Differences in Hatching of Heterodera glycines Egg-mass and Encysted Eggs in vitro

Hatching studies with Heterodera glycines typically have been conducted with a mixture of egg-mass and encysted eggs. Laboratory research was conducted to compare hatching of H. glycines eggs from external egg masses with that of eggs extracted from within females and cysts (encysted eggs). Egg-mass eggs were collected by soaking infected soybean roots in 0.5% sodium hypochlorite, and encysted eggs were collected from females and cysts dislodged from the same roots with a stream of water. Eggs were incubated at 25 °C in deionized water, 3.0 mM ZnSO₄solution, or one of three synthetic H. glycines hatch inhibitors, mad hatched juveniles were counted every other day for 22 days. Samples of eggs collected at the beginning and end of all experiments were analyzed to determine extent of embryo development. Egg-mass eggs hatched more rapidly than encysted eggs during the first 16 days, but not thereafter. Throughout the experiments, hatch of egg-mass eggs in deionized water was greater than that of encysted eggs. From day 8 to day 22, egg-mass eggs were less sensitive than encysted eggs to the hatch inhibitor 2-(2''-carboxyethyl)-5-[carboxy(hydroxy)methylidenyl]cyclopentanone. A greater proportion of egg-mass eggs contained vermiform juveniles than did encysted eggs at the beginning of the experiments, but not at the end. Results indicated that H. glycines egg-mass and encysted eggs have different hatching behaviors that cannot be explained entirely by differences in embryological development.     

Plant-Induced Hatching of Eggs of the Soybean Cyst Nematode Heterodera glycines

Root diffusate from soybean plants caused greater hatching of Heterodera glycines eggs during vegetative growth of the host, but the activity declined with plant senescence. Chelation of the root diffusate with ethylenediamine tetraacetic acid (EDTA) significantly increased hatching activity for H. glycines eggs. Diffusate from leafless plants caused little hatching, whereas treatment of intact plants with the growth regulators gibberellin and kinetin had no effect on the hatching activity of root diffusate. Treating H. glycines eggs with zinc chloride and root diffusate reduced egg hatching from zinc chloride alone. Levels of zinc in the root diffusate were insufficient to induce egg hatch, based on analysis by atomic absorption spectrophotometry. The enzymatic activity of leucine aminopeptidase in H. glycines eggs was not altered by treatment with chelated or nonchelated root diffusate.     

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.     

Recovery and Longevity of Egg Masses of Meloidogyne incognita during Simulated Winter Survival

Effects of soil matrix potential on longevity of egg masses of Meloidogyne incognita were determined during simulated winter conditions. Egg masses were recovered from isolated root fragments incubated in field soil at matrix potentials of -0.1, -0.3, -1.0, and -4.0 bars throughout winter survival periods of 10 weeks for tomato roots and 12 weeks for cotton roots. Egg masses were more superficial on cotton roots than on tomato roots and were more easily dislodged from cotton roots during recovery of root fragments by elutriation. The rate of decline in numbers of eggs and J2 per egg mass was greater in wet as compared to dry soils (P = 0.01), with the relationship between numbers of eggs and J2 per egg mass and time being best described by quadratic models. Percentage hatch of recovered eggs declines linearly with time at soil matrix potentials of -0.1 and -0.3 bars, but at -1.0 and -4.0 bars the percentage hatch of recovered eggs increased before declining. Effects of soil matrix potential on numbers of eggs per egg mass and percentage hatch of recovered eggs were consistent with previous reports that low soil moisture inhibits egg hatch before affecting egg development. Estimations of egg population densities during winter survival periods will be affected by ability to recover infected root fragments from the soil without dislodging associated egg masses. There is a need for procedures for extraction of egg masses not attached to roots from the soil.     

Heterodera glycines Cyst Components and Surface Disinfestants Affect H. glycines Hatching

We investigated the effects of Heterodera glycines cyst components and surface disinfestants on hatching of H. glycines eggs in vitro. Eggs were incubated in either H. glycines cyst wall fragments, cyst wall and egg rinsate, egg homogenate, or control solutions of soybean root diffusate, sterile distilled water, or zinc sulfate. Hatch in cyst wall and egg rinsate, and egg homogenate, was greater (α = 0.05) than hatch in sterile distilled water; however, it was not different from hatch in zinc sulfate according to Dunnett''s test. Hatch in cyst wall fragments was similar to hatch in sterile distilled water. To determine whether surface disinfestants affected hatch, eggs were treated first with chlorhexidine diacetate, mercuric chloride, sodium hypochlorite, or streptomycin sulfate and then incubated in H. glycines egg homogenate, soybean root diffusate, sterile distilled water, or zinc sulfate. Hatch of eggs treated with chlorhexidine diacetate, mercuric chloride, and streptomycin sulfate was reduced (α = 0.05), and hatch of eggs treated with sodium hypochlorite was increased (α = 0.05) relative to hatch of nontreated eggs in all incubation solutions except zinc sulfate according to Dunnett''s Test. Hatch in zinc sulfate was similar among all surface disinfestants except mercuric chloride, where hatch was reduced relative to hatch of nontreated and other surface disinfestant-treated eggs.     

In-vitro Assays of Meloidogyne incognita and Heterodera glycines for Detection of Nematode-antagonistic Fungal Compounds

In-vitro methods were developed to test fungi for production of metabolites affecting nematode egg hatch and mobility of second-stage juveniles. Separate assays were developed for two nematodes: root-knot nematode (Meloidogyne incognita) and soybean cyst nematode (Heterodera glycines). For egg hatch to be successfully assayed, eggs must first be surface-disinfested to avoid the confounding effects of incidental microbial growth facilitated by the fungal culture medium. Sodium hypochlorite was more effective than chlorhexidine diacetate or formaldehyde solutions at surface-disinfesting soybean cyst nematode eggs from greenhouse cultures. Subsequent rinsing with sodium thiosulfate to remove residual chlorine from disinfested eggs did not improve either soybean cyst nematode hatch or juvenile mobility. Soybean cyst nematode hatch in all culture media was lower than in water. Sodium hypochlorite was also used to surface-disinfest root-knot nematode eggs. In contrast to soybean cyst nematode hatch, root-knot nematode hatch was higher in potato dextrose broth medium than in water. Broth of the fungus Fusarium equiseti inhibited root-knot nematode egg hatch and was investigated in more detail. Broth extract and its chemical fractions not only inhibited egg hatch but also immobilized second-stage juveniles that did hatch, confirming that the fungus secretes nematode-antagonistic metabolites.     

Relationship Between Egg Viability and Population Densities of Meloidogyne incognita on Cotton

Cotton seedlings were inoculated with a range of initial populations (Pi) of Meloidogyne incognita in greenhouse experiments to test the relationship between nematode population densities and egg viability. In two of three experiments, a significant (P < 0.05) negative linear relationship was detected between percentage of hatch of first generation eggs and log Pi. A similar relationship between hatch and root-gall index was observed. In two experiments numbers of eggs judged to be nonviable based on appearance were significantly greater (P < 0.05) in the highest Pi (60,000 eggs/seedling) treatments than in treatments with lower Pi (600-6,000 eggs/seedling). It was concluded that Pi affects egg viability measured as percentage of hatch and that this relationship may play a role in the density-dependent winter survival rates of Meloidogyne species.     

Life Cycle and Mating Behavior of Belonolaimus longicaudatus in Gnotobiotic Culture

The life cycle of Belonolaimus longicaudatus was observed in vitro on excised roots of Zea mays. Roots were cultured on Gamborg''s B5 medium in petri dishes with 1.5% agar adjusted to pH 5.8 and incubated at 28 °C in darkness. Second-stage juveniles (J2) fed on the roots and started the second molt (M2) to the third-stage juveniles 2 days after inoculation (DAI). The third molt (M3) to the fourth-stage juveniles occurred 7 DAI, followed by the fourth molt (M4) to males 13 DAI or to females 14 DAI. Nematode gender differences were observed by the end of the fourth molt. The first male appeared 15 DAI and the first female 17 DAI, after which mating occurred. Males were attracted to females, and mating was observed. Mating was required for reproduction. Fertilized females began to lay eggs 19 DAI and continued egg laying without the further presence of males during a 90-day observation. All of the eggs hatched. Unfertilized females rarely laid eggs, and none of the eggs were able to hatch. Feeding took place between each molt and before egg deposition occurred. The first-stage juveniles molted in the eggs 4 days after deposition, and J2 hatched from eggs 5 days after egg deposition. The life cycle from J2 to J2 was completed in 24 days.     

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.     

Effects of Zinc Fertilization of Corn on Hatching of Heterodera glycines in Soil

Experiments were conducted to determine the effects of zinc fertilizers on hatching and soil population densities of Heterodera glycines. In vitro egg hatching in solutions of reagent-grade zinc sulfate and zinc chloride and fertilizer-grade zinc sulfate was significantly greater than hatching in deionized water, whereas zinc chelate fertilizer significantly inhibited egg hatching relative to deionized water. In greenhouse experiments, no differences in cumulative percentage egg hatch were detected in soil naturally infested with H. glycines amended with fertilizer-grade zinc sulfate and zinc chelate at rates equivalent to 0, 1.12, 11.2, and 112 kg Zn/ha and subsequently planted with corn (Zea mays L.). In a field experiment, no significant differences in H. glycines egg population densities and corn yields were detected among plots fertilized with 0, 11.2, and 22.4 kg Zn/ha rates of zinc chelate. Yields of H. glycines-susceptible soybean planted in plots 1 year after zinc fertilization of corn plots also were not significantly affected. Zinc compounds significandy affected H. glycines egg hatching in vitro, but had no effect on hatching in natural soils.     

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.     

Zur fortpflanzungsbiologie von mesostoma ehrenbergii (Focke, 1836) (Turbellaria)

1. At temperature levels from 10 to 25°C animals from resting eggs produce subitaneous eggs independent on temperature. In contrast animals from subitaneous eggs produce subitaneous eggs dependent on temperature. At a high rate subitaneous eggs are only formed at temperature levels above 20°C.
2. Below 10°C no development occurs in the juveniles. At temperatures of 30/22°C (24.7°C) the first subitaneous eggs are formed after 6–9 days, at 14/9°C (10.7°C) they are formed after 34 days. At different temperature levels the developmental rate of the young is from 10.5 to 42 days. One generation extends over 16.5 (30/22°C) to 75 days (14/9°C). The average egg production is 10–20 subitaneous eggs or 30–60 resting eggs. The maximum egg production of one individual is 50 subitaneous eggs or 84 resting eggs. 50% of the animals have just formed resting eggs, before the juveniles are hatched. Resting eggs in the first egg-batch are formed 6–20 days later than subitaneous eggs. The duration of life is between 65 (30/22°C) and 140 days (19/13°C).
3. Young worms in resting eggs have a dormance period of at least 15–30 days.

At room temperatures (20°C) no juvenile in resting eggs hatches from water. By combining room and refrigerator (3.5°C) temperatures the hatching rate increases to a maximum of 85%. To reach a hatching rate of 50–65% the influence of low temperatures must be at least 30 days. At room temperatures 60% of the young in resting eggs hatch from mud covered with water. Combining high and low temperatures the hatching success is between 67 and 81%, where the highest percentage of the young may hatch at room temperature. Up to 90 days low temperatures cause a maximum hatching rate of 79%. It decreases to approximately 30% after 180 days. At high temperatures resting eggs preserved in 100% moist mud, survive for two months. By adding a period of low temperatures the hatching rate increases to a maximum of 52%. Low temperatures are survived for more than 6 months. Up to 30 days preservation at 3.5°C causes a maximum hatching rate of 61%, up to 12o days it decreases to 30%. At room temperature the young in resting eggs are not resistant against air-dried mud (30–40% rel. air moisture). Combining high and low temperatures air-dried mud is endured 1 month (hatching rate 5–14%). Preservation of 30–120 days at 3.5°C and 70% rel. air moisture result in a hatching rate of 43–61%. li]4. In the open air in Middle-Europe there occur 5–6 generations of M. ehrenbergii per life-cycle. The first generation hatches from resting eggs in May, where the production of subitaneous eggs is independent on temperature. All other generations up to October hatch from subitaneous eggs. The egg-production of those worms is dependent on environmental factors. In summer subitaneous egg production prevails, in autumn resting egg production. The abundance during the life-cycle is dependent on the number of animals which produce subitaneous eggs. Resting eggs are predestinated to endure periods of dryness and cold. The life-cycles of the species M. lingua and M. productum are different from those of M. ehrenbergii in length and in the number of generations. In both species 7 generations occur over 8 to 8.5 respectively 5.5 months. M. nigrirostrum only forms resting eggs. The life-cycle consists of one generation from February/March to May/June.     

Effect of storage temperature and duration on viability of eggs of Helicoverpa armigera (Lepidoptera: Noctuidae)

The ability to store different insect stadia for prolonged periods provides considerable flexibility and ability to conduct experiments properly. Therefore, studies were undertaken to determine the effect of storage temperature and duration on viability of eggs of Helicoverpa armigera (Hübner). The percentage egg hatch and incubation period were significantly (P=0.01) influenced by egg age, storage temperature, and storage duration. Egg hatch ranged from 0.0 to 96.8% across temperatures and storage durations. None of the eggs hatched when stored at -20 and 0 degrees C. The regression model with the optimum Mallow Cp statistic for any of the identified linear and quadratic terms did not improve the precision of prediction in egg hatch beyond 67.0%. Forecasting of incubation period based on egg age, storage duration, and durationxtemperature was quite effective (R2=84.2%). Day degrees required for egg hatching decreased with an increase in temperature from 10 to 27 degrees C, and egg age from 0 to 3 days. The day degree requirements were highest for 0-day-old eggs at 10 degrees C, and lowest at 27 degrees C. Although the incubation period was higher, the hatchability was lower for 0- and 1-day-old eggs stored at constant 10 degrees C, these eggs can be stored for 10 days at 10 degrees C, with a hatchability of >75.0%. It was safer to store the H. armigera eggs for 10 days at 10 degrees C, which will hatch within 1.6 to 2.0 days after restoration at 27 degrees C with a hatchability of >75.0%. This information will be useful in planning and execution of experiments involving H. armigera on various aspects of research in entomology.     

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.     

Degree-day Models for Predicting Egg Hatch and Population Increase of Criconemella xenoplax

A degree-day model was derived to predict egg hatch for Criconemella xenoplax. Eggs collected from gravid females were incubated in distilled water at constant temperatures of 10-35 C. Sixty-six percent of all eggs hatched between 13 and 32 C, and 42% hatched at 10 C. All eggs aborted above 32.5 C. Between 25 and 32 C, 8.5 ± 0.5 days were required for egg hatch. Degree-day requirement for egg hatch at 10-30 C was estimated to be 154 ± 5 with a base of 9.03 ± 0.04 C. This base of 9 C was adopted in studies of the relationship between degree-days and nematode population increase on Prunus seedlings grown 9-11 weeks in a greenhouse. Degree-day accumulations were based upon daily averages from maximum and minimum air temperatures. Ratios of final to initial population densities exhibited an exponential pattern in relation to degree-day accumulations with proportionate doubling increment of 0.100 ± 0.049 every 139 ± 8 degree-days. These results provide a means of predicting nematode population increase under greenhouse conditions and a basis for choosing sampling intervals when evaluating nematode multiplication.     

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.     

The hatching time of Locusta migratoria under outdoor conditions: role of temperature and adaptive significance

The present study investigates the time of hatching of the migratory locust Locusta migratoria using egg pods that are artificially buried in the soil under outdoor conditions. Most eggs hatch in the mid‐morning, with a peak between 11.00 and 12.00 h, and none hatch before 09.00 or after 16.00 h. Furthermore, most egg pods complete hatching within a day, although some take 2 or 3 days, and egg hatching is interrupted by rain. There are no large differences in hatching time from May to September. Laboratory experiments in which the eggs are exposed to temperatures simulating outdoor conditions show that soil temperature is the main factor controlling hatching activity. The increase in temperature in the morning appears to trigger egg hatching, as confirmed by laboratory experiments, which may explain the similar hatching times between seasons. The seasonal patterns of temperature variation and hatching time suggest that the hatching time of L. migratoria eggs may be adjusted to allow the hatchlings to be exposed to high temperatures in the afternoon so that they can harden their bodies quickly.     

Influence of Rhizoctonia solani on Egg Hatching and Infectivity of Rotylenchulus reniformis

The effects of culture filtrates of Rhizoctonia solani and root exudates of R. solani-infected cotton (Gossypium hirsutum) seedlings on hatching of eggs and infectivity of females of Rotylenchulus reniformis were evaluated in an attempt to account for the enhanced nematode reproduction observed in the presence of this fungus. Crude filtrates of R. solani cultures growing over sterile, deionized distilled water did not affect egg hatching. Exudates from roots of cotton seedlings increased hatching of R. reniformis eggs over that observed in water controls. Exudates from cotton seedling roots not infected or infected with R. solani did not differ in their effect on egg hatching. However, infection of cotton seedlings by reniform females was increased in the presence of R. solani, resulting in the augmented egg production and juvenile population densities in soil observed in greenhouse studies.     

Hatching rhythms in the desert locust, Schistocerca gregaria

ABSTRACT. The eggs of Schistocerca gregaria (Orthoptera, Acrididae) incubated under natural conditions hatch only within a few of hours on either side of dawn. This gated hatching is controlled by a circadian clock that is phase set by the diel fluctuations in the temperature of the soil surrounding the eggs. There is a circadian fluctuation in haemolymph sugar concentration which is initiated at least 4 days before hatching. However, eggs hatch arrhythmically unless given cycled temperature incubation for at least 10 days of a 12–13-day incubation. Increase in acetylcholine esterase content of the brain during the penultimate day suggests that increased hatching rhythmicity occurring at this time is the result of increased neural organization. Embryonic activity and respiration show no circadian rhythm but do provide confirmatory evidence of a quiescent phase prior to hatching. This quiescent phase is an integral part of gated hatching behaviour.     

Larval hatching in the horseshoe crab, Limulus polyphemus: facilitation by environmental cues

Female horseshoe crabs, Limulus polyphemus (Linnaeus), lay their eggs in nests on sandy beaches near the high water line. Embryos develop within the sand, hatch into trilobite larvae, and enter the water column when the nest is inundated. Given the diversity of tidal and shoreline inundation patterns that populations of L. polyphemus experience throughout their range (semidiurnal and diurnal tides, microtidal, and nontidal), hatching may also be facilitated by environmental triggers that serve to synchronize hatching and larval emergence with periods of high water. The objective of this study was to determine if larval hatching in L. polyphemus is triggered or facilitated by environmental cues. Stage 21 embryos were subjected to one of seven different treatments that simulated conditions experienced during inundation: (1) hydration, (2) agitation, (3) hydration and agitation, (4) hydration and agitation with sand, (5) osmotic shock, (6) terrestrial hypoxia, and (7) aquatic hypoxia. Hatching rates increased significantly under all simulated tidal conditions compared to controls and were highest (96%) for eggs simultaneously exposed to both hydration and agitation with sand. Measurements of the osmolarity of the perivitelline fluid of developing eggs collected from the field indicated that it is hyperosmotic to the ambient seawater and porewater. Thus, when inundated, eggs also experience a hypoosmotic shock, which would likely facilitate hatching by causing the eggs to swell, rupturing the egg membrane and thereby increasing the likelihood that larvae would hatch and enter the water column during periods of high water.