Incubation temperature affects multiple measures of immunocompetence in young wood ducks (Aix Sponsa)

Parental effects play a vital role in shaping offspring phenotype. In birds, incubation behaviour is a critical parental effect because it influences the early developmental environment and can therefore have lifelong consequences for offspring phenotype. Recent studies that manipulated incubation temperature found effects on hatchling body composition, condition and growth, suggesting that incubation temperature could also affect energetically costly physiological processes of young birds that are important to survival (e.g. immune responses). We artificially incubated wood duck (Aix sponsa) eggs at three biologically relevant temperatures. Following incubation, we used two immunoassays to measure acquired immune responses of ducklings. Ducklings incubated at the lowest temperature had reduced growth, body condition and responses to both of our immune challenges, compared with those from the higher temperatures. Our results show that incubation temperatures can be an important driver of phenotypic variation in avian populations.     

Embryonic developmental patterns and energy expenditure are affected by incubation temperature in wood ducks (Aix sponsa)

Recent research in birds has demonstrated that incubation temperature influences a suite of traits important for hatchling development and survival. We explored a possible mechanism for the effects on hatchling quality by determining whether incubation temperature influences embryonic energy expenditure of wood ducks (Aix sponsa). Because avian embryos are ectothermic, we hypothesized that eggs incubated at higher temperatures would have greater energy expenditure at any given day of incubation. However, because eggs incubated at lower temperatures take longer to hatch than embryos incubated at higher temperatures, we hypothesized that the former would expend more energy during incubation. We incubated eggs at three temperatures (35.0°, 35.9°, and 37.0°C) that fall within the range of temperatures of naturally incubated wood duck nests. We then measured the respiration of embryos every 3 d during incubation, immediately after ducks externally pipped, and immediately after hatching. As predicted, embryos incubated at the highest temperature had the highest metabolic rates on most days of incubation, and they exhibited faster rates of development. Yet, because of greater energy expended during the hatching process, embryos incubated at the lowest temperature expended 20%-37% more energy during incubation than did embryos incubated at the higher temperatures. Slower developmental rates and greater embryonic energy expenditure of embryos incubated at the lowest temperature could contribute to their poor physiological performance as ducklings compared with ducklings that hatch from eggs incubated at higher temperatures.     

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 manipulation during the middle incubation stage has a repressive effect on the immune organ development of Peking ducklings

Avian embryos are easily influenced by their environment during incubation. Previous studies have demonstrated that incubation temperature changes could influence muscle development and body weight, which subsequently determine the adult phenotype. The objective of this study was to investigate whether the development of immune organs in ducklings could be influenced by thermal manipulation during the middle stage of incubation. To evaluate this hypothesis, a control group was incubated under a normal temperature from E11 to E24, while the incubation temperature of the experimental group was increased by 1 °C. Our results indicated that slight changes in the incubation temperature significantly repressed the bursa of Fabricius index of the duck embryo on E25 (F1, 58=122.51, P<0.0001) and significantly repressed the spleen index of neonatal ducklings (F1, 58=74.38, P<0.0001). At 0 day posthatching (dph) and 14 dph, ducklings hatched from eggs incubated under the higher temperature had a lower percentage of globulin than the control group (F1, 10=19.97, P=0.0111; F1, 10=9.8, P=0.0352). The IFN-γ concentration of ducklings at 14 dph displayed the same trend (F1, 10=284.49, P<0.0001). These results suggested that thermal manipulation during the middle stage of incubation had a repressive effect on the development of immune organs and reduced the concentrations of serum globulin and IFN-γ. These results demonstrated that the subtle alteration of incubation temperature may weaken ducklings' immunity.     

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。     

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。     

Age of the incubating parents affects nestling survival: an experimental study of the herring gull Larus argentatus

The quality of conditions provided by avian parents will have consequences for both parental and offspring fitness. While many components of avian reproduction appear to vary with parental age, the effect of age on incubation has largely been ignored so far. In this study, we tested whether young herring gulls provide a different incubation environment from mature ones and whether this has consequences for offspring performance. Laying and rearing conditions were standardised using a cross-fostering protocol. Egg predation rates tended to be higher in the nests of young parents. However, nest site, nest construction and egg temperature during incubation did not vary with parental age. Overall, the duration of incubation was shorter in young compared to mature birds and this reflected the later laying date of the former, since incubation duration generally decreased across the season. However, male eggs incubated by young parents had longer incubation periods than predicted for their laying dates. In contrast, incubation length of female eggs incubated by young pairs, and of male and female eggs incubated by mature birds did not deviate from the expected for any given laying date. Offspring that had been incubated by young parents had considerably poorer survival than those incubated by mature pairs, despite being reared under standardized, favourable conditions (singly, by mature parents). This was due to increased mortality among female chicks that had been incubated by young parents. The chicks incubated as eggs by young and mature birds, which survived until fledging, did not differ in body mass and size growth, or body condition. The results of this study demonstrate that parental age can influence offspring performance via variation in incubation environment, and that females are more susceptible than males to conditions experienced during embryonic development.     

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.     

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.     

孵化温度对王锦蛇胚胎代谢和幼体适合度的影响

孵化温度所驱动的爬行动物的表型变异是生理生态学研究的热点。本研究以王锦蛇(Elaphe carinata)为实验动物,检验了24℃和28℃孵化温度对王锦蛇胚胎代谢速率、孵化过程中的卵重量、孵出幼体代谢和行为的影响。研究结果显示:卵重和胚胎的呼吸代谢均与孵化时间呈正相关;28℃下胚胎代谢速率大于24℃;幼蛇孵出15 d内体重随着生长时间的延长而减小,24℃孵出幼体的代谢速率大于28℃孵出幼体,两温度下孵出幼体的呼吸代谢速率和生长时间无显著关系;28℃孵出幼体的疾游速和吐信频次均大于24℃;两孵化温度孵出幼体的选择体温无显著差异,但在消耗完体内的剩余卵黄后28℃孵出幼体有60%的个体摄食,而24℃孵出幼体无摄食个体。总体而言,王锦蛇28℃孵出幼体适合度优于24℃孵出幼体。     

The effects of incubation temperature on the morphology and composition of Australian Brush-turkey (<Emphasis Type="Italic">Alectura lathami</Emphasis>) chicks

Environmental heterogeneity during embryonic development generates an important source of variation in offspring phenotypes and can influence the evolution of life histories. The effects of incubation temperature on offspring phenotypes in reptiles has been well documented but remains relatively unexplored in birds as their embryos typically develop over a narrow range of temperatures. Megapode birds (Order Galliformes; Family Megapodiidae) are unique in that their embryos tolerate and develop over a wide range of incubation temperatures, yet little is known of the effect that temperature has on hatchling morphology and composition. Australian Brush-turkey eggs collected on the day of laying were incubated in the laboratory under constant temperatures of 32, 34 and 36°C until hatching in order to determine the influence of temperature on hatchling mass, size and composition. The dry mass of the yolk-free body and residual yolk of hatchlings were temperature dependent, such that higher temperatures produced chicks of lesser yolk-free body mass and greater residual yolk mass than chicks incubated at lower temperatures. However the overall size (linear dimensions) and lipid, protein and ash content of chicks were independent of temperature.     

Potential targets for selection during the evolution of viviparity in cold-climate reptiles

Viviparity (live-bearing) has evolved from oviparity (egg-laying) in more than 100 lineages of squamate reptiles (lizards and snakes). This transition generally has occurred in cool climates, where thermal differentials between eggs in the (cool) nest versus the (warm) maternal oviduct influence embryonic development, in ways that may enhance offspring fitness. To identify specific traits potentially under selection, we incubated eggs of a montane scincid lizard at conditions simulating natural nests, maternal body temperatures, and an intermediate stage (2-week uterine retention of eggs prior to laying). Incubation at maternal temperatures throughout incubation affected the hatchling lizard’s activity level and boldness, as well as its developmental rate, morphology, and locomotor ability. A treatment that mimicked the initial stages of the transition toward viviparity had a major effect on some hatchling traits (locomotor speeds), a minor effect on others (tail length, total incubation period) and no effect on yet others (offspring behaviors). More generally, different aspects of the phenotype are sensitive to incubation conditions at different stages of development; thus, the evolution of reptilian viviparity may have been driven by a succession of advantages that accrued at different stages of embryogenesis.     

When to be born? Prolonged pregnancy or incubation enhances locomotor performance in neonatal lizards (Scincidae)

The degree of offspring development at hatching (or birth) varies among species within most major vertebrate lineages; altricial vs. precocial birds offer the clearest example of a trade-off between early hatching and the degree of locomotor development of the hatchling. No such diversity has been reported for reptiles, but we suggest that natural selection may fine-tune the time of hatching (in oviparous species) or birth (in viviparous species) to optimize offspring phenotypes and hence, maximize fitness. This hypothesis predicts enhanced neonatal performance after more prolonged incubation or gestation, within as well as among populations. Both published and original data on Australian scincid lizards support this prediction. In a field study, viviparous alpine skinks (Niveoscincus microlepidotus) that gave birth later in the season had faster-running offspring, that had a higher probability of surviving through the first year of life. The enhanced performance and survival were not secondary results of larger offspring size. After controlling for effects of mean incubation temperature, prolonged development also correlated with enhanced locomotor performance in hatchlings from eggs of an oviparous skink (Bassiana duperreyi) incubated at warm temperatures (> 20 degrees C) but not at cooler temperatures (< 20 degrees C). We suggest that embryonic reptiles control their date of hatching or birth and thus, their stage of development at this critical life-history transition.     

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.     

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。     

安徽宿州山地麻蜥卵孵化温度效应

用3个恒定温度(27、30、33℃)和波动温度(14.0～37.5℃)孵化山地麻蜥(Eremias brenchleyi)卵。结果表明,各温度处理下卵孵化成功率差异不显著,但温度对孵化期、孵出幼体表型特征及疾跑速度有显著影响;27、30℃和波动温度下孵出幼体的SVL、重量及躯干干重比33℃的要大,33℃孵出幼体的运动能力比其他3个温度处理弱。波动温度处理下山地麻蜥卵虽短期经历潜在致死的极端温度,但对孵化成功率、孵出幼体表型特征和运动表现均无负效应。     

Influence of incubation temperature on morphology, locomotor performance, and early growth of hatchling wall lizards (Podarcis muralis)

Eggs of wall lizards (Podarcis muralis) were incubated at three temperatures approaching the upper limit of viability for embryonic development in this species (26, 29, and 32 degrees C) to assess the influence of temperature on various aspects of hatchling phenotype likely affecting fitness. The thermal environment affected size and several morphometric characteristics of hatchling lizards. Hatchlings from eggs incubated at 32 degrees C were smaller (snout-vent length, SVL) than those from 26 and 29 degrees C and had smaller mass residuals (from the regression on SVL) as well as shorter tail, head, and femur relative to SVL. Variation in the level of fluctuating asymmetry in meristic and morphometric traits associated with incubation temperatures was quite high but not clearly consistent with the prediction that environmental stress associated with the highest incubation temperatures might produce the highest level of asymmetry. When tested for locomotor capacity in trials developed at body temperatures of 32 and 35 degrees C, hatchlings from the 32 degrees C incubation treatment exhibited the worst performance in any aspect considered (burst speed, maximal length, and number of stops in the complete run). Repeated measures ANCOVAs (with initial egg mass as covariate) of snout-vent length and mass of lizards at days 0 and 20 revealed significant effects of incubation temperature only for mass, being again the hatchlings from eggs incubated at 32 degrees C those exhibiting the smallest final size. All together, our results evidenced a pervasive effect of thermal regime during incubation (and hence of nest site selection) on hatchling phenotypes. However, incubation temperature does not affect hatchling phenotypes in a continuous way; for most of the analysed traits a critical threshold seems to exist between 29 and 32 degrees C, so that hatchlings incubated at 32 degrees C exhibited major detrimental effects. J. Exp. Zool. 286:422-433, 2000.     

Influence of water availability during incubation on hatchling size, body composition, desiccation tolerance, and terrestrial locomotor performance in the snapping turtle Chelydra serpentina

The effects of water availability during incubation on the water contents of neonatal snapping turtles at hatching were examined, along with the influence of hatchling water content on desiccation tolerance and terrestrial locomotor performance. The water contents of hatchlings from eggs incubated on wet substrates were both absolutely and proportionally greater than were those of hatchlings from eggs incubated on dry substrates. Hatchlings with greater water contents at hatching were able to survive longer and to lose more water before physiological performance was adversely affected by desiccation. Increased water contents in hatchlings with greater water availability during incubation may enhance survival by increasing the amount of water the animal can afford to lose before dehydration begins to adversely affect whole animal performance.     

Geographic variation in offspring size of a widespread lizard (Takydromus septentrionalis): importance of maternal investment

Geographic variation in offspring size is widespread, but the proximate causes of this variation have not yet been explicitly determined. We compared egg size and egg contents among five populations of a lizard (Takydromus septentrionalis, Günther, 1864) along a latitudinal gradient, and incubated eggs at two temperatures to determine the influence of maternal investment and incubation temperature on offspring size. The mean values for female size and egg size were both greater in the two northern populations (Chuzhou and Anji) than in the three southern populations (Lishui, Dongtou, and Ningde). The larger eggs were entirely attributable to the body size of females in the Anji population, but their increased size also stemmed from further enlargement of egg size relative to female body size in Chuzhou, the northernmost population sampled in this study. Eggs of the Chuzhou population contained more yolk and less water than those of southern populations. Despite the lower lipid content in the yolk, eggs from the Chuzhou population had higher energy contents than those from the two southern populations, owing to the larger egg size and increased volume of yolk. Hatchling size was not affected by incubation temperature, but differed significantly among populations, with hatchlings being larger in the Chuzhou population than in the other populations. Our data provide an inference that oviparous reptiles from cold climates may produce larger offspring, not only by increasing egg size but also by investing more energy into their eggs. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 101 , 59–67.     

The effects of thermal and hydric environments on hatching success, embryonic use of energy and hatchling traits in a colubrid snake, Elaphe carinata.

We examined the effects of thermal and hydric environments on hatching success, the embryonic use of energy and hatchling traits in a colubrid snake, Elaphe carinata. The eggs were incubated at four temperatures ranging from 24 to 32 degrees C on substrates with water potentials of 0 and -220 kPa using a 4x2 factorial design. Both thermal and hydric environments affected the water exchange between eggs and their surroundings. Eggs incubated in wetter substrates gained mass throughout the course of incubation, whereas eggs in drier substrates gained mass during the first half of incubation and lost mass thereafter. Hatching success was noticeably higher at 26 and 30 degrees C than at 24 and 32 degrees C, but among treatments, differences in hatching success were not significant. Temperature significantly affected the duration of incubation and most hatchling traits examined. Deformed hatchlings were found in all temperature treatments, with more deformities observed at 32 degrees C. Hatchlings from eggs incubated at different temperatures differed in wet body mass, but the differences stemmed mainly from variation in water contents. Embryos at different temperatures completed development at nearly the same expenditure of energy and catabolized nearly the same amount of lipids, but hatchlings from different temperatures differed in the development condition of carcass at hatching. Hatchlings from eggs incubated at 26 degrees C were larger in SVL than those from other higher or lower incubation temperatures, characteristically having larger carcasses; hatchlings from 32 degrees C eggs were smaller in SVL and had smaller carcasses but larger residual yolks than those from lower incubation temperatures. Hatchlings from eggs incubated at 24 degrees C were shorter in tail length but greater in size (SVL)-specific body wet mass than those from higher incubation temperatures. Within the range from -220 to 0 kPa, the substrate water potential did not affect hatching success, the embryonic use of energy and all hatchling traits examined, and the effects of temperature were independent of the effects of substrate water potential. Therefore, our data add evidence showing that embryonic development in reptiles with pliable-shelled eggs is relatively insensitive to variation in hydric environments during incubation.