Effect of food concentration on the production and viability of resting eggs of the rotifer Brachionus: implications for the timing of sexual reproduction

1. Life‐table experiments with Brachionus calyciflorus test several hypotheses related to the idea that sexual reproduction in monogonont rotifers should occur when food resources are favourable. 2. The food concentration necessary for a fertilised mictic female to produce one phenotypically normal resting egg was higher than that for an amictic female to produce one daughter. At the lowest concentration of Cryptomonas erosa (1.25 × 10³ cells mL^?1), the lifetime fecundity of these two types of females was 0.9 and 1.4, respectively. 3. The lifetime fecundity of both fertilised mictic females and amictic females increased with food concentration to 3.4 resting eggs and 15.2 daughters female^?1, respectively. The approach to maximal fecundity with increasing food concentration was more rapid for fertilised mictic females, such that their lifetime fecundity relative to that of amictic females gradually decreased from 0.64 (at 1.25 × 10³ C. erosa mL^?1) to 0.22 (at 2.5 × 10⁴ C. erosa mL^?1). 4. The probability of a fertilised mictic female producing one or more abnormal resting eggs during her lifetime was high (approximately 75%). The mean proportion of abnormal eggs produced per female varied among the different food‐concentration treatments (26–38%) but was not higher at the low food concentrations. 5. The proportion of normal resting eggs that hatched was high (51–71%); those produced at low food concentrations were no less likely to hatch than those produced at high food concentrations. No abnormal resting eggs hatched. 6. The probability of a fertilised mictic female producing an abnormal resting egg increased rapidly with her age at all food concentrations. The probability of a normal resting egg hatching declined with maternal age at the low food concentration in one of two experiments. 7. The results support the idea that induction of mictic females should occur when food resources are good. Coincidence of sexual reproduction with low food availability risks low production of resting eggs for several reasons. Population size may be small, with a low probability of encounters between young mictic females and males, and fertilised mictic females may be unable to mature and produce resting eggs.     

Use of swimming speed and egg ratio as predictors of the status of rotifer cultures in aquaculture

This study evaluated the use of egg ratio (eggs rotifer^–1) and swimming speed (mm min^–1) as prediction criteria for production and culture quality in mass cultures of the rotifer Brachionus plicatilis. Egg ratio was determined to be a suitable predictor of rotifer growth and production in the cultures. Low egg ratios (i.e., 0–0.17 eggs rotifer^–1) indicate reduced rotifer population over time (i.e., negative net population growth rates). However, at this time egg ratio dynamics are not suitably understood to predict in advance a sudden population collapse.Swimming speed of reproductive, egg-carrying females in the exponential growth phase was 40–45 mm min^–1. During exponential growth swimming speed was independent of the food used. Lower swimming speeds were obtained in late stationary phase (10–25 mm min^–1) when yeast was used as a food source. Both environmental factors (e.g., accumulating metabolites) and changes in nutritional state of the rotifers may have affected the swimming speed, but environmental factors appear to be the most important. We believe that swimming speed has the potential of becoming an accurate predictor of culture quality in mass cultures of rotifers.     

An individual-based population model for the prediction of rotifer population dynamics and resting egg production

Most species of rotifers have a combination of sexual and asexual reproduction, with sexual reproduction resulting in resting eggs, which can lay dormant for long periods. The occurrence of sexual reproduction affects population dynamics through the temporary presence of male rotifers, and a reduction in the growth of the number of female rotifers. A previously published, individual-based model used dynamic energy budget theory to describe rotifer food intake, growth, egg production, and mortality, but assumed asexual reproduction only. In the current study, we have expanded the model to describe the entire reproductive cycle of the rotifers, making it usable for investigating relationships, such as those between the signal triggering mictic egg production, and the timing and number of resting eggs produced. The model is intended for use in predicting the specific future development of cultures, for instance, as a process model in rotifer or resting egg production for aquaculture. Guest editors: S. S. S. Sarma, R. D. Gulati, R. L. Wallace, S. Nandini, H. J. Dumont & R. Rico-Martínez Advances in Rotifer Research     

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

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

Reproductive trade-off of the copepod Acartia tonsa in a hypersaline estuary of the Southwestern Atlantic. Temporal variations in the morphology of eggs

The purpose of the present work was to study the seasonal variations in egg production, morphology and hatching success in the cryptic species Acartia tonsa, taking into account variations in female size, population abundance and environmental factors in a turbid and hypersaline estuary. Sampling was performed during the austral warm (18–23°C and 32–36 salinity) and cold seasons (5–7°C; 32–38) in Bahía Blanca Estuary (BBE), Argentina, during 2007 and 2009. Field-collected females were incubated in the laboratory simulating in situ environmental conditions, and specimens from fixed samples were measured using optical and scanning electronic microscopy. Acartia tonsa’s marked seasonality in its reproductive traits was found to ensure its permanence in the water column all over the year. During the warm season, small-sized females were observed to invest their energy in the production of subitaneous eggs with high hatching success and smooth appearance (12.95?±?2.38 eggs f^?1 day^?1 and specific egg production rate (SEP) of 16.57%C f^?1 day^?1). During the cold season, females invested C in body mass as well as in the production of resting eggs of three different morphotypes (6.56?±?3.2 eggs f^?1 day^?1 and SEP of 7.37%C f^?1 day^?1). Although these morphotypes were found to show differences in surface ornamentation, they exhibited the same delayed hatching behaviour. The eggs with shorter spines were found to integrate the resting egg bank in BBE. Our findings confirming a delayed egg hatching behaviour and a great tolerance to low temperatures and high salinities in the A. tonsa population in BBE suggest that this possible strain is a valuable phenotype for aquaculture.     

Influence of ethanol and ethylene on the seed germination of three nymphaeid water plants

1. We offered Scenedesmus obliquus in five densities, from 0.5 to 8 ± 10⁶ cells ml^?1, to the rotifer Anuraeopsis fissa. Growth rates (r) during the exponential phase (first 7 days) were significantly and positively related to food density. The r values (mean ± SD) varied between 0.454 ± 0.067 and 0,856 ± 0.090, from the lowest to the highest food concentration, respectively. Population growth went through a phase of exponential increase which lasted from 7 to 10 days before a plateau and, in some cases, a decrease occurred. 2. There was a linear relation between food density and rotifer plateau density. At the highest food density, a peak abundance (mean ± SD) of 2312 ± 226 individuals ml^?1 was reached, while at the lowest food density there was no identifiable single peak but a plateau, at a density of 361 ± 62 ind. ml^?1. 3. The egg ratio decreased with increasing population density. The ratio of loose eggs to eggs attached to females indicated that more eggs became detached at higher food densities. 4. As in many other rotifer species studied so far, population density of A. fissa was less stable at higher algal food concentrations. Numerically, A. fissa could be grown at twice the density achieved in Brachionus.     

Females from resting eggs and parthenogenetic eggs in the rotifer Brachionus calyciflorus: lipid droplets, starvation resistance and reproduction

1. In the heterogonic life cycle of monogonont rotifers, amictic (female‐producing) females develop from two types of eggs: fertilised resting (diapausing) eggs and parthenogenetic subitaneous eggs. Females hatched from resting eggs initiate clonal populations by female parthenogenesis and are called stem females. This study compares females from resting and parthenogenetic eggs that were produced under identical culture conditions and were of similar birth order. 2. Newborn stem females had many more lipid droplets in their tissues than similar‐sized, newborn females from parthenogenetic eggs. When neonates were stained with Nile Red and viewed under epifluorescent illumination, these droplets were shown to be sites of neutral‐lipid storage products. 3. Stem females had no posterolateral spines and short anterior spines, while their mothers and offspring in subsequent, parthenogenetic generations typically had long posterolateral spines and elongated anterior spines. 4. Newborn stem females survived starvation significantly longer than newborn females from parthenogenetic eggs. 5. When females from resting and parthenogenetic eggs were cultured from birth to death at a high food concentration, the reproductive potential (r day^?1) of the stem females was significantly higher (0.82–0.88 versus 0.70), primarily because of egg production at an earlier age. The mean lifetime fecundity (R_o) of stem females was significantly greater than that of females from parthenogenetic eggs. 6. Extensive lipid reserves should increase the ability of stem females to colonise new habitats. Firstly, compared with females from parthenogenetic eggs, stem females are more likely to experience starvation or food limitation. Resting eggs hatch in response to physical and chemical factors that are not directly related to food availability, and from sediments that may be far from food‐rich surface waters. Secondly, when food is abundant, stem females have a greater reproductive potential.     

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.     

食物浓度和品系对萼花臂尾轮虫生殖期历时和混交雌体形成的影响

应用单个体培养方法比较研究了藻类食物浓度(2.0、5.0、8.0和11.0×106cells/mL)和品系对采自广州、青岛和芜湖等地的萼花臂尾轮虫生殖期历时、后代数和后代中混交雌体百分率的影响。结果表明,食物浓度对轮虫的生殖期历时和轮虫全部后代中的混交雌体百分率有显著的影响,但对轮虫的总后代数无显著的影响。品系对轮虫的生殖期历时、总后代数和全部后代中的混交雌体百分率均有显著影响。3品系间,广州品系轮虫的生殖期历时最长,为(4.53±0.19)d;总后代数最多达(18.75±0.26)个;全部后代中的混交雌体百分率最低,仅为4.32%±0.51%。而青岛品系轮虫全部后代中的混交雌体百分率最高,达49.90%±2.15%;其生殖期历时和总后代数与芜湖品系间均无显著的差异。食物浓度和品系间的交互作用仅对轮虫全部后代中的混交雌体百分率有显著影响。3品系中,仅芜湖品系轮虫的生殖期历时、总后代数和全部后代中的混交雌体百分率受食物浓度的显著影响;母体年龄对轮虫每天所产后代中的混交雌体百分率的影响因轮虫品系的不同和培养时所用的食物浓度的不同而异。     

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.     

Population growth rate of the parasitic rotifer Proales gigantea,and susceptibility to parasitization in the snail Lymnaea acuminata at different stages of embryonic development

Developing eggs of the host snail Lymnaea acuminata were experimentally parasitized with the parasitic rotifer Proales gigantea to study the population growth rate of the parasite within the snail egg capsule and the susceptibility of the host eggs at different stages of embryonic development. The population growth rate of P. gigantea was 0.46 ± 0.07 individual^–1 day^–1 at the ambient temperature of 18–22 °C. Snail eggs were most susceptible to rotifer attack during the initial stages of development, becoming progressively more resistant after the hippo stage. Yet, regardless of the stage of development, the host embryo was doomed to die without hatching even if one individual rotifer gained entry inside the egg capsule. The presence of P. gigantea within the parasitized egg capsules or in the mucilage had no effect on the developmental rates and hatching success of non-parasitized eggs within the same egg mass.     

休眠卵孵化对策与食物浓度对萼花臂尾轮虫(Brachionus calyciflorus)两品系干雌体种群增长的影响

周期性孤雌繁殖的轮虫靠休眠卵度过不良环境,等环境适宜时休眠卵孵化出干雌体,再次通过孤雌繁殖建立种群。通常休眠卵要经历一段休眠期再孵化,但也有些休眠卵生成后很快孵化,称为早孵化现象。有关休眠卵不同孵化对策如何影响其干雌体克隆种群的增长,目前尚不清楚。分别观测了萼花臂尾轮虫（Brachionus calyciflorus）的两个品系H1（窄温度生态位）和D1（宽温度生态位）的休眠卵在不同孵化对策和不同食物浓度下所生产的干雌体克隆群的种群增长差异。孵化对策包括早孵化（early hatching,EH）和晚孵化（late hatching,LH）;食物浓度包括高食物浓度（high food concentration,HF：2×10⁶个细胞/mL）和低食物浓度（low food concentration,LF：5×10⁵个细胞/mL）,每个品系下各设置4个实验组（LH-HF、LH-LF、EH-HF、EH-LF）,每组10个重复。结果发现,对D1品系来说：孵化对策和食物浓度对最大种群数量具有显著的综合影响（P=0.002）,但两因素间不存在交互作用（P=0.911）;早孵化的干雌体种群在最大种群数量上显著高于晚孵化的干雌体种群（P=0.001）。对H1品系：孵化对策和食物浓度对最大种群数量综合影响显著（P<0.001）,且两个因素之间存在交互作用（P<0.001）;高食物浓度下,EH干雌体克隆群的最大种群数量显著低于LH干雌体克隆群（P<0.001）。高食物浓度下干雌体克隆群开始有性生殖的密度阈值,D1品系EH组显著高于LH组（P=0.041）;而H1品系EH组却显著低于LH组（P=0.022）。最高种群密度下,H1品系的有性生殖率在两种孵化对策之间存在显著差异（P=0.044）,EH种群的有性生殖率低于LH种群,而D品系却未见有性生殖率在不同孵化对策间存在明显差异。本研究结果显示休眠卵的孵化对策会影响其后代干雌体种群的增长特性,其影响的结果可能与种群的生境适应相关。     

Induction of diapause egg production in Eurytemora affinis by their own metabolites

Chemically mediated crowding effect on induction of diapause egg production in the calanoid copepod Eurytemora affinis was investigated. Two types of eggs, i.e. subitaneous and diapause eggs, produced by females that were reared in crowded culture conditions or the water from crowded culture under excess food supply were examined. 2.3% of the females that were reared at a population density of 5 or 6 animals per 10 ml of filtered lake water (FLW) produced diapause eggs. Females that were reared individually in 2-ml containers with FLW did not produce diapause eggs while 72.7% of females that were individually reared in 2-ml containers, but with water from a crowded culture (500 ind. l^–1, culture medium was changed once a week), produced diapause eggs. These results demonstrate that accumulation of their own metabolic products in the medium induces diapause egg production by E. affinis.     

Disturbances due to increased salinity and the resilience of zooplankton communities: the potential role of the resting egg bank

When faced with disturbances such as increased salinity, aquatic communities inhabiting inland coastal systems change and may or may not be resilient after salinity decreases. The purpose of this study was to assess the potential role of the resting egg bank for zooplankton community resilience. We predicted that (1) hatching of resting eggs is inhibited by increased salinities and (2) resting eggs remain viable when exposed to salinity and hatch when returned to freshwater. At the community level, we evaluated the hatching responses and the short-term viability of resting eggs exposed to a salinity gradient. The hatching of resting eggs was inhibited at higher salinities (16.0 and 32.0 g l^?1). However, some resting eggs remained viable and hatched when returned to freshwater. Additionally, combining our experimental results to previously published field data, we observed that the pattern of hatching during exposure to salinity matches the temporal succession observed in the zooplankton community at our model system, after increased salinity. The recovery of zooplankton communities after disturbances involving increased salinity is likely facilitated by the presence of an egg bank. This finding has important implications for the recovery of zooplankton communities and the management of aquatic systems vulnerable to salinization worldwide.     

Reduced investment in sex as a cost of inducible defence in Brachionus calyciflorus (Rotifera)

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Effect of two microalgae concentrations on body size and egg size of the rotifer <Emphasis Type="Italic">Brachionus calyciflorus</Emphasis>

The rotifer, Brachionus calyciflorus, was grown with two algae species (Chlorella sp. and Scenedesmus obliquus) at different concentrations (0.1, 1 and 10 × 10⁶ cells ml⁻¹). The body size (lorica biovolume) of individual rotifer and their egg size were measured when the populations were roughly in the exponential phase of population growth. The body size of the rotifers differed significantly (P < 0.05) among the two algae species used, however this effect was not observed for egg size. The body size of rotifers fed on higher densities of Chlorella sp. (10 × 10⁶ cells ml⁻¹) was significantly larger than for those fed on lower and medium densities (0.1 and 1 × 10⁶ cells ml⁻¹). Body size and egg size of rotifers fed with different amounts of Scenedesmus did not differ significantly. The egg size was significantly larger at higher food level of Chlorella. A significantly positive correlation was observed between the adult rotifer body size and their egg size.     

Reproductive investment of several rotifer species

A comparative study was carried out on the seasonal variation in egg and body volumes of four species of rotifers, Brachionus calyciflorus, Brachionus angularis, Keratella quadrata and Anuraeopsis fissa, from ponds near Sevilla and in Doñana National Park. Temperature and food concentration significantly affected egg volume in all four species; clutch size significantly affected egg volume in A. fissa, B. calyciflorus and B. angularis, but not in K. quadrata. As temperature increased, egg and body volumes declined in A. fissa and B. angularis but increased in K. quadrata and B. calyciflorus due, perhaps, to complex interactions between temperature, food level and clutch size.All four rotifer species responded similarly to changes in food concentration: below a certain food level, egg and body volumes were small but increased to a maximal size, which differed in the four species, as the food concentration was raised. At higher food levels, there was a reduction in egg and body volumes. The food levels for maximum egg and body volumes are interpreted as optimal ones; below these, food limitation reduces the size of the reproducing adult and, consequently, the size of eggs. Above this optimal level, higher food levels accelerated the rotifer life cycles, resulting in adults maturing at smaller sizes with a larger number of smaller eggs.     

Polymorphism in stem females and successive parthenogenetic generations in Brachionus calyciflorus Pallas

We have described the polymorphism in the hatchlings of resting eggs and the morphological variations between the stem females hatched from resting eggs and their successive parthenogenetic generations in the rotifer Brachionus calyciflorus. We hatched resting eggs of B. calyciflorus in two different culture mediums: unconditioned medium (IOM) and Asplanchna-conditioned (ACM). The hatching rate of resting eggs in IOM and ACM were 32.5% and 28.5%, respectively, and showed no significant difference. Stem females hatching from these resting eggs had three morphotypes (unspined, single short-spined, and two short-spined) and over 80% of these females were spineless ones. Moreover, the frequency of each morphotype stem females showed similar tendency in IOM and ACM. The production of a variety of morphotypes among stem mothers—rather than all unspined as has been previously reported for this species—may be regarded as a form of bet-hedging in this population of B. calyciflorus. Phenotypic changes in morphology between the stem females hatched from resting eggs and their successive parthenogenetic generations in B. calyciflorus were found. The possible mechanisms, responsible for high proportion of spineless phenotype at early generations from resting eggs and increased spined phenotype in successive parthenogenetic generations, were discussed in the article.     

Induction of diapausing amictic eggs in Synchaeta pectinata

Amictic females of a clone of S. pectinata from Star Lake (Norwich, Vermont) may produce diapausing as well as non-diapausing (subitaneous) eggs. The proportion of diapausing eggs produced in cultures was unaffected by temperature (12 vs 19 °C) or rotifer population density (minima of 0.33 vs 3 ind. ml^–1) at 19 °C. However, at 19 °C this proportion was higher in cultures maintained at a low food level suppressing reproduction (5 × 10³ cells ml^–1 Cryptomonas erosa) than in those maintained at a high food level (2 × 10⁴ cells ml^–1); the treatment effect was marginally significant (p=0.067). Consistent with the effect of low food availability, a period of starvation was very effective in inducing the development of diapausing eggs. None of 19 females cultured individually from hatching at 19 °C on C. erosa (2 × 10⁴ cells ml^–1) in 1-ml volumes produced any diapausing eggs in 4 days (0 out of 349 eggs), while 13 out of 16 females subjected to a 15-hour starvation period 6 hours after birth produced one or more diapausing eggs during that time (34% of the 158 eggs produced by the 16 females were diapausing). Diapausing eggs produced and left at 19 °C hatched after 4 to 13 days. Those produced in cultures with a low food level took significantly longer to hatch (9.7 days) than those produced in cultures with a high food level (8.1 days) (p=0.022). In natural communities, S. pectinata should be able to respond directly and rapidly to poor food conditions by producing eggs that undergo an obligatory dormant period before resuming development.     

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.