Cloning,expression and cellular localization of Daphnia pulex senescence-associated protein,DpSAP

Daphnia (water fleas) are small crustaceans that undergo an unusual switch from asexual to sexual reproduction that is dependent on environmental conditions. In this study, a senescence-associated protein (SAP) from the common freshwater species Daphnia pulex was cloned using primers based on homologous sequences and rapid amplification of cDNA ends (RACE). Real-time PCR was employed to quantify the expression of D. pulex SAP (DpSAP) in individual organisms. The role of DpSAP in the reproductive transformation was further investigated in both parthenogenetic and sexual females by using digoxin-labeled SAP RNA probes and RNA whole-mount in situ hybridization. DpSAP was more highly expressed in sexual females, indicating a role in growth and reproduction. Cellular localization studies using RNA whole-mount in situ hybridization showed specific expression in the second tentacle joints. These expression patterns suggest an important role for DpSAP in the reproductive transformation of D. pulex.     

Effects of temperature and salinity on the standard metabolic rate (SMR) of the caridean shrimp Palaemon peringueyi

The standard metabolic rate (SMR) of the caridean shrimp Palaemon peringueyi to changes in temperature (15-30 °C), salinity (0-45‰) and a combination thereof was investigated. The rate of oxygen consumption of the shrimp was determined using a YSI oxygen meter. At a constant salinity of 35‰ the respiration rate of P. peringueyi increased with an increase in temperature and ranged between 0.260 and 0.982 μl O₂ mg wwt^− 1 h^− 1. The Q₁₀ value over the temperature range 15-25 °C was estimated at 3.13. At a constant temperature of 15 °C the respiration rate of P. peringueyi also increased with an increase in salinity and ranged between 0.231 and 0.860 μl O₂ mg wwt^− 1 h^− 1. For combination experiments the absence of any significant difference in the respiration rate of P. peringueyi at the four temperatures over the salinity range 15-35‰ suggests that the shrimp is well adapted to inhabiting environments characterised by variations in salinity and temperature such as those encountered within the middle and lower reaches of permanently open estuaries with substantial freshwater inflow. On the other hand, the total mortality of the shrimp recorded at salinities < 5‰ at all four temperatures suggests that the upper distribution of the shrimp may reflect physiological constraints. Similarly, the increase in the respiration rate of the shrimp at the four temperatures at salinities > 35‰ suggests that the shrimp may experience osmotic stress in freshwater deprived permanently open and intermittently open estuaries where hypersaline conditions may develop.     

Influence of oxygen and temperature on growth and metabolic performance of Paralichthys lethostigma (Pleuronectiformes: Paralichthyidae)

In this study, we apply Fry's classification of environmental factors to demonstrate the limiting effects of oxygen and its interaction with temperature on the growth of juvenile P. lethostigma. We also evaluated the properties of two metabolic indices, marginal metabolic scope (MMS) and limiting oxygen concentration (LOC), as indicators of metabolic scope. We found that oxygen limitation has its greatest impact near the optimum temperature for growth of the species. At 29 °C a reduction from 6.00 mg/L to 4.00 mg/L caused a 50% reduction in growth rate while at 27 °C the reduction had no significant effect on growth rate. The results are particularly relevant because these temperatures and oxygen concentrations are commonly observed in nursery areas during summer months. At all temperatures fish from the lowest oxygen treatment (1.75 mg/L) had negative growth rates. Comparisons between daily oscillating oxygen treatments and constant treatments failed to demonstrate significant effects. At temperatures past the optimum, growth rates between the 6.00 mg/L and 4.00 mg/L treatments were not statistically different. LOC was significantly affected by temperature, oxygen, and their interaction. Estimates were positively correlated with oxygen treatment (R² > 0.71) and negatively correlated with temperature at moderate and low oxygen concentrations (R² > − 0.84). MMS was significantly affected by temperature and oxygen and was significantly correlated with oxygen treatment (R² > − 0.91), but correlations with temperature were not as clear. In conclusion, oxygen and temperature interactions have significant effects on metabolic scope and growth rates of fish, well above the accepted hypoxia threshold of 2.00 mg/L and MMS has proved a useful estimator of the metabolic scope of the organism within an environment.     

Effects of temperature, salinity, and air exposure on development of the estuarine pulmonate gastropod Amphibola crenata

The primitive pulmonate snail Amphibola crenata embeds embryos within a smooth mud collar on exposed estuarine mudflats in New Zealand. Development through hatching of free-swimming veliger larvae was monitored at 15 salinity and temperature combinations covering the range of 2-30 ppt salinity and 15-25 °C. The effect of exposure to air on developmental rate was also assessed. There were approximately 18,000 embryos in each egg collar. The total number of veligers released from standard-sized egg collar fragments varied with both temperature and salinity: embryonic survival was generally higher at 15 and 20 °C than at 25 °C; moreover, survival was generally highest at intermediate salinities, and greatly reduced at 2 ppt salinity regardless of temperature. Even at 2 ppt salinity, however, about one-third of embryos were able to develop successfully to hatching. Embryonic tolerance to low salinity was apparently a property of the embryos themselves, or of the surrounding egg capsules; there was no indication that the egg collars protected embryos from exposure to environmental stress. Mean hatching times ranged between 7 and 22 days, with reduced developmental rates both at lower temperature and lower salinity. At each salinity tested, developmental rate to hatching was similar at 20 and 25 °C. At 15 °C, time to hatching was approximately double that recorded at the two higher exposure temperatures. Exposing the egg collars to air for 6-9 h each day at 20 °C (20 ppt salinity) accelerated hatching by about 24 h, suggesting that developmental rate in this species is limited by the rates at which oxygen or wastes can diffuse into and from intact collars, respectively. Similarly, veligers from egg capsules that were artificially separated from egg collars at 20 °C developed faster than those within intact egg collars. The remarkable ability of embryos of A. crenata to hatch over such a wide range of temperatures and salinities, and to tolerate a considerable degree of exposure to air, explains the successful colonization of this species far up into New Zealand estuaries.     

温度、盐度和pH对马氏珠母贝稚贝清滤率的联合效应

清滤率(Clearance rate, CR)与滤食性贝类生长发育密切相关,采用Box-Behnken设计(BBD)和响应曲面法,在实验室条件下研究了温度(18-34℃)、盐度(20-40)和pH(6.5-9.5)对马氏珠母贝(Pinctada martensii)稚贝清滤率(CR)的联合效应,旨在建立温度、盐度和pH对马氏珠母贝稚贝清滤率的定量关系模型,并通过统计优化方法得出温度、盐度和pH的最佳组合。结果表明:温度的一次效应、温度和pH的互作效应、盐度和pH的互作效应以及温度、盐度和pH的二次效应对马氏珠母贝稚贝清滤率的影响均极显著(P<0.01);盐度的一次效应、pH的一次效应以及温度和盐度的互作效应对清滤率无显著影响 (P>0.05)。实验得出的清滤率模型决定系数为0.9950,预测决定系数为0.9284,表明该模型建立有效并可用于预测马氏珠母贝稚贝的清滤率。通过采用优化方法得出,在温度26.95℃,盐度29.69,pH8.09时,稚贝清滤率达到最大,最大值为1.4894×10^-3L/h,满意度为0.9886。研究结果可为马氏珠母贝滤食生理研究及稚贝培育提供理论依据。     

Effects of food concentration on clearance rate and energy budget of the Arctic bivalve Hiatella arctica (L) at subzero temperature

The influence of food concentration on clearance rate, respiration, assimilation, and excretion at −1.3 °C was studied on individuals of the bivalve Hiatella arctica (L.) from Young Sound, NE Greenland. Clearance rate, assimilation efficiency, respiration, and excretion rates were determined over a range of food concentrations using the microalga Rhodomonas baltica as food source. Physiological rates were generally low but responded significantly to increased food levels. Clearance rates and assimilation efficiency were reduced at increased food levels, whereas respiration and excretion increased. Assimilation efficiency was generally high, which may be an adaptation to the low food concentration during most of the year in NE Greenland. Low filtration rates limited ingestion rates and resulted in a low maximum assimilation of 3 J h⁻¹. Despite the low food intake, very low food concentrations were required for individual specimens to obtain a positive energy budget. Predicted growth based on rates of assimilation and respiration were compared to published estimates of annual growth in Young Sound. We estimate that 3 weeks of growth in the laboratory under optimal food conditions could match annual growth in situ. We interpret this as evidence that food limitation is the primary impediment to growth in this Arctic population.     

Heat tolerance, behavioural temperature selection and temperature-dependent respiration in larval Octopus huttoni

This study reports temperature effects on paralarvae from a benthic octopus species, Octopus huttoni, found throughout New Zealand and temperate Australia. We quantified the thermal tolerance, thermal preference and temperature-dependent respiration rates in 1-5 days old paralarvae. Thermal stress (1 °C increase h⁻¹) and thermal selection (∼10-24 °C vertical gradient) experiments were conducted with paralarvae reared for 4 days at 16 °C. In addition, measurement of oxygen consumption at 10, 15, 20 and 25 °C was made for paralarvae aged 1, 4 and 5 days using microrespirometry. Onset of spasms, rigour (CT_max) and mortality (upper lethal limit) occurred for 50% of experimental animals at, respectively, 26.0±0.2 °C, 27.8±0.2 °C and 31.4±0.1 °C. The upper, 23.1±0.2 °C, and lower, 15.0±1.7 °C, temperatures actively avoided by paralarvae correspond with the temperature range over which normal behaviours were observed in the thermal stress experiments. Over the temperature range of 10 °C-25 °C, respiration rates, standardized for an individual larva, increased with age, from 54.0 to 165.2 nmol larvae⁻¹ h⁻¹ in one-day old larvae to 40.1-99.4 nmol h⁻¹ at five days. Older larvae showed a lesser response to increased temperature: the effect of increasing temperature from 20 to 25 °C (Q₁₀) on 5 days old larvae (Q₁₀=1.35) was lower when compared with the 1 day old larvae (Q₁₀=1.68). The lower Q₁₀ in older larvae may reflect age-related changes in metabolic processes or a greater scope of older larvae to respond to thermal stress such as by reducing activity. Collectively, our data indicate that temperatures >25 °C may be a critical temperature. Further studies on the population-level variation in thermal tolerance in this species are warranted to predict how continued increases in ocean temperature will limit O. huttoni at early larval stages across the range of this species.     

Seasonal variability in feeding behaviour, metabolic rates and carbon and nitrogen balances in the Sydney oyster, Saccostrea glomerata (Gould)

To establish if nutrients limit the growth of bivalves requires information not only on the quality of food available, but also the animals' feeding behaviour and endogenous metabolic demands. We hypothesized that growth of the Sydney rock oyster (Saccostrea glomerata) would vary in response to seasonal changes in food quality rather than quantity. We also predicted that the oysters would show feeding preferences for nitrogen over carbon, and this behaviour would result in carbon/nitrogen (C/N) ratios for ingested and absorbed matter that would be lower than the C/N of both the seston and the oysters' estimated metabolic maintenance requirements. The experiments were done in two phases under natural conditions. In phase 1, feeding behaviour was assessed on a single occasion and the results used to pose hypotheses for testing in phase 2, which included measurements made on three occasions encompassing autumn, winter and spring conditions. Growth rates varied with changes in ambient food quality and not with the concentration of total suspended matter. Feeding behaviour responded to food quality and, in most cases, resulted in nitrogen enrichment. For example, when nitrogen was potentially limiting to growth and/or maintenance, due to high food C/N (July) or high nitrogen demand (March), pre-ingestive selection ensured nitrogen enrichment of ingested matter and C/N ratios of ingested matter which were below the maintenance requirement. However, in November, when endogenous demands indicated an increased requirement for carbon, feeding behaviour resulted in carbon enrichment, an increase in carbon conversion efficiency, and ingested C/N ratios greater than the maintenance requirement. The results support the assertion of variable feeding physiology in oysters, responsive to both exogenous (seasonal differences in carbon and nitrogen availability) and endogenous (cycles of reproduction and growth) factors.     

Comparison of low salinity tolerance in Callinectes sapidus Rathbun and Callinectes similis Williams postlarvae upon entry into an estuary

Planktonic larvae of estuarine crabs are commonly exported to the continental shelf for development and then return to coastal and estuarine areas as postlarvae (megalopae). Megalopae returning to estuaries must be adapted to survive in brackish water whereas those of coastally distributed species should not need such adaptations. We investigated 1) whether megalopae of the estuarine crab Callinectes sapidus and the coastal crab Callinectes similis undergo changes in salinity tolerance upon entry into an estuary and 2) what factors induce those changes. Megalopae were collected at a coastal site and a nearby estuarine site and exposed to a range of salinities (5, 10, 15, 20 and 30) for 6 h. Percent survival was determined after 24 h reintroduction to the collection site water. We also investigated 1) whether increased salinity tolerance was induced by reduced salinity or estuarine chemical cues, 2) the time to acclimation and 3) the salinity necessary for acclimation. C. sapidus megalopae from the estuarine site were more likely to survive exposure to low salinities than those from the coastal site. C. sapidus megalopae from the coastal site exhibited increased survival after acclimation to salinities of 27 and 23 for 12 h. Estuarine chemical cues had no effect on salinity tolerance. C. similis megalopae were less likely to survive at low salinities and did not exhibit an acclimation response upon exposure to reduced salinities. These results suggest that megalopae of C. sapidus are physiologically adapted to recruit to estuaries whereas megalopae of C. similis are unable to acclimate to low salinity conditions.     

Thermopreference,tolerance and metabolic rate of early stages juvenile Octopus maya acclimated to different temperatures

Thermopreference, tolerance and oxygen consumption rates of early juveniles Octopus maya (O. maya; weight range 0.38–0.78 g) were determined after acclimating the octopuses to temperatures (18, 22, 26, and 30 °C) for 20 days. The results indicated a direct relationship between preferred temperature (PT) and acclimated temperature, the PT was 23.4 °C. Critical Thermal Maxima, (CTMax; 31.8±1.2, 32.7±0.9, 34.8±1.4 and 36.5±1.0) and Critical Thermal Minima, (CTMin; 11.6±0.2, 12.8±0.6, 13.7±1.0, 19.00±0.9) increased significantly (P<0.05) with increasing acclimation temperatures. The endpoint for CTMax was ink release and for CTMin was tentacles curled, respectively. A thermal tolerance polygon over the range of 18–30 °C resulted in a calculated area of 210.0 °C². The oxygen consumption rate increased significantly α=0.05 with increasing acclimation temperatures between 18 and 30 °C. Maximum and minimum temperature quotients (Q₁₀) were observed between 26–30 °C and 22–26 °C as 3.03 and 1.71, respectively. These results suggest that O. maya has an increased capability for adapting to moderate temperatures, and suggest increased culture potential in subtropical regions southeast of México.     

体重和盐度对中国蛤蜊耗氧率和排氨率的影响

采用室内实验生态方法研究了不同软体部干重((1.022 ±0.821)、(0.557±0.266)、(0.303±0.277) g)和盐度(13、18、23、28、33)对中国蛤蜊(Mactra chinensis Philippi)耗氧率和排氨率的影响。结果表明:盐度、个体大小对中国蛤蜊耗氧率的影响极显著(P＜0.01),二者的交互作用对中国蛤蜊耗氧率影响显著(P＜0.05);中国蛤蜊单位体重耗氧率(R₀)与软体部干重(W)负相关,符合幂函数方程R₀=aW^-b, 其中a值的取值范围是0.695-1.762,平均值为1.449,b值的取值范围是0.446-0.587,平均值为0.542。盐度、个体大小对中国蛤蜊排氨率影响也极显著(P＜0.01);随着中国蛤蜊个体的增大,其单位体重排氨率逐渐降低;排氨率与其软体部干重呈负相关,它们之间可以用幂函数R_N=a₀W^-b₀表示。单位体重耗氧率和排氨率与盐度(S)、软体部干重(W)的二元线性回归方程分别为: R_O=2.111-1.817W+0.49S (R²=0.546, F=34.294, P＜0.001);R_N=168.186-120.589W+1.734S (R²=0.561, F=36.418, P＜0.001)。     

Differential thermal tolerance and energetic trajectories during ontogeny in porcelain crabs,genus Petrolisthes

Thermal tolerance limits of marine intertidal zone organisms are elevated compared to subtidal species, but are typically just slightly higher than maximal habitat temperatures. The small thermal safety margins maintained by intertidal zone organisms suggest that high thermal tolerance is associated with a physiological cost. If true, we hypothesize that species that transition between intertidal zone and planktonic habitats during ontogeny, will adjust their thermal tolerance accordingly to capitalize upon potential energy savings while in a thermally benign habitat. We tested this hypothesis in porcelain crabs that transition between the thermally stressful, intertidal zone as embryos, to the thermally benign pelagic zone as larvae, and back at settlement. We found the more thermally tolerant, mid-intertidal zone species, Petrolisthes cinctipes, and the less thermally tolerant, subtidal zone species, Petrolisthes manimacilis, exhibited reduced thermal tolerance (LT₅₀) in the transition from embryos to larvae. This was associated with an increased oxygen consumption rate in both species, though P. cinctipes exhibited a significantly greater increase in oxygen consumption. P. cinctipes also showed an increase in thermal tolerance in settled juveniles compared to pelagic zoea I larvae, resulting in an overall V-shaped thermal tolerance relationship during ontogeny, while in P. manimaculis thermal tolerance was significantly lower in juveniles compared to zoea I. In neither species were these changes (zoea I to juvenile) associated with a significant change in metabolism. While embryos and juveniles of P. cinctipes have thermal tolerance limits near intertidal habitat thermal maxima (∼32.5 °C), all three life-history stages in P. manimaculis (especially embryos and larvae) exhibit considerable thermal safety margins. The mechanisms underlying this “excess” thermal tolerance in P. manimacilis embryos are unknown, but suggest that patterns of thermal tolerance in early life history stages are species-specific.     

温度、盐度及交互作用对僧帽牡蛎(Saccostrea cucullata)呼吸排泄的影响

采用实验生态学方法研究了温度(13、18、23、28、31、34℃)和盐度(18、23、28、33)对僧帽牡蛎(Saccostrea cueullata)耗氧率和排氨率的影响,并分析了二者交互作用对呼吸排泄的影响。结果表明,不同温度条件下,僧帽牡蛎的耗氧率变化范围为0.43-3.15mg g~(-1)h~(-1),排氨率的变化范围为72.62-1245.77μg g~(-1)h~(-1),方差分析显示,温度和盐度对僧帽牡蛎耗氧率(F=60.440,P0.01)和排氨率(F=61.083,P0.01)的影响均极显著;二者的交互作用对僧帽牡蛎耗氧率(F=4.907,P0.01)和排氨率(F=11.412,P0.01)影响也极显著,耗氧率(Y)、排氨率(Y)与温度(T)和盐度(S)之间的二元线性回归方程为y=a+bT+cS+dT×S。在一定温度范围内(13-31℃)僧帽牡蛎单位体重耗氧率(Y)与温度(T)呈正相关符合指数方程Y=ae~(bT),其中a的取值范围为0.1378-0.2117,b的取值范围为0.0757-0.0912;单位体重排氨率也与温度呈正相关,符合上述指数方程,其中a的取值范围为20.364-30.634,b的取值范围为0.1004-0.1341。不同温度和盐度条件下,僧帽牡蛎呼吸排泄的氧氮比在2.20-7.78之间,平均值为3.81。     

Osmotic regulation and salinity tolerance of the stonefly nymph, Paragnetina media

Nymphs of Paragnetina media were able to tolerate a range of salinities in the external media up 1.1% NaCl (～ 354 mOsm/l.) Nymphs maintained in distilled water showed excellent osmoregulation by producing hyposmotic urine. In a range of salinities, 0.4–0.95% NaCl, nymphs maintained their baemolymph concentration hyperosmotic to the external media. A breakdown in osmotic regulation occurred at 1.1% NaCl. There was no significant change (P > 0.01) in the oxygen consumption, indicating that metabolic energy demand for the osmoregulation was not affected.     

Zooid size and growth rate of the bryozoan Cryptosula pallasiana Moll in relation to temperature, in culture and in its natural environment

The size of cheilostome bryozoan zooids has been widely discussed for its potential in inferring palaeotemperatures, based on correlations between zooid size and temperature. Studies in both the natural environment and under experimental laboratory conditions have shown that an increase in temperature significantly decreases zooid size in a range of bryozoan taxa. In order to test the effect of temperature on zooid size, the cheilostome bryozoan Cryptosula pallasiana was for the first time successfully cultured under laboratory conditions. C. pallasiana was grown at 14 °C and 18 °C using Rhodomonas sp. as a food organism. Zooid size, tentacle number and growth rate were measured over a period of 26 days. For comparison, zooids from colonies of C. pallasiana collected from the natural environment were measured in winter and summer months. Results showed that colonies grown in laboratory culture had significantly longer and wider zooids at 14 °C than at 18 °C. The specific growth rate of C. pallasiana doubled from 14 °C to 18 °C. Comparison of tentacle number in culture showed a significantly higher value at lower temperatures. This may be related to differing food availability, longer polypide life spans, or a shift of energy use at colder temperatures. In nature the zooids were significantly longer in colonies sampled in July than in January, a clear difference from laboratory results. The utility of cheilostome Bryozoa as indicators of environmental change and their potential for studies of paleotemperature are highlighted.     

虎纹蛙选择体温和热耐受性在个体发育过程中的变化

体温是影响变温动物表现的最重要生理学变量。检测了国家二级保护动物虎纹蛙的雌性亚成体、雄性亚成体、幼体和蝌蚪这4个发育阶段的选择体温和热耐受性。单因子方差分析表明,虎纹蛙选择体温、耐受低温、耐受高温和温度耐受范围的组间差异均显著,幼体的选择体温(24.13℃)显著低于雌性亚成体(28.06℃)、雄性亚成体(29.27℃)和蝌蚪(28.23℃),雌性亚成体、雄性亚成体和蝌蚪之间差异不显著;幼体的耐受低温(13.85℃)显著高于雌性亚成体(11.27℃)、雄性亚成体(10.84℃)和蝌蚪(10.74℃),雌性亚成体、雄性亚成体和蝌蚪之间差异不显著;幼体具有显著低的耐受高温(35.48℃)、蝌蚪具有显著高的耐受高温(43.31℃),雌性亚成体(39.55℃)和雄性亚成体(39.02℃)的耐受高温差异不显著;幼体(21.62℃)具有显著小的温度耐受范围、蝌蚪(32.58℃)具有显著大的温度耐受范围,雌性亚成体(28.28℃)和雄性亚成体(28.18℃)的温度耐受范围差异不显著。虎纹蛙幼体和亚成体体温和水温之间在降温速度和升温速度的相关关系均显著。用回归剩余值去除水温变化速度对体温变化的影响,双因子方差分析(降温和升温速度为重复检验设置)表明,幼体的体温变化速度显著大于亚成体,两性亚成体间差异不显著;温度变化类型(降温和升温)和两因子的交互作用对体温变化的影响不显著。基本热生态位分离和体温调节能力的发育限制是形成上述现象的最可能的原因。     

Responses of the saltmarsh rush Juncus kraussii to salinity and waterlogging

Juncus kraussii Hochst., an important saltmarsh macrophyte, is intensively harvested for many commercially orientated products and current populations are under threat of overexploitation. In saline, intertidal mud banks, this species occurs on higher ground, suggesting that it is adapted to lower salinities and less frequent inundation. The objectives of this study were to determine biomass accumulation, as well as morphological and physiological adaptations of J. kraussii to salinity and waterlogging stresses. Plants collected from the field were subjected to 0.2, 10, 30, 50 and 70% seawater under drained or flooded conditions for three months. Measurements were made of biomass accumulation, CO₂ exchange, chlorophyll fluorescence, ion and water relations. Furthermore, seed germination responses to a range of salinities were investigated. Total dry biomass accumulation, as well as the number and height of culms, decreased with increase in salinity under both flooded and drained conditions. Generally, CO₂ exchange, stomatal conductance, Photosystem II (PSII) quantum yield and electron transport rate (ETR) through PSII declined with increase in salinity in both the flooded and drained treatments. Predawn and midday ψ in culms decreased with increase in salinity, being lower under drained than flooded conditions. Inorganic solute concentrations in culms increased with increase in salinity, with Na⁺ and Cl⁻ being the predominant ions. Na⁺/K⁺ ratios in culms increased significantly with increase in salinity. Proline concentrations in roots and culms, which increased with salinity, were considerably higher under drained than flooded conditions. Germination was best at salinities less than 20% seawater and decreased significantly with further increase in salinity to 110% seawater. Transfer of ungerminated salt-treated seeds to distilled water stimulated germination. This study has demonstrated that J. kraussii is a highly salt and flood tolerant species, being able to grow and survive in salinities up to 70% seawater, under both drained and flooded conditions. Maximal growth occurred at low salinities (<10% seawater) under flooded condition.