Heat Stress Experiment Research Articles

Broiler chicken vocalization analysis during a medium-scale heat stress experiment

The poultry industry in Flanders, Belgium, is characterized by highly efficient production mechanisms. To improve awareness for animal health and welfare, the use of neural network-based broiler vocalization detectors has been recently proposed in scientific literature. However, these tools have not yet been tested when used continuously in large-scale deployments. This paper describes the deployment of the broiler vocalization detector to analyze the sonic environment during four experimental rounds, each containing 560 broilers, in a medium scale housing at ILVO, Belgium. Half the broilers were subjected to periods of heat stress (ambient temperature increased to 32°C for 6h/day for 10 days during the last weeks of their lives) and the effect on the diurnal broiler vocalization pattern and occurrence of warbles, pleasure notes, short beeps and distress calls is discussed. While these four different types of vocalizations are common vocalizations in young birds, new types of vocalizations are emerging at an older age. A clustering has been performed to further analyze these new types of vocalizations occurring.

Climate adaptive loci revealed by seascape genomics correlate with phenotypic variation in heat tolerance of the coral Acropora millepora

One of the main challenges in coral reef conservation and restoration is the identification of coral populations resilient under global warming. Seascape genomics is a powerful tool to uncover genetic markers potentially involved in heat tolerance among large populations without prior information on phenotypes. Here, we aimed to provide first insights on the role of candidate heat associated loci identified using seascape genomics in driving the phenotypic response of Acropora millepora from New Caledonia to thermal stress. We subjected 7 colonies to a long-term ex-situ heat stress assay (4 °C above the maximum monthly mean) and investigated their physiological response along with their Symbiodiniaceae communities and genotypes. Despite sharing similar thermal histories and associated symbionts, these conspecific individuals differed greatly in their tolerance to heat stress. More importantly, the clustering of individuals based on their genotype at heat-associated loci matched the phenotypic variation in heat tolerance. Colonies that sustained on average lower mortality, higher Symbiodiniaceae/chlorophyll concentrations and photosynthetic efficiency under prolonged heat stress were also the closest based on their genotypes, although the low sample size prevented testing loci predictive accuracy. Together these preliminary results support the relevance of coupling seascape genomics and long-term heat stress experiments in the future, to evaluate the effect size of candidate heat associated loci and pave the way for genomic predictive models of corals heat tolerance.

AMPK-mediated regulation of cardiac energy metabolism: Implications for thermotolerance in Argopecten irradians irradians

AMPK is a key regulator of metabolism in eukaryotes across various pathways related to energy regulation. Although extensive investigations of AMPK have been conducted in mammals and some model organisms, research on AMPK in scallops is comparatively limited. In this study, three AMPK family genes (AiAMPKα, AiAMPKβ and AiAMPKγ) in scallop Argopecten irradians irradians were identified through genome scanning. Structure prediction and phylogenetic analyses of AiAMPKs were performed to determine their structural features and evolutionary relationships. Spatiotemporal expression patterns of AiAMPKs at different developmental stages and in healthy adult tissues were analyzed to elucidate the function of AiAMPKs in bay scallops’ growth and development. The spatiotemporally specific expression of AiAMPKs implied their important roles in growth and development of bay scallops. Heat stress experiment was performed to determine the regulations of AiAMPKs in four kinds of thermosensitive tissues. Expression profiles revealed distinct molecular mechanisms of AiAMPKs in different tissues in response to heat stress: significant down-regulations in mobile hemocytes, but dominant up-regulations occurring in stationary gills, mantles and hearts. Functional verification including knock-down of AiAMPKα and inhibition of AiAMPK was separately conducted in the thermotolerant tissue heart at the post-transcription and translation levels. The thermotolerant index Arrhenius break temperature (ABT) showed a significant decrease and the rate-amplitude product (RAP) peaked earlier in the individuals after RNAi targeting AiAMPKα, displaying an earlier transition to anaerobic metabolism under heat stress, indicating an impairing ability of aerobic metabolism. After AiAMPK inhibition, widespread down-regulations of genes in key energy metabolism pathways, RNA polymerase II-mediated transcription, and aminoacyl-tRNA synthesis pathways were obviously observed, revealing the post-translational inhibition of AiAMPK hindered cardiac energy metabolism, basal transcription and translation. Overall, our findings provide evidences for exploring the molecular mechanisms of energy regulation in thermotolerant traits in bay scallops under ongoing global warming.

Host starvation and in hospite degradation of algal symbionts shape the heat stress response of the Cassiopea-Symbiodiniaceae symbiosis

BackgroundGlobal warming is causing large-scale disruption of cnidarian-Symbiodiniaceae symbioses fundamental to major marine ecosystems, such as coral reefs. However, the mechanisms by which heat stress perturbs these symbiotic partnerships remain poorly understood. In this context, the upside-down jellyfish Cassiopea has emerged as a powerful experimental model system.ResultsWe combined a controlled heat stress experiment with isotope labeling and correlative SEM-NanoSIMS imaging to show that host starvation is a central component in the chain of events that ultimately leads to the collapse of the Cassiopea holobiont. Heat stress caused an increase in catabolic activity and a depletion of carbon reserves in the unfed host, concurrent with a reduction in the supply of photosynthates from its algal symbionts. This state of host starvation was accompanied by pronounced in hospite degradation of algal symbionts, which may be a distinct feature of the heat stress response of Cassiopea. Interestingly, this loss of symbionts by degradation was concealed by body shrinkage of the starving animals, resulting in what could be referred to as “invisible” bleaching.ConclusionsOverall, our study highlights the importance of the nutritional status in the heat stress response of the Cassiopea holobiont. Compared with other symbiotic cnidarians, the large mesoglea of Cassiopea, with its structural sugar and protein content, may constitute an energy reservoir capable of delaying starvation. It seems plausible that this anatomical feature at least partly contributes to the relatively high stress tolerance of these animals in rapidly warming oceans.7jQYTVpxtJyVhU5epTFb61Video

Multi-omics reveals phenol-based coordinated defense of Sparganium stoloniferum rhizoma

Sparganium stoloniferum is a widely distributed aquatic medicinal plant that exhibits exceptional tolerance to environmental stress. Interestingly, the phenols in S. stoloniferum which have medicinal properties, are defensive compounds. Studies integrating transcriptome, proteome, and metabolome analyses have highlighted the phenylpropanoid and flavonoid biosynthesis pathways, which are related to the defense mechanisms of S. stoloniferum. During the growth process, S. stoloniferum rhizoma (SL) accumulates lignin, forming a physical defense, and phenylpropanoids and flavonoids derived during the lignin biosynthesis process provide chemical defense. In this study, a map of the molecular processes involved in the coordinated defense of S. stoloniferum was generated. The expression patterns of genes involved in the coordinated defense of SL were consistent with the omics results. The cDNA of the phenylalanine ammonia-lyase (PAL) gene from S. stoloniferum (SsPAL) was cloned. The recombinant protein SsPAL showed good affinity and catalytic activity for L-phenylalanine. Heat stress experiments indicated that genes related to phenol biosynthesis were upregulated when S. stoloniferum responded to stress. Therefore, phenols played a crucial defense role when S. stoloniferum was at an early developmental stage or responded to stress in a short time. In contrast, the physical defense mainly relied on lignin formation, which was the main defense mechanism of SL in the late growth stage. Based on these results, we discuss the causes of phenolic pharmacological substance formation in medicinal aquatic plants. These results provide new insights into the defense strategies of aquatic plants.

Heat-tolerant intertidal rock pool coral Porites lutea can potentially adapt to future warming.

The growing threat of global warming on coral reefs underscores the urgency of identifying heat-tolerant corals and discovering their adaptation mechanisms to high temperatures. Corals growing in intertidal rock pools that vary markedly in daily temperature may have improved heat tolerance. In this study, heat stress experiments were performed on scleractinian coral Porites lutea from subtidal habitat and intertidal rock pool of Weizhou Island in the northern South China Sea. Thermotolerance differences in corals from the two habitats and their mechanisms were explored through phenotype, physiological indicators, ITS2, 16S rRNA, and RNA sequencing. At the extremely high temperature of 34°C, rock pool P. lutea had a stronger heat tolerance than those in the subtidal habitat. The strong antioxidant capacity of the coral host and its microbial partners was important in the resistance of rock pool corals to high temperatures. The host of rock pool corals at 34°C had stronger immune and apoptotic regulation, downregulated host metabolism and disease-infection-related pathways compared to the subtidal habitat. P. lutea, in this habitat, upregulated Cladocopium C15 (Symbiodiniaceae) photosynthetic efficiency and photoprotection, and significantly increased bacterial diversity and coral probiotics, including ABY1, Ruegeria, and Alteromonas. These findings indicate that rock pool corals can tolerate high temperatures through the integrated response of coral holobionts. These corals may be 'touchstones' for future warming. Our research provides new insights into the complex mechanisms by which corals resist global warming and the theoretical basis for coral reef ecosystem restoration and selection of stress-resistant coral populations.

Genome-Wide Association Analysis of Heat Tolerance in F2 Progeny from the Hybridization between Two Congeneric Oyster Species.

As the world's largest farmed marine animal, oysters have enormous economic and ecological value. However, mass summer mortality caused by high temperature poses a significant threat to the oyster industry. To investigate the molecular mechanisms underlying heat adaptation and improve the heat tolerance ability in the oyster, we conducted genome-wide association analysis (GWAS) analysis on the F2 generation derived from the hybridization of relatively heat-tolerant Crassostrea angulata ♀ and heat-sensitive Crassostrea gigas ♂, which are the dominant cultured species in southern and northern China, respectively. Acute heat stress experiment (semi-lethal temperature 42 °C) demonstrated that the F2 population showed differentiation in heat tolerance, leading to extremely differentiated individuals (approximately 20% of individuals die within the first four days with 10% survival after 14 days). Genome resequencing and GWAS of the two divergent groups had identified 18 significant SNPs associated with heat tolerance, with 26 candidate genes located near these SNPs. Eleven candidate genes that may associate with the thermal resistance were identified, which were classified into five categories: temperature sensor (Trpm2), transcriptional factor (Gata3), protein ubiquitination (Ube2h, Usp50, Uchl3), heat shock subfamily (Dnajc17, Dnaja1), and transporters (Slc16a9, Slc16a14, Slc16a9, Slc16a2). The expressional differentiation of the above genes between C. gigas and C. angulata under sublethal temperature (37 °C) further supports their crucial role in coping with high temperature. Our results will contribute to understanding the molecular mechanisms underlying heat tolerance, and provide genetic markers for heat-resistance breeding in the oyster industry.

PSI-A-7 Prediction Models for Estimating Metabolizable Energy Intake of Pigs Based on Body Weight and Ambient Temperature

Abstract Prediction models for estimating feed intake (FI) or metabolizable energy intake (MEI) of pigs are available in the literature. During the recent two decades, several reports have addressed the influence of heat stress on FI. The objectives of the present work were to validate the accuracy of prediction equations for estimating FI or MEI of pigs in the literature using data from heat stress experiments and to develop novel equations for estimating FI and MEI of pigs under thermoneutral or heat stress environment. A total of 123 data from 25 research papers measuring the effects of heat stress on FI of pigs published between 2001 and 2021 were used for the validation of previous equations and the development of novel equations. In the 25 papers, the thermoneutral temperature (T) ranged from 17 to 24℃ and the heat stress T ranged from 21 to 35℃. The body weight (BW) of pigs ranged from 19 to 119 kg. In the first validation study, the equation for estimating FI was evaluated: FI, g/day = –1,264 + 73.6×BW – 0.26×BW2 + 117×T – 2.40×T2 – 0.95×T×BW. The slope representing a linear bias did not differ from 0 but the intercept representing a mean bias was greater than 0 (109 ± 36; P < 0.01), meaning that the predicted mean underestimated the FI. In the second validation study, the equation for estimating MEI using equations for lower critical temperature (LCT) and fraction of MEI: LCT, ℃ = 17.9 – 0.0375×BW and fraction of MEI = 1 – 0.012914 × [T – (LCT + 3)] – 0.001179 × [T – (LCT + 3)]2. The slope representing a linear bias did not differ from 0 but the intercept representing a mean bias was greater than 0 (731 ± 68; P < 0.001), meaning that the predicted mean underestimated the MEI. A novel equation for estimating MEI under thermoneutral or heat stress environment was developed: MEI, kcal/day = 2,603 + 137×BW – 0.189×BW2 – 38.4×T – 0.556×T2 – 1.165×T×BW (P < 0.001; R2 = 0.83). An equation for estimating fraction of MEI under thermoneutral or heat stress environment was also developed: fraction of MEI = 1 – 0.00677 × [T – (LCT + 3)] – 0.000668 × [T – (LCT + 3)]2 (P < 0.001; R2 = 0.98). In conclusion, previously published equations underestimate FI and MEI and novel equations for estimating MEI under thermoneutral or heat stress environment have been developed in the present work. However, the novel equations have not been validated under cold stress.

The roles of heating rate, intensity, and duration on the response of corals and their endosymbiotic algae to thermal stress

Anthropogenic ocean warming is one of the biggest threats to marine organisms worldwide. However, it remains unclear how the duration and intensity of thermal anomalies affect organismal stress responses and thermal thresholds. We used detailed tracking of coral endosymbiont and host physiology and dose-response analyses to compare the effects of multiple heating rates, intensities, and exposure durations on two reef-building corals, Acropora hemprichii and Porites lobata, from adjacent sides of a reef (protected vs. exposed) in the Central Red Sea known to differ in high-frequency (< 24 h) temperature variability. Corals were exposed to acute heat exposures (18 h) with four target temperatures (32 °C, 35 °C, 36.5 °C, and 38 °C), versus prolonged heat exposures lasting 7–15 days where temperatures were raised 0.5 and 1.5 °C day−1 to four target temperatures (32 °C, 33.5 °C, 35 °C, and 36.5 °C). In the prolonged experiment, dose-response curves assessing algal endosymbiont Fv/Fm revealed little initial effect of temperature, before an exponential decline above 34 °C for both species. Temperature at time of measurement and degree heating hours above 34 °C (DHH34) were the variables most strongly associated with declines in Fv/Fm. The Fv/Fm thermal thresholds for P. lobata from the high-variability protected site were higher than the exposed site in the faster heating, prolonged heat stress experiment despite minimal differences in endosymbiont density, chlorophyll-a, and host protein between sites. Together, our dose-response analysis revealed complex effects of DHH34, heating rate, and species-specific differences in the influence of local thermal histories shaping thermotolerance limits for these corals.

Wet-Bulb Temperature or Heat Index: Which Better Predicts Fatal Heat in a Warming Climate?

The wet-bulb temperature of 35 °C has been widely used as the threshold for human survivability in the field of climate science. However, recent experiments have shown that a person's core temperature starts to rise at a range of critical wet-bulb temperatures well below this threshold. It is shown here why this state of severe heat stress cannot be predicted using the wet-bulb temperature. Instead, it is shown that the recently extended heat index can explain nearly all of the variance in the observed critical combinations of temperature and humidity in those heat-stress experiments. For light and moderate exertion in an indoor setting, the heat index predicts that the critical wet-bulb temperature ranges from 20 to 32 °C, depending on the relative humidity, consistent with experimental results. For the same setting and exertion, the heat index predicts fatal wet-bulb temperatures ranging from 24 to 37 °C. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Differences in heat tolerance, water use efficiency and growth among Douglas-fir families and varieties evidenced by GWAS and common garden studies.

Severe and frequent heat and drought events challenge the survival and development of long-generation trees. In this study, we investigated the genomic basis of heat tolerance, water use efficiency and growth by performing genome-wide association studies in coastal Douglas-fir (Pseudotsuga menziesii) and intervarietal (menziesii × glauca) hybrid seedlings. GWAS results identified 32 candidate genes involved in primary and secondary metabolism, abiotic stress and signaling, among other functions. Water use efficiency (inferred from carbon isotope discrimination), photosynthetic capacity (inferred from %N), height and heat tolerance (inferred from electrolyte leakage in a heat stress experiment) were significantly different among Douglas-fir families and varieties. High-elevation seed sources had increased water use efficiency, which could be a result of higher photosynthetic capacity. Similarly, families with greater heat tolerance also had higher water use efficiency and slower growth, suggesting a conservative growth strategy. Intervarietal hybrids showed increased heat tolerance (lower electrolyte leakage at 50 and 55 °C) and higher water use efficiency compared with coastal families, suggesting that hybridization might be a source of pre-adapted alleles to warming climates and should be considered for large-scale reforestation projects under increasingly arid conditions.

Predicting fatal heat and humidity using the heat index model.

A unique wet-bulb temperature of 35°C is often used as the threshold for human survivability, but recent experiments have shown that a person's core temperature starts to rise at a wide range of critical wet-bulb temperatures. Here, it is shown that the model underlying the heat index correctly predicts those critical wet-bulb temperatures, explaining 95% of the variance in the values observed in laboratory heat-stress experiments. This is the first time the heat-index model has been validated against physiological data from laboratory experiments. For light and moderate exertion in an indoor setting, the heat index model predicts that the critical wet-bulb temperature ranges from 20°C to 32°C, depending on the relative humidity, consistent with experimental results. For the same setting and exertion, the heat index model predicts fatal wet-bulb temperatures ranging from 24°C to 37°C.NEW & NOTEWORTHY Recent experiments have identified the critical combinations of heat and humidity, in an indoor setting, above which an individual is unable to maintain a standard core temperature, indicating severe heat stress. It is shown here why this state of severe heat stress cannot be predicted using the wet-bulb temperature. Instead, it is shown that the recently extended heat index model can explain nearly all of the variance in the observed critical combinations of temperature and humidity, and can be used to calculate fatal combinations.

Infrared Thermal Imaging and Morpho-Physiological Indices Used for Wheat Genotypes Screening under Drought and Heat Stress

Bread wheat, one of the largest broadacre crops, often experiences various environmental stresses during critical growth stages. Terminal drought and heat stress are the primary causes of wheat yield reduction worldwide. This study aimed to determine the drought and heat stress tolerance level of a group of 46 diverse wheat genotypes procured from the Australian Grains Gene Bank, Horsham, VIC Australia. Two separate drought stress (DS) and heat stress (HS) pot experiments were conducted in separate growth chambers. Ten days after complete anthesis, drought (40 ± 3% field capacity for 14 days) and heat stress (36/22 °C for three consecutive days) were induced. A significant genotype × environment interaction was observed and explained by various morpho-physiological traits, including rapid, non-destructive infrared thermal imaging for computational water stress indices. Except for a spike length in DS and harvest index in HS, the analysis of variance showed significant differences for all the recorded traits. Results showed grains per spike, grains weight per spike, spike fertility, delayed flag leaf senescence, and cooler canopy temperature were positively associated with grain yield under DS and HS. The flag leaf senescence and chlorophyll fluorescence were used to measure each genotype's stay-green phenotype and photosystem II activity after DS and HS. This study identified the top ten best and five lowest-performing genotypes from drought and heat stress experiments based on their overall performance. Results suggest that if heat or drought adaptive traits are brought together in a single genotype, grain yield can be improved further, particularly in a rainfed cropping environment.

The effects of marine heatwaves on acute heat tolerance in corals.

Scleractinian coral populations are increasingly exposed to conditions above their upper thermal limits due to marine heatwaves, contributing to global declines of coral reef ecosystem health. However, historic mass bleaching events indicate there is considerable inter- and intra-specific variation in thermal tolerance whereby species, individual coral colonies and populations show differential susceptibility to exposure to elevated temperatures. Despite this, we lack a clear understanding of how heat tolerance varies across large contemporary and historical environmental gradients, or the selective pressures that underpin this variation. Here we conducted standardised acute heat stress experiments to identify variation in heat tolerance among species and isolated reefs spanning a large environmental gradient across the Coral Sea Marine Park. We quantified the photochemical yield (Fv /Fm ) of coral samples in three coral species, Acropora cf humilis, Pocillopora meandrina, and Pocillopora verrucosa, following exposure to four temperature treatments (local ambient temperatures, and + 3°C, +6°C and + 9°C above local maximum monthly mean). We quantified the temperature at which Fv /Fm decreased by 50% (termed ED50) and used derived values to directly compare acute heat tolerance across reefs and species. The ED50 for Acropora was 0.4-0.7°C lower than either Pocillopora species, with a 0.3°C difference between the two Pocillopora species. We also recorded 0.9°C to 1.9°C phenotypic variation in heat tolerance among reefs within species, indicating spatial heterogeneity in heat tolerance across broad environmental gradients. Acute heat tolerance had a strong positive relationship to mild heatwave exposure over the past 35 years (since 1986) but was negatively related to recent severe heatwaves (2016-2020). Phenotypic variation associated with mild thermal history in local environments provides supportive evidence that marine heatwaves are selecting for tolerant individuals and populations; however, this adaptive potential may be compromised by the exposure to recent severe heatwaves.

Persistence of phenotypic responses to short-term heat stress in the tabletop coral Acropora hyacinthus.

Widespread mapping of coral thermal resilience is essential for developing effective management strategies and requires replicable and rapid multi-location assays of heat resistance and recovery. One- or two-day short-term heat stress experiments have been previously employed to assess heat resistance, followed by single assays of bleaching condition. We tested the reliability of short-term heat stress resistance, and linked resistance and recovery assays, by monitoring the phenotypic response of fragments from 101 Acropora hyacinthus colonies located in Palau (Micronesia) to short-term heat stress. Following short-term heat stress, bleaching and mortality were recorded after 16 hours, daily for seven days, and after one and two months of recovery. To follow corals over time, we utilized a qualitative, non-destructive visual bleaching score metric that correlated with standard symbiont retention assays. The bleaching state of coral fragments 16 hours post-heat stress was highly indicative of their state over the next 7 days, suggesting that symbiont population sizes within corals may quickly stabilize post-heat stress. Bleaching 16 hours post-heat stress predicted likelihood of mortality over the subsequent 3-5 days, after which there was little additional mortality. Together, bleaching and mortality suggested that rapid assays of the phenotypic response following short-term heat stress were good metrics of the total heat treatment effect. Additionally, our data confirm geographic patterns of intraspecific variation in Palau and show that bleaching severity among colonies was highly correlated with mortality over the first week post-stress. We found high survival (98%) and visible recovery (100%) two months after heat stress among coral fragments that survived the first week post-stress. These findings help simplify rapid, widespread surveys of heat sensitivity in Acropora hyacinthus by showing that standardized short-term experiments can be confidently assayed after 16 hours, and that bleaching sensitivity may be linked to subsequent survival using experimental assessments.

Understanding Knowledge Level of The Thermal Implication and Its Effects Towards Physiological State in Construction Sector

Heat stress normally known as a hidden cause of accidents in construction sectors. To ensure the productivity and health of workers in the construction site, it is necessary to evaluate the effects of temperature and relative humidity on the workers’ physiology under hot conditions. Hence, the aims of this paper are: (i) to investigate the knowledge of heat stress and workers perceptions on workers performance in construction site, (ii) to identify the environment factor of heat stress in the construction site and (iii) explore the measurement physiology parameters for heat stress. Heat stress questionnaires and experiment test were combined to extract useful information. An online survey was undertaken with a representative sample (N=292) from Malaysia construction sector. While, the experiment was carried out in a well-controlled climate chamber to obtain datasets with four conditions combining air temperature and relative humidity (32 °C/70 %, 34 °C/92, 34 °C/74% and 38 °C/83%). At a climate chamber, the subjects doing a job such as lifting and carry the 10 kg workload were exposed to different combinations of air temperatures and relative humidity. The subject’s physiological responses to the environment were then investigated. The survey’s finding showed 71.9% of the workers understand about heat stress. 22.6% of the workers perceive that the temperature is hot and quite hot and relative humidity result showed that 50.9 % of the workers perceived that part of their mouth and throat are dry while working. Besides, the experiment study showed that workers physical demands varies according to their work task with a combination of the influences from individual and environmental factors.

Ocean Warming and Heat Stress Impact Molecules of Keystone Significance in a Predatory Marine Gastropod

Water temperature is a major abiotic driver, controlling the rates and nature of biochemical reactions and subsequently affecting the physiology of marine organisms. However, relatively little is known about the implications of heat stress or predicted ocean climate change on marine secondary metabolites. The predatory gastropod Dicathais orbita is a useful model organism for climate change and natural product studies. Here we determine the upper thermal limit (CTMax) of D. orbita and investigate the effects of thermal stress on the bioactive compounds stored in their biosynthetic organ, the hypobranchial gland. Two CTMax experiments were undertaken, along with a static heat stress experiment where whelks were exposed to an elevated temperature of 30°C for one week, compared to a 20°C seawater control. An additional 35-day ocean climate change experiment used combinations of temperature (ambient: 23°C and future: 25°C) and pCO2 (ambient: ~380 ppm and future: ~765 ppm). The impacts on secondary metabolites in all experiments were assessed using liquid chromatography-mass spectrometry. The mean CTMax of the whelks, from the northern limit of their distribution, was found to be 35.2°C using a rapid temperature increase rate of 1°C/1 h, but was only 30.6°C when a gradual heating rate of 1°C/12 h was used. The overall composition of the secondary metabolites was significantly affected by heat stress in all four experiments, but not by elevated pCO2 in the ocean climate change experiment. The proportion of the choline ester murexine was significantly reduced in heat-stressed snails compared to the controls. Tyrindoxyl sulphate was significantly reduced under prolonged exposure to future temperature, whereas the relative abundance of the oxidation product, 6-bromoisatin significantly increased with elevated temperature exposure. Despite the fact that intertidal gastropods like D. orbita might be able to buffer the impact of external temperatures within the predicted future range, this study provides evidence that ocean warming could have significant implications for secondary metabolite production and/or storage in marine invertebrates. Impacts on bioactive molecules with multifunctional ecological roles could have implications for predator populations with possible flow on effects in some marine communities.

High-resolution in situ thermal metrics coupled with acute heat stress experiments reveal differential coral bleaching susceptibility

High-resolution in situ thermal metrics coupled with acute heat stress experiments reveal differential coral bleaching susceptibility

Identification of functional features underlying heat stress response in Sprague–Dawley rats using mixed linear models

Since global temperature is expected to rise by 2 °C in 2050 heat stress may become the most severe environmental factor. In the study, we illustrate the application of mixed linear models for the analysis of whole transcriptome expression in livers and adrenal tissues of Sprague–Dawley rats obtained by a heat stress experiment. By applying those models, we considered four sources of variation in transcript expression, comprising transcripts (1), genes (2), Gene Ontology terms (3), and Reactome pathways (4) and focussed on accounting for the similarity within each source, which was expressed as a covariance matrix. Models based on transcripts or genes levels explained a larger proportion of log2 fold change than models fitting the functional components of Gene Ontology terms or Reactome pathways. In the liver, among the most significant genes were PNKD and TRIP12. In the adrenal tissue, one transcript of the SUCO gene was expressed more strongly in the control group than in the heat-stress group. PLEC had two transcripts, which were significantly overexpressed in the heat-stress group. PER3 was significant only on gene level. Moving to the functional scale, five Gene Ontologies and one Reactome pathway were significant in the liver. They can be grouped into ontologies related to DNA repair, histone ubiquitination, the regulation of embryonic development and cytoplasmic translation. Linear mixed models are valuable tools for the analysis of high-throughput biological data. Their main advantages are the possibility to incorporate information on covariance between observations and circumventing the problem of multiple testing.

Genome-wide scan for selection signatures and genes related to heat tolerance in domestic chickens in the tropical and temperate regions in Asia

Heat stress is one of the major environmental stressors challenging the global poultry industry. Identifying the genes responsible for heat tolerance is fundamentally important for direct breeding programs. To uncover the genetic basis underlying the ambient temperature adaptation of chickens, we analyzed a total of 59 whole genomes from indigenous chickens that inhabit South Asian tropical regions and temperate regions from Northern China. We applied FST and π-ratio to scan selective sweeps and identified 34 genes with a signature of positive selection in chickens from tropical regions. Several of these genes are functionally implicated in metabolism (FABP2, RAMP3, SUGCT, and TSHR) and vascular smooth muscle contractility (CAMK2), and they may be associated with adaptation to tropical regions. In particular, we found a missense mutation in thyroid-stimulating hormone receptor (41020238:G>A) that shows significant differences in allele frequency between the chicken populations of the two regions. To evaluate whether the missense mutation in TSHR could enhance the heat tolerance of chickens, we constructed segregated chicken populations and conducted heat stress experiments using homozygous mutations (AA) and wild-type (GG) chickens. We found that GG chickens exhibited significantly higher concentrations of alanine aminotransferase, lactate dehydrogenase, and creatine kinase than AA chickens under heat stress (35 ± 1°C) conditions (P < 0.05). These results suggest that TSHR (41020238:G>A) can facilitate heat tolerance and adaptation to higher ambient temperature conditions in tropical climates. Overall, our results provide potential candidate genes for molecular breeding of heat-tolerant chickens.