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.     

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.     

Diapause dynamics of two diaptomid copepod species in a large lake

Skistodiaptomus oregonensis and Leptodiaptomus minutus produce diapausing eggs in a large lake, Oneida Lake, in New York State. The timing of the switch from production of subitaneous (immediately hatching) eggs to diapausing eggs for both species is in October. This timing is consistent with a pattern, reviewed here, for other populations of diaptomid copepods: populations living in large lakes tend to begin production of diapausing eggs later in the season than those living in small lakes. Populations living in temporary ponds tend to switch still earlier in the season. All populations reviewed here live in the north temperate zone. The sediments of Oneida Lake contain densities of diaptomid diapausing eggs on the order of 10⁵ m^–2 per cm below the sediment surface down to 5 cm. Below this sediment depth, egg densities decline. The highest egg densities were found in sediments under the deepest water. Diapausing eggs of L. minutus survive in the sediments at least two years, as shown by the recovery of the population after a year in which no new diapausing eggs were produced, and probably for two or more decades. Long-term dormancy can have the effect of ensuring the continuation of a population through periods of poor recruitment, and can help create conditions for the coexistence of competing species. Other investigators have suggested that S. oregonensis and L. minutus are competitors in other lakes when they co-occur.     

Morphological differences between subitaneous and diapause eggs of Boeckella triarticulata (Copepoda: Calanoida)

1. Many calanoid copepods produce subitaneous eggs that hatch immediately and diapause eggs that remain dormant for long periods of time. Boeckella triarticulata (Copepoda: Calanoida) produces diapause eggs that hatch asynchronously over an extended period of time and appear identical under light microscopy to those that hatch immediately.
2. We compared the internal morphology of subitaneous and diapause eggs of B. triarticulata that had been prepared for transmission electron microscopy by four methods.
3. Subitaneous eggs had a thin, single-layered shell whereas diapause eggs had a thick, three-layered shell that appeared consistent over 3 months of diapause. Boeckella triarticulata appears to enter diapause at the multicellular stage.
4. Consistent morphological differences between subitaneous and diapause eggs of B. triarticulata suggest that these two egg types also have distinct differences in physiology, and are not merely extremes of a continuum from subitaneous to diapause eggs.     

Finding copepod footprints: a protocol for molecular identification of diapausing eggs in lake sediments

Even though calanoid copepods produce diapausing eggs that stay alive in lake sediments, these eggs have rarely been used paleolimnologically, as they lack diagnostic morphological features. In this study, we developed a method to identify copepod diapausing eggs in Japan as a clue toward reconstructing past plankton populations. We first determined a 28S ribosomal DNA (rDNA) (i.e., nc28S) regional sequence library (240 bp) of various calanoid copepod species using ethanol-fixed plankton samples collected from across the Japanese archipelago. Then we applied the UltraSHOT method to extract DNA from an individual diapausing egg. Finally, the nc28S region of diapausing eggs collected from various lakes was sequenced and compared with the regional library for species identification. In total, 21 haplotypes of the nc28S region were recovered from planktonic samples of 11 Japanese freshwater calanoid copepod species. Despite the short length of this region, no identical haplotypes were shared among the species analyzed, including the Acanthodiaptomus pacificus complex treated as a species. Even different lineages of A. pacificus could be separated. These results indicate that the nc28S region can be used as a barcode in Japan. A total of 112 diapausing eggs collected from various lakes and ponds was processed, and the nc28S region of each was successfully sequenced. All of these egg sequences matched one or the other of the nc28S haplotypes in the regional library mentioned above. The set of protocols we applied (i.e., preparing a comprehensive regional sequence library and sequencing egg DNA) is thus useful for involving copepod diapausing eggs in paleolimnological studies in lakes. The nc28S region treated in this study has a strong potential to uncover the paleodiversity of copepods, at least in Japan.     

Seasonal Variation in Diel Production Pattern of Centropages tenuiremis in Xiamen Waters, China

The seasonal variations in diel production pattern, and egg hatching time of the copepod Centropages tenuiremis in Xiamen waters, China, were investigated between January and June of 2003. The results show that in the winter–spring (January to early May) the adult females tended to lay subitaneous eggs at night, while in the beginning of summer (late May–June) they tended to lay diapause eggs as well as subitaneous eggs during the daytime. The egg hatching time negatively correlated with water temperature. These results, combined with the fact that C. tenuiremis migrates vertically in Xiamen waters, may explain the reproductive strategy of this dominant species in winter–spring.     

Predation and the distribution and abundance of a pulmonate pond snail

Summary The abundances of a freshwater pulmonate snail, Lymnaea elodes were studied in a temporary pond and a permanent, more productive pond in northeastern Indiana, USA. When snails from both populations were reared in each of the ponds in containers excluding predators, snails grew to be 1.3 to 2 times as large in the more productive pond, and laid 9 times as many eggs. However, field sampling data showed adults to be more abundant in the temporary pond. The only obvious difference between the two ponds was the presence of the molluscivorous central mudminnow (Umbra limi) in the permanent pond. These fish fed upon L. elodes when eggs and juvenile snails were abundant. In an experiment in the temporary pond, addition of mudminnows lowered egg and juvenile snail survival in pens where snail abundances had been increased. We suggest that vertebrate predators like the mudminnow can be significant sources of mortality for thin shelled species like L. elodes, possibly excluding them from habitats like lakes and rivers.Address for offprint requests     

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.     

Dissolved solids do not induce diapause in the calanoid copepod Aglaodiaptomus leptopus

The effect of different total dissolved solid levels (TDS) was investigated as a predictor of diapause induction in the calanoid copepod Aglaodiaptomus leptopus. We collected adult copepods in the field and monitored seasonal changes in diapause egg production. We determined total dissolved solid levels, conductivity and temperature values from five ponds. In experimental work, females bearing clutches, collected from the field, were monitored under laboratory conditions to determine the extent of diapause induced. Females without clutches were placed with males and then subjected to low, medium and high TDS levels. After six days, females bearing clutches were isolated to determine the proportion of clutches that were diapausing. The relationship between TDS and food levels was calculated using Williamson and Butlers food limitation index (f). We concluded that neither TDS nor temperature are predictors of the induction of diapause in this study. The obtained results suggest, however, that density possibly plays a role in predicting diapause induction.     

Structure, development and induction of a new diapause stage in rotifers

1. This study shows that females of Synchaeta pectinata Ehrenberg can produce two morphologically and physiologically distinct kinds of amictic eggs that develop parthenogenetically into females. One kind, until now the only one known in monogonont rotifers, is a thin-shelled (about 1.4 μm) subitaneous egg that develops without arrest. The other kind is a thick-shelled (about 9 μm) egg that enters obligatory diapause after 1–3 cleavage divisions before resuming development. The thicker shell of the diapausing egg is due to an expansion of the outer sublayer of the single-layered shell. 2. The mean duration of the diapause at 19 °C (L : D 16 : 8) is about 14 days. This diapause is not broken by low temperature (5 °C), and it is not greatly extended by storage at low temperature (5 °C). 3. Diapausing eggs are induced immediately after a brief starvation period, and seem to be produced at no additional energetic cost. However, a population producing a high proportion of diapausing eggs has a much reduced reproductive potential. 4. Production of diapausing amictic eggs may be a strategy to increase the ability of clones to survive food limitation. 5. Diapausing amictic eggs differ markedly from fertilized resting eggs produced following bisexual reproduction during favourable conditions.     

Ecological and evolutionary significance of resting eggs in marine copepods: past, present, and future studies

The occurrence of a resting egg phase in the life cycle of marine and freshwater planktonic copepods is well documented and receiving increasing attention by investigators. The species generally occur in coastal marine waters, freshwater ponds and lakes in areas that undergo strong seasonal fluctuations, though examples have been reported for tropical and sub-tropical areas not subject to such extreme fluctuations. Typically such species disappear from the water column for portions of the year, but remain in the region as benthic resting eggs. Studies to date have focused on the conditions that promote the occurrence of resting eggs, the factors that affect their survival and hatching from sediments, the existence of egg banks in sediments, and the impact of resting eggs on plankton community structure. Benthic resting eggs of copepods include diapause eggs as well as subitaneous (non-diapause) eggs that are quiescent due to conditions in the sediments. As with other groups of organisms the resting egg phase is viewed as being critical for the perpetuation of species year after year, especially those that disappear from the water column for portions of the year. Some data indicate that eggs can survive for many years in sediments which would expand their influence to evolutionary time scales. This paper summarizes our understanding of embryonic dormancy in marine copepods.     

Patterns in rotifer diapausing egg banks: Density and viability

Here we present a quantitative study on the density, age and viability of the diapausing egg banks of the rotifer species complex Brachionus plicatilis in the sediments of 15 water bodies from Eastern Spain. Sampled ponds, located in coastal and inland areas, varied in salinity and ranged in size, depth and permanence. By identifying ‘hatched’, ‘deteriorated’ and ‘viable’ diapausing eggs in the sediment samples, we estimated production, hatching and deterioration in relation to the habitat properties of each pond. Our results indicate the presence of large numbers of diapausing eggs in the sediments of almost all of the ponds studied (2-115 eggs cm^− 2). Inland ponds tended to have higher densities than coastal lagoons. The vertical distribution of eggs in the sediments frequently showed a non-decreasing pattern, which suggested a high among-year variation in egg production. Despite maximum age of eggs of 60-80 years, the median age (3-30 years) suggests that rotifer egg banks are young in the studied ponds. Egg senescence is suggested by the declining abundance of ‘healthy-looking’ eggs with depth. The proportion of ‘deteriorated’ eggs ranged 75-99% suggesting that deterioration rates in the sediments are high and vary among habitats. Hatching and deterioration rates, as estimated from the counts of ‘hatched’, ‘deteriorated’ and ‘healthy-looking’ eggs in the sediments, largely varied among ponds. An association between hatching and deterioration rates is suggested by our data. This is in agreement with the hypothesis that hatching rates of diapausing eggs depended not only on the risks associated with the water column, as initially expected by the general theory on diapause, but they are also related to the incidence of deterioration processes in the sediment.     

Reproduction strategies of <Emphasis Type="Italic">Daphnia pulicaria</Emphasis> population in a high mountain lake of Southern Spain

For three consecutive years, a population study of Daphnia pulicaria was undertaken in Río Seco Lake, a small high-mountain lake, in order to elucidate the reproductive strategies adopted by Daphnia in this system. Daphnia appears to colonize this lake every spring by hatching from ephippia and reproduce by means of subitaneous (non-diapausing) and ephippial (diapausing) eggs. D. pulicaria in this lake is an obligate parthenogenetic population. There is a short time period for subitaneous egg production and a much longer period for ephippial egg production. The contribution of subitaneous eggs to Daphnia population density and structure appears to be low. Diapause onset showed a high temporal synchronization in the three studied years in Río Seco Lake, and day-length emerged as the main cue triggering diapause onset and the main explanatory factor for the proportion of ephippial females observed. The development and reproduction of D. pulicaria in Río Seco Lake involves taking a gamble on resting forms to guarantee inter-annual Daphnia persistence in the lake, giving priority to investment in future generations. Guest editor: Piet Spaak Cladocera: Proceedings of the 7th International Symposium on Cladocera     

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.     

Tadpole competition in a desert anuran community

Summary Five interspecific competition experiments performed in artificial ponds under temporary pond-like conditions established that temporary and permanent pond tadpoles interact. Temporary pond species typically lowered survivorship and reduced progress towards metamorphosis of permanent pond tadpoles. These results suggest that competitive interactions may partially determine the local distribution of tadpoles in the Chihuahuan Desert by preventing some permanent pond anurans from using temporary ponds. Predation by Scaphiopus multiplicatus tadpoles (a temporary pond species) may also prevent permanent pond tadpoles from using temporary ponds.     

Fine structure of the subitaneous eggshell of the sessile rotifer Stephanoceros millsii (Monogononta) with observations on vesicle trafficking in the integument during ontogeny

Rotifers that engage in cyclical parthenogenesis produce two types of eggs: subitaneous eggs that hatch as clonal females and meiotic eggs that hatch as haploid males, or if fertilized, as females after a period of diapause (resting eggs). The ultrastructure of resting eggshells is known for some motile species, but there are limited data on subitaneous eggshells, and no data on any eggshells of sessile rotifers. Here, we investigated the ultrastructure of the subitaneous eggshell of the sessile rotifer Stephanoceros millsii and its potential origins of secretion, the maternal vitellarium and embryonic integument. We also explored secretory activity in the larval and adult integuments to determine whether activity changes during ontogeny. The eggshell consists of a single layer with two sublayers: an external granular sublayer apparently derived from the maternal vitellarium, and an internal flocculent sublayer secreted by the embryonic integument that may form a hatching membrane or glycocalyx. Secretory activity remains high in both the larva and adult and appears to be the source of the thickening glycocalyx. Altogether, the subitaneous eggshell of S. millsii is the thinnest among monogonont rotifers. Thin eggshells may have evolved in response to the added protection provided by the mother’s extracorporeal tube.     

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.     

Invertebrate communities and turnover in wetland ponds affected by drought

1. Changes to the macroinvertebrate fauna found in small ponds on a freshwater marsh (Aberlady Bay, Scotland) in 1986, 1987 and 1992 were used to assess the impact of the 1988–92 drought on taxon turnover in pond communities. 2. Permanent ponds accumulated taxa over the study period. Ponds that were wet throughout 1986–87 but dried in 1992 lost some of the many taxa associated with permanent water but acquired a reduced fauna typical of temporary ponds. Ponds that were temporary in 1986–87 were dry during most of 1992 and lost almost all aquatic taxa. 3. Faunal turnover was considerable even in permanent ponds. Extinction rates were high for taxa typical of permanent or temporary ponds. Colonization rates were poor for the taxa from permanent water, but high for the taxa from temporary ponds. 4. The importance of colonization and extinction rates as main predictors of the distribution of species between the ponds was looked at using metapopulation incidence functions, where observed incidence of a taxon can be used to predict colonization and extinction rates. Predicted rates were then compared with observed rates. Incidence functions gave reasonable predictions of observed colonization rates but were poor predictors of extinction, even for taxa that appeared likely to be true metapopulations. 5. For the pond fauna, including fugitive species adapted to temporary ponds, whilst colonization may well depend on environmental stochasricity (how long a pond holds water), subsequent survival depends on other, demographic, processes (e.g. finding a mate, predation) rather than the pond drying out.     

Induction of embryonic dormancy in the Calanoid copepod Acartia hudsonica: proximal cues and variation among individuals

In this study of Acartia hudsonica, the change in egg types from subitaneous eggs to dormant eggs was investigated. Temperature and photoperiod were hypothesized to act as the proximal environmental cues inducing this population to produce dormant eggs, and the variation in the dormant egg responses of individual females was quantified. Some field-collected females produced some dormant eggs early in the spring (March and April), but most of the switch to dormant eggs occurred in late June. Short-term experiments showed that increased temperatures increased the percentage of dormant eggs produced, whereas increased length of day did not. Long-term experiments showed a different pattern from short-term experiments, indicating that duration of exposure to particular temperatures or photoperiods may influence dormant egg production. Under certain conditions, it appeared that nearly all copepods were capable of producing dormant eggs, although identical experimental conditions did not affect all individuals similarly. In several experiments that revealed a significant treatment effect, between 42% and 85% of the variance detected could be attributed to variation among individuals. Also, the fraction of individuals producing dormant eggs generally increased with increasing water temperature. In Narragansett Bay, A. hudsonica appears to produce dormant eggs primarily in response to increased water temperatures although an effect of photoperiod could not be ruled out. Furthermore, individual responses vary greatly. Appreciation of such variation in an important life history trait, such as embryonic dormancy, is important in understanding how this population may be adapted to its environment.