Hatching mechanism and delayed hatching of the eggs of three broadcast spawning euphausiid species under laboratory conditions

Three different egg hatching mechanisms were observed underlaboratory conditions in Euphausia pacifica Hansen, Thysanoessaspinifera Holmes and Thysanoessa inspinata Nemoto: backward,forward and flipping. Like all broadcast spawning euphausiids,these species usually hatch as nauplius 1 (N1). Some hours beforehatching the vitelline membrane breaks and the embryo is freelysuspended within the chorion; later the embryo takes on a slightlyoval shape. When ready to hatch, the N1 pushes against the chorionwith the posterior part of the abdomen producing a protuberance.No spine or egg tooth is present to break the chorion. The pressurebreaks the chorion, and the nauplius pushes itself backwardswith the first and second antennae and mandible to slide fromthe chorion. After about three quarters of the body is outside,the nauplius brings all the appendages together to move backwardswithout becoming stuck in the chorion. This is the backwardhatching mechanism. The vitelline membrane remains within theegg after the nauplius leaves the chorion. Hatching takes 5–20s, and most of the eggs in a clutch hatch during <2 h. Severaleggs of E. pacifica hatched as meta-nauplii (MN) (>200 hafter spawning) or as calyptopis 1 (C1) stage (>232 h), ratherthan as N1. Delayed hatching of embryos also was observed inT. spinifera as nauplius 2 (N2) (>120 h) or as MN stage (>180h), and in T. inspinata as N2 (106 h) after spawning. Eggs withlarvae in stages of development beyond N1 have not been observedfrom preserved zooplankton samples. However, eggs spawned inthe field and incubated in the laboratory also had extendeddevelopment and late hatching but with low frequency (<0.06%).It is proposed that, if the backward hatching mechanism fails,alternate hatching mechanisms can be used by the euphausiid.There is high flexibility in their hatching modes. The N2 andMN break the chorion with the first and second antennae, hatchingforwards, and the C1 breaks it with the telson spines and byflipping of the abdomen, resembling the decapod hatching mechanism.Delayed hatching using the forward and flipping mechanisms wereassociated with low hatching success in comparison with thebackward hatching mechanism.     

Embryonic, early larval development time, hatching mechanism and interbrood period of the sac-spawning euphausiid Nyctiphanes simplex Hansen

Females of the sac-spawning euphausiid Nyctiphanes simplex Hansenwere incubated under shipboard laboratory conditions to observethe embryonic and larval development time and hatching mechanism.Females ready to spawn have a pale pink ovary that extends fromthe back of the stomach to the first abdominal segment, fillingmost of the haemocoel. This species usually behaves as a totalspawner (produces one batch of oöcytes per cycle of theovary) leaving an ‘empty’ space in the cephalothoraxwhere the spent ovary is located. After spawning, the youngoöcytes mature and turn pale pink. The eggs do not havea measurable perivitelline space (PVS) in any of the embryonicstages (6.6 x magnification). The embryos hatch as nauplius(80–91 h after spawning, 16°C ± 1°C). Theyfurther develop into pseudometanauplii (PMN, 90–105 hafter spawning) and metanauplii (MN, 92–140 h after spawning)inside the ovigerous sac. The nauplius breaks the thin and fragilechorion by increasing the volume of the body and by using thefirst and second antennae. We call this an ‘expansion’hatching mechanism, the fifth distinct hatching mechanism observedso far among euphausiids. N. simplex larvae escape from theovigerous sac late in the MN stage (5 days after spawning),just a few hours before molting into calyptopis 1 (C1) (0.5–4h). This delayed release extends protection by the female, likelydecreasing the risk of predation or early cannibalism. Additionally,this may save energy by not swimming independently increasingthe time of not return if the calyptopis does not find favorablefeeding conditions. Females are not ready to spawn again untilat least two days after the previous batch of embryos leavesthe ovigerous sac. The interbrood period (IBP) observed rangedbetween 7 and 15 days at 16–18°C. This IBP is aboutone-fourth to half than was previously assumed for this speciessuggesting a significant underestimation of the fecundity ofthis species. N. simplex hatching success usually was 100%,except for a few females with all of their embryos dying duringembryonic development. Other females either molted before releasingthe embryos, or the oöcytes were spawned unfertilized (0%hatching success), particularly during winter conditions. Efficienthatching and late free-swimming strategy may partially explainwhy this species is the most abundant neritic euphausiid inthe southern part of the California Current System (CCS) andin the Gulf of California.     

Structure and enzymatic removal of the chorion of embryos of the Nile tilapia

Dechorionation or partial digestion of the egg chorion is necessary for introducing embryonic stem cells into blastulas for chimera production and for harvesting blastulas. Several methods to digest the Nile tilapia Oreochromis niloticus chorion were tried and it was found that the chorions of most clutches of eggs were digested in <3 h using hatching medium [produced by allowing embryos to hatch in Hanks balanced salt solution (HBSS) in an incubator], or 2 mg ml⁻¹ pronase P6911 in 10% Ca/Mg free HBSS. The chorion of Nile tilapia possesses multiple lamellae as found in most teleost species that have been studied. It was found to be thinner than that of medaka Oryzias latipes and thicker than that of zebrafish Danio rerio . During natural hatching the chorion was digested from the inner surface, and tail movements helped to break the remaining chorion; however, chorion digestion has to be complete for experimental dechorionation, because digestion starts at the external surface. The zona radiata externa remained intact after experimental digestion with hatching media but was disrupted by pronase. Embryos dechorionated at the cleavage or blastula stage only survived for 2 or 3 days, but some dechorionated at the gastrula stage or early segmentation stage developed until the natural hatching time. If the chorion was partially digested at the cleavage or blastula stage, some embryos survived to hatch.     

Corticosterone stimulates hatching of late-term tree lizard embryos

The regulation of hatching in oviparous animals is important for successful reproduction and survival, but is poorly understood. We unexpectedly found that RU-486, a progesterone and glucocorticoid antagonist, interferes with hatching of viable tree lizard (Urosaurus ornatus) embryos in a dose-dependent manner and hypothesized that embryonic glucocorticoids regulate hatching. To test this hypothesis, we treated eggs with corticosterone (CORT) or vehicle on Day 30 (85%) of incubation, left other eggs untreated, and observed relative hatch order and hatch time. In one study, the CORT egg hatched first in 9 of 11 clutches. In a second study, the CORT egg hatched first in 9 of 12 clutches, before vehicle-treated eggs in 10 of 12 clutches, and before untreated eggs in 7 of 9 clutches. On average, CORT eggs hatched 18.2 h before vehicle-treated eggs and 11.6 h before untreated eggs. Thus, CORT accelerates hatching of near-term embryos and RU-486 appears to block this effect. CORT may mobilize energy substrates that fuel hatching and/or accelerate lung development, and may provide a mechanism by which stressed embryos escape environmental stressors.     

新疆石河子棉蚜越冬卵的发育规律

棉蚜雌性蚜交配个体平均产卵量为 6 3± 1 1粒 ;未交配个体也产卵 ,卵亦变色 ,但卵量极少。越冬卵经 5 %NaOH浸泡 2 4h后卵壳透明 ,镜下观察可见受精卵胚胎发育到体躯分节期开始越冬。冬后胚胎发育仅处在颗粒团状的饱满卵应为无效卵 ,不会孵化 ;颜色发亮的卵处理后胚胎外貌清晰可见 ,此种卵当天即可破壳。棉蚜卵为滞育卵 ,春季降水对孵化有促进作用 ;室外棉蚜卵的饱满率和饱满卵的孵化率较低。该地区秋冬气候特点可能是石河子地区棉蚜卵低温条件下安全越冬的重要因素     

The effect of light and darkness on hatching in the pomacentrid Abudefduf saxatilis

Synopsis Reports that pomacentrid embryos hatch after dusk are confirmed by photic manipulation of sergeant major eggs. Embryos placed in the dark for 20 minutes or longer prior to their normal hatching after sunset hatched, whereas controls held in light did not hatch. Percent of hatched embryos correlated with increasing exposure to darkness up to one hour after which no further improvement in hatching was observed. Embryos maintained in continuous light during their normal twilight hatching period did not hatch. Also, embryos exposed to 60 minutes of darkness, if interrupted by one minute of light every 10 minutes did not hatch. The percent hatch in dark treatments varied significantly between nests and, in some treatments, correlated negatively with the size of the egg clumps (number of eggs per clump) tested. To initiate hatching in the presence of light required intensities of 0.03 lux or less. These low intensities are not reached until about 20 minutes after sunset on the reef where the embryos occur. We conclude that hatching for some embryos occurs about 30 minutes after sunset but for most is not completed until at least one hour after sunset. Hatching therefore takes place at a time long after potential diurnal fish predators have refuged in the reef structure.     

Patterns in the abundance,phenology, and hatching of the resting egg stage of the invasive zooplankter <Emphasis Type="Italic">Bythotrephes longimanus</Emphasis>: implications for establishment

To examine how dormancy contributes to the establishment and persistence of Bythotrephes longimanus, we investigated resting egg production and hatching in relation to the demography of the planktonic stage and environmental conditions in Island Lake Reservoir (USA). During a 3-year study, the largest contribution to the egg bank occurred in autumn and most eggs hatched in spring, but we also detected some resting egg production and hatching in summer. The difference between summer and late autumn densities of eggs in sediments averaged 47% (range 0–98%) for 18 sites throughout the reservoir, which was similar to experimental estimates of in situ hatching fraction of 67% for eggs in the spring and summer following their production. Based on emergence traps, neonates hatch in the field during May and June. We estimated mortality rates of 64% for resting eggs and embryos, and 59% for newly emerged neonates. Although hatching fraction saturated at the same level, eggs incubated offshore hatched later than those nearshore where water temperature was warmer and light was detectable at the sediment surface. Low dissolved oxygen concentration did not significantly reduce hatching fraction but resulted in some eggs that initiated development but failed to hatch. Collectively, our results demonstrate substantial annual turnover in the resting egg bank of B. longimanus and high mortality of resting eggs during recruitment from the egg to the first molt of the planktonic stage. These patterns suggest that propagule pressure in the form of resting eggs requires large numbers for establishment, and that considerable post-establishment resting egg production is necessary for inter-annual persistence.     

Plasticity of hatching in amphibians: evolution, trade-offs, cues and mechanisms

Many species of frogs and salamanders, in at least 12 families, alter their timing of hatching in response to conditions affecting mortality of eggs or larvae. Some terrestrially laid or stranded embryos wait to hatch until they are submerged in water. Some embryos laid above water accelerate hatching if the eggs are dehydrating; others hatch early if flooded. Embryos can hatch early in response to predators and pathogens of eggs or delay hatching in response to predators of larvae; some species do both. The phylogenetic pattern of environmentally cued hatching suggests that similar responses have evolved convergently in multiple amphibian lineages. The use of similar cues, including hypoxia and physical disturbance, in multiple contexts suggests potential shared mechanisms underlying the capacity of embryos to respond to environmental conditions. Shifts in the timing of hatching often have clear benefits, but we know less about the trade-offs that favor plasticity, the mechanisms that enable it, and its evolutionary history. Some potentially important types of cued hatching, such as those involving embryo-parent interactions, are relatively unexplored. I discuss promising directions for research and the opportunities that the hatching of amphibians offers for integrative studies of the mechanisms, ecology and evolution of a critical transition between life-history stages.     

Temperature effects on embryonic development of Paratlanticus ussuriensis (Orthoptera: Tettigoniidae) in relation to its prolonged diapause

Paratlanticus ussuriensis eggs overwinter by entering diapause, which can be prolonged to more than 1 year depending on environmental conditions. To determine temperature effects on diapause duration of P. ussuriensis eggs, the rates of embryonic development and hatching were compared at various temperatures conditions by measuring embryonic stages and egg weights. Most eggs stayed in a very young stage (blastoderm formation, stage 4) when reared at 15 and 20 °C, 10–30% eggs developed into middle or late stages when reared at 25 °C, and most embryos developed fully (stage 23/24) when reared at 30 °C. Egg weight at 30 °C was 1.5 times higher than those reared at 20 °C. Chilling induced hatching in embryos at stage 23/24. Chilling caused stage 4 embryos to develop into stage 24, but they failed to hatch in response to a second warm period. Thus, P. ussuriensis eggs can overwinter either as young embryos (initial diapause) or as fully-developed embryos (final diapause). Eggs that experience an initial diapause overwinter again the second year in a final stage diapause. The post-diapause period was shorter when embryos overwintered in a final stage diapause. The hatching rate was highest in a temperature range of 7.5–15 °C. Our results suggest that temperature is an important environmental factor for the control of prolonged diapause in P. ussuriensis and initial diapause plays an important role in the control of its life cycle.     

Differences in egg size, shell thickness, pore density, pore diameter and water vapour conductance between first and second eggs of Snares Penguins Eudyptes robustus and their influence on hatching asynchrony

Brood reduction in birds is frequently induced by hatching asynchrony. Crested penguins (genus Eudyptes) are obligate brood reducers, but in contrast to most other birds, first‐laid eggs are considerably smaller in size than second‐laid eggs; furthermore, first‐laid eggs hatch after their siblings. The mechanisms underlying this reversal in size and hatching order remain unclear. In this study, we tested whether the second‐laid eggs of Snares Penguins Eudyptes robustus have a higher eggshell porosity allowing them to maintain a higher metabolic rate throughout incubation and to hatch before their first‐laid siblings. We investigated differences in egg size, shell thickness, pore density, pore diameter and water vapour conductance between first and second eggs within clutches and examined the influence of these shell characteristics on hatching asynchrony. First‐laid eggs of Snares Penguins were approximately 78% of the size of the larger second eggs. Second‐laid eggs had considerably thicker shells and more pores per cm² than first eggs, whereas pore diameter did not differ between eggs. Water vapour conductance was greater in second‐ (16.8 mg/day/torr) than in first‐laid eggs (14.9 mg/day/torr). The difference in water vapour conductance between first‐ and second‐laid eggs within clutches was related to hatching patterns. In nests where second eggs hatched before first‐laid eggs, second eggs had a considerably greater water conductance than their sibling, whereas in nests where both eggs hatched on the same day, the difference in water conductance between eggs was very small, and in a few nests where small first eggs hatched before their larger sibling, they had a greater water conductance than their larger second‐laid nestmate. Surprisingly few studies have investigated differences in shell characteristics between eggs within clutches and associated effects on hatching asynchrony. This study has demonstrated that such differences exist between eggs within clutches and that they can influence hatching patterns.     

Embryonic Development and Rates of Metabolic Activity in Early and Late Hatching Eggs of the Major Malaria Vector Anopheles gambiae

Anopheles gambiae eggs generally hatch at the completion of embryo development; two-three days post oviposition. However, staggered or delayed hatching has been observed whereby a single batch of eggs shows marked variation in time-to-hatch, with some eggs hatching 18 days post oviposition or later. The mechanism enabling delayed hatch has not been clearly elucidated but is likely mediated by environmental and genetic factors that either induce diapause or slow embryo development. This study aimed to compare metabolic activity and embryonic development between eggs collected from sub-colonies of the baseline Anopheles gambiae GAH colony previously selected for early or late time-to-hatch. Egg batches from early and late hatch sub-colonies as well as from the baseline colony were monitored for hatching. For both time-to-hatch selected sub-colonies and the baseline colony the majority of eggs hatched on day two post oviposition. Nevertheless, eggs produced by the late hatch sub-colony showed a significantly longer mean time to hatch than those produced by the early hatch sub-colony. The overall proportions that hatched were similar for all egg batches. CO₂ output between eggs from early and late hatch sub-colonies showed significant differences only at 3 and 7 days post oviposition where eggs from the early hatch and the late hatch sub-colony were more metabolically active, respectively. No qualitative differences were observed in embryo development between the sub-colonies. It is concluded that all viable embryos develop to maturity at the same rate and that a small proportion then enter a state of diapause enabling them to hatch later. As it has previously been shown that it is possible to at least partially select for late hatch, this characteristic is likely to involve genetic as well as environmental factors. Delayed hatching in An. gambiae is likely an adaptation to maximise reproductive output despite the increased risk of desiccation in an unstable aquatic environment.     

Wasp predation and wasp-induced hatching of red-eyed treefrog eggs

Eggs often suffer high levels of predation and, compared with older animals, embryos have few options available for antipredator defence. None the less, hatchlings can escape from many predators to which eggs are vulnerable. I studied early hatching as an antipredator defence of red-eyed treefrog embryos, Agalychnis callidryas, in response to egg predation by social wasps (Polybia rejecta). Red-eyed treefrogs attach their eggs to vegetation overhanging water, where they are exposed to arboreal and aerial predators. Wasps attacked half the egg clutches and killed almost a quarter of the eggs I monitored at a natural breeding site in Panama. Hatching tadpoles fall into the water, where they face aquatic predators. As predicted from improved survival of older hatchlings with aquatic predators, most undisturbed eggs hatched relatively late. However, many younger embryos directly attacked by wasps hatched immediately. Embryos attacked by wasps hatched as much as a third younger than the peak undisturbed hatching age, and most hatching embryos escaped. Thus hatching is an effective defence against wasp predation, and plasticity in hatching stage allows embryos to balance risks from stage-specific egg and larval predators. Wasp-induced hatching is behaviourally similar to the snake-induced hatching previously described in A. callidryas, but occurs in fewer eggs at a time, congruent with the scale of the risk. Individual embryos hatch in response to wasps, which take single eggs, whereas whole clutches hatch in response to snakes, which consume entire clutches. Embryos of A. callidryas thus respond appropriately to graded variation in mortality risks. Copyright 2000 The Association for the Study of Animal Behaviour.     

Embryonic communication in the nest: metabolic responses of reptilian embryos to developmental rates of siblings

Incubation temperature affects developmental rates and defines many phenotypes and fitness characteristics of reptilian embryos. In turtles, eggs are deposited in layers within the nest, such that thermal gradients create independent developmental conditions for each egg. Despite differences in developmental rate, several studies have revealed unexpected synchronicity in hatching, however, the mechanisms through which synchrony are achieved may be different between species. Here, we examine the phenomenon of synchronous hatching in turtles by assessing proximate mechanisms in an Australian freshwater turtle (Emydura macquarii). We tested whether embryos hatch prematurely or developmentally compensate in response to more advanced embryos in a clutch. We established developmental asynchrony within a clutch of turtle eggs and assessed both metabolic and heart rates throughout incubation in constant and fluctuating temperatures. Turtles appeared to hatch at similar developmental stages, with less-developed embryos in experimental groups responding to the presence of more developed eggs in a clutch by increasing both metabolic and heart rates. Early hatching did not appear to reduce neuromuscular ability at hatching. These results support developmental adjustment mechanisms of the 'catch-up hypothesis' for synchronous hatching in E. macquarii and implies some level of embryo-embryo communication. The group environment of a nest strongly supports the development of adaptive communication mechanisms between siblings and the evolution of environmentally cued hatching.     

Laboratory studies of factors affecting egg hatch of triops longicaudatus (LeConte) (Notostraca : Triopsidae)

Egg hatch was greatest (78.33%) for eggs not previously desiccated. A reduction in numbers hatched occurred as the relative humidity at which they were dried decreased. Some eggs hatched (0.67–79.33%) at pH levels of 3.10–10.01 with the highest hatch at pH 5.60. Water temperature greatly affected egg hatch. No hatch occurred until temperatures were above 14°C. A constant 29°C significantly inhibited hatching. Egg hatch increased 13.00 to 43.42% as salinity decreased from 2200 to 9.24 micromhos/cm. As little as 13 mm of flooded soil covering the eggs prevented them from hatching for 14 days. Eighteen percent hatch resulted when soil and eggs were redistributed to a 1 mm soil layer. Egg samples from the same parent, even though treated similarly, often hatched at greatly varying rates and only rarely was hatching 100% within a replication.     

Methylmercury teratogenesis in the killifish, Fundulus heteroclitus.

Exposure of developing eggs of the killifish, Fundulus heteroclitus, to 0.03 or 0.04 mg/l of methylmercuric chloride resulted in a variety of abnormalities. Percentage of axis formation was reduced somewhat, and many embryos developed cyclopia or intermediate conditions leading to cyclopia, reflecting interference with induction of the forebrain. Defects in the cardiovascular system also appeared in the form of failure of the heart to differentiate properly into chambers. The heart was a thin, feebly beating tube, incapable of causing the blood to circulate. Other tissues, however, continued developing fairly normally, and embryos showed spontaneous movement comparable to controls. Embryos with severe cardiovascular or optic defects did not hatch. Upon hatching, some embryos which had previously appeared normal were found to have skeletal malformations in the form of vertebral bends or the inability to uncurl from the position which they had while still inside the chorion. Exposure to the toxicant for shorter periods of time (6, 12, or 24 hours) reduced the incidence of abnormalities. The second day of development was found to be the most sensitive period.     

Effect of Storage Environment on Hatching of the Cyst Nematode Globodera ellingtonae

Globodera spp. eggs go through a diapause, which remains dormant until favorable hatching conditions are reached. Because of the regulatory concerns with cyst nematodes, it is often only possible to rear eggs for research in the greenhouse. However, hatch is often lower for greenhouse-produced eggs than for eggs obtained from the field. The goal of this research was to determine storage conditions for Globodera ellingtonae eggs produced in the greenhouse that would increase percentage hatch. Over 3 yr, G. ellingtonae greenhouse-produced eggs were stored in different environments (−20°C, 4°C, room temperature, and the field) in either dry or moist soil. Percentage hatch after exposure to the different environments was determined in potato root diffusate. Across two experiments, field-produced eggs had higher hatch rates (65.2%) than greenhouse-produced eggs (10.4%). Temperature did not have an appreciable influence on hatch of eggs stored dry in two experiments (2.8% to 8.4% and 3.8% to 8.6%), but hatch of eggs stored in moist soil was significantly higher than in dry soil at all temperatures except −20°C (26.8% and 28.7%). However, the ability of G. ellingtonae greenhouse-, microplot-, and field-produced eggs to reproduce on potato in field microplots was not different. Although it may not be possible to produce G. ellingtonae eggs in the greenhouse that have the magnitude of hatch as those produced in the field, hatching can be greatly increased by storing eggs in moist soil at either 4°C or room temperature.     

Improvement of egg hatch of Ceratitis capitata (Diptera: Tephritidae) for enhanced output

Embryonated eggs of Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) genetic sexing strain (GSS), VIENNA 8 were treated with low concentrations of five disinfectants-formaldehyde, iodine, sodium hypochlorite, peracetic acid, and quaternary ammonium--for decontamination and egg hatch improvement. The newly laid eggs were successfully treated with formaldehyde at 100 ppm for 1 min with 74.2% hatching and with quaternary ammonium at 150 ppm for 1 and 2 min with 70.4 and 69% hatching, respectively. Increased formaldehyde concentration may have affected the embryos, because it resulted in a decrease in the hatching percentage. However, egg viability was not impaired and hatch was not affected by quaternary ammonium treatment compared with controls and eggs treated with other disinfectants. Quaternary ammonium shows promise for decontaminating eggs and improving egg hatch.     

Captive breeding and embryonic development of Chiloscyllium punctatum Muller & Henle, 1838 (Elasmobranchii: Hemiscyllidae)

The fecundity, embryonic development, growth and viability of captive Chiloscyllium punctatum were investigated over two breeding and laying seasons. Chiloscyllium punctatum is an oviparous species, which lays rectangular eggs from late July to February. The six adult female C. punctatum in the open ocean display tank at UnderWater World, Mooloolaba, Australia laid a total of 692 eggs between July 2004 and February 2006, with 228 of those being viable. This equates to each female laying 115·3 eggs over the two seasons, with 38 viable. The embryos are first visible to the naked eye at c . 18 days post deposition (dpd), while pre-hatching is first seen c . 35 dpd, and the embryos hatch at an average of 153 dpd (temperature 21–25° C). Embryonic growth is most rapid from 99 dpd until hatching. An average of 21·4% of the eggs hatched over the two seasons.     

Desiccation tolerance in embryonic stages of the tardigrade

Desiccation tolerance commonly found among tardigrades allows them to cope with temporal variation of available water. Although the long-term survival of adults has been demonstrated in several species, desiccation tolerance of eggs and embryos is less well studied, however it is an important aspect from an ecological and evolutionary point of view. For the first time we evaluated the desiccation tolerance and subsequent hatching success of five different developmental stages of the tardigrade species Milnesium tardigradum , when rehydrated following drying at eight different humidity levels (10, 20, 31, 40, 54, 59, 72, 81%). Humidity level and developmental stage are significant factors in determining successful hatch rates. The results showed that the less developed stages were quite sensitive to desiccation. Low humidity levels during the first 3 days of development lead to a decrease in hatch rates following rehydration. Later developmental stages showed higher hatch rates than embryos dried at earlier stages. However, fast drying at low humidity levels resulted in delayed development and lower hatch rates following rehydration. In general, further developed embryos exhibit a better survival capacity compared with younger stages.     

Desiccation tolerance in fully developed embryos of two cicadas, Cryptotympana facialis and Graptopsaltria nigrofuscata

In two Japanese cicadas, Cryptotympana facialis and Graptopsaltria nigrofuscata , with different habitat distributions, fully developed embryos hatch in response to high humidity due to rainfall. Despite the advantage of hatching on rainy days, this trait burdens embryos with an extra period of desiccation until the unpredictable advent of rain. We compared the ability of the fully developed embryos of these cicadas to endure periods of low humidity. Eggs were exposed to a combination of different humidities (43% and 75% relative humidity, RH) and durations (0–15 days), and then transferred to an environment with 100% RH to stimulate hatching. In both species, total hatching rates decreased as duration increased, although there was no significant effect of humidity. In C. facialis , a considerable proportion of the eggs hatched during the desiccation period, and the hatching rate was higher at 75% RH than at 43% RH. After transfer to 100% RH, most hatching occurred within a day regardless of the desiccation level. In G. nigrofuscata , no nymphs hatched during the desiccation periods. However, more eggs required more than a day after transfer to 100% RH to hatch after desiccation at 43% RH than at 75% RH. Consequently, the overall proportion of timely hatching of eggs (eggs hatching within a day of moisture supply) was higher after desiccation at 43% RH in C. facialis , but it was higher after desiccation at 75% RH in G. nigrofuscata . These different physiological responses of the two species may reflect adaptation to habitat dryness.