did not show significant differences among these groups. The queens were among the older females in the colony and were always inseminated, while the age of the workers varied according to the stage of colony development.     

Formation of lipid reserves in fat body and eggs of the yellow fever mosquito, Aedes aegypti

We examined the accumulation of lipids in adult females of the mosquito, Aedes aegypti. Females emerged with about 100 μg lipid in the fat body. With access to sugar water lipids increased over seven days to 300 μg. After a blood meal on day five, sugar-fed females accumulated 120-140 μg of lipids in their ovaries within 2 days. At the same time the lipid content of the fat body decreased by 100 μg, indicating transfer of lipids from fat body to oocytes. Experiments in which fat body lipids were prelabelled support this conclusion. Label was transferred to oocytes: in mature oocytes the specific radioactivity of lipids was 80% of the specific radioactivity of prelabeled fat body lipids. Components of blood meals are also used to synthesize oocyte lipids. Fat bodies of females starved for four days had only 27 μg of lipids left. When these females were given a blood meal, they matured oocytes, although the number of ooyctes was reduced and ovaries contained only half the amount of lipids found in ovaries of females which had first fed on sugar water. Fat body lipids of these females had only slightly increased to 36 μg. This demonstrates that female Ae. aegypti use sugar to synthesize lipids, but they can also use components of blood for this purpose.     

The relationship between radiation-induced and transposon-induced genetic damage during Drosophila oogenesis

The combined effect of X-irradiation and transposon mobility on the frequencies of X-linked recessive lethals and dominant lethals was investigated in female hybrids in the P-M system of hybrid dysgenesis. X-linked lethals were measured in G2 hybrid dysgenic females whose X chromosome was derived from the M X P cross. To test for additivity or synergism, the mutation rate in irradiated dysgenic females was compared to that of unirradiated females as well as to irradiated nondysgenic hybrid females derived from M X M crosses. Eggs collected for 2 days after irradiation, were represented by the more radiation-sensitive A and B oocytes (about 75%) and the least sensitive C oocytes (about 25%). The production of X-linked lethal events in X-irradiated dysgenic females was 8.1%, as compared to 4.5% in dysgenic controls and 3.4% in irradiated, nondysgenic controls, demonstrating an additive effect of radiation and dysgenesis-induced genetic damage. The effect of irradiation on sterility of dysgenic hybrid females was a negative one, resulting in 20% less sterility than expected from an additive effect. The combined effect of radiation and dysgenesis on dominant lethality tested in A, B and C oocytes of the same hybrid females was synergistic. Egg broods collected for 3.5 days after irradiation showed that significantly more damage was induced in the presence of ionizing radiation in dysgenic females than in their nondysgenic counterparts. This effect was most obvious in B and C oocytes. The synergism observed may be related to the inability of cells to repair the increased number of chromosome breaks induced both by radiation and transposon mobility.     

Profile of the ecdysteroid hormone and its receptor in the salivary gland of the adult female tick, Amblyomma hebraeum

We examined the hemolymph ecdysteroid titer (by radioimmunoassay) and profile of the ecdysteroid receptor (EcR/USP; by [3H]ponasterone A binding, gel mobility shift assay, Western blot) in the salivary gland of the ixodid tick, Amblyomma hebraeum Koch (Acari: Ixodidae) throughout the tick feeding period and first 6 days post-engorgement. Throughout the slow phase of feeding, the hemolymph ecdysteroid titer was approximately 18 pg/microliter. The titer peaked at approximately 52 pg/microliter during the rapid phase of feeding, falling back to approximately 22 pg/microliter on the day of engorgement. Ecdysteroid titer rose again to approximately 750 pg/microliter by day 6 post-engorgement. EcR was undetectable by any of the three assays in unfed ticks. Following the onset of feeding, there appeared both specific ponasterone A binding and two major EcR bands detected by Western blot analysis. Both measurements were sustained throughout the feeding period, but declined after detachment when the salivary glands were degenerating. After ticks reached about 100 mg (by which time most females are mated), a discrete DNA-binding band was shown by gel mobility shift assay using Drosophila hsp27 EcRE as a probe. Moreover, the band intensified when hemolymph ecdysteroid titer reached its peak during the rapid phase of feeding; it declined along with decreasing EcR/USP levels, and with specific ligand binding activity following engorgement. This study suggests a role for the small hemolymph ecdysteroid peak during the rapid phase of feeding in initiating salivary gland degeneration.