Reproduction of an inbred strain of Culex pipiens prevented by loss of Wolbachia pipientis

The development time for an autogenous strain of Culex pipiens was prolonged after the mosquitoes were “cured” of their symbiotic Wolbachia pipients with tetracycline. Eggs produced by aposymbiotic females caged with either aposymbiotic or normal males failed to hatch. Neither growth rate nor reproduction was improved by feeding the larvae a commensal flagellate, bovine blood cells, or adenylic acid.     

Cultivation of the mosquito Culex pipiens pipiens f. molestus (Diptera, Culicidae) without blood feeding

Optimal photoperiod and temperature conditions for rearing of the mosquito Culex pipiens pipiens f. molestus without blood feeding (through autogenous eggs) were experimentally substantiated. This method may be used in order to obtain relatively small quantities of mosquitoes or for preservation of a laboratory colony. Optimal conditions of rearing are the short-day photoperiod (12 h of light per day) at 20 degrees C, when 87 % of the females produce autogenous eggs rafts consisting of 52 eggs on the average. It should be emphasized that larval food must be adequate and abundant, because the quality and quantity of it has a pronounced effect on the autogeny rate.     

Is the expression of autogeny by Culex molestus Forskal (Diptera: Culicidae) influenced by larval nutrition or by adult mating,sugar feeding,or blood feeding?

Culex molestus Forskal is suspected to have been introduced into southern Australia during the 1940s. Investigations to determine factors influencing the expression of autogeny, the response of this mosquito to potential blood meals, and the subsequent influence on oviposition were undertaken. Immature mosquitoes raised at five feeding regimes had mortality rates, development rates, wing length, and autogenous egg raft size measured. All surviving female mosquitoes laid autogenous eggs but there was a significant difference between the mean number of eggs per raft. For mosquitoes raised at each of the feeding regimes, there was a significant linear relationship between the number of eggs per autogenous egg raft and wing length. Newly emerged mosquitoes were offered a blood meal (i.e., rodent) daily but no blood feeding occurred until the autogenous egg raft was laid. There was no statistical difference in the rate of autogenous oviposition or post-oviposition blood feeding between control or treatment groups. The results of this study indicate that Cx. molestus is perfectly adapted to subterranean habitats in close association with human habitation, but their preference to delay blood feeding until up to day 8 following emergence may reduce their relative importance as a vector of arboviruses.     

Effects of blood meal sources on the biological characteristics of Aedes aegypti and Culex pipiens (Diptera: Culicidae)

Mosquitoes transmit many diseases to humans and animals e.g., malaria, yellow fever, dengue, filariasis and encephalitis. The fundamental target of this search was to study the effect of three different blood meal sources (human; rabbit and pigeon) on some biological and behavioral properties of Aedes aegypti and Culex pipiens. The results have assured that the females of the mosquito Ae. aegypti that were fed on human blood meal has registered the highest feeding activity from feeding on the blood meal whereas the females of the other mosquito Cx. pipiens have shown the highest feeding activity after being fed on pigeons when compared with its feeding on other factors. The results have shown non-significant variation in the average time necessary to digest the blood meal on both mosquito species Ae. aegypti and Cx. pipiens that were fed on vertebrate hosts under laboratory conditions. Furthermore, results assured that the difference in blood meal sources has yielded distinct variation in the reproductive capacity and efficiency of both female mosquitoes under investigation where both species Ae. aegypti and Cx. pipiens already fed on human blood meal have yielded a pronounced distinctive increase in egg production (oviposition) when compared with females that were fed on pigeon or rabbit blood meal respectively. Moreover, feeding of the female mosquitoes under lab conditions on different blood meal sources did not affect the level of the hatching eggs that were laid by both mosquito females.     

Facultative blood-feeding in the crabhole mosquito, Deinocerites cancer

1. Most of the female crabhole mosquitoes Deinocerites cancer Theobald entering bait traps had undeveloped ovaries. 2. Nearly half of the D. cancer females that were captured in bait traps, but denied blood, were gravid following a 1 week holding period; they developed relatively small egg clutches autogenously (mean = 7.4 eggs per gravid female). 3. Although both parous and nulliparous D. cancer females were captured exiting from crabholes, only parous females were attracted to and collected in bait traps. 4. Much larger egg clutches (mean = 41.8 eggs per female) were produced anautogenously by mosquitoes that blood-fed on chickens. 5. All D. cancer females from field collected pupae developed eggs autogenously when allowed to mate and sugar feed in the laboratory. These mosquitoes produced an initial egg clutch with a mean size equivalent to that found in blood-fed, parous mosquitoes from bait traps. 6. In the field, mating and sugar feeding rather than blood-feeding seem to stimulate production of the initial egg clutch. 7. D. cancer blood-feeding appears to be delayed until after the first egg clutch has been laid; some parous crabhole mosquitoes are facultative blood feeders and can produce an egg clutch either with or without a blood meal.     

The etiological agent of cytoplasmic incompatibility in Culex pipiens

All individuals of Culex pipiens (wide sense) which have been examined were infected with Wolbachia pipientis. Larvae reared in tetracycline were freed of these symbiotes and remained free (aposymbiotic) in future generations. When males were freed of their symbiotes, they no longer displayed incompatibility. Aposymbiotic males were compatible with all females, whether infected or not. Aposymbiotic females, on the other hand, laid fertile eggs after mating with aposymbiotic males, but not after mating with normal males. Most eggs laid by aposymbiotic females after mating with normal males showed no development at all, even though the females had been inseminated.     

Reduction in egg hatch after a sublethal dose of chlorfluazuron to larvae of the common cutworm, Spodoptera litura

Abstract.  The effects of a sublethal dose of chlorfluazuron on egg hatching in Spodoptera litura are examined under laboratory conditions. When LD₁₀ of chlorfluazuron is topically applied to newly moulted fifth-instar larvae of either sex, a significant reduction in both the number of eggs laid and subsequent hatching is observed after mating but no significant differences in daily of oviposition are observed when compared with the controls. In addition, examination of the unhatched eggs reveals that the number of unfertilized eggs is greater than those that were fertilized but there are significantly more unfertilized eggs laid by treated insects. Interference by chlorfluazuron, transferred by copulation through sperm fluids or ova, appears improbable. All the fertilized unhatched eggs in the treated crosses die at an earlier stage than those of the controls. In the female-treated crosses, the egg size is significantly reduced compared with the control or male-treated crosses. During mating, the treated-males transferred significantly lower-weight spermatophores into the females. The weight of spermatophores transferred by untreated males is the same to both treated and untreated females. The duration of mating is not affected by chlorfluazuron treatment.     

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.     

Enteric release of vasoactive intestinal peptide after a peptone meal in the dog

Vasoactive intestinal peptide (VIP) concentrations were measured by radioimmunoassay in plasma from portal and peripheral venous blood obtained from six alert, non-anesthetized dogs before and after gastric infusion of a 10% peptone meal. Mean basal portal and cephalic vein plasma VIP concentrations were $\text{42 ± 11.7}$ and $\text{42 ± 8.0}\text{(S.E.M.) pg/ml}$, respectively. No significant changes in peripheral venous plasma VIP concentrations were noted after the peptone meal throughout the duration of the collection period. In contrast, however, the mean VIP concentration in portal plasma increased promptly after the peptone meal with a peak of $\text{79 ± 8.2}\text{pg/ml}$ ($\text{P < 0.02}$) occurring 8 min after infusion of the meal. This was followed by a gradual decline in portal plasma VIP levels, with a return to prefeeding concentrations at 60 min ($\text{44 ± 6.3}\text{pg/ml}$). Results of these studies demonstrate that following gastric infusion of a peptone meal in the dog, portal, but not peripheral, plasma VIP concentrations increase significantly. Failure to detect augmentation of peripheral vein VIP levels after the meal is probably due to hepatic clearance of VIP.     

Search for a sensitive stage for aposymbiosis induction in Culex pipiens pipiens by antibiotic treatment of immature stages

The elimination of symbiotic Wolbachia pipientis from Culex pipiens pipiens was studied by exposing the larvae or pupae to chlortetracycline hydrochloride for different lengths of time and for different stages in the life cycle. The effect of the antibiotic was cumulative. The longer the immature stages were treated, the poorer was the hatching rate of their eggs when they became adults. Larvae reared in antibiotic from the beginning of first, second, or third instar developed into adults with 100% hatching failure. If the insects were reared in the antibiotic from the beginning of the fourth instar only, about one-third of their eggs hatched. Rearing the immatures in antibiotic for single instars demonstrated that the ultimate deleterious effects were essentially the same for all stages, indicating that there is no uniquely sensitive stage for destroying the rickettsia. Many of the unhatched eggs contained fully developed, but dead, embryos.     

Two stages of juvenile hormone-mediated growth of secondary follicles in Culex pipiens

To investigate the effect of juvenile hormone deprivation on the growth of secondary follicles during the second gonotrophic cycle, female mosquitoes, Culex pipiens L., were allatectomized daily after the first blood meal. Allatectomy on days 1–3 suppressed growth of secondary follicles indicating that juvenile hormone was required for a second gonotrophic cycle. When allatectomy was performed 4 days or more after the first blood meal, secondary follicles grew, indicating the presence of juvenile hormone. However, if mosquitoes were allatectomized before oviposition, only 25% developed a second batch of eggs after a second blood meal.Allatectomies performed 1 and 24 h after oviposition indicated that additional juvenile hormone was released after deposition of the first batch of eggs, and that this second release was needed for secondary follicles to complete previtellogenic growth. Thus, in C. pipiens, secondary follicles undergo two periods of juvenile hormone-mediated growth-one before and one after oviposition.     

Eggs of the Australian saltmarsh mosquito,Aedes camptorhynchus,survive for long periods and hatch in instalments: implications for biosecurity in New Zealand

The Australian saltmarsh mosquito, Aedes camptorhynchus (Diptera: Culicidae), is a significant biting pest and disease vector and is the subject of an eradication programme in New Zealand (NZ), where it has been resident for more than 10 years. To better understand the ecology of this common and widespread pest, we studied egg longevity and hatching patterns in the laboratory. By regularly testing for the presence of viable embryos, we found that eggs may last more than 15 months when stored dry (13% viable at this time). Eggs display instalment hatching, with no more than 56% of a batch hatching upon first inundation. Further hatching may occur for at least six inundations and some unhatched eggs may remain viable even after this. Variation in hatching rates can be observed using different water types, with weaker hatching media stimulating lower hatching rates spread over more inundations. By applying average hatching rates to a non‐linear model of natural egg attrition, we showed that egg batches exposed to three inundations should be exhausted (zero live eggs present) in approximately 11 months at the conditions tested here. These findings have implications for the current eradication programme for Ae. camptorhynchus in NZ and for our understanding of the ecology of a widespread and common disease vector in Australia.     

广聚萤叶甲成虫交配时长对产卵量与孵化率的影响

【目的】明确不同交配时长对广聚萤叶甲雌虫产卵量和卵的孵化率的影响。【方法】在室内条件下,对不同交配时长下广聚萤叶甲的雌虫产卵量和卵的孵化率进行观察:(1)选取羽化第3d的广聚萤叶甲雌雄虫随机配对,观察24 h,记录交配情况和时长;(2)在交配开始1、5、15、30、60 min时,强行分开雌雄成虫,然后将不同交配时长的雌虫进行单独饲养,以正常交配一次的雌虫作为对照,每个处理选取23组;(3)将15～30 cm健壮豚草小苗插入注满水的塑料小瓶内,将配对的一组雌雄成虫和豚草小苗放入养虫盒中饲养,每天更换带卵的小苗并记录叶片上的产卵量;(4)将上述带卵的小苗至于适宜条件下培养,记录5～7 d内卵块孵化的情况。【结果】广聚萤叶甲正常交配一次的对照组的产卵水平显著高于各处理组,单雌产卵为889粒,交配时间15 min以下各组雌虫的产卵量明显低于交配30 min以上的各组雌虫。同时交配时长5 min以下雌虫产的卵基本不能孵化,而交配时间达到30 min以上的各组卵块的孵化率明显提高。【结论】雄虫转移雌虫受精所需精子量需要耗费的时间为30 min左右,且雄虫有延长交配时间的趋性。该结果为研究广聚萤叶甲的生态特性以及优化种群繁殖提供科学依据。     

Fluid transport across the ducts of the salivary glands of a mosquito

We compared the volumes of fluid expelled from the salivary stylets of mosquitoes having ducts transected at different levels. An oil-filled apparatus was devised to measure salivary output. About $\text{1}\text{10}$ of normal salivary volume was produced by mosquitoes when ducts were transected and externalized just anterior to the salivary glands. Such mosquitoes ejected more saliva than could be contained within the walls of the ducts. Transection posterior to the pump prevented salivation entirely. Skin reactive antigen was not detected (as a wheal) when mosquitoes with transected ducts fed on man. We conclude that fluid is transported across the walls of the ducts that drain the salivary glands of a mosquito.     

Egg hatching of mosquitoes Aedes caspius and Ae. vittatus stimulated by water vibrations

Simple laboratory experiments with eggs of wild-caught Aedes caspius (Pallas) and Aedes vittatus (Bigot) mosquitoes (Diptera: Culicidae) demonstrated that egg hatch rates (i.e. percentage hatching daily) increased significantly in response to water vibrations caused by finger-tip drumming on containers of submerged eggs. When disturbed by daily vibrations for only 30 s, eggs hatched sooner than when they remained flooded or were periodically drained and reflooded without agitation. For autogenous Ae. caspius from saltmarsh, egg hatch rates were three- to four-fold greater for groups of 50 eggs than for solitary eggs, possibly due to chemicals (functional kairomones) emanating from other eggs or larvae. For anautogenous Ae. vittatus from freshwater rock-pools, tapping the container daily (for 30 s each 2 h in 8 h) hatched 93% of eggs, compared with only 42% hatch of eggs agitated only when first flooded or 55% hatch of eggs dried and reflooded three times during 17 days without other disturbance. It is concluded that, apart from flooding and other factors, vibrations - simulating the patter of rainfall provide a very significant hatching stimulus for Aedes eggs.     

Effect of mating period and time‐of‐day for bloodmeal on rearing of Asian tiger mosquito (Aedes albopictus) in laboratory conditions

Colonization and maintenance of mosquitoes in the laboratory is required to study physiology, ecology, and behavior of mosquitoes and interactions between mosquito and pathogens. Artificial blood feeding systems have been widely used to maintain the laboratory colony of Aedes albopictus. In this study, we investigated the effects of mating period (1, 3, 6, and 10 days) and time‐of‐day for bloodmeal (08:00, 13:00, and 18:00) in the use of an artificial feeding system on blood‐feeding rate, female fecundity, egg hatching rate, and developmental time of the Asian tiger mosquito, A. albopictus. Younger females mated for three or fewer days reproduced more eggs compared to those of oldest females mated for ten days. Similar to the result for eggs laid, the mean egg‐hatching rate was significantly higher from the offspring of younger females than from those of older females. However, mating period and time‐of‐day for bloodmeal had no effect on the blood feeding rate and developmental time. Taken together, we suggest that three‐day mating with bloodmeal at 18:00 is optimal for maintaining colonies of A. albopictus in laboratory conditions.     

Age modifies the effect of body size on fecundity in Culex quinquefasciatus Say (Diptera: Culicidae)

Fecundity of mosquitoes can vary with many factors and can have a strong effect on population growth. This study reports the effects of body size, blood meal size, and age on the reproductive output of nulliparous Culex quinquefasciatus, a vector of arboviruses and other pathogens. Mated adult female mosquitoes from a colony were reared under standard conditions and fed on chickens at different ages post‐eclosion. Blood meal size and wing length were recorded, as well as the number of eggs in the first‐cycle egg raft. Each of these factors had a significant influence on fecundity considered in a simple regression context. Multiple regression analysis revealed a significant interaction effect between age and body size on fecundity. Up to 13 days of age, fecundity was positively correlated with body size, but in mosquitoes older than 13 days, this relationship was not significantly different from zero. These results are discussed in terms of the known physiology of this and other species.     

Cytoplasmic factors in the gonads of Culex pipiens complex mosquitoes

Electron microscopic examination of the gonads of six species of the $\text{Culex pipiens}$ complex revealed rickettsia-like, intracellular symbionts in $\text{C.pipiens}$, $\text{C.molestus}$, $\text{C.pallens}$ and $\text{C.fatigans}$.¹ In the two endemic Australian members, $\text{C.a}$$\text{C.globocoxitus}$, no rickettsia-like symbionts were found, but virus-like particles were seen in the cytoplasm of the ovaries and certain associated tissues.     

Effect of body size and sugar meals on oviposition of the yellow fever mosquito,Aedes aegypti (Diptera: Culicidae)

The effects of dietary sugar and body size on the oviposition of Ae. aegypti were studied under laboratory conditions. In female mosquitoes provided with sugar, the start of maximum fecundity was significantly delayed and the oviposition period was longer than in females provided with water. The peak of oviposition was also delayed in sugar‐fed females. Large females oviposited more eggs per day than small females at maximum fecundity and during eight days of observations. Large females also visited significantly more water‐containing cups in their cages per day than small females at maximum fecundity. During the eight days of observations, large females and sugar‐fed females visited more water‐containing cups in their cages than water‐fed small females. Both large females and sugar‐fed females oviposited their eggs at sites higher above the water line than water‐fed small females. These results suggested that large and sugar‐fed female Ae. aegypti mosquitoes had more energy reserves and oviposited their eggs at higher sites, which would lead to a time lag in hatching.     

Enzymes in the exsheathing fluid of nematodes and their biological significance

Rogers W. P. 1982. Enzymes in the exsheathing fluid of nematodes and their biological significance. International Journal for Parasitology12: 495–502. The characteristics of an enzyme which hydrolysed denatured collagen and a lipase in exsheathing (ecdysal) fluid are described. A highly purified collagenase from Clostridium histolytica attacked isolated sheaths and reacted to additives in the same way as exsheathing fluid. However, relative to their activities with Azocoll or $\text{p-}\text{phenylazobenzyloxycarbonyl}\text{-}\text{L}\text{-}\text{Pro}\text{-}\text{L}\text{-}\text{Leu}\text{-}\text{Gly}\text{-}\text{L}\text{-}\text{Pro}\text{-}\text{D}\text{-}\text{Arg}$ as substrates, the enzyme of exsheathing fluid was >400 times as active as the bacterial collagenase in its action on isolated sheaths.It is suggested that the lipase in exsheathing and hatching fluids may, in association with the pseudocollagenase (and sometimes with chitinase also) have a function in the hatching of eggs. The pseudocollagenase alone may serve as the exsheathing enzyme. The leucine aminopeptidase in hatching and exsheathing fluids may be concerned in the breakdown of the excretory cell and the release of the fluids.