Influence of incubation temperature on hatchling phenotype in reptiles

Incubation temperature influences hatchling phenotypes such as sex, size, shape, color, behavior, and locomotor performance in many reptiles, and there is growing concern that global warming might adversely affect reptile populations by altering frequencies of hatchling phenotypes. Here I overview a recent theoretical model used to predict hatchling sex of reptiles with temperature-dependent sex determination. This model predicts that sex ratios will be fairly robust to moderate global warming as long as eggs experience substantial daily cyclic fluctuations in incubation temperatures so that embryos are exposed to temperatures that inhibit embryonic development for part of the day. I also review studies that examine the influence of incubation temperature on posthatch locomotion performance and growth because these are the traits that are likely to have the greatest effect on hatchling fitness. The majority of these studies used artificial constant-temperature incubation, but some have addressed fluctuating incubation temperature regimes. Although the number of studies is small, it appears that fluctuating temperatures may enhance hatchling locomotor performance. This finding should not be surprising, given that the majority of natural reptile nests are relatively shallow and therefore experience daily fluctuations in incubation temperature.     

Longterm effects of incubation temperatures on the morphology and locomotor performance of hatchling lizards (Bassiana duperreyi, Scincidae)

The phenotypes of hatchling reptiles are known to be affected by the thermal environments they experience during incubation, but the evolutionary and ecological significance of this phenotypic plasticity remains unclear. Crucial issues include: (i) the magnitude of effects elicited by thermal regimes in natural nests (as opposed to constant-temperature incubation); (ii) the persistence of these effects during ontogeny; and (iii) the consistency of these effects across different test conditions (does the thermal regime during embryogenesis simply shift the hatchling's thermal optimum for performance, or actually modify overall performance ability regardless of temperature?). We examined these questions by incubating eggs of scincid lizards (Bassiana duperreyi) from montane southeastern Australia, under two fluctuating-temperature regimes that simulated ‘cold’ and ‘hot’ natural nests. These thermal regimes substantially modified hatchling morphology (mass, body length, tail length, and the relationship between these variables), locomotor performance (running speeds over distances of 25 cm and lm), anti-predator ‘tactics’ and survival rates. The differences in locomotor performance persisted throughout the 20 weeks of our experiment. Lizards that emerged after ‘hot’ incubation were faster runners than their ‘cold’-incubated siblings under all thermal conditions that we tested. Thus, incubation temperatures modified overall locomotor ability, with only a minor effect on the set-point for optimum performance. The magnitude, persistence and consistency of these incubation-induced phenotypic modifications suggest that they may play an important role in evolutionary and ecological processes within lizard populations.     

Effects of constant and fluctuating incubation temperatures on hatching success and hatchling traits in the diamondback terrapin (Malaclemys terrapin) in the context of the warming climate

As global temperatures continue to rise, so too will the nest temperatures of many species of turtles. Yet for most turtle species, including the estuarine diamondback terrapin (Malaclemys terrapin), there is limited information on embryonic sensitivity to elevated temperature. We incubated eggs of M. terrapin at three, mean temperatures (31, 34, 37 °C) under two thermal exposure regimes (constant or semi-naturally fluctuating temperature) and measured hatching success, developmental rate, and hatchling size. Hatching success was 100% at 31 °C and 67% at 34 °C, respectively; at 37 °C, all eggs failed early in the incubation period. These values were unaffected by exposure regime. The modeled LT₅₀ (temperature that was lethal to 50% of the test population) was 34.0 °C in the constant and 34.2 °C in the fluctuating thermal regime, reflecting a steep decline in survival between 33 and 35 °C. Hatchlings having been incubated at a constant 34 °C hatched sooner than those incubated at 31 °C under either constant or fluctuating temperature. Hatchlings were smaller in straight carapace length (CL) and width after having been incubated at 34 °C compared to 31 °C. Larger (CL) hatchlings resulted from fluctuating temperature conditions relative to constant temperature conditions, regardless of mean temperature. Based upon recent temperatures in natural nests, the M. terrapin population studied here appears to possess resiliency to several degrees of elevated mean nest temperatures, beyond which, embryonic mortality will likely sharply increase. When considered within the mosaic of challenges that Maryland's M. terrapin face as the climate warms, including ongoing habitat losses due to sea level rise and impending thermal impacts on bioenergetics and offspring sex ratios, a future increase in embryonic mortality could be a critical factor for a population already experiencing ecological and physiological challenges due to climate change.     

恒定和波动温度下丽斑麻蜥孵出幼体的表型变异

作者以丽斑麻蜥(Eremias argus)为模型动物研究恒定和波动孵化温度对孵化成功率和孵出幼体表型的影响。卵在四个恒定[24 ,27 ,30 and 33 (±0·3)℃]、一个波动温度下孵化。不同温度处理下的孵化成功率相同,但孵出幼体表型不同。孵化期随孵化温度升高呈指数式缩短;在相同平均温度下,波动温度孵化卵的孵化期比恒温孵化卵长。在所有被检表型特征中,幼体的干重、剩余卵黄干重和运动表现更易受孵化温度影响。总体而言,低温(24℃、27℃)孵出幼体运动表现最佳,高温(33℃)孵出幼体最差、温和温度(30℃和波动温度)孵出幼体居中。本文研究数据显示: (1)丽斑麻蜥卵每日短期暴露于潜在致死的极端温度下对孵化成功率和孵出幼体形态特征无明显的不利效应; (2)温度波动对孵出幼体运动表现无促进作用,对孵化期的影响则不同于平均值相同的恒定温度。     

Experimental evolution in fluctuating environments: tolerance measurements at constant temperatures incorrectly predict the ability to tolerate fluctuating temperatures

The ability to predict the consequences of fluctuating environments on species distribution and extinction often relies on determining the tolerances of species or genotypes in different constant environments (i.e. determining tolerance curves). However, very little is known about the suitability of measurements made in constant environments to predict the level of adaptation to rapidly fluctuating environments. To explore this question, we used bacterial clones adapted to constant or fluctuating temperatures and found that measurements across a range of constant temperatures did not indicate any adaptation to fluctuating temperatures. However, adaptation to fluctuating temperatures was only apparent if growth was measured during thermal fluctuation. Thus, tolerance curves based on measurements in constant environments can be misleading in predicting the ability to tolerate fast environmental fluctuations. Such complications could lead to false estimates of the genetic merits of genotypes and extinction risks of species due to climate change‐induced thermal fluctuations.     

Incubation temperature fluctuation does not affect incubation length and hatchling phenotype in the Chinese skink Plestiodon chinensis

Studies examining the effects of incubation temperature fluctuation on the phenotype of hatchling reptiles have shown species variation. To examine whether incubation temperature fluctuation has a key role in influencing the phenotype of hatchling Chinese skinks (Plestiodon chinensis), we incubated eggs produced by 20 females under five thermal regimes (treatments). Eggs in three treatments were incubated in three incubators, one set constant at 27 °C and two ramp-programmed at 27±3 °C and 27±5 °C on a cycle of 12 h (+) and 12 h (−). The remaining eggs were incubated in two chambers: one inside a room where temperatures varied from 23.0 to 31.1 °C, with a mean of 27.0 °C; the other outside the room where temperatures varied from 20.2 to 35.3 °C, with a mean of 26.1 °C. We found that: (1) for eggs at a given embryonic stage at ovipositon, the mean rather than the variance of incubation temperatures determined the length of incubation; (2) most (egg mass, embryonic stage at oviposition, incubation length and all examined hatchling traits except tail length and locomotor performance) of the examined variables were affected by clutch; and (3) body mass was the only hatchling trait that differed among the five treatments, but the differences were tiny. These findings suggest that incubation temperature fluctuation has no direct role in influencing incubation length and hatchling phenotype in P. chinensis.     

滑鼠蛇的繁殖输出及孵化热环境对孵出幼体表型特征的影响

用15条2002年6月中旬捕自浙江丽水的怀卵滑鼠蛇研究繁殖输出及孵化热环境对孵出幼体表型特征的影响。母体在捕后3周内各产一窝柔性卵。窝卵数、窝卵重和卵大小均随母体体长增加而增大,平均值分别为13．3(枚卵)、332．4g和24．8g。窝卵数和卵大小的变异系数分别为0．18和0．13。窝卵数与产后母体状态呈正相关,卵数量和大小无关。每窝部分可孵卵分别用恒温(24、27、30、33℃)和21．0-39．0℃(平均28．3℃)范围内的波动温度孵化,每隔5d记录恒温孵化卵的重量。孵化热环境对卵与环境之间的水分交换有显著影响,并影响孵化卵重量的时间变化。24、27、30、33℃和波动温度的平均孵化期分别为105．4、78．0、57．8、51．3和58．6d。不同热环境下的孵化成功率和幼体畸形率有一定差别,但统计上不显著。24℃和30℃孵出幼体雄性比例较高,27℃、33℃和波动温度孵出幼体雌性比例较高,但没有证据表明孵化温度能决定滑鼠蛇性别。除孵出幼体灰分含量外,孵化温度对其它幼体特征均有显著影响。33℃孵出幼体SVL较小,但剩余卵黄和其中的灰分含量大于其它温度孵出的幼体。24℃和30℃孵出幼体的总干重、总能量、躯干干重、脂肪体干重总体上小于27℃和波动温度孵出幼体。24℃孵出幼体特征性具有最小的剩余卵黄,33℃孵出幼体则特征性地具有最大的剩余卵黄。24℃和30℃孵出幼体特征较为接近,27℃和波动温度孵出幼体较为接近。对9个幼体特征变量做主成分分析发现,第一和第二主成分共解释81．2％的变异。幼体SVL、湿重、干重、脂肪含量、能量、躯干干重和脂肪体干重在第一主成分有较高的正负载系数(解释56．9％变异),剩余卵黄干重在第二主成分有较高的正负载系数(解释24．3％变异),第一和第二主成分的分值均差异显著。主成分分析进一步显示,24、30和33℃孵出幼体总体上小于27℃和波动温度孵出幼体[动物学报50(4)：541-550,2004]。     

Effects of constant and fluctuating temperatures on egg survival and hatchling traits in the northern grass lizard (Takydromus septentrionalis, Lacertidae)

To understand how nest temperatures influence phenotypic traits of reptilian hatchlings, the effects of fluctuating temperature on hatchling traits must be known. Most investigations, however, have only considered the effects of constant temperatures. We incubated eggs of Takydromus septentrionalis (Lacertidae) at constant (24 degrees C, 27 degrees C, 30 degrees C and 33 degrees C) and fluctuating temperatures to determine the effects of these thermal regimes on incubation duration, hatching success and hatchling traits (morphology and locomotor performance). Hatching success at 24 degrees C and 27 degrees C was higher, and hatchlings derived from these two temperatures were larger and performed better than their counterparts from 30 degrees C and 33 degrees C. Eggs incubated at fluctuating temperatures exhibited surprisingly high hatching success and also produced large and well-performed hatchlings in spite of the extremely wide range of temperatures (11.6-36.2 degrees C) they experienced. This means that exposure of eggs to adversely low or high temperatures for short periods does not increase embryonic mortality. The variance of fluctuating temperatures affected hatchling morphology and locomotor performance more evidently than did the mean of the temperatures in this case. The head size and sprint speed of the hatchlings increased with increasing variances of fluctuating temperatures. These results suggest that thermal variances significantly affect embryonic development and phenotypic traits of hatchling reptiles and are therefore ecologically meaningful.     

Influence of incubation recess patterns on incubation period and hatchling traits in wood ducks Aix sponsa

Parental effects are influential sources of phenotypic variation in offspring. Incubation temperature in birds, which is largely driven by parental behavior and physiology, affects a suite of phenotypic traits in offspring including growth, immune function, stress endocrinology, and sex ratios. The importance of average incubation temperature on offspring phenotype has recently been described in birds, but parental incubation behaviors like the duration and frequency of recesses from the nest can be variable. There are few studies describing how or if thermal variation as a result of variable incubation affects offspring phenotype. We incubated wood duck Aix sponsa eggs under three different incubation regimes, based on patterns that occur in nature, which varied in off‐bout duration and/or temperature. We measured incubation period, morphometrics at hatching, and monitored growth and body condition for nine days post hatch. When average incubation temperature was allowed to drop from 35.9°C to 35.5°C as a result of doubled off‐bout duration, we found a significant 2 d extension in incubation period, but no effects on duckling hatch mass, or growth and body condition up to nine days post hatch. However, when average incubation temperatures were equivalent (35.9°C), doubling the duration of the simulated off‐bouts did not influence incubation period or any post hatch parameters. Our results suggest that if incubating parents can maintain favorable thermal environments in the nest via altered behavior (e.g. manipulating nest insulation) and/or physiology (e.g. heat production), parents may be able to avoid the costs of longer incubation periods resulting from increased off‐bout duration.     

孵化温度对赤链蛇胚胎代谢和幼体行为的影响

研究了赤链蛇(Dinodon rufozonatum)在孵化过程中卵的生长、孵化期、胚胎代谢和孵出幼体行为表现的热依赖性。结果显示:孵化温度对孵化期、卵增重、孵化过程中消耗的总能量和孵出幼体的运动表现有显著影响,但不影响胚胎代谢率、孵化成功率和幼体吐信频次。孵化期随着孵化温度的升高而缩短,孵化过程中,24℃终末卵重和胚胎代谢率显著大于30℃,而27℃与其他两个温度没有差异;27℃孵出幼体游速较24℃快,30℃孵出幼体与其他两个温度孵出幼体的游速无显著差异。上述结果显示:24—30℃是赤链蛇适合的孵化温度范围,与赤链蛇所处的生境温度相近。     

升温对北草蜥雌体繁殖、卵孵化及幼体特征的影响

在围栏条件下,比较升温和对照处理北草蜥(Takydromus septentrionalis)繁殖、卵孵化及幼体特征的差异,以揭示升温对其繁殖生活史特征的作用。升温处理对北草蜥母体体温有显著影响,但并不影响其繁殖输出。升温显著影响卵孵化期和幼体的运动能力,但不影响幼体大小等形态特征。升温条件下孵出的幼体运动能力较弱。结果表明,北草蜥母体能耐受短期的环境增温,维持相对恒定的繁殖输出;升温能影响幼体的功能表现,进而可能改变后代适合度。     

Phenotypic responses of hatchlings to constant versus fluctuating incubation temperatures in the multi-banded krait, Bungarus multicintus (Elapidae)

Most studies on egg incubation in reptiles have relied on constant temperature incubation in the laboratory rather than on simulations of thermal regimes in natural nests. The thermal effects on embryos in constant-temperature studies often do not realistically reflect what occurs in nature. Recent studies have increasingly recognized the importance of simulating natural nest temperatures rather than applying constant-temperature regimes. We incubated Bungarus multicintus eggs under three constant and one fluctuating-temperature regimes to evaluate the effects of constant versus fluctuating incubation temperatures on hatching success and hatchling phenotypes. Hatching success did not differ among the four treatments, and incubation temperature did not affect the sexual phenotype of hatchlings. Incubation length decreased as incubation temperature increased, but eggs incubated at fluctuating temperatures did not differ from eggs incubated at constant temperatures with approximately the same mean in incubation length. Of the hatchling phenotypes examined, residual yolk, fat bodies and locomotor performance were more likely affected by incubation temperature. The maximal locomotor speed was fastest in the fluctuating-temperature and 30 degrees C treatments and slowest in the 24 degrees C treatment, with the 27 degrees C treatment in between. The maximal locomotor length was longest in the fluctuating-temperature treatment and shortest in the 24 degrees C and 27 degrees C treatments, with the 30 degrees C treatment in between. Our results show that fluctuating incubation temperatures do not influence hatching success and hatchling size and morphology any differently than constant temperatures with approximately the same mean, but have a positive effect on locomotor performance of hatchlings.     

Thermal regimes during incubation do not affect mean selected temperatures of hatchling lizards (Bassiana duperreyi, Scincidae)

Incubation temperatures profoundly affect many phenotypic traits of squamate reptiles, and mean selected body temperatures of such animals also are plastic in response to environmental factors. Plausibly, then, incubation temperatures might affect hatchling thermoregulation, either via adaptation (i.e., populations that historically experience different nest conditions, also will diverge in hatchling thermoregulatory behaviour) or phenotypic plasticity (incubation temperatures directly modify hatchling behaviours). We tested this hypothesis with a montane scincid lizard (Bassiana duperreyi), using thermal-imaging methods to quantify temperatures (of both head and body) selected by hatchling lizards. The young lizards kept their heads cooler than their bodies, but mean selected temperatures did not differ among hatchlings from three populations with differing thermal regimes in natural nests, nor were they affected by thermal conditions during incubation. The conservatism of mean selected temperatures stands in strong contrast to the lability of many other phenotypic traits in response to incubation temperatures in this species.     

Swimming performance of hatchling green turtles is affected by incubation temperature

In an experiment repeated for two separate years, incubation temperature was found to affect the body size and swimming performance of hatchling green turtles (Chelonia mydas). In the first year, hatchlings from eggs incubated at 26°C were larger in size than hatchlings from 28 and 30°C, whilst in the second year hatchlings from 25.5°C were similar in size to hatchings from 30°C. Clutch of origin influenced the size of hatchlings at all incubation temperatures even when differences in egg size were taken into account. In laboratory measurements of swimming performance, in seawater at 28°C, hatchlings from eggs incubated at 25.5 and 26°C had a lower stroke rate frequency and lower force output than hatchlings from 28 and 30°C. These differences appeared to be caused by the muscles of hatchlings from cooler temperatures fatiguing at a faster rate. Clutch of origin did not influence swimming performance. This finding that hatchling males incubated at lower temperature had reduced swimming ability may affect their survival whilst running the gauntlet of predators in shallow near-shore waters, prior to reaching the relative safety of the open sea.     

The selective basis for increased egg retention: early incubation temperature determines hatchling phenotype in wall lizards (Podarcis muralis)

The thermal environment during development influences many aspects of the phenotype of hatchling reptiles. We hypothesized that temperature should differentially affect early incubation stages, in which differentiation dominates over growth, and late incubation stages, characterized by high growth rates. To test this idea, we incubated eggs of wall lizard ( Podarcis muralis ) under three regimes with the same mean temperature (29 °C), one constant and two variable with opposite sequences: first cold (25 °C) and then hot (32 °C), and vice versa. Hatchlings incubated at high temperature during the initial period had shorter hindlimbs and tails than those incubated under the other two temperature regimes and shorter heads than those incubated initially at low temperature. Thus, temperature experienced by embryos during the early external incubation period produced similar phenotypic responses compared to those reported in previous studies for the same constant temperature applied over the whole incubation period. Because female wall lizards select lower body temperatures during pregnancy, an increase of intrauterine retention would extend the time of exposure of developing embryos to suitable temperatures. Diminution of body temperature during pregnancy is contrary to the expected pattern under the hypothesis that egg retention has evolved to accelerate development, as proposed by the cold-climate model for evolution of viviparity in squamates, and the results of the present study support the alternative hypothesis of developmental optimization as a special case of the broader maternal manipulation view.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 92 , 441–447.     

Geographic variation in hatchling size in an oviparous skink: effects of maternal investment and incubation thermal environment

Geographic variation in offspring size can be viewed as an adaptive response to local environmental conditions, but the causes of such variation remain unclear. Here, we compared the size and composition of eggs laid by female Chinese skinks (Plestiodon chinensis) from six geographically distinct populations in southeastern China to evaluate geographic variation in hatchling size. We also incubated eggs from these six populations at three constant temperatures (24, 28 and 32 °C) to evaluate the combined effects of incubation temperature and population source on hatchling size. Egg mass and composition varied among populations, and interpopulation differences in yolk dry mass and energy content were still evident after accounting for egg mass. Population mean egg mass and thus hatchling mass were greater in the colder localities. Females from three northern populations increased offspring size by laying larger eggs relative to their own size. Females from an inland population in Rongjiang could increase offspring size by investing relatively more dry materials and thus more energy into individual eggs without enlarging the size of their eggs. The degree of embryonic development at oviposition was almost the same across the six populations, so was the rate of embryonic development and thus incubation length at any given temperature. Both incubation temperature and population source affected hatchling traits examined, but the relative importance of these two factors varied between traits. Our data show that in P. chinensis hatchling traits reflecting overall body size (body mass, snout‐vent length and tail length) are more profoundly affected by population source. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 113 , 283–296.     

Fluctuations in the incubation moisture environment affect growth but not survival of hatchling lizards

Few studies have collected longitudinal data that follow the complete microevolutionary path of an organism linking sources of variation (e.g. environmental versus genetic) to a trait and its subsequent relationship with fitness. Identifying the links within this pathway is imperative for understanding the ecological relevance of effects found at the phenotypic level. Furthermore, experimental studies that examine parts of the pathway in ectothermic organisms often fail to mimic the complexities of the natural developmental environment. Temperature and moisture conditions in reptile nests, for example, can fluctuate greatly on a seasonal and daily basis. Despite the potential effects of fluctuating environments, the vast majority of studies have held environmental treatments constant during the developmental period. We investigated the effects of fluctuating moisture regimes during incubation on eggs, hatchling phenotypes, and subsequent survival in the eastern fence lizard Sceloporus undulatus. Moisture fluctuations during embryonic development caused water absorption by eggs to follow the environmental availability of moisture. Initial hatchling tail length was affected by the pattern of moisture fluctuations, and hatchling growth rates in fluctuating treatments were significantly faster than those in a constant treatment, resulting in larger hatchlings after 4 weeks. A release–recapture experiment conducted in the field did not detect a treatment effect on survival despite the larger body sizes. In summary, although fluctuations affected water absorption by eggs and some hatchling traits, these effects did not have subsequent fitness consequences. The results obtained suggest that egg and hatchling survival are buffered against natural soil moisture fluctuations during incubation, even when egg and hatchling traits are significantly affected. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 100 , 89–102.