Genetic variability and transgenerational regulation of investment in sex in the monogonont rotifer Brachionus plicatilis

In cyclical parthenogens such as aphids, cladocerans and rotifers, the coupling between sexual reproduction and the production of resting stages (diapausing eggs) imposes strong constraints on the timing of sex. Whereas induction of sex is generally triggered by environmental cues, the response to such cues may vary across individuals according to genetic and nongenetic factors. In this study, we explored genetic and epigenetic causes of variation for the propensity for sex using a collection of strains from a Spanish population of monogonont rotifers (Brachionus plicatilis) in which variation for the threshold population density at which sex is induced (mixis threshold) had been documented previously. Our results show significant variation for the mixis threshold among 20 clones maintained under controlled conditions for 15 asexual generations. The effect of the number of clonal generations since hatching of the diapausing egg on the mixis ratio (proportion of sexual offspring produced) was tested on 4 clones with contrasted mixis thresholds. The results show a negative correlation between the mixis threshold and mixis ratio, as well as a significant effect of the number of clonal generations since fertilization, sex being repressed during the first few generations after hatching of the diapausing egg.     

Low crowding threshold for induction of sexual reproduction and diapause in a Patagonian rotifer

1. This study investigates the basis and ecological significance of the extremely high propensity for mixis (sexual reproduction) observed in laboratory populations of Brachionus calyciflorus from a temporary pond in Patagonia. 2. Experiments with stem females hatched from resting eggs showed that these females were exclusively amictic but produced mictic daughters even at very low population densities. In six experiments, newly hatched stem females were cultured individually in different volumes (1.5, 12, 40 and 150 mL). The percentage of mictic daughters (mixis ratio) was high in the smaller volumes (e.g. 44–83% in 1.5 mL) and lower in the largest volume (6–21% in 150 mL). A regression analysis combining the data from these experiments showed a highly significant decrease in mixis ratio with volume and indicated that the lowest population density at which mixis still occurs (mixis threshold) would be 3.4 females L^?1 (95% CL 2.9–4.0 females L^?1). This value is considerably lower than mixis thresholds for other rotifers (25–250 females L^?1 for many species and 9000–477 000 females L^?1 for some Spanish Brachionus plicatilis). 3. In three additional experiments, stem females and their amictic daughters were cultured individually in 150 mL. The percentage of mictic daughters produced by these two generations of females was not significantly different, showing that the mixis response to crowding is not inhibited in the stem‐female generation. 4. Laboratory experiments showed that two common predators of the Patagonian B. calyciflorus (the calanoid copepod Parabroteas sarsi and the backswimmer Notonecta vereertbruggheni) each ate 60–70 B. calyciflorus predator^?1 per day and cleared all rotifers from c. 250 mL per day. Thus, a very low mixis threshold and high maximal mixis ratio may ensure production of some resting eggs soon after colonisation of the pond and before complete removal from the plankton. 5. Two laboratory experiments showed that resting eggs of the Patagonian B. calyciflorus hatched at variable rates (28 and 81%) after a brief diapause when kept in the conditions under which they were produced and oviposited (20–21 °C; L: D 16: 8). Early hatching of resting eggs from pond sediment may allow multiple periods of colonisation and resting egg production in a season. This may offset the fitness cost of limited population growth through female parthenogenesis in the face of unpredictable and abrupt risk of extinction because of predators.     

Effect of diazinon on life stages and resting egg hatchability of rotifer Brachionus plicatilis

The effects of organophosphate pesticide, diazinon, on life history parameters and hatchability of resting eggs of rotifer Brachionus plicalitis were assessed. Newly hatched (<1 h-old) neonates were individually cultured in six varying concentrations (0/control, 0.1, 1.0, 2.5, 5.0 and 10.0 mg/l) of diazinon. The life history parameters such as time (h) the rotifers bear first egg and release first neonate, reproductive period, net reproductive rate, mixis, intrinsic rate of population increase, and life span were evaluated. Results showed that among the life history parameters, the time the rotifers took to release neonates is the most sensitive, giving the lowest EC₅₀ value of 1.24 mg/l. The fecundity of maternal females, amictic and mictic daughters was also investigated. Rotifers exposed to 10.0 mg/l produced significantly fewer amictic daughters, and at this concentration, rotifers did not produce any mictic daughter. At 5.0 mg/l, the number of male offspring was significantly lower than the control. Furthermore, the hatchability of resting eggs produced by the rotifers was evaluated when exposed to diazinon: from birth until they produced resting eggs (early development); during late developmental stage of resting eggs (before diapause); and during diapausing stage. The hatchability of the resting eggs was not affected when exposure was timed at late developmental and diapausing stages. Overall results showed that even though amictic females reproduced normally in the presence of low-concentration of diazinon, sexual reproduction is severely affected, especially the hatchability of resting eggs when the exposure was timed on its early developmental stages. Guest editors: S. S. S. Sarma, R. D. Gulati, R. L. Wallace, S. Nandini, H. J. Dumont & R. Rico-Martínez Advances in Rotifer Research     

Sexual reproduction and diapause of Hexarthra sp. (Rotifera) in short-lived ponds in the Chihuahuan Desert

1. In the life cycle of monogonont rotifers it is generally assumed that diapausing eggs invariably hatch into amictic stem females which produce female offspring parthenogenetically. Diapausing eggs are only produced by later generations after sexual reproduction has been induced by environmental cues. 2. We show that populations of an undescribed Hexarthra species inhabiting small temporary ponds in the Chihuahuan Desert deviate from this life cycle pattern. These ponds may dry within days and up to 85% of females were mictic. Females producing male offspring and diapausing eggs were observed 1 or 2 days, respectively, after ponds had filled with water. 3. Under laboratory conditions, 7–46% of females hatching from re‐hydrated sediments were sexual. Male offspring of these females can fertilise other mictic stem females leading to diapausing egg formation. In laboratory experiments, females produced fully developed diapausing eggs within 1.9 days at 20 °C and 1.2 days at 30 °C. 4. In addition, embryonic development time (1.1–0.3 days at temperatures between 12 and 30 °C) and juvenile period (2.1–0.5 days for the same temperature range) are shorter than those of other rotifer species. In short‐lived habitats, the potential for rapid population development and production of new diapausing eggs may be crucial in the long‐term survival of populations.     

Factors influencing the induction of diapausing egg production in the calanoid copepod Diaptomus leptopus

Field and laboratory studies were carried out between 1995 and 1997 on four populations of Diaptomus leptopus found in seasonally temporary, occasionally temporary, and a permanent environment to assess the relative importance of photoperiod and temperature regimes versus other proximate local cues in inducing diapause egg production. Patterns of diapausing and subitaneous egg production were determined by observation of individual females bearing clutches that were produced in the field. A laboratory common-garden experiment was performed to assess the effects of four different regimes of temperature and photoperiod on the induction of diapause. Patterns of diapausing egg production differed among ponds: diapause occurred early in the seasonally temporary environment and occurred rarely in the permanent environment. In the common-garden experiment, populations exhibited substantial changes in the onset of diapause when compared to patterns found under field conditions. These results provide indirect evidence that the different populations respond to available cues of environmental change in different ways in nature, or that environmental cues vary among habitats.     

Effect of incubation and preservation on resting egg hatching and mixis in the derived clones of the rotifer Brachionus plicatilis

The marine rotifer Brachionus plicatilis typicus (Clone 8105A, Univ. of Tokyo) was cultured in 500 ml beakers to form resting eggs. Tetraselmis tetrathele was used as a culture food. Just after formation, resting eggs were exposed to various temperature (5–25 °C) and light regimes (24L: OD and OL : 24D). When eggs were exposed to light just after formation, the eggs hatched sporadically over a month. No hatching was observed for six months when eggs were preserved under dark conditions regardless of the temperature. These eggs hatched simultaneously after being exposed to light and eggs preserved at 5 °C showed twice as high hatching rate (40%) as that of eggs preserved at 15–25 °C (24%). Clones from resting eggs that were kept under different temperature and light regimes were reared individually to the third generation. Incubation at 25 °C with lighting produced the highest (5.4% and 5.2 %) rate of mictic females during their 2nd and 3rd generations, respectively. The lowest rates (0 and 1.5%) were found when the eggs were kept at 5 °C in total darkness for six months. A lower rate of amictic female production was found in clones with higher rates of mixis.     

Nondormanz-Entwicklung prospektiver Diapause-Eier vonBombyx mori mit oder ohne Chorion unter Paraffinöl

Summary Prospective diapause eggs ofBombyx mori become permeable to water in or after contact with paraffin, and they die before reaching diapause. If the eggs are hindered from drying up, their germ anlagen develop to diapausing germ bands. Prospective diapause eggs opened in paraffin, always develop without egg diapause, but their degree of organogenesis depends on the chorion: When eggs are completely dechorionized most of them develop to hatching or nearly hatching larvae; when eggs are completely dechorionized, but their chorion remains in the paraffin, the degree of organogenes is issignificantly lower; and when the opened eggs keep 1/2–3/4 of the chorion, their organogenesis is significantly lower than in the two previous experiments.It is discussed that nondormancy development ofprospective diapause eggs in vitro is not a specific effect of culture media containing water but is possible in paraffin oil too. It is not only possible to stop but also to prevent egg diapause. Nothing, as yet, can be said about the prospective significance of the chorion's reducing factor in organogenesis.     

Life cycle patterns of rotifers in Lake Peipsi

Life cycle strategies of rotifers in Lake Peipsi (Estonia) were examined. Bisexual (mictic) reproduction was detected in 26 species. Life cycle patterns were determined for 17 species. All three basic life cycle patterns were represented, with some evidence of intraspecific variability. The midcycle pattern, with mixis occurring near the population maximum, prevailed. Extended mixis was observed in littoral populations of Polyarthra luminosa and P. remata. Most species probably are monocyclic. Polyarthra dolichoptera and Synchaeta oblonga may be dicyclic. Mictic periods were not detected in Conochilus hippocrepis and C. unicornis, which probably are acyclic in L. Peipsi. Mictic ratios and egg ratios were calculated for some of the dominant species. The highest amictic egg ratios were observed slightly before or at the population maxima. High mictic ratios (1.0) indicated very intense bisexual reproduction in populations of A. fissa, P. dolichoptera and S. verrucosa. Definite periods of bisexual reproduction could not be distinguished in the rotifer community. During May–October, the spread of mictic reproduction merely reflects the general seasonal distribution of rotifers. Their life cycle strategies represent specific adaptations to unpredictability of their habitat. The results of the study confirm that mixis is an anticipatory event, and not a response to environment deterioration, or an ending of a rotifer population cycle.     

Induction of sexual reproduction and resting egg production in Brachionus Plicatilis reared in sea water

Brachionus plicatilis raised in our laboratory in sea water reproduces asexually even under high crowding conditions (at least 40 individuals per ml). Amictic females were induced to produce mictic females, males and resting eggs by reducing the concentration of the sea water culture medium. Mictic females and males appeared predominantly among the progeny produced by the amictic females during 4 days following their transfer into 25% sea water. Resting eggs appeared first 5–12 days after the onset of the experiment. Following the disappearance of males, the culture consisted of amictic females.Resting eggs produced by the method described above may be preserved for at least three months at –14°C or by desiccation at room temperature. Under the appropriate experimental conditions, resting eggs hatch into amictic females. Since B. plicatilis is one of the most commonly used food sources of fish larvae in aquaculture, the methods reported here may offer an easy and versatile way of preserving rotifer culture stock to be used on demand.     

Optimal rates of bisexual reproduction in cyclical parthenogens with density-dependent growth

This work explores theoretical patterns of reproduction that maximize the production of resting eggs and the long-term fitness of genotypes in cyclical parthenogens. Our focus is on density-dependent reproduction as it influences the consequences of a trade-off between producing amictic daughters – which reproduce parthenogenetically and subitaneously – and producing mictic daughters – which undergo meiosis and bisexual reproduction. Amictic females increase competitive ability and allow the population to achieve a larger size; mictic females directly contribute to population survival through harsh periods by producing resting eggs. Although morphologically indistinguishable, the two types of females differ greatly in their ecological and reproductive roles. What factors underlie the differential allocation of resources to produce amictic and mictic females? Using a demographic model based on readily accessible parameters we demonstrate the existence of a frequency of mictic females that will maximize the population's long-term fitness. This frequency, termed the optimal mictic ratio, m_o, is 1 ? (q/b)^1/2, where q is the mortality rate and b is the maximum birth rate. Using computer simulation we compared the fitness of a population with this constant mictic ratio with populations having multiple switches from complete parthenogenetic growth to complete allocation in mixis (mictic ratio either 0 or 1). Two important conclusions for optimal mixis in density-dependent growth conditions are: (1) intermediate mictic ratios are optimal, and (2) optimal mictic ratios are higher when habitat conditions are better. Physiological cues responding to differences in birth and death rates are common so that it is possible that populations may adjust their relative rates of mictic and amictic female production in response to environmentally induced changes to the optimum mictic ratio. Our analysis demonstrates that different patterns of mixis are expected in different type of habitats. Since the optimal mictic ratio is sensitive to the effects of a variety of environmental challenges, our model makes possible a new means to evaluate life history evolution in cyclical parthenogens.     

The influence of processing and temperature conditions on hatching of resting eggs of Streptocephalus proboscideus (Crustacea: Branchiopoda: Anostraca)

In large freshwater branchiopods, erratic hatching success of resting eggs is a major obstacle to various applications. Lack of knowledge of the diapause-regulating processes makes control of hatching difficult. In the Sudanese fairy shrimp Streptocephalus proboscideus cysts are considered to include a diapausing and a quiescent fraction. To effect hatching, diapausing cysts have to be activated, while the quiescent portion has to be triggered by suitable environmental conditions. Of several attempts to control hatching with varying production, processing, and incubation conditions, only a few treatments proved consistently successful. Cysts were produced in an indoor culture system under controlled conditions and were harvested, washed, and dried according to defined procedures before processing (if any) and incubation. Hatchability (first-day and cumulative) was consistently higher in chemically decapsulated than in untreated (non-decapsulated) cysts at 25 °C. At a temperature of 28 °C, hatching was comparable in untreated and in decapsulated cysts and was significantly higher than at 25 °C. Pre-treatment with 7.5% NaOCl for 5–10 minutes, resulted in higher hatching than other decapsulation procedures and durations. It is believed that the decapsulation and temperature treatments were only effective in triggering quiescent cysts but did not activate the diapausing fraction.     

The Insect Growth Regulator Pyriproxyfen Terminates Egg Diapause in the Asian Tiger Mosquito,Aedes albopictus

The Asian tiger mosquito, Aedes albopictus, is a highly invasive mosquito species that transmits chikungunya and dengue. This species overwinters as diapausing eggs in temperate climates. Early diapause termination may be a beneficial strategy for winter mosquito control; however, a mechanism to terminate the diapause process using chemicals is not known. We tested the hypothesis that a hormonal imbalance caused by the administration of juvenile hormone analog would terminate egg diapause in A. albopictus. We tested the insect growth regulator pyriproxyfen on all developmental stages to identify a susceptible stage for diapause termination. We found that pyriproxyfen treatment of mosquito eggs terminated embryonic diapause. The highest rates of diapause termination were recorded in newly deposited (78.9%) and fully embryonated (74.7%) eggs at 0.1 and 1 ppm, respectively. Hatching was completed earlier in newly deposited eggs (25–30 days) compared to fully embryonated eggs (71–80 days). The combined mortality from premature diapause termination and ovicidal activity was 98.2% in newly deposited and >98.9% in fully embryonated eggs at 1 ppm. The control diapause eggs did not hatch under diapausing conditions. Pyriproxyfen exposure to larvae, pupae and adults did not prevent the females from ovipositing diapausing eggs. There was no effect of pyriproxyfen on diapausing egg embryonic developmental time. We also observed mortality in diapausing eggs laid by females exposed to pyriproxyfen immediately after blood feeding. There was no mortality in eggs laid by females that survived larval and pupal exposures. In conclusion, diapausing eggs were the more susceptible to pyriproxyfen diapause termination compared to other life stages. This is the first report of diapause termination in A. albopictus with a juvenile hormone analog. We believe our findings will be useful in developing a new control strategy against overwintering mosquito populations.     

Temperature conditions enhancing resting egg production of the euryhaline rotifer Brachionus plicatilis O. F. Müller (Kamiura strain)

We examined the effect of low temperature treatment(12°C), followed by transfer to highertemperature (25°C), on resting egg formation ofthe rotifer Brachionus plicatilis, Kamiurastrain. This strain has been mass cultured as livefeed at Kamiura Station (Japan Sea FarmingAssociation) for 9 years at 20°C without theappearance of sexual reproductive stages.After preculture in 20 l of 27 seawater at 12°C for 0, 10, 20, and 30 days,rotifers were inoculated into 0.5 l mass cultures andcultured at 25°C for 7–9 days. The inoculationdensities were changed from 20 to 400 ind. ml^–1,depending on mixis rate. Condensed and frozen Nannochloropsis oculata was fed to rotifers at thefeeding rate of 0.14 µg (dry weight)rotifer^–1day^–1. The control was cultured at12°C for the entire 36 day experiment. No mixisappeared and no resting eggs were produced when thelow temperature treatment was 0 or 10 days. However,mixis rates reached 50-60% after 20 or 30 days ofexposure to 12°C. The number of resting eggsproduced in these treatments reached 25,500 about 13 times higher than the control. Our resultssuggest that low temperature stimulated mictic femaleproduction and the transfer to the high temperatureaccelerated resting egg formation. This method may beuseful for producing resting eggs of rotifer strainsthat lack sexual reproduction in the common culturecondition at larval rearing facilities.     

Experimental evolution of bet hedging in rotifer diapause traits as a response to environmental unpredictability

The adaptive response of organisms to unpredictable environments is increasingly recognized as a central topic in fundamental and applied evolutionary ecology. Selection due to environmental unpredictability can act on multiple traits of an organism's life cycle to reduce the impact of high environmental variance. The aim of this research was to study how unpredictability selects for diapause traits: 1) the timing of sex (a proxy of the timing of diapausing egg production), and 2) the diapausing egg hatching fraction (a proxy of diapause duration). We used an experimental evolution approach with the facultative sexual rotifer Brachionus plicatilis. Laboratory populations experiencing two contrasting regimes of environmental fluctuation (predictable versus unpredictable) evolved divergently over a short time span (< 77 days). The populations under the unpredictable regime showed an earlier initiation of sexual reproduction and a lower hatching fraction of diapausing eggs than populations under the predictable regime. These findings demonstrate empirically the existence of bet‐hedging strategies in B. plicatilis regarding both traits, consistent with theoretical predictions of bet‐hedging evolution under conditions of unpredictable environmental variance. Given that scenarios of increased environmental variability are expected to occur in the near future, a comprehensive understanding of the role of bet‐hedging strategies is necessary for predicting population responses to environmental change.     

Delayed mixis in rotifers: an adaptive response to the effects of density-dependent sex on population growth

In most cyclically parthenogenetic life cycles, sex is neededto produce resting stages. In several species of cyclicallyparthenogenetic rotifers, some generations of clones are notresponsive to a density-dependent signal that triggers sexualfemale production. These unresponsive rotifers hatch from restingeggs and typically pass 8–12 generations of female parthenogenesisbefore becoming receptive to the mixis signal. We addressedthe selection for mixis delay using a simulation model. A delayof sexual reproduction could increase population growth throughparthenogenesis and thus the number of resting eggs ultimatelyproduced. In a monomorphic population without mixis delay, wedetermined the optimal ratio of mictic to amictic females (mixisratio) to be 45%, and the optimal population density thresholdfor induction of mictic females (mixis threshold) to be 82 rotifersL^–1. This mixis pattern, however, was not an evolutionarilystable strategy. A mixis ratio of 14% and threshold of 70 rotifersL^–1 proved to be resistant to invasion by other mixispatterns. When we gave this phenotype a mixis delay of 8–10days, it could invade a population with the same mixis pattern,but lacking a mixis delay. The advantage of delaying mixis wasrelatively small, suggesting that a polymorphism is possible.     

Effect of Some Pesticides on Reproduction of Rotifer <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> Müller

Pesticides have been major contributors to environmental pollution and they are now widely distributed in aquatic environments. Zooplankters are frequently used as test animals to detect aquatic contaminants because of their sensitivity and ecological importance. We investigated the effect of a 7-day exposure to four commonly used pesticides (diazinon, fenitrothion, methoprene and isoprothiolane) on reproduction of the rotifer Brachionus plicatilis. Pesticide concentrations of 3–7 times lower than the 24-h 50% lethal concentration (24-h LC₅₀) were tested to determine the ‚no observed effect’ concentration (NOEC), ‚lowest observed effect’ concentration (LOEC), and the ‚50% effective’ concentration (EC₅₀) on specific growth rate (r), sexual reproduction, fertilization, resting egg production, and hatchability of resting eggs. Results showed that the lowest EC₅₀ value of r, mixis, fertilization, and resting egg production of 1.4 μM for diazinon was 63 times lower than its 24-h LC₅₀ of 88.4 μM, while for fenitrothion it was 66 times (3.5 and 229.8 μM, respectively). For isoprothiolane, the lowest EC₅₀ value of r, mixis, fertilization, and resting egg production of 8.9 μM was 25 times lower than its 24-h LC₅₀ of 220.7 μM, while for methoprene was 37 times (2.7 and 100.8 μM, respectively). In all pesticides, hatching rate of resting eggs consistently gave the lowest EC₅₀ values which is about 2–40 times lower than the lowest EC₅₀ of r, mixis, fertilization, and resting egg production. Hatchability of resting eggs therefore is the most sensitive parameter in detecting effects of pesticides exposure in rotifer B. plicatilis.     

Population differences in the timing of diapause: a test of hypotheses

Summary The reproductive phenology of the freshwater copepod Diaptomus sanguineus differs markedly between populations residing in two Rhode Island ponds. In a permanent pond the population switches abruptly from making subitaneous (immediately hatching) eggs to diapausing eggs at the end of March each year. In contrast, a temporary pond population switches egg types in May, returns to production of subitaneous eggs in June, and concludes the reproductive season by making diapausing eggs in July. An ESS model suggests that the pattern of diapause expected of a copepod population is a function of annual variation in the onset of harsh conditions (catastrophe date). When variation is relatively low, the superior strategy is for diapause to begin a constant period before the mean catastrophe date. When variation is high, females should make first subitaneous eggs and then diapausing eggs irrespective of the expected catastrophe date. With discrete generations, such a population would alternate between egg types. In the permanent pond, variation of catastrophe date the spring onset of planktivory by sunfish is low, whereas in the temporary pond variation of the catastrophe (pond drying) is high. The model predicts well the phenology of the two copepod populations.In the research reported here, we tested the hypothesis that copepods from the permanent pond, which switch to diapause at the same time every year, are cued by the environment to begin diapause (i.e. by photoperiod, temperature, or both), whereas those from the temporary pond make both egg types regardless of environmental conditions. In opposition to our hypothesis, experimental results indicate that diapause in both populations is cued by the environment. The distinct reproductive phenologies documented in the two populations apparently result from the copepods responding to different environmental cues, rather than one being responsive to the environment while the other is not.     

Resting eggs of the marine rotifer Brachionus plicatilis Müller: development,and effect of irradiation on hatching

Diapause and hatching of Brachionus plicatilis Müller resting eggs were examined through histological and optical approaches. Compound microscope observations on 1% toluidine blue-stained embryo sections suggests that the total number of nuclei in an embryo during the internal diapause period increased from 22 on Day 2 to 39 (each n = 1) on Day 6. The outer layer of embryo membrane gradually thickens from 1.2 (Day 0) to 4.0 µm (Day 8) (each n = 10).Resting eggs that have completed maturation and are in the external diapause period require light for hatching. The threshold of light (halogen lamp) intensity for hatching was estimated to be 4400 lux for 30 min. Hatching rate decreased with longer wavelength irradiation (mercury lamp). Irradiation at more than 350 nm caused 1–25% hatching, but it reached 50–60% at 250–310 nm light. The addition of hydrogen peroxide or prostaglandins (E ₁, E ₂ or F ₂) caused resting egg hatching even in darkness. The production of peroxide in seawater caused by light as well as the oxidation of fatty acid to prostaglandins inside the embryo is a possible mechanism of resting egg hatching.     

Diapause duration, survival in relation to desiccation and egg-pod morphology of the Senegalese grasshopper, Oedaleus senegalensis

Abstract. The Senegalese grasshopper, Oedaleus senegalensis (Krauss) (Orthoptera: Acrididae), is periodically a devastating pest of subsistence crops in the sahelian zone of West Africa.Egg diapause has been suggested as an important component of the mechanism acting to generate outbreaks and therefore diapause duration, egg survival in relation to desiccation and egg-pod morphology were investigated in this study.
Three months after oviposition, 50% of initially diapausing egg pods maintained in the laboratory exhibited 'partial emergence' and produced hoppers.No difference in diapause duration was evident between egg pods maintained at a constant 30 ± 1°C and those maintained under alternating temperature conditions of 23 ± 1°C and 35 ± 1°C.Laboratory maintained eggs showed significant variation in diapause duration with diapause lasting between 2 months and 4 years.Diapause duration was shorter in the field with only c. 1.4% of eggs remaining in diapause after a single dry season (6–7 months).Survival of eggs in relation to desiccation was high in the field and low in the laboratory, and field eggs lost only c. 20% of their body weight over the dry season.Survival of diapausing and non-diapausing egg pods in the laboratory was similar, indicating that diapause in this species is primarily a mechanism for preventing emergence at an inappropriate time of year, rather than conferring any additional resistance to desiccation.Egg pods oviposited under diapause-inducing conditions (LD 14:10 h, 25°C) were significantly shorter than those laid under non-diapausing conditions (LD 10:14 h, 40°C).These results are discussed both in relation to possible O.senegalensis 'bet-hedging' strategies and to the probable mechanism operating behind major outbreaks of this species.     

Spawning Behavior and Biparental Egg Care of the Crosshatch Triggerfish, Xanthichthys mento (Balistidae)

Reproductive ecology of the crosshatch triggerfish, Xanthichthys mento (Balistidae) was studied at Hachijojima, Izu Islands, Japan. Males established territories and repeatedly chased females passing nearby. There were 1–3 females in each male's territory before spawning and during egg care. This species spawned in pairs on the sandy bottom. Eggs were scattered and attached to sand particles. Females care for the eggs by blowing water on them and guarding them against intruders, while males helped in guarding. Thus, biparental egg care was observed for 2 days until hatching. Both the males and females disappeared from the territories after the egg care. The reproductive ecology of this species is compared with that of other balistids and the unique features of X. mento are described.