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.     

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.     

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.     

A mathematical model for the phase of sexual reproduction in monogonont rotifers

Recently, the optimal sex allocation in monogonont rotifers is studied in [1], and, as a closely related question, the relative frequencies of the relevant types of mictic females. The authors focus on the evolution of the age at which young mictic females lose their fertilization susceptibility and they address the threshold age of fertilization that maximizes resting egg production. Assuming that a stationary population is achieved, with stable age distribution, they obtain their results, without knowing the stationary population. Our aim is to study this problem in the framework of the theory of nonlinear age-dependent population dynamics developed by G. F. Webb in [13], which is more appropriate from the mathematical point of view and permits to us to obtain analytically the stationary population and consequently it is analytically shown that a threshold age of fertilization equal to the age of maturation is not an ESS, despite the fact that then the production of resting eggs is maximum, which has been obtained by simulation in [1]. Received: 30 June 1998 / Revised version: 17 May 1999 / Published on line: 5 May 2000     

Isolation of mixis-related genes from the rotifer <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> using subtractive hybridization

The monogonont rotifer Brachionus plicatilis produces resting eggs through sexual reproduction (mixis), which is affected by external and internal factors. We collected resting eggs from rotifers cultured at 15 and 25°C and hatched them with and without 14-day dormancy in the dark. Stem females hatched from both conditions were further cultured at 15, 20 and 25°C. We obtained two phenotypes, one with high mixis (more than 50%), which was hatched from resting eggs formed at 15°C without dormancy, and another in which sexual reproduction occurred at 25°C and resting eggs were formed at 15°C with a 14-day dormancy. In the latter phenotype, mictic females appeared at 15°C, but not at 25°C. Using subtractive hybridization, we isolated one gene from the latter phenotype of females that had no significant similarity to known genes in BLAST searches. We propose that this gene is unique to rotifer mictic reproduction. Ongoing characterization of this gene attempts understand its role in mixis.     

Influence of the age of algae fed to rotifers (Brachionus plicatilis O.F. Müller) on the expression of mixis in their progenies

Summary The sequence of the appearance of mixis in the rotifer Brachionus plicatilis was followed among the descendents of amictic rotifers transferred from a high salinity media (40 S) to a low one (9 S). All the neonates that hatched from the amictic eggs, after being transferred to a low salinity, were amictic. Each one of these neonates was cultured individually and its offspring removed periodically every 8–10 h. It was observed that throughout their reproductive phase, these parental females retained their potential to produce either mictic or amictic offspring. All the first produced neonates developed into amictic females, but among those produced later, three patterns were prevalent. The prevalent pattern (type A) was one in which the probability of a neonate being mictic increased towards the middle of the parents' reproductive phase and was followed by a slow decline. In the second pattern (type B), the probability of a daughter being mictic was constant throughout the parents' reproductive phase. It is suspected that the quality of food supplied to the rotifers determines the appearance of patterns, A, B or C. It is postulated that the innate capacity of rotifers to undergo mixis is genetically controlled, while its expression is modulated by environmental conditions.     

Endogenous regulation of environmentally induced sexuality in a rotifer: a multigenerational parental effect induced by fertilisation

SUMMARY 1. Sexual reproduction in the heterogonic life cycle of many rotifers occurs when amictic females, which produce diploid eggs developing parthenogenetically into females, are environmentally induced to produce mictic females. Mictic females produce haploid eggs which develop parthenogenetically into males or, if fertilised, into resting eggs – encysted embryos which develop into amictic females after an obligatory diapause. 2. A Florida strain of Brachionus calyciflorus was used to test the prediction that amictic females hatching from resting eggs (Generation 1), and those from the next few parthenogenetic generations, have a lower propensity to produce mictic daughters in response to crowding than those from later parthenogenetic generations. In 10 replicate clones, populations initiated by amictic females from generations 1, 5, 8, 12 and 18 were exposed to a standardised crowding stimulus, and the proportion of mictic females in the populations was determined. These proportions varied significantly across generations and clones. They were very low in the early generations and gradually increased to a mean of about 0.5 at Generation 12. 3. The mechanism for the transgenerational plasticity in response to crowding is not known. One possibility is that resting eggs contain an agent from their fertilised mictic mother's yolk gland that prevents development into mictic females and is transmitted in increasingly low concentrations through successive parthenogenetic generations of amictic females. 4. This parental effect may contribute to clonal fitness by ensuring that a clone developing from a resting egg will attain a higher population size through female parthenogenesis before maximising its commitment to sexual reproduction, even in the presence of a crowding stimulus from a high population density of other clones. Therefore, the number of resting eggs to which a clone contributes its genes should be maximised. 5. The clonal variation in propensity to produce mictic females in this strain indicates genetic variation in the trade‐off between maximising population growth via female parthenogenesis and increasing the probability of producing at least some resting eggs before local extinction from the plankton.     

Effects of three organophosphorus pesticides on population growth and sexual reproduction of rotifer Brachionus calyciflorus Pallas

Effects of organophosphorus pesticides including dichlorvos, triazophos and chlorpyrifos on population growth and sexual reproduction of freshwater rotifer Brachionus calyciflorus were studied by 3-d population growth and 4-d resting egg production tests. The results showed that all the three organophosphorus pesticides influenced significantly the population growth rate, the ratio ovigerous females/non-ovigerous females in the rotifer populations and the resting egg production of the rotifers. Both dichlorvos and chlorpyrifos influenced markedly the mictic rate of the rotifers, but triazophos did not. Compared to the controls, both dichlorvos at 10.0–1000.0 μg/L and chlorpyrifos at 0.01–100.0 μg/L increased the population growth rate, but the reverse was also true for dichlorvos and triazophos both at 10000.0 μg/L. Chlorpyrifos at 10000.0 μg/L made the rotifers dead after 24-hr exposure. Dichlorvos and triazophos both at 10000.0 μg/L, and chlorpyrifos at 1000.0 μg/L all increased the ratio ovigerous females/non-ovigerous females. Both dichlorvos at 10000.0 μg/L and chlorpyrifos at 0.1–100.0 μg/L increased the mictic rate. Dichlorvos at 10.0 μg/L and 100.0 μg/L, and triazophos and chlorpyrifos both at 0.1–100.0 μg/L increased the resting egg production. Both population growth rate and ratio ovigerous females/non-ovigerous females are suitable endpoints for assessing the effects of dichlorvos, triazophos and chlorpyrifos, and mictic rate is a suitable endpoint for monitoring the effects of dichlorvos and chlorpyrifos on the reproduction of the rotifers. Both population growth rate and ratio ovigerous females/non-ovigerous females are more sensitive to dichlorvos and chlorpyrifos than mictic rate.     

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     

Virus morphological diversity and relationship to bacteria and chlorophyll across a freshwater trophic gradient in the Lake Michigan watershed

The effect of starvation on sexual reproduction in cyclic parthenogenetic rotifers has been studied using life history experiment. Short-time starvation of rotifers that experienced starvation immediately after hatching from resting eggs can cause high induction of sexual reproduction up to the 10th generation. However, it is not clear whether the induction of sexual reproduction can occur beyond the 10th generation. To investigate this phenomenon, we conducted a sex induction study using the monogonont rotifer Brachionus manjavacas. Newborn stem females were starved for 12 h, while controls were supplied with 7.0 × 10⁶ cells ml^?1 of Nannochloropsis oculata. In a life history experiment, the rotifers were individually cultured in 96-well microplates containing 0.2 ml of seawater (22 ppt) in each well at 25 °C with daily feeding thereafter. Mixis induction in offspring from starved stem females was significantly higher than in those from non-starved stem females up to the 40th generation. The effect of accumulative generations increased mixis induction up to the 20th generation. Effect on future generations of the rise in mixis ratio by the starvation to stem females may facilitate colonization by favoring population growth via female parthenogenesis and by decreasing food requirements for survival and reproduction.     

Effects of three organophosphorus pesticides on population growth and sexual reproduction of rotifer Brachionus calyciflorus Pallas

Effects of organophosphorus pesticides including dichlorvos, triazophos and chlorpyrifos on population growth and sexual reproduction of freshwater rotifer Brachionus calyciflorus were studied by 3-d population growth and 4-d resting egg production tests. The results showed that all the three organophosphorus pesticides influenced significantly the population growth rate, the ratio ovigerous females/non-ovigerous females in the rotifer populations and the resting egg production of the rotifers. Both dichlorvos and chlorpyrifos influenced markedly the mictic rate of the rotifers, but triazophos did not. Compared to the controls, both dichlorvos at 10.0–1000.0 μg/L and chlorpyrifos at 0.01–100.0 μg/L increased the population growth rate, but the reverse was also true for dichlorvos and triazophos both at 10000.0 μg/L. Chlorpyrifos at 10000.0 μg/L made the rotifers dead after 24-hr exposure. Dichlorvos and triazophos both at 10000.0 μg/L, and chlorpyrifos at 1000.0 μg/L all increased the ratio ovigerous females/non-ovigerous females. Both dichlorvos at 10000.0 μg/L and chlorpyrifos at 0.1–100.0 μg/L increased the mictic rate. Dichlorvos at 10.0 μg/L and 100.0 μg/L, and triazophos and chlorpyrifos both at 0.1–100.0 μg/L increased the resting egg production. Both population growth rate and ratio ovigerous females/non-ovigerous females are suitable endpoints for assessing the effects of dichlorvos, triazophos and chlorpyrifos, and mictic rate is a suitable endpoint for monitoring the effects of dichlorvos and chlorpyrifos on the reproduction of the rotifers. Both population growth rate and ratio ovigerous females/non-ovigerous females are more sensitive to dichlorvos and chlorpyrifos than mictic rate.     

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.     

Environmental and endogenous control of sexuality in a rotifer life cycle: developmental and population biology

Induction of mictic females, and hence initiation of sexuality, in the life cycle of some Brachionus requires an environmental stimulus associated with crowding. The inducing stimulus appears to be a taxonomically specific chemical released into the environment by the rotifers. Oocytes are induced to develop into mictic females before they are oviposited by their amictic mothers and begin cleavage divisions. Thus, the inducer affects the oocyte in the maternal body cavity either directly or indirectly by altering the physiology of its mother. The level of sexual reproduction expressed in populations of a Florida strain of B. calyciflorus is controlled by two types of endogenous factors and by the degree of crowding. First, some fraction of genetically identical oocytes in a clonal population fails to respond to even extreme crowding conditions, thus ensuring some potential for continued population growth by female parthenogenesis. Second, the propensity of amictic females to produce mictic daughters is extremely low when they hatch from fertilized resting eggs and then gradually increases to an asymptote after about 12 parthenogenetic generations. This multigenerational parental effect likely is due to a cytoplasmic factor in fertilized eggs that inhibits expression of the mictic-female phenotype and that is gradually diluted in successive parthenogenetic generations. The effect may increase a clone's genetic contribution to the resting-egg bank by increasing its population size through parthenogenetic generations before mictic females are induced.     

甲胺磷和乙酰甲胺磷对萼花臂尾轮虫实验种群动态的影响

应用3天种群增长实验方法,在(25±1)℃、无光照、以3.0×106 cells/mL的斜生栅藻为轮虫的食物等条件下,研究了亚致死浓度(0.01、0.1、1.0、10.0、100.0、1000.0和10000.0 µg/L)的甲胺磷和乙酰甲胺磷对萼花臂尾轮虫实验种群动态的影响。结果显示,甲胺磷和乙酰甲胺磷显著地影响萼花臂尾轮虫的种群增长率、混交雌体数与非混交雌体数的比值和混交率。甲胺磷显著地影响萼花臂尾轮虫种群中的带卵雌体数与不带卵雌体数的比值,但乙酰甲胺磷对其无显著的影响。和对照组相比,浓度为100.0 µg/L 的甲胺磷和浓度为1.0－10,000.0 µg/L 的乙酰甲胺磷均使轮虫种群增长率显著增大,而浓度为1000.0 µg/L和10000.0 µg/L的甲胺磷却使之显著减小;1000.0 µg/L 的甲胺磷使轮虫种群中的带卵雌体数与不带卵雌体数的比值显著上升,0.1 µg/L的甲胺磷和10.0－10000.0 µg/L的乙酰甲胺磷均使轮虫种群中的混交雌体数与非混交雌体数的比值显著上升,0.1µg/L 的甲胺磷和10 µg/L的乙酰甲胺磷均使轮虫的混交率显著增大,10.0－0000.0 µg/L 的乙酰甲胺磷使轮虫休眠卵产量显著提高。上述结果表明,亚致死浓度的甲胺磷和乙酰甲胺磷对萼花臂尾轮虫实验种群动态具有显著的影响。     

Mictic patterns of the rotifer Brachionus plicatilis Müller in small ponds

Populations of the rotifer Brachionus plicatilis were monitored in three small ponds in a marsh on the Mediterranean coast. Samples were taken approximately every three weeks from July 1992 to November 1993. Salinity, temperature, conductivity, pH and oxygen concentration were measured in the field. Population density was determined from preserved quantitative samples. Individuals were classified as mictic females, amictic females, non-ovigerous females, and males, differentiating between two morphotypes (S and L). From these counts, a level of mixis was calculated. We also determined the proportion of mictic females in natural populations by culturing females isolated from fresh samples. From these data, mictic patterns over time and correlation between levels of mixis and environmental and population parameters were analyzed. From a previous study S and L morphotypes were known to correspond to genetically different clonal groups. Our data showed that reproduction was predominantly parthenogenetic in these clonal groups, but mictic females were found in most samples, the proportion of mictic females ranging from 0 to 29%. The clonal groups showed different patterns of mixis. L clonal group presented a continuous sexual reproductive pattern. In contrast, S clones showed a rather punctuated mictic pattern. A positive correlation between levels of sexual reproduction and population density was found for S and L groups. However, they differed in their density threshold for mictic reproduction. The adaptive meaning of these patterns and their implications in maintaining genetic diversity within and between populations are discussed.     

Analysing threshold effects in the sexual dynamics of cyclically parthenogenetic rotifer populations

Sex in some cyclically parthenogenetic rotifers is triggered when a threshold population density is achieved. In Brachionus, accumulation of a mixis-inducing protein (MIP) that the rotifers excrete into the medium is the proximal signal. Models explicitly accounting for MIP dynamics suggest that either (1) a positive feedback of MIP concentration on MIP production or (2) a switching relationship between the proportion of sexual females and MIP concentration is needed to produce a threshold response of mixis to population density. In order to examine both possibilities, experiments were conducted to estimate the sexual response to a dilution series of medium containing the MIP. The resulting sexual response patterns are consistent with the existence of a threshold response of mixis, but are insufficient to reject one hypothesized mechanism for the threshold response in favor of the other. However, the observed smooth increase of sexual offspring with MIP concentration is consistent with the hypothesis of positive feedback on MIP production, and deserves further investigation.     

Effects of Juvenile Hormone and Precocene on the Reproduction of Brachionus calyciflorus

We studied the effects of juvenile hormone and precocene on reproduction of the rotifer Brachionus calyciflorus. Amictic females of B. calyciflorus that were 2‐4 hours old were exposed to different concentrations of juvenile hormone (0.004, 0.02, 0.1, 0.5, 2.5, 12.5 mg/L) and/or precocene (0.05, 0.25, 0.75, 3.75, 7.5 mg/L) for 24 h. They were then transferred to a new medium without hormone and checked every 2 h during the next 48 h, and thereafter monitored daily until the individual died. Precocene had no effects on the length of the rotifer juvenile period, hatching time of the first neonate, lifetime reproduction, or the mixis ratio. In contrast, juvenile hormone at 0.5, 2.5, and 12.5 mg/L significantly prolonged the juvenile period by 6.1, 9.2, and 8.6%, respectively. When 26‐28‐h‐old amictic females were exposed to the same concentration series of juvenile hormone or precocene, precocene at 3.75 mg/L resulted in an increase in lifetime reproduction of 30.39%. However, at 0.75 and 3.75 mg/L precocene, a significantly lower percentage of mictic females was found, whereas juvenile hormone had no effect on the lifetime reproduction or mixis ratio. The population growth test showed that juvenile hormone had significant effects on the population growth rate and mixis ratio, but no effect on resting egg production. In comparison, precocene had no effect on any of these parameters. (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Maternal Effect by Stem Females in <Emphasis Type="Italic">Brachionus plicatilis</Emphasis>: Effect of Starvation on Mixis Induction in Offspring

We examined whether starvation during the initial period of life in stem females affected reproductive characteristics of the offspring. Starvation treatment had different effects on rotifers hatched from resting eggs and those hatched from amictic eggs. When stem females experienced starvation after hatching, this induced a higher percentage of mixis in their offspring. When the same starvation treatment was applied to rotifers hatched from amictic eggs, there was no effect on the induction of mixis. It is probable that stem females hatched from resting eggs have specific features that are vulnerable to unfavorable environmental conditions, and that these features can be inherited by their offspring through the maternal cytoplasm.     

Hopf bifurcation in a structured population model for the sexual phase of monogonont rotifers

 We are studying a population of monogonont rotifers in the context of non-linear age-dependent models. In the sexual phase of their reproductive cycle we consider the population structured by age, and composed of three subclasses: virgin mictic females, mated mictic females, and haploid males. The model system has a unique stationary population density which is stable as long as a parameter, related to male-female encounter rate, remains below a critical value. When the parameter increases beyond this critical value, the stationary solution becomes unstable and a stable limit cycle (isolated periodic orbit) appears. The occurrence of this supercritical Hopf bifurcation is shown analytically. Received: 2 August 2001 / Revised version: 3 January 2002 / Published online: 26 June 2002     

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.