Deterioration patterns in diapausing egg banks of Brachionus (Müller, 1786) rotifer species

Rotifers are cyclical parthenogens that produce sexual diapausing eggs at some stage in their life cycle. These eggs are encysted embryos that remain viable for extended periods in lake and pond sediments, thus acting as an egg bank with many ecological and evolutionary consequences. Despite its importance to rotifer evolution, there are no studies on resting egg deterioration and associated processes in natural environments. In this study, more than 4000 diapausing eggs of species from the Brachionus plicatilis complex, which includes several closely related cryptic species, were collected from different sediment depths in 15 ponds in eastern Spain and were classified according to three features thought to be related to their viability: shell integrity, embryo size, and embryo colour. A positive association was found between embryo size and hatching success in those eggs having an intact shell. Diapausing eggs that showed good shell integrity and no more than a 25% reduction of multinuclear embryo maximum size were classified as healthy and 98.9% hatched. Darkening of diapausing egg embryo was an indicator of viability loss. A decreasing frequency of healthy-looking diapausing eggs was observed with increasing sediment depth, although some exceptions were found.     

Barcoding rotifer biodiversity in Mediterranean ponds using diapausing egg banks

The biodiversity of Mediterranean freshwater bodies is among the most threatened worldwide; therefore, its accurate estimation is an urgent issue. However, traditional methods are likely to underestimate freshwater zooplankton biodiversity due to its high species seasonality and cryptic diversity. We test the value of applying DNA barcoding to diapausing egg banks, in combination with the creation of a reference collection of DNA barcodes using adult individual samples, to characterize rotifer communities. We use monogonont rotifers from two lakes in Doñana National Park and one from Ruidera Natural Park in Spain as models to create a reference collection of DNA barcodes for taxonomically diagnosed adult individuals sampled from the water column, to compare with the sequences obtained from individual eggs from the diapausing egg banks. We apply two different approaches to carry out DNA taxonomy analyses, the generalized mixed Yule coalescent method (GMYC) and the Automatic Barcode Gap Discovery (ABGD), to the obtained sequences and to publicly available rotifer sequences. We obtained a total of 210 new rotifer COI sequences from all three locations (151 diapausing eggs and 59 adults). Both GMYC and ABGD generated the same 35 operational taxonomic units (OTUs), revealing four potential cryptic species. Most sequences obtained from diapausing eggs (85%) clustered with sequences obtained from morphologically diagnosed adults. Our approach, based on a single sediment sample, retrieved estimates of rotifer biodiversity higher than or similar to those of previous studies based on a number of seasonal samples. This study shows that DNA barcoding of diapausing egg banks is an effective aid to characterize rotifer diversity in Mediterranean freshwater bodies.     

Hatching and viability of rotifer diapausing eggs collected from pond sediments

1. Planktonic rotifers inhabiting variable environments produce diapausing eggs that accumulate in the sediment of lakes and ponds, forming egg banks that may withstand adverse periods. A common assumption in zooplankton diapausing egg bank studies is to count as viable all eggs in the sediment that look healthy. This assumption should be challenged by asking how effectively ‘healthy‐looking’ eggs represent viable eggs. 2. In this study, viability of more than 1100 ‘healthy‐looking’ diapausing eggs belonging to the Brachionus plicatilis species complex was assessed in a laboratory hatching experiment. Eggs were collected at different depths from sediment cores obtained from 15 ponds located in coastal and inland areas of Eastern Spain. 3. Only approximately one half of the ‘healthy‐looking’ diapausing eggs hatched after incubation in experimental conditions. Almost all the hatchlings (99.4%) survived to maturity. The proportion of ‘healthy‐looking’ diapausing eggs that hatched varied among areas and among ponds within area, and substantially declined with sediment depth. Most of the hatchlings (88%) were obtained from the uppermost 2 cm of sediment. ‘Healthy‐looking’ eggs from upper sediment layers hatched after significantly shorter incubation times than eggs recovered from deeper layers. 4. Both decreased hatching success and increased incubation time for hatching with sediment depth suggest that older ‘healthy‐looking’ eggs are less responsive to hatching stimuli and could become unviable. However, the strong correlation found between the number of ‘healthy‐looking’ eggs and the number of hatchlings indicates that the abundance of ‘healthy‐looking’ eggs is a good index of egg bank viability.     

Resting egg banks can facilitate recovery of zooplankton communities after extended exposure to saline conditions

1. Salinisation has had a major effect on the diversity of biota associated with freshwater wetlands. However, there is no information available about whether elements of the biotic communities would be able to recover if the concentration of salts within secondarily salinised wetlands was lowered to levels more typical of freshwater wetlands. 2. We tested the hypothesis that dormant eggs of zooplankton are able to persist in wetlands with elevated salinities for extended periods of time by using zooplankton communities that had developed in mesocosms exposed to either salt concentrations of 13 500 mg L^?1 or freshwater (<300 mg L^?1) for a period of 22 months. We measured the response of the zooplankton community as concentration was reduced along a gradient of decreasing salinity from 13 500 mg L^?1 to freshwater. 3. In the freshwater mesocosms, the zooplankton community was abundant and taxon rich. In comparison at the start of the experiment in the high salinity mesocosms, the zooplankton community had low abundances and very few taxa. Numbers remained low in these mesocosms until salinity was reduced to <2500 mg L^?1. Below this, there was a rapid increase in the abundance, and richness of zooplankton and communities became similar to the communities in the freshwater mesocosms. 4. These results indicate that dormant eggs of zooplankton are able to persist in wetlands exposed to high salinity levels for up to 22 months and provide a means for zooplankton communities to rapidly respond once a wetland returns to freshwater. 5. It is likely that if the underlying causes of secondary salinisation in wetlands are addressed, it will be possible to undertake restoration activities that allow the rapid return of some components of their biotic communities.     

镜湖不同湖区沉积物中轮虫休眠卵萌发的比较研究

实验室内对镜湖沉积物中的轮虫休眠卵进行了萌发,共孵出轮虫47种,隶属于10科19属;其中大湖区沉积物中萌发出的轮虫40种,小湖区41种,两湖区共同种类34种.每毫升大湖区沉积物中休眠卵萌发出的轮虫平均数量为(1.1±0.1)个,显著高于小湖区的(0.4±0.0)个.在两湖区沉积物中,以3种孵化方式所萌发出的平均密度超过4.0ind./200mL的轮虫种类总计有6种,它们是多须伪前翼轮虫(Proalides tentaculates)、裂痕龟纹轮虫(Anuraeopsis fissa)、螺形龟甲轮虫(Keratella cochlearis)、裂足臂尾轮虫(Brachionus diversicornis)、臂三肢轮虫(Filinia brachiata)和萼花臂尾轮虫(Brachionus calyciflorus);其中前三种轮虫为两湖区所共有.共有的3种轮虫的孵出情况在两湖区沉积物间也存在着差异.在小湖区沉积物中,3种孵化方式下孵出的平均密度大于0.05ind./200mL的轮虫属是臂尾轮属、伪前翼轮属、龟甲轮属和龟纹轮属,而大湖区沉积物中孵出的平均密度较高的轮虫属还包括三肢轮属、巨头轮属(Cephalodella)和异尾轮属(Trichocerca);水体营养程度较高的大湖区沉积物中孵出的臂尾轮属、伪前翼轮属和龟甲轮属轮虫的密度均高于营养程度较低的小湖区.     

Growth pattern of four strains of a Bdelloid rotifer species: egg size and numbers

Life-history traits and growth patterns of four strains of Macrotrachela quadricornifera (Rotifera, Bdelloidea) were studied to assess the influence of maternal traits on egg size. There were two small (S, Va) and two large strains (H, G) with similar patterns of life cycle and body growth. They allocated to reproduction similar relative amounts of resources partitioned into eggs of similar relative size. All strains started reproduction while growing and, although their final sizes differed, at maturity had similar large or small sizes but different ages. Their egg sizes were unaffected by the clutch size, but were positively correlated with mother's body size.     

Wind-borne short-range egg dispersal in anostracans (Crustacea: Branchiopoda)

Evidence is provided for the first time that at least part of the egg bank of the southern African fairy shrimp Branchipodopsis wolfi is dispersed over short distances by wind. A total of 423 sticky surfaces were mounted around and between individual basins at three rock pool sites, to trap any dispersing eggs during a 3 day period. Eight viable eggs were found in different egg traps (1.9% of the total), seven of which were located at one site with shallow basins. Given the proximity of vast egg banks to the egg traps, the low observed dispersal rate cautions against overestimating the importance of wind dispersal for population genetic processes. By generating egg banks to hedge against drought catastrophes, and producing egg types with different potential dispersibility, B. wolfi is a strong bet-hedger which has established a means of escaping temporal and spatial stress. These life history functions enable populations to persist in small desert rock-pools, which are the most extreme of temporary habitats.     

Egg banks in freshwater zooplankton: evolutionary and ecological archives in the sediment

Many representatives of freshwater zooplankton produce at some stage in their life cycle resting stages. A variable portion of the eggs of the previous growing period will hatch at the next occasion while the remaining ones are added to a persistent egg bank, where they can remain viable for decades or longer. The importance of the study of resting eggs and egg banks in general for such different disciplines as taxonomy, ecological biogeography, paleolimnology, nature conservation, evolutionary ecology and community and population ecology is generally appreciated. The major current and expected future developments in this rapidly expanding field of research are presented here. The structure and dynamics of the egg bank are determined by the life history characteristics of the species (or local population), the hatching phenology of their resting stages, and the characteristics of the habitat. The horizontal distribution of dormant stages is generally patchy, with a greater density in the deeper and/or windward parts of a pond or lake. In sediment cores, most viable (responsive) eggs occur in the upper centimeters, although vertical variation related to the history of fish predation or water quality occurs. The accumulation of resting stages of different species, generations and genotypes with variable regeneration niches results in a mixed egg bank with greater potential biodiversity than the active community sampled at any one moment. Through the benthic–pelagic coupling, this dormant reservoir may have considerable impact on the evolutionary potential of the organisms, the ecological dynamics of the community and the distribution of species. Egg banks can be considered the archive of the local habitat, since the pattern of changes in species assemblage and genotypes from the past up to the present reflect changes due to natural or anthropogenic impact that can be used to reconstruct evolutionary processes or even to restore the local habitat. Overlooking the egg bank as an important component of zooplankton communities may lead to erroneous interpretations in the analysis of community and population genetic structure. This review integrates technical and scientific information needed in the study of the structure and function of egg banks in zooplankton with special focus on the fascinating latest developments in the field.     

Local and landscape effects on temporary pond zooplankton egg banks: conservation implications

Understanding landscape correlates of local habitat integrity and community structure and the identification of spatial scales at which these associations operate are relevant for management and conservation of unique but globally threatened temporary ponds. We use a multivariate variance decomposition approach to determine taxon-specific associations of zooplankton communities banked in dry pond soils (rotifers, cladocerans, copepods) with local habitat features and landscape characteristics across four spatial scales (100 buffer strip, 1, 5, and 10 km scales). Results show similar degrees of correlation between rotifer and cladoceran communities with local habitat conditions (chiefly water quality). This is interpreted according to life-history traits of component species of wetland propagule banks. Associations with landscape features varied between communities with rotifers correlating with landscape structural features only at the buffer scale while cladocerans showed no significant correlations with landscape characteristics across all scales. Copepods were neither significantly associated with local nor landscape characteristics. The results of this study contrast strikingly with our previous population-based study, where populations of Triops cancriformis and Branchinecta orientales were significantly correlated with landuse features at the broadest scale. The combined results suggest that a three-way management scheme could be useful for conservation of zooplankton resting egg banks in this remnant wetland complex. These schemes could focus on the restoration of wetland water quality, the establishment of vegetated buffer strips around the ponds to counteract degradation resulting from runoff, and reforestation and/or the creation of hedgerows in agricultural catchments to avoid impacts resulting from broad-scale diffuse pollution fluxes. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

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     

Phylogeography and regional endemism of a passively dispersing zooplankter: mitochondrial DNA variation in rotifer resting egg banks.

We investigated the phylogeography of the salt water rotifer Brachionus plicatilis, a cyclical parthenogen with passive dispersal mechanisms, using resting eggs recovered from saline lake sediments. Individual resting eggs were obtained from a large selection of lakes which were representative of five endorheic basins and the chain of coastal ponds in the Iberian Peninsula. The novel use of resting eggs allows the integration of seasonal and annual variations as well as the impact of stochastic effects such as drift and local extinction. A 653 bp fragment of the mitochondrial cytochrome oxidase subunit I (COI) gene was sequenced from 98 eggs. Our results revealed a deep phylogeographical structure in this species, with a division into two main lineages with distinct geographical distributions, which probably diverged at the beginning of the Pleistocene period. Most of the mitochondrial DNA haplotypes were restricted to single lakes. Nested clade analysis supported Early Pleistocene fragmentation of populations, low gene flow and some long-distance colonization. These conclusions contrast strongly with previous ideas on rotifer biogeography and this pattern is consistent with a recolonization of the Iberian Peninsula from two glacial refugia. The results provide new insights into the processes responsible for the genetic diversification of passive dispersers, a life-history trait typical of zooplanktonic biotas.     

The interplay between colonization history and gene flow in passively dispersing zooplankton: microsatellite analysis of rotifer resting egg banks

Zooplanktonic organisms that disperse passively as diapausing eggs often exhibit surprisingly strong population subdivision given their high colonization ability. Here we attempt to disentangle the impacts of colonization history and gene flow on these organisms by studying the population genetic structure of the rotifer Brachionus plicatilis. The resting egg banks of B. plicatilis in fourteen salt lake populations in the Iberian Peninsula were examined using seven microsatellite loci. A remarkably high degree of geographical structuring was found (F_st=0.43), with a significant pattern of isolation by distance. Microsatellite loci were in genetic equilibrium, ruling out inbreeding as an important force in population structuring. Comparisons are drawn with previously published phylogeographical data. Surprisingly, introgression of nuclear genes was detected in neighbouring populations with divergent mtDNA haplotypes. These results stress the long lasting impact of colonization history and the modulating effect of gene flow at local scales in these organisms.     

Some peculiar elements in the rotifer fauna of the atlantic sahara and of the atlas mountains

In addition to ubiquitous forms, the Adrar of Mauretania has a number of rare tropicopolitan species, which occur here in relative abundance. In the Moroccon Atlas mountains, West-African forms meet boreo-alpine forms.A population of Hexarthra from Atar, Mauretania, was intermediate between the fennica-group and the jenkinae-group.Contribution n° 28 from project Limnology of the Sahara, under contract n° 2.0009/75 with the Fonds voor Kollektief Fundamenteel Onderzoek, Belgium     

Desiccation and egg viability of the ragwort flea beetle,Longitarsus flavicornis (Coleoptera: Chrysomelidae)

A study was conducted to investigate the effect of desiccation on the survival of eggs of Longitarsus flavicornis. Eclosion of L. flavicornis eggs in laboratory trials decreased with increasing desiccation time between 0 days (93% hatching) and 42 days (no egg hatching) at 50±2% relative humidity and 23±2°C. Probit analysis indicated that 25, 50 and 99% mortality of L. flavicornis eggs occurred after 5.7, 9.3 and 50.4 days desiccation, respectively. Egg development varied between a minimum of 8 days at 7 days desiccation to a maximum of 15 days at 28 days desiccation. Hatching span did not differ between treatments with all eggs hatching within 12 days of each other. A relative humidity of 88–100% was measured under ragwort rosettes in non-drought field conditions. This would be expected to facilitate successful egg eclosion. However, the occurrence of summer drought could be detrimental to egg survival.     

萼花臂尾轮虫有性生殖、种群增长和休眠卵产量间的关系

利用种群指数增长和Logistic增长模型,通过计算机模拟研究了萼花臂尾轮虫有性生殖发生的频率,后代中的混交雌体发率对种群增长和休眠卵产量的影响。在所模拟的参数范围内,随有性生殖发生频率由100％减小到20％,获得了大休眠卵产量所需的混交雌体百分率由9％增大到69％,随密度制约作用的增大（环境容纳量K值由1000减小到100）,该混交雌体百分率由18％增大到69％,休眠卵产量由1072．10降低至133．67,种群的内禀增长率与获得最大休眠卵产量所需的混交雌体百分率间呈曲线相关。当有性生殖发生得越频繁,种群增长所受的密度制约作用较小时,后代中10％－30％的个体为混交雌体时的种群中休眠卵产量较大。     

Microbial infection affects egg viability and incubation behavior in a tropical passerine

Many avian species initiate incubation before clutch completion,which causes eggs to hatch asynchronously. This influences broodcompetitive dynamics and often results in nestling mortality.The prevailing hypotheses contend that parents incubate earlybecause asynchronous hatching provides fitness benefits to parentsor surviving offspring. An alternative idea is that early incubationis the best of a bad job because of the costs of delaying incubationto the viability of first-laid eggs. To explore this, we examinedthe potential for microbial infection, and the relative effectsof infection and suboptimal development temperatures on theviability of pearly-eyed thrasher (Margarops fuscatus) eggs.We exposed newly laid eggs for 5 days at either end of a tropicalaltitudinal gradient and cleaned shells of half the eggs toreduce microbial growth. Uncleaned eggs were infected more thanwere cleaned eggs, and infection was greater for eggs exposedat the cool, humid site than at the hot, less humid site. Parentallyincubated eggs, however, were not infected, suggesting thatincubation limits infection. The consequence of exposure toinfection and high ambient temperatures was a dramatic reductionin viability; cleaned eggs held at the cool site had the highesthatching success, which was significantly greater than for uncleanedeggs at this site and for cleaned eggs held at the hot site.This provides the first evidence that microbes can infect unincubatedeggs of a wild bird, and that infection and ambient temperatureact independently to reduce hatching success. These factorscould affect avian life-history strategies in diverse habitats.     

In vitro storage of unfertilized eggs of the Eurasian perch and its effect on egg viability rates and the occurrence of larval malformations

Ova ageing is the most important factor affecting fish egg quality after ovulation. Long-term storage of fish ova, using cryopreservation and vitrification techniques, has been unsuccessful to date. Instead, short-term in vitro ova storage has been used successfully and optimized in some cultured fish species. In vitro ova storage can drastically improve mass production of larvae and juveniles in the hatcheries by providing the possibility of the synchronous artificial fertilization for different females. To study how long unfertilized eggs of Eurasian perch (Perca fluviatilis L.) can retain their fertilizing ability after stripping, eggs were stored at temperatures of 4°C, 8°C and 12°C for 72 h post-stripping (HPS). The stored eggs of four female perch were separately fertilized at 0 h (i.e. control eggs fertilized before storage) and at 6-hour intervals during the experimental period of 72 h. The embryos reaching the eyed-egg and hatched-larvae stages, eyed-egg mortality and larval malformation rates were recorded as indices of egg quality. The results indicated that the maximum eyed eggs and hatched larvae (86% and 63%, respectively) were observed for eggs fertilized immediately after stripping, whereas the storage of the eggs at 4°C for 48 HPS decreased the eyed-egg and hatched-larvae rates to 46% and 17%, respectively. The use of a higher storage temperature resulted in a more rapid decrease in egg viability: eyed-egg and hatched-larvae rates of 23% and 9%, respectively, were obtained after 48 HPS storage at 8°C and 2% and 1% for eggs stored at 12°C. Eyed-egg mortality and larval malformation rates were not significantly affected by post-stripping ova ageing for at least up to 36 h. Thereafter, both values increased significantly and were measured to be the highest in the most aged ova. The present study demonstrated that stripped Eurasian perch eggs can be stored for at least 12 h at 4°C to 12°C without a significant reduction in their quality.     

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.     

Necator americanus: Activity patterns in the egg and the mechanism of hatching

Necator americanus developed normally in distilled water and in solutions of up to 4% NaCl, hatching however, occurred only in the lower tonicities. Together with other observations, this led to the conclusion that, as with many other nematodes, osmotic changes were critical for emergence. Behavioural movements of the larva within the egg have been described, and it has been suggested that the larva ‘feeds’ within the egg, and this involves head waving and oesophageal pumping into the intestine. Enzymes are thus flushed into the egg, causing membrane changes, an influx of water, and a considerable distention of the egg. The pressure is finally released by the rotary movements of the stoma against the egg membranes, causing a break in the egg through which the larva immediately passes.     

The evolution of egg clustering in butterflies: A test of the egg desiccation hypothesis

Females of many insect species cluster their eggs. Egg clustering by lepidopteran species usually results in aggregation of larvae that are more often conspicuously coloured and apparently distasteful or unpalatable than larvae of solitary species. While the costs and benefits of aggregation in terms of larval survival and growth are well documented, the evolutionary ecology of egg clustering has been long debated and is still unresolved. We tested the egg desiccation hypothesis, first proposed by Stamp (1980), which to our knowledge has never been examined experimentally. The egg desiccation hypothesis proposes that egg clustering is adaptive per se (i.e. increases fitness of females) by reducing egg mortality via desiccation.We tested this hypothesis for the Nymphalid butterfly, Chlosyne lacinia, an egg-clustering species on its sunflower host plant, Helianthus annuus. We first documented natural variation in batch size for this butterfly. We then tested experimentally hatch success of varying batch sizes and egg-layering arrangements under controlled humidity levels. Hatch success was positively related to relative humidity. Eggs in larger groups with greater number of layers had greater hatch success than smaller, monolayered egg batches, especially when relative humidity was low. Our results indicate that, not only number of eggs, but also the arrangement (i.e. layering and density), increase batch survival by protecting eggs from desiccation. However, despite increased hatch success in dense, multilayered clusters, we found wide variation in layering and density in natural populations of C. lacinia. This variation is probably maintained by trade-offs in egg survival, such as increased cannibalism of eggs by siblings, in dense clusters. Nevertheless, protection from egg desiccation provides an alternative explanation for the origin and maintenance of egg clustering in lepidopterans and possibly other insects. The pattern of egg deposition in the Nymphalidae supports this hypothesis, since most North American species cluster their eggs tightly, whereas most species in tropical regions lay eggs singly or in loose monolayers.