Hatching of cladoceran resting eggs: temperature and photoperiod

Summary 1. We identified temperature and photoperiod conditions under which the hatching of 45 cladoceran species could be elicited. Identification of appropriate hatching cues is of primary importance for the exploration of the ties between active and diapausing stages.
2. Incubation temperature affected the hatching success of resting eggs isolated from Danish, Belgian/Dutch and Spanish sediments. In general, most hatchlings and species were retrieved at 15 °C. Danish and Belgian/Dutch resting eggs hatched more successfully under a long day photoperiod than in continuous illumination.
3. Most species could be retrieved after incubation of resting eggs isolated from a limited amount of sediment (0.4 kg) under a single, well chosen combination of temperature and photoperiod. Processing additional sediment samples under seven more incubation regimes only allowed detection of 21% (Spain) to 34% (Denmark) more species.
4. The incubation period for resting eggs to hatch was strongly influenced by incubation temperature. Our results show that hatching experiments aimed at assessing cladoceran species richness and conducted at 15 °C should be continued for a period of at least 2 weeks, after which a random subset of hatchlings (e.g. n  = 100) can be selected from the total hatchling assemblage.     

Benthic resting eggs of calanoid copepods in Norwegian enclosures used in mariculture: abundance, species composition and hatching

Abundance, species composition and viability of benthic resting eggs of calanoid copepods were investigated in the sediments of seven Norwegian enclosed pond systems used in mariculture as well as in the sediments of the nearby open sea. The ponds represented different types of morphometry and zooplankton communities and were located from approximately 60 to 70 °N. Differences in environmental conditions in deep water also typical with anoxia and the presence of hydrogen sulphide at most of the localities. The lowest and highest egg densities in the enclosures were 7.5 ± 6.4 × 10⁴ and 5.5 ± 2.5 × 10⁶ m^-2, respectively, higher than in the sediments of the nearby open sea. Highest densities were found at larger or median depths. Altogether, egg densities higher than 10³ m^-2 were found of only four calanoid species; Eurytemora affinis, Acartia teclae, Acartia clausi, Temora longicornis. Hatching success was variable (0–92%), and no significant correlation to the presence of anoxia and hydrogen sulphide was found. This high hatching success was achieved with eggs exposed to hydrogen sulphide for several months. The recruitment potential to the plankton of nauplii was estimated from 4.8 to 1682 nauplii l^-1 for the different localities. The results showed that resting egg production is a common life history characteristic of some neritic calanoid species in Norwegian waters. In addition to copepod eggs, low densities (< 10³ m^-2) of eggs from rotifers and cladocerans were found in most of the localities.     

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

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

Molecular genetic analysis of Daphnia in the English Lake District: species identity, hybridisation and resting egg banks

• 1 Taxonomic issues in many Daphnia species complexes are often confused by a high degree of phenotypic plasticity and by interspecific hybridisation. Here, we employ molecular genetic tools to confirm the species composition and incidence of hybridisation in extant and resting egg populations of Daphnia from Windermere and Esthwaite Water in the English Lake District.
• 2 A combination of species‐diagnostic allozymes and mtDNA, confirms that contemporary populations are dominated by a single species, D. galeata.
• 3 We present the first account of genetic characterisation of dated ‘resting’ eggs using microsatellites and mtDNA, employing PCR‐based DNA recovery, thus providing a temporal dimension to taxonomic patterns. Thirty years ago, two species were present in Esthwaite, D. galeata and D. hyalina, but Windermere populations were dominated by D. galeata only.
• 4 The use of PCR‐based mtDNA RFLP analysis as a species‐diagnostic tool, and microsatellites to monitor clonal diversity, provide a valuable approach for long‐term studies, especially in populations free from the complicating effect of frequent hybridisation. The detailed limnological records available for many large lakes, and associated changes in land‐use, pollutants and climate, combined with long‐term ephippial molecular genetic data, provide opportunities for exploring natural and anthropogenic impacts on genetic and community structure.

     

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.     

Variation in ephippial buoyancy in Daphnia pulicaria

1. Freshwater zooplankton often produce diapausing eggs to survive environmental stress. The diapausing eggs of Daphnia (Crustacea, Cladocera) are encased in an ephippium that either floats at the surface or sinks to the sediment. These two types of ephippia may represent different strategies between spatial (floaters) and temporal (sinkers) dispersal of offspring. 2. We observed floating and sinking characteristics of ephippia obtained from eight lakes. We then conducted an experiment with 26 Daphnia pulicaria clones obtained from six of these lakes and observed the production of buoyant and non‐buoyant ephippia under high and low food conditions. 3. Ephippia were more often non‐buoyant than buoyant both from females caught in nature and those reared in the laboratory. The experiment revealed that each clone was able to produce both types of ephippia, but that there was considerable among‐clone variation in the percentage of non‐buoyant ephippia produced. 4. We conclude that production of non‐buoyant versus buoyant ephippia may be driven by both genetic and environmental factors.     

Size variation in Brachionus plicatilis resting eggs

The effect of temperature and salinity on resting egg size of two Brachionus plicatilis (Rotifers) clones was investigated. Clones were selected according to their different behaviour in laying resting eggs: one clone ejects them, whereas they remain inside the females body in the other clone. The difference in resting eggs size between the two clones is noticeable, although the difference is not as great as that between female body size. An important temperature-salinity interaction on resting egg size has been observed. The general inverse relationship between size and temperature is only true at lower temperatures. At high temperatures size varies around the mean although could be greater than at intermediate temperatures. This is more evident at the intermediate salinity tested which is considered to be the closest to the optimum in our experiments. This pattern of variation suggests that mean size is bigger than expected, in relation to temperature and salinity, when these factors have values close to the extremes of their range, normally found in nature, and to which adaptative mechanisms can evolve. Size is bigger at the salinity — temperature low - low and high - high combinations which are the most commonly found in the temperate environments.     

The significance of mating processes for the genetics and for the formation of resting eggs in monogonont rotifers

Crossbreeding experiments with three geographically distinct strains (E, S, and L) of the rotifer Brachionus plicatilis have been carried out in an attempt to elucidate the apparent male sterility of strain L, which is unable to produce resting eggs.The 9 crossing possibilities of the three strains have been investigated in 27 experiments. The results refute the concept of male sterility. L-males copulate successfully with mictic as well as with amictic females of strains E and S. Fertilized amictic E and S females produce defective resting eggs, which have only one thin shell and, which disintegrate after deposition. L-females cannot, as a rule, be fertilized. Moreover, crosses between strain E and S are succesful only in one direction; the reciprocal crosses failed.To explain the present results a hypothesis is suggested that the thickness of the body wall of newborn females differs in the three strains, and between mictic and amictic individuals. A sequence of gradually increasing body wall thickness of all types of females involved, together with a comparably increasing penetration ability of the males of strain E, S, and L explains the success or failure of all crosses, including the unilateral cross E × S.     

广东流溪河水库表层沉积物中枝角类休眠卵的多样性及空间分布

为了解华南地区深水水库中枝角类休眠卵在底泥中的种群特征,于2009年2月对广东省流溪河水库沉积物中的枝角类休眠卵种类、数量和分布进行研究。在湖泊区、过渡区和河流区共设置8个采样点,采集表层15cm柱状底泥,经600、200和35μm三种孔径的网筛分离休眠卵后显微镜下计数。15cm样品根据柱状底泥的纹层确定为18～24年的沉积物。共检出12种枝角类休眠卵,河流区采样点未检出,两条入库河流汇合区种类数为10,其余采样点有4至5种。15cm沉积物中休眠卵的平均密度为1.3×10⁵ ind.·m^-3。休眠卵的多样性及密度分布特征与生境相关,过渡区底泥中的多样性高于湖泊区,但湖泊区底泥中密度高。卵库中的优势种类为微型裸腹溞(Moina micrura)、船卵溞属(Scapholeberis sp.)和溞属(Daphnia sp.),均为逃避能力弱的种类。流溪河位于热带北缘,水温常年较高,多数枝角类可通过孤雌生殖终年存在于水体中,导致休眠卵的种类数量与密度均明显地低于温带湖泊。     

The resting eggs of the Ctenopoda (Crustacea: Branchiopoda): a review

The resting eggs of the Ctenopoda have been studied since the second half of the 19th century. By now they are known in 21 species of the group but only in 4 species (Sida crystallina, Diaphanosoma brachyurum, Penilia avistoris and Holopedium gibberum) have they been investigated thoroughly. Gross and internal structure of eggs are characterized by considerable diversity. In general their structure has much in common with resting eggs of the Anostraca, Notostraca, Conchostraca and Onychopoda as well as with that of some other remotely related groups of freshwater invertebrates (Rotifera, Hydra). The taxonomic value of resting eggs is discussed. Probably they play a significant role in determining geographical distribution. Waterfowl are the main agent of their dispersal.     

Effect of incubation temperature on the hatching of rotifer resting eggs collected from sediments

Sediment samples from Loch Leven, Scotland, were incubated at 5°C, 10°C and 15°C to induce hatching of rotifer resting eggs. The emergent animals were identified and counted. The temperature which induced hatching varied among the nine species studied. These results are discussed in relation to seasonality and temperature preferences previously recorded for the most abundant species. Resting egg densities of 2.2–13.9 eggs cm^–3 were recorded in the upper 5 cm of sediment.     

Long-term droughts change the hatching patterns of zooplankton resting eggs from permanent and temporary lakes

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The effect of temperature on the development times of eggs,naupliar and copepodite stages of five species of cyclopoid copepods

The duration times of eggs, combined naupliar instars and of the different copepodite stages of five species of cyclopoid copepods — Acanthocyclops robustus, Cyclops vicinus, Diacyclops bicuspidatus, Mesocyclops leuckarti, and Thermocyclops crassus — were investigated at five different temperatures. The five species can be divided in two groups: two species, C. vicinus and D. bicuspidatus, adapted to cold water conditions and three species, A. robustus, M. leuckarti and T. crassus adapted to warm water conditions. The cold water species showed a faster egg development than M. leuckarti and T. crassus at 5–15 °C. The eggs and instars of the warm water species M. leuckarti tend to develop faster than those of the former two species at higher temperatures. A. robustus showed the shortest egg and instar development at 10–25 °C. The warm water species T. crassus produced no eggs at 10 °C and temperatures below. At higher temperatures (20, 25 °C) the egg and instar duration times were similar or longer than those of the other species. When cultured in total darkness a great part of the C_IV respectively C_V copepodites of the summer forms entered arrest and the percentage of copepodites that showed an arrest of development was highest at lowest temperatures. The present results are compared with data from literature and differences are discussed.     

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.     

Fluctuating asymmetry and developmental instability in evolutionary biology: past, present and future

The role of developmental instability (DI), as measured by fluctuating asymmetry (FA), in evolutionary biology has been the focus of a wealth of research for more than half a century. In spite of this long period and many published papers, our current state of knowledge reviewed here only allows us to conclude that patterns are heterogeneous and that very little is known about the underlying causes of this heterogeneity. In addition, the statistical properties of FA as a measure of DI are only poorly grasped because of a general lack of understanding of the underlying mechanisms that drive DI. If we want to avoid that this area of research becomes abandoned, more efforts should be made to understand the observed heterogeneity, and attempts should be made to develop a unifying statistical protocol. More specifically, and perhaps most importantly, it is argued here that more attention should be paid to the usefulness of FA as a measure of DI since many factors might blur this relationship. Furthermore, the genetic architecture, associations with fitness and the importance of compensatory growth should be investigated under a variety of stress situations. In addition, more focus should be directed to the underlying mechanisms of DI as well as how these processes map to the observable phenotype. These insights could yield more efficient statistical models and a unified approach to the analysis of patterns in FA and DI. The study of both DI and canalization is indispensable to obtain better insights in their possible common origin, especially because both have been suggested to play a role in both micro- and macro-evolutionary processes.     

池塘桡足类休眠卵对浮游种群的潜在补充及其影响因素

休眠卵是桡足类的重要生存策略,其在沉积物中的丰度对水体种群的补充具有重要意义.干塘是池塘养殖管理的重要措施,对池塘生态系统具有重要影响.2014年2月15日(干塘前)和3月2日(干塘后),分别采集广州市南沙区养殖池塘内表层沉积物,研究沉积物中桡足类休眠卵对浮游种群的潜在补充及影响因素.结果表明: 干塘前未冷藏沉积物样品在盐度20时的潜在补充量和孵化率最高,4 ℃冷藏4周后最高值出现在盐度为15的处理组.干塘后未冷藏沉积物样品在盐度15时的潜在补充量和孵化率最高,冷藏4周后最高值出现在盐度为20的处理组.这说明干塘前桡足类休眠卵的潜在补充量显著高于干塘后,且干塘对沉积物中休眠卵的持续孵化时间有影响.     

Structural differences between the resting eggs of Brachionus plicatilis and Brachionus rotundiformis (Rotifers, Brachionidae): an electron microscopic study

Surface topography and layering of resting eggs of Brachionus plicatilis and B. rotundiformis are described, based on scanning and transmission electron microscope studies. In B. plicatilis, the resting eggs are spherical with smooth wavy ridges on the surface, whereas these ridges are small and condensed in B. rotundiformis. The distribution of pores on the egg surface clearly distinguishes the two species.Ultrastructure of the egg membranes of both species varies greatly with regard to size, shape and sculpturing of each membrane. The alveolar and dense sublayers, which constitute the outer egg membrane (S1) are very thick (10–12 µm) in B. rotundiformis compared to B. plicatilis (4–5 µm). Thus, each species has a characteristic surface and membrane architecture. The functional roles of these membranes, during the resting phase of rotifers, are discussed.     

家蚕滞育卵与非滞育卵中几种关键酶活性的比较

家蚕Bombyx mori是卵滞育的昆虫, 在滞育期间无形态变化, 也不存在器官发育和组织分化, 然而其生理代谢过程仍在进行。为进一步研究家蚕滞育的机制, 本研究测定了家蚕滞育卵、 即时浸酸处理的滞育卵及非滞育卵在胚胎发育过程中的超氧化物歧化酶(superoxide dismutase, SOD, EC 1.15.1.1)、 过氧化氢酶(catalase, CAT, EC 1.11.1.6)、 丙酮酸激酶(pyruvate kinase, PK, EC 2.7.1.40)、 乙酰胆碱酯酶(acetylcholine esterase, AchE, EC 3.1.1.7)和乳酸脱氢酶(lactate dehydrogenase, LDH, EC 1.1.1.28) 的活性变化。结果表明： 处理后1-7 d, 即时浸酸处理的滞育卵, SOD活性由56 517.00 U/g提高到81 986.94 U/g, CAT活性由14.98 U/g提高到106.90 U/g, PK活性由25.19 U/g提高到181.70 U/g, AChE活性由17.88 U/g提高到287.86 U/g, 而LDH活性由169.96 U/g下降到122.82 U/g。 而在非滞育卵中, SOD活性由86 417.99 U/g下降到66 024.19 U/g, LDH活性由169.07 U/g下降到135.02 U/g; CAT活性由1.47 U/g提高到44.37 U/g, PK活性由20.56 U/g提高到92.09 U/g, AChE活性由21.40 U/g提高到99.17 U/g。在滞育卵中, SOD和AChE活性较稳定; CAT活性随发育上升, 而LDH活性随发育而下降; PK活性在胚胎发育的前 4 d呈上升趋势, 随后基本保持稳定。通过了解家蚕滞育卵、 非滞育卵与即时浸酸卵的相关酶活性在胚胎发育过程中存在的变化, 有助于进一步揭示家蚕滞育的机理。     

Chitosomes: past, present and future

José Ruiz-Herrera's discovery that chitin microfibrils could be made by a fungal extract paved the way for elucidating the intracellular location of chitin synthetase. In collaboration with Charles Bracker, chitosomes were identified as the major reservoir of chitin synthetase in fungi. Unique in size, buoyant density, and membrane thickness, chitosomes were found in a wide range of fungi. Their reversible dissociation into 16S subunits is another unique property of chitosomes. These 16S subunits are the smallest molecular entities known to retain chitin synthetase activity. Further dissociation leads to complete loss of activity. From studies with secretory mutants, yeast researchers concluded that chitosomes were components of the endocytosis pathway. However, key structural and enzymatic characteristics argue in favor of the chitosome being poised for exocytotic delivery rather than endocytotic recycling. The chitosome represents the main vehicle for delivering chitin synthetase to the cell surface. An immediate challenge is to elucidate chitosome ontogeny and the role of proteins encoded by the reported chitin synthetase genes in the structure or function of chitosomes. The ultimate challenge would be to understand how the chitosome integrates with the cell surface to construct the organized microfibrillar skeleton of the fungal cell wall.