Influence of temperature on female,embryonic and hatchling traits in syntopic newts,Ichthyosaura alpestris and Lissotriton vulgaris

Amphibian populations have been declining globally for the last several decades, and climate change is often regarded as one of the most important factors driving these declines. Amphibians are particularly sensitive to climatic changes due to their physiological, ecological and behavioral characteristics. Here we performed a laboratory experiment to investigate how temperature affects ovipositing females, eggs and hatchlings in two syntopic populations of alpine newts, Ichthyosaura alpestris, and smooth newts, Lissotriton vulgaris. Female newts were assigned to two different oviposition temperatures (11 °C and 14 °C) for the duration of their oviposition period. Deposited eggs were equally divided and assigned to three different incubation temperatures (11 °C, 14 °C and 17 °C). We hypothesized that oviposition will be affected by temperature, that the combination of different oviposition and incubation temperatures may have an effect on embryonic and hatchling traits (embryonic mortality, days to hatch and hatchling length), and that these effects might differ between the two newt species. Temperature affected the number of deposited eggs in smooth newts, but not in alpine newts. Larval hatching success was not affected by oviposition or incubation temperature. Temperature effects on hatching time and hatchling length differed between the two species. These results suggest that temperature changes may have disparate effects on amphibian reproduction, even in syntopic taxa.     

孵育时间在二氧化钛选择性富集磷酸化肽中的影响

目的:旨在探索孵育时间在二氧化钛选择性富集磷酸化肽中的影响。方法:以大肠杆菌BL21(DE3)为材料,预实验确定二氧化钛磁珠与肽段之间的最适比例,之后在此比例下继续探究不同孵育时间条件下的富集效果,以观察孵育时间在二氧化钛富集体系中的影响。结果:预实验确定磁珠与肽段之间最适比例为3 :1,在此比例下发现随着孵育时间的增加其富集效率会逐渐降低,最后确定大肠杆菌中最适孵育时间为5min。结论:孵育时间在大肠杆菌二氧化钛富集体系中对富集效果有明显的影响,并且呈负相关。推测不同样品的最佳孵育时间与样品种属有关。     

Do temperature fluctuations during incubation always play an important role in shaping the phenotype of hatchling reptiles?

Fluctuating temperatures (FTs) influence hatchling phenotypes differently from constant temperatures (CTs) in some reptiles, but not in others. This inconsistency raises a question of whether thermal fluctuations during incubation always play an important role in shaping the phenotype of hatchlings. To answer this question, we incubated eggs of Naja atra under one CT (28 °C, CT), two temperature-shift [cold first (CF) and hot first (HF) in which eggs were first incubated at 24 or 32 °C and then at the other, each for 20 days, and finally at 28 °C until hatching], and one FT thermal regimes. Female hatchlings were larger in snout–vent length but smaller in tail length, head size than male hatchlings from the same-sized egg; female hatchlings had more ventral scales than did male hatchlings. The FT and HF treatments resulted in shorter incubation lengths. Tail length was greatest in the CT treatment and smallest in the FT treatment, with the CF and HF treatments in between; head width was greater in the CT treatment than in the other three treatments. Other examined hatchling traits did not differ among treatments. The observed morphological modifications cannot be attributed to the effect of thermal fluctuations but to the effect of temperatures close to the upper and lower viable limits for the species. Our results therefore support the hypothesis that hatchling phenotype is not altered by thermal fluctuation in species with no phenotypic response to incubation temperature within some thresholds.     

高寒草甸常见雀形目鸟类的卵壳特征

作为胚胎发育的屏障,卵壳特征如卵壳厚度和气孔密度具有重要的生态学意义。本文研究了高寒草甸繁殖的7种代表性雀形目鸟类角百灵（Eremophila alpestris）、小云雀（Alauda gulgula）、黄头鹡鸰（Motacilla citreola）、树麻雀（Passer montanus）、粉红胸鹨（Anthus roseatus）、黄嘴朱顶雀（Carduelis flavirostris）、赭红尾鸲（Phoenicurus ochruros）的卵与卵壳特征,以探索在高寒缺氧环境下,不同科鸟类的适应性调节。结果发现,7种鸟类中,除小云雀和角百灵之外,其他鸟类的卵与卵壳特征均有显著的种间差异（除卵壳厚度和气孔直径之外）;对7种鸟类卵壳厚度、气孔密度、卵体积、卵重之间的线性拟合显示,卵壳厚度、卵体积与气孔密度均无显著相关性（卵壳厚度：P = 0.11,卵体积：P = 0.09）,卵重、卵体积与卵壳厚度呈显著正相关（卵重：r2 = 0.46,P ＜ 0.001;卵体积：r2 = 0.44,P ＜ 0.001）;对7种鸟类卵的数据与繁殖期雌鸟平均体重的线性拟合结果显示：平均卵重、卵壳厚度的差异与雌鸟体重成正相关（卵重：r2 = 0.66,P = 0.03;卵壳厚度：r2 = 0.92,P ＜ 0.01）;对6种鸟类（缺乏粉红胸鹨窝卵数数据）卵的数据与孵卵相关变量的线性拟合结果显示：卵壳厚度及气孔率与窝卵数成负相关（卵壳厚度：r2 = 0.64,P = 0.056;气孔率：r2 = 0.87,P ＜ 0.01）,6种鸟类（缺乏树麻雀巢杯指数数据）气孔率与巢杯指数或巢型无显著相关性（巢杯指数：P = 0.49,巢型：P = 0.435）。卵表面积和总气孔数解释了大部分气孔率差异（87%）,卵重和气孔率与孵卵期无显著相关（P = 0.77）,气孔率显著低于预期气孔率（P＜0.001）。这些结果表明,种间的遗传性（如成鸟的形态、窝卵数等）决定了大部分卵与卵壳特性的差异,但是为适应高寒低氧的气候特征,不同种的鸟都具有相对厚的卵壳和低的气孔率特征,说明环境因素同样影响卵的进化。另外,单纯的卵特性（卵体积和气孔率）并不能决定孵卵期的长短,亲鸟的孵卵行为同样具有重要的影响,角百灵虽然离巢频率相对较高,但是孵卵期却相对较短,这可能与其较高的气孔率有关。     

Does shade cover availability limit nest-site choice in two populations of a turtle with temperature-dependent sex determination?

Shifts in resource use may be an important mechanism by which organisms can adjust to novel environmental conditions, such as those imposed by climate change. However, for such shifts to be possible, environmental space must exist into which organisms can move. Habitat that ensures successful survival and reproduction is one such critical resource. We studied resource selection of shade cover over nest sites by painted turtles in populations in Illinois (center of range) and New Mexico (southern edge of range). We targeted this habitat feature because shade can influence hatching success and offspring phenotype (including sex in the study species) by affecting nest microenvironments. We found that while turtles in both populations selected nest sites that were shadier than average available sites, overall resource selection differed between the populations. This disparity may have been due to differences in structure of vegetation that provides shade at each site, because areas with high shade cover in New Mexico (low dense thickets) were much more difficult for turtles to access than those in Illinois (dense tree canopy cover). Further, shade cover predicted different parameters of incubation regime at each site, suggesting that turtles must assess dissimilar components of shade cover in order to choose nest sites and predict their future incubation regimes. Our results suggest that shade cover within nesting areas is a key component of painted turtle habitat, and that accessible, highly-shaded nest sites may be limited at the New Mexico site. Maintaining a range of shade cover from which turtles can select nest sites would permit plasticity in nest-site choice to be expressed, which may be important in preventing sex ratio skews due to climate change.     

Relationship between the dinoflagellate cyst Spiniferites pachydermus and Gonyaulax ellegaardiae sp. nov. from Izmir Bay,Turkey, and molecular characterization

Here, we established the cyst‐motile stage relation‐ship for Spiniferites pachydermus through incubation of cysts with a characteristically microreticulate/perforate surface isolated from Izmir Bay in the eastern Aegean Sea of the eastern Mediterranean. The morphology of the motile stage was similar to Gonyaulax spinifera but had a different size, overhang, displacement and reticulations. Based on the distinct morphology of the cyst and morphological differences in motile cells, we assigned S. pachydermus from Izmir Bay to the new species Gonyaulax ellegaardiae. We elucidate the phylogenetic relationship of G. ellegaardiae through large and small subunit ribosomal DNA and show that it forms a clade with other species that belong to the G. spinifera complex.     

孵化水热环境对渔异色蛇孵化卵和孵出幼体的影响

渔异色蛇卵孵化时能从环境中吸收水分导致质量增加,卵质量的增加与初始卵质量和孵化基质湿度有关。较大幅度的孵化基质湿度变化对孵化期、孵化成功率、胚胎动用孵内物质和能量、孵出幼体的性比、大小和质量无显著影响。孵化期随温度升高而缩短,并显示极强的窝间差异。温度对孵出幼体的性别无影响,但显著影响孵化成功率、胚胎对卵内物质和能量的动用、幼体的大小和质量、躯干和剩余卵黄的质量。孵出幼体总长的两性差异不显著,但雌体体长大于雄体而尾长小于雄体。３２℃不适于孵化渔异色蛇卵,该温度下孵出的幼体躯干发育不良,剩余孵黄较多,尾部均呈畸形,孵化过程中能量转化率较低。２４℃和２６℃中孵出的幼体躯干发育良好,孵化过程中能量转化率较高,各项被测定的幼体特征指标均极相似。     

温度对中华鳖卵孵化和胚胎发育的影响

在25℃,27．5℃,30℃,32．5℃和35℃条件下,分别研究了中华鳖（Trionyx sinensis)卵的孵化和胚胎的发育。卵黄和卵壳的湿重分别占卵重的85．10％和14．90％,含水量分别为76．63％和21．01％。中华鳖胚胎重随孵化时间呈逻辑斯谛曲线增长,孵化温度越高,胚胎发育速度r越大。25℃－30℃的胚胎发育仅表现为‘S’型生长曲线的前,中段,在孵化后期发育速度未受到明显的抑制,32．5℃和35℃温度条件下,后期发育速度受到了较大的影响。孵出稚鳖湿重和干重均与初始孵重存在极显著的经性相关关系。孵化温度对孵出稚鳖湿重没有显著影响,但对孵出稚鳖干重的影响极显著。随孵化温度的升高,孵出稚鳖干重吾‘U’形变化,在适中温度下,胚胎的生长效率最低。卵黄干重随孵化时间呈指数递减,温度越高,孵黄的利用速度越快。在孵化过程中,卵重,卵干重和卵内水总量都随孵化时间的延长,孵化温度的上升而略有下降。孵化期随温度升高呈双曲线缩短,其相关关系可用方程I＝30．6＋109／（T－22．5）来表示。中华鳖卵的最适孵化温度在32．5℃左右,胚胎孵化后期比早期对温度的需求可能要低一些。     

Sex ratios in naturally incubating Macrochelys temminckii nests,a species with type II temperature-dependent sex determination

The alligator snapping turtle, Macrochelys temminckii, exhibits type II temperature-dependent sex determination (TSD), wherein females are produced at high and low incubation temperatures. This TSD pattern is well studied at constant temperatures, but little work has focused on sex ratios in natural nests that experience daily and seasonal temperature fluctuations. We monitored nesting activity of reintroduced Macrochelys temminckii at Tishomingo National Wildlife Refuge in 2010–2011. Nests located prior to predation were excavated to determine clutch size and the eggs were reburied with a temperature data logger to collect nest temperatures. Overall, 24% of nests were protected with wire mesh prior to predation, and the average clutch size in intact nests was 22.4 eggs. Nest predation rates in the study population will likely approach 100% if nest protection efforts do not continue. Temperature profiles were used to compare estimated sex ratios using two methods—mean nest temperature during middle third of incubation and the degree-day model—to actual sex ratios in naturally incubated Macrochelys temminckii nests. The sex ratio in all 2010 recruits was female-biased (91.8% female); 2011 nests did not produce any hatchlings, likely the result of severe drought. The predicted sex ratios based on mean nest temperature and the degree-day model matched actual sex ratios in the warmer nests (0% male), but the degree-day model estimate proved more accurate in the cooler nest. A strongly skewed population sex ratio could become a threat to this reintroduced population if the strongly female-biased sex ratio in 2010 reflects a long-term trend.     

A laboratory test for screening iron chelates for use in alkaline soils

Summary A soil incubation test is described for screening Fe-chelates for use under alkaline conditions. Ten grams of air-dry soil were mixed with 4 ml of an aqueous solution containing the appropriate amount of iron of the test chelate adding to the soil 24 ppm of iron (soil air-dry weight basis). The moistened soil was incubated in stoppered plastic vials at 24°C. On day 4 from the start of incubation, and at weekly intervals thereafter, the contents of triplicate vials were extracted with 16 ml of DTPA solution prepared according to the method of Lindsay and Norvell. The Fe extracted in this way was considered to be a measure of the stability of the chelates tested in comparison to Sequestrene 138.