Metabolic heating in Mediterranean loggerhead sea turtle clutches

Offspring sex ratio is an important demographic parameter and, given its determination by incubation temperature in sea turtles, might be a key factor for their conservation under climate warming. An appealing approach to estimate hatchling sex ratios is to measure sand temperatures at nest depth and deduce hatchling sex ratios from a beforehand-established relationship of hatchling sex ratio and sand temperature. Such estimates will only be accurate though if metabolic heat produced by the embryos is considered. Judging whether metabolic heating has a potential effect on hatchling sex ratios without actually measuring temperature within clutches would greatly facilitate monitoring protocols. We tested for a relationship between the amount of metabolic heating and the number of developed embryos as well as clutch size in the largest known loggerhead sea turtle (Caretta caretta) population of the Mediterranean on Zakynthos (Greece). Temperatures were measured within 20 nests as well as at a reference site in the sand at nest depth. Metabolic heating was detected, but only during the last third of the incubation period did nests heat up considerably (1.6 °C on average) above the temperature of the surrounding sand. During the middle third of incubation, when sex is determined, the amount of metabolic heating was negligible. The amount of metabolic heating during the last third of the incubation duration was significantly correlated to the number of offspring developed to at least about 75% of incubation duration. This factor explained nearly 50% of variation in metabolic heating. Metabolic heating was also significantly correlated to clutch size. Given that clutch size within the Mediterranean is largest in Zakynthos loggerheads, we conclude that metabolic heating can be ignored in the estimate of hatchling sex ratios in Mediterranean loggerhead populations. These results thus provide the basis for a feasible monitoring of hatchling sex ratios in the loggerhead sea turtle in the Mediterranean.     

Metabolic heating and the prediction of sex ratios for green turtles (Chelonia mydas)

We compared incubation temperatures in nests (n=32) of the green turtle (Chelonia mydas) on Ascension Island in relation to sand temperatures of control sites at nest depth. Intrabeach thermal variation was low, whereas interbeach thermal variation was high in both control and nest sites. A marked rise in temperature was recorded in nests from 30% to 40% of the way through the incubation period and attributed to metabolic heating. Over the entire incubation period, metabolic heating accounted for a mean rise in temperature of between 0.07 degrees and 2.86 degrees C within nests. During the middle third of incubation, when sex is thought to be determined, this rise in temperature ranged between 0.07 degrees and 2.61 degrees C. Metabolic heating was related to both the number of eggs laid and the total number of hatchlings/embryos produced in a clutch. For 32 clutches in which temperature was recorded, we estimate that metabolic heating accounted for a rise of up to 30% in the proportion of females produced within different clutches. Previous studies have dismissed any effect of metabolic heating on the sex ratio of marine turtle hatchlings. Our results imply that metabolic heating needs to be considered when estimating green turtle hatchling sex ratios.     

Microclimate of American crocodile nests in Banco Chinchorro biosphere reserve, Mexico: Effect on incubation length, embryos survival and hatchlings sex

Crocodilians have temperature-dependent sex determination (TSD) in which incubation temperature determines sex of embryo. Global warming is expected to alter hatchling sex ratio, leading to the extinction of small populations. Regional climate influence on crocodile nest microclimate and hatchlings' characteristics is poorly known. Here, microclimate in natural nests of American crocodile (Crocodylus acutus) and its relation with incubation length, hatchling sex and nesting success was studied in Banco Chinchorro Biosphere Reserve (Mexico) from 2007 to 2010. Temperature and relative humidity in different locations within and outside the nests were registered by data loggers. Incident solar radiation above nest was calculated from hemispheric photographs. Incubation length, proportion of hatchling reaching complete development and hatchling sex were determined at hatching. Nest temperatures exhibited a cyclic daily fluctuation due to solar radiation, which is the major heat source for nests. Clutch temperature was relatively stable and its daily amplitude was negatively correlated with clutch depth and size. Rainfall was the major source of clutch temperature decrease. Clutch and metabolic temperatures increased significantly during incubation. A small sample size failed to demonstrate a statistical relationship between length of incubation and mean clutch temperature. Proportion of embryos reaching complete development depended on maximum and minimum clutch temperature, maximum daily amplitude of clutch temperature and maximum decrease in clutch temperature on a period ≤4 day. Results confirmed a Female-Male-Female TSD pattern for C. acutus, with 31 and 32.5 °C as possible pivotal temperatures. Population and hatchling sex ratios were male-biased and fate of crocodiles of Banco Chinchorro could depend on the magnitude of temperature increase in the future.     

Sand and nest temperatures and an estimate of hatchling sex ratio from the Heron Island green turtle (Chelonia mydas) rookery, Southern Great Barrier Reef

Sand and nest temperatures were monitored during the 2002–2003 nesting season of the green turtle, Chelonia mydas, at Heron Island, Great Barrier Reef, Australia. Sand temperatures increased from ∼ 24°C early in the season to 27–29°C in the middle, before decreasing again. Beach orientation affected sand temperature at nest depth throughout the season; the north facing beach remained 0.7°C warmer than the east, which was 0.9°C warmer than the south, but monitored nest temperatures were similar across all beaches. Sand temperature at 100 cm depth was cooler than at 40 cm early in the season, but this reversed at the end. Nest temperatures increased 2–4°C above sand temperatures during the later half of incubation due to metabolic heating. Hatchling sex ratio inferred from nest temperature profiles indicated a strong female bias.     

Temperature and genotype affect asparagus somatic embryogenesis

Summary Calluses from five asparagus genotypes G14, G32, G171, G203, and G447 and hybrid Jersey Giant (JG) were incubated at three temperature regimes (24, 27, and 30°C) on embryo induction medium to assess somatic embryo development and conversion to plantlets. The calluses from three genotypes (G14, G32, and G171) were not responsive, failing to produce somatic embryos at any temperature regime. For three responsive genotypes (G203, G447, and JG), both incubation temperature and genotype significantly affected the numbers of somatic embryos produced. The calluses produced the most and the least numbers of total, bipolar, and globular embryos when incubated at 27°C and 24°C, respectively. When incubated at 27°C, G203 produced the highest numbers of total and globular embryos, 178 g⁻¹ callus and 142 g⁻¹ callus, respectively while G447 produced the highest number of bipolar embryos, 77 g⁻¹ callus. Incubation temperature but not genotype significantly affected the conversion of somatic embryos to plantlets. The somatic embryos recovered from the three responsive genotypes incubated at 27°C also converted to plantlets at the highest frequencies, 60–63% of the bipolar embryos and 42–43% of the globular embryos converted to plantlets, while the somatic embryos recovered from the calluses incubated at 24°C converted to plantlets at the lowest frequencies.     

Volume contents volume 38 (1985)

Isolated microspores from six cultivars of Brassica napus and one of B. carinata were cultured in modified Nitsch and Nitsch (NN) medium supplemented with 13% (W/V) sucrose, 0.05 mg/l benzyladenine (BA) and 1.00 mg/l nahpthaleneacetic acid (NAA). Embryogenic responses were observed at cultured temperatures ranging from 22 to 32°C. For most genotypes tested, the highest frequency of embryos occurred at 30°C and 7–54 embryos per anther (approx. 17 000 microspores per anther) developed. Although incubation at 30°C produced the highest frequency of embryos, lower culture temperatures induced better quality embryos. A split temperature culture regime of incubation at 32°C for 3 days followed by incubation at 25°C resulted in both high embryo yields and a high percentage of normal embryos. Plantlet development from microspore-derived embryos appeared to be influenced by both genotype and medium.     

The influence of maternal thermal environments on reproductive traits and hatchling traits in a Lacertid lizard,Takydromus septentrionalis

The thermal environment can induce substantial variation in important life-history traits. Experimental manipulation of the thermal environment can help researchers determine the contribution of this factor to phenotypic variation in life-history traits. During the reproductive season, we kept female northern grass lizards, Takydromus septentrionalis (Lacertidae), in three temperature-controlled rooms (25, 28 and 32 °C) to measure the effect of the maternal thermal environment on reproductive traits. Maternal thermal environment remarkably affected reproductive frequency and thereby seasonal reproductive output, but had little effect on reproductive traits per clutch or hatchling traits. Females kept at 32 °C produced more clutches and thus had shorter clutch intervals than females from 28 to 25 °C. Clutch size, clutch mass, relative clutch mass, egg size and hatchling traits did not vary among the three treatments. The eggs produced by the females were incubated at 27 °C and the traits of hatchlings were measured. The result that egg (offspring) size was independent of maternal thermal environments is consistent with the prediction of the optimal egg size (offspring) theory. The eggs produced by low temperature females (28 and 25 °C) took longer time to complete their post-oviposition development than did eggs produced by high temperature females (32 °C). This suggests that the eggs from low temperatures might have been laid when the embryos were at relatively early stages. Therefore, maternal thermal environment prior to oviposition could affect post-oviposition development in T. septentrionalis.     

Embryonic communication in the nest: metabolic responses of reptilian embryos to developmental rates of siblings

Incubation temperature affects developmental rates and defines many phenotypes and fitness characteristics of reptilian embryos. In turtles, eggs are deposited in layers within the nest, such that thermal gradients create independent developmental conditions for each egg. Despite differences in developmental rate, several studies have revealed unexpected synchronicity in hatching, however, the mechanisms through which synchrony are achieved may be different between species. Here, we examine the phenomenon of synchronous hatching in turtles by assessing proximate mechanisms in an Australian freshwater turtle (Emydura macquarii). We tested whether embryos hatch prematurely or developmentally compensate in response to more advanced embryos in a clutch. We established developmental asynchrony within a clutch of turtle eggs and assessed both metabolic and heart rates throughout incubation in constant and fluctuating temperatures. Turtles appeared to hatch at similar developmental stages, with less-developed embryos in experimental groups responding to the presence of more developed eggs in a clutch by increasing both metabolic and heart rates. Early hatching did not appear to reduce neuromuscular ability at hatching. These results support developmental adjustment mechanisms of the 'catch-up hypothesis' for synchronous hatching in E. macquarii and implies some level of embryo-embryo communication. The group environment of a nest strongly supports the development of adaptive communication mechanisms between siblings and the evolution of environmentally cued hatching.     

The effect of temperature on embryo survival and development in pallid sturgeon Scaphirhynchus albus (Forbes & Richardson 1905) and shovelnose sturgeon S. platorynchus (Rafinesque, 1820)

The thermal response of pallid sturgeon Scaphirhynchus albus and shovelnose sturgeon S. platorynchus embryos was determined at incubation temperatures from 8 to 26°C and 8 to 28°C, respectively. The upper and lower temperatures with 100% (LT₁₀₀) embryo mortality were 8 and 26°C for pallid sturgeon and 8 and 28°C for shovelnose sturgeon. It was concluded that 12–24°C is the approximate thermal niche for embryos of both species. Generalized additive and additive‐mixed models were used to analyze survival, developmental rate and dry weight data, and predict an optimal temperature for embryo incubation. Pallid sturgeon and shovelnose sturgeon embryo survival rates were different in intermediate and extreme temperatures. The estimated optimal temperature for embryo survival was 17–18°C for both species. A significant interaction between rate of development and temperature was found in each species. No evidence was found for a difference in timing of blastopore, neural tube closure, or formation of an S‐shaped heart between species at similar temperatures. The estimated effects of temperature on developmental rate ranged from linear to exponential shapes. The relationship for rate of development to temperature was relatively linear from 12°C to 20°C and increasingly curvilinear at temperatures exceeding 20°C, suggesting an optimal temperature near 20°C. Though significant differences in mean dry weights between species were observed, both predicted maximum weights occurred at approximately 18°C, suggesting a temperature optimum near 18°C for metabolic processes. Using thermal optimums and tolerances of embryos as a proxy to estimate spawning distributions of adults in a river with a naturally vernalized thermal regime, it is predicted that pallid sturgeon and shovelnose sturgeon spawn in the wild from 12°C to 24°C, with mass spawning likely occurring from 16°C to 20°C and with fewer individuals spawning from 12 to 15°C and 21 to 24°C. Hypolimnetic releases from Missouri River dams were examined; it was concluded that the cooler water has the potential to inhibit and delay sturgeon spawning and impede embryo incubation in areas downstream of the dams. Further investigations into this area, including potential mitigative solutions, are warranted.     

Effect of plant growth conditions, plating density, and genotype on the anther culture response of soft white spring wheat hybrids

Ten soft white spring wheat (Triticum aestivum L.) F₁ hybrids were grown under three temperature regimes, and anthers were cultured at two plating densities to investigate the effect of plant growth conditions, plating density, and genotype on embryo induction and plant regeneration. Anthers from plants grown at high temperature (25 °/18 °C) or from plants transferred from low (15 °/12 °C) to high temperature generally produced more embryos and green shoots, with a lower frequency of albinos, than did anthers from plants grown at low temperature. However, plating densities of 10 versus 20 anthers per milliliter, had little effect on anther response. Four of the five hybrids with `Fielder' as the female parent produced more embryos and green shoots than did hybrids with `AC Reed' as the female parent. Received: 12 July 1996 / Revision received: 1 April 1997 / Accepted: 30 April 1997     

Influence of temperature on the interaction of Concanavalin A with chick fibroblasts during embryo development

The interaction between Concanavalin A and chick embryo fibroblasts was studied. Cells from younger and older embryos had the same number of lectin receptor sites per cell at 4°, 21° and 37°C but the affinity constants increased with increasing temperature. Analysis of the binding data according to Scatchard showed that the apparent changes in binding as a function of temperature might be related to thermodynamic properties. The lectin binding sites on the cell surface proved homogeneous with regard to binding properties.The age-related differences noted in the affinities of the cells to bind Concanavalin A could be related to differences in the degree of rearrangement of the cell surface components and/or to a change in the structure of cell surface glycoconjugates, and may serve to explain the differences in the effect of Concanavalin A on cell growth.     

Anther culture in Brussels sprouts (Brassica oleracea var. gemmifera)

In Brussels sprouts, yields of up to 357 embryos per 100 anthers cultured were obtained using a thermal shock treatment of 16 h at 35°C at the start of the culture period. Treatments of 48 h at 35°C and 14 days at 30°C gave no embryos. The F₁ hybrid cv. Gower consistently gave higher embryo yields than the F₁ hybrid cv. Nym, the differences being 3 to 10-fold. Differences in embryo yield of 3-fold or less were usually not statistically significant because of great variation within a treatment. This variation was less with donor plants raised in a growth room than with those raised in a glasshouse, where temperature and light intensity could not be so accurately controlled. From 842 embryos cultured, 270 plants were regenerated, mostly via hypocotyl explants, which developed from the anther-derived embryos. Most of the regenerants were haploid or diploid, with a few of higher ploidy.     

Maternal effect of low temperature on handedness determination in C. elegans embryos

C. elegans embryos, larvae, and adults exhibit several left-right asymmetries with an invariant dextral handedness, which first becomes evident in the embryo at the 6-cell stage. Reversed (sinistral) handedness was not observed among > 10,000 N2 adults reared at 16°C or 20°C under standard conditions. However, among the progeny of adults reproducing at 10°C, the frequency of animals with sinistral handedness was increased to ∼0.5%. Cold pulse experiments indicated that the critical period for this increase was in early oogenesis, several hours before the first appearance of left-right asymmetry in the embryo. Hermaphrodites reared at 10°C and mated with males reared at 20°C produced sinistral outcross as well as sinistral self-progeny, indicating that the low temperature effect on oocytes was sufficient to cause reversals. Increased frequency of reversal was also observed among animals developed from embryos lacking the egg shell. Possible mechanisms for the control of embryonic handedness are discussed in the context of these results, including the hypothesis that handedness could be dictated by the chirality of a gametic component. © 1996 Wiley-Liss, Inc.     

Nest temperatures in a loggerhead nesting beach in Turkey is more determined by sea surface than air temperature

While climate change is now fully recognised as a reality, its impact on biodiversity is still not completely understood. To predict its impact, proxies coherent with the studied ecosystem or species are thus required. Marine turtles are threatened worldwide (though some populations are recovering) as they are particularly sensitive to temperature throughout their entire life cycle. This is especially true at the embryo stage when temperature affects both growth rates and sex determination. Nest temperature is thus of prime importance to understand the persistence of populations in the context of climate change. We analysed the nest temperature of 21 loggerheads (Caretta caretta) originating from Dalyan Beach in Turkey using day-lagged generalised mixed models with autocorrelation. Surprisingly, the selected model for nest temperature includes an effect for sea surface temperature 4-times higher than for air temperature. We also detected a very significant effect of metabolic heating during development compatible with what is already known about marine turtle nests. Our new methodology allows the prediction of marine turtle nest temperatures with good precision based on a combination of air temperature measured at beach level and sea surface temperature in front of the beach. These data are available in public databases for most of the beaches worldwide.     

Intraspecific Variation of Reproductive Traits in a Mediterranean Lizard: Clutch, Population, and Lineage Effects

Widely distributed terrestrial ectotherms from the southern European peninsulas show patterns of subdivision (related to isolation in temperate refugia) that allow us to test the relative importance of phylogeographic lineage, population of origin and familial effects as sources of variation for life-history traits. We collected gravid females from 15 geographically separated populations of the lacertid lizard Psammodromus algirus, a widely distributed species with well differentiated eastern and western lineages. We incubated eggs under two treatments of constant (28°C) and fluctuating (28 ± 4°C) temperature, and we examined clutch, population, and lineage effects on several traits of females, eggs, and hatchlings. Incubation time was mainly explained by differences between lineages, but it was also influenced by population and female effects. Within each lineage, incubation was shorter at cooler and wetter sites, and for a given climate it was shorter for eastern than for western populations, suggesting that countergradient variation has evolved independently in the two lineages. Female size, clutch size, and relative fecundity were primarily influenced by inter-population differences, a pattern that seemed attributable to environmental differences in productivity, because mean female size was positively correlated with a gradient of increasing precipitation and decreasing temperature. Clutch size was mainly, but not entirely, dependent on female SVL, suggesting both a proximate effect of local conditions and intrinsic differences among populations. Females from drier and warmer sites produced larger hatchlings. Mean egg mass was mainly determined by familial effects. Eggs incubated at a constant temperature hatched earlier than did their siblings incubated at fluctuating temperatures, a fact that could be explained by considering that in Mediterranean environments developmental rate might increase at a lower speed above average incubation temperature than it does decrease below it.     

Determinants of reproductive success and offspring sex in a turtle with environmental sex determination

Despite the importance of maternal effects in evolution, and knowledge of links among nest site choice, timing of nesting, offspring sex, and reproductive success in animals with environmental sex determination, these attributes have not been rigorously studied in a combined and natural context. To address this need we studied the relationships between three maternal traits (nest site choice, lay date, and nest depth) and two fitness‐related attributes of offspring (hatchling sex and embryonic survival) in the riverine turtle Carettochelys insculpta, a species with temperature‐dependent sex determination, for four years. Predation and flooding were the major sources of embryonic mortality in 191 nests. Embryonic survival was influenced by both lay date and nest site choice: in one year when nesting began later than average, nests laid later and at lower elevations were destroyed by early wet season river rises. In other years early nesting precluded flood mortality. However, turtles did not nest at the highest available elevations, and a field experiment confirmed that turtles were constrained to nest at lower elevations where they could construct a nest chamber. The principal determinant of hatchling sex in 140 nests was lay date, which in turn was apparently related to the magnitude of the previous wet season(s). Clutches laid earlier in the season (a female's first clutch) produced mainly males, while later clutches (her second clutch) yielded mostly females, due to seasonal increases in air temperatures. Accordingly, later nesting produced female‐biased hatchling sex ratios in 1996, while earlier nesting resulted in sex ratios near unity in the other years. However, all‐female nests were more likely to be flooded than mixed‐sex or all‐male nests in years when nesting was late. In conclusion, we found evidence that the position of two maternal trait distributions (elevation of the nest site and lay date), associated with the reproductive strategy of C. insculpta, reflect a combination of natural selection, physical constraints, and phenotypic plasticity. © 2004 The Linnean Society of London, Biological Journal of the Linnean Society, 2004, 81 , 1–16.     

Sperm limitation affects sex allocation in a parasitoid wasp Nasonia vitripennis

Insect reproduction is influenced by various external factors including temperature, a well-studied constraint. We investigated to what extent different levels of sperm limitation of males exposed to different heat stresses (34 and 36℃) afFect fem ales' offspring production and sex allocation in Nasonia vitripennis. In this haplodiploid parasitoid wasp attacking different species of pest flies, we investigated the effect of the quantity of sperm females received and stored in their spermatheca on their sperm use decisions, hence sex allocation, over successive ovipositions. In particular, we compared the sex allocation of females presenting three levels of sperm limitation (i.e.,mated with control, 34 ℃ heat-stressed or 36℃heat-stressed males) on each host they parasitized. To disentangle the potential reduction of sperm quality after a heat stress exposure from that of sperm quantity, we also explored the clutch size and sex ratio produced by fem ales that were partially sperm limited after copulating with multiply mated males. Independently of their sperm numbers, all types of fem ales produced a similar total number of offspring, but the more limited ones had fewer daughters. Sperm limitation further affected the distribution of daughters' production across time.In addition to constraints acting on female physiology, male fertility should therefore be considered in studies measuring reproductive outputs of insects submitted to heat stresses.     

High temperatures influence sexual development differentially in male and female tadpoles of the Indian skipper frog, <Emphasis Type="Italic">Euphlyctis cyanophlyctis</Emphasis>

Although sex determination in amphibians is believed to be a genetic process, environmental factors such as temperature are known to influence the sex differentiation and development. Extremely low and high temperatures influence gonadal development and sex ratio in amphibians but the mechanism of action is not known. In the present study, effect of different temperatures on gonadal development, sex ratio and metamorphosis was studied in the Indian skipper frog, Euphlyctis cyanophlyctis. The embryos of Gosner stage 7 were exposed to 20, 22, 24, 26, 28, 30 and 32°C up to tadpole stage 42. The embryos (stage 7) were also exposed to 20 and 32°C up to tadpole stage 25 (non-feeding stages). Tadpoles of stage 25 were reared at 20 and 32°C up to stage 42 (feeding stages). The results show that exposure to higher temperatures (28, 30 and 32°C) during stages 7–42 produced male-biased sex ratio. Rearing of tadpoles at 32°C during stages 25–42 produced male-biased sex ratio, while exposure during stages 7–25 did not affect sex ratio. Embryos and tadpoles exposed to lower temperatures (20 and 22°C) died during the early stages. High temperatures stimulated testis development, and disturbed ovary development. Exposure to high temperatures resulted in the early metamorphosis of tadpoles with reduced body size. These results demonstrated that high temperatures influence gonadal development differently in male and female tadpoles, leading to male-biased sex ratio. These results suggest that high temperature probably acts through stress hormones and favours the small-sized sex.     

The effect of a desert climate on the lactation and on the fertility of the nursing rat

Forty white rats were raised and mated in a controlled environment at 22°C. Twelve hours after parturition they were mated and separated into two equal groups. One group (control) remained at 22°C whilst the second group(desert) was kept in an open shed during July (daily temperature range: 18 to 37°C). The rats were sacrificed at 18 days post partum. Control rats ate 25% more food than desert rats; water consumption was the same in both groups. Body weights were initially similar, but from 12 days post partum onwards, the control rats increased their body weight at a significantly higher rate than did the desert rats. Milk production, assessed by litter weight gains, was higher in control group rats. Body composition and organ weights were similar except for the gut of the control group which was heavier. The number of CORPORA LUTEA was significantly lower in the desert group as were conception rates and embryo numbers; the weight of individual embryos and the weight of embryo per dam were, however, significantly higher in the desert group. It was postulated that during early pregnancy, maternal weight and mammary tissue may have preference over the uterus for nutritional sources, whilst in later pregnancy the embryos may have preference.