Heat Stress Experiment Research Articles

Liver Transcriptome Response to Heat Stress in Beijing You Chickens and Guang Ming Broilers.

Heat stress is one of the most prevalent issues in poultry production that reduces performance, robustness, and economic gains. Previous studies have demonstrated that native chickens are more tolerant of heat than commercial breeds. However, the underlying mechanisms of the heat tolerance observed in native chicken breeds remain unelucidated. Therefore, we performed a phenotypical, physiological, liver transcriptome comparative analysis and WGCNA in response to heat stress in one native (Beijing You, BY) and one commercial (Guang Ming, GM) chicken breed. The objective of this study was to evaluate the heat tolerance and identify the potential driver and hub genes related to heat stress in these two genetically distinct chicken breeds. In brief, 80 BY and 60 GM, 21 days old chickens were submitted to a heat stress experiment for 5 days (33 °C, 8 h/day). Each breed was divided into experimental groups of control (Ctl) and heat stress (HS). The results showed that BY chickens were less affected by heat stress and displayed reduced DEGs than GM chickens, 365 DEGs and 382 DEGs, respectively. The transcriptome analysis showed that BY chickens exhibited enriched pathways related to metabolism activity, meanwhile GM chickens’ pathways were related to inflammatory reactions. CPT1A and ANGPTL4 for BY chickens, and HSP90B1 and HSPA5 for GM chickens were identified as potential candidate genes associated with HS. The WGCNA revealed TLR7, AR, BAG3 genes as hub genes, which could play an important role in HS. The results generated in this study provide valuable resources for studying liver transcriptome in response to heat stress in native and commercial chicken lines.

Impacts of Air Velocity Treatments under Summer Condition: Part I-Heavy Broiler's Surface Temperature Response.

Simple SummaryThe surface temperature variation of heavy broilers (42–61 d age) under heat stress is an important indicator of thermal comfort, but it is not well studied and reported yet. This study examined the variation of surface temperatures of broilers through two dynamic air velocity treatments under hot summer conditions. It was discovered that the surface temperatures varied over age, daytime, and environmental factors (air temperature, relative humidity, and temperature humidity index). A simple linear regression model to predict the surface temperature of heavy broilers was developed. The findings from this study will enhance knowledge to understand the broilers’ responses under heat stress, which will be helpful in providing necessary management decisions to create a comfortable thermal environment.Heavy broilers exposed to hot summer conditions experience fluctuations in surface temperatures due to heat stress, which leads to decreased performance. Maintaining a bird’s homeostasis depends on several environmental factors (temperature, relative humidity, and air velocity). It is important to understand the responses of birds to environmental factors and the amount of heat loss to the surrounding environment to create thermal comfort for the heavy broilers for improved performances and welfare. This study investigates the variation in surface temperatures of heavy broilers under high and low air velocity treatments. Daytime, age and bird location’s effect on the surface temperature variation was also examined. The experiment was carried out in the poultry engineering laboratory of North Carolina State University during summers of 2017, 2018, and 2019 as a part of a comprehensive study on the effectiveness of wind chill application to mitigate heat stress on heavy broilers. This live broiler heat stress experiment was conducted under two dynamic air velocity treatments (high and low) with three chambers per treatment and 44 birds per chamber. Surface temperatures of the birds were recorded periodically through the experimental treatment cycles (flocks, 35–61 d) with infrared thermography in the morning, noon, evening, and nighttime. The overall mean surface temperature of the broilers under two treatments was found to be 35.89 ± 2.37 °C. The variation in surface temperature happened due to air temperature, thermal index, air velocity, bird’s age, daytime, and position of birds inside the experimental chambers. The surface temperatures were found lower under high air velocity treatment and higher under low air velocity treatment. During the afternoon time, the broilers’ surface temperatures were higher than other times of the day. It was also found that the birds’ surface temperature increased with age and temperature humidity indices. Based upon the experimental data of five flocks, a simple linear regression model was developed to predict surface temperature from the birds’ age, thermal indices, and air velocity. It will help assess heavy broilers’ thermal comfort under heat stress, which is essential to provide a comfortable environment for them.

Alteration of hemoglobin ß gene expression in mucosal tissues of Japanese flounder, Paralichthys olivaceus, in response to heat stress, Edwardsiella piscicida infection, and immunostimulants administration

Hemoglobin beta (Hbß) is a heme-binding protein capable of oxygen delivery. The oligopeptides derived from Hbβ in fish mucus are active against a variety of gram-negative bacteria and protozoa. To gain information on the physiological and immunological roles of Hbβ in the mucosal tissues of fish, we analyzed changes in Hbß gene expression levels in the epidermis, gills, and intestine of Japanese flounder, Paralichthys olivaceus, in response to heat stress, Edwardsiella piscicida infection, and trial feeding of immunostimulants, high-concentration ascorbic acid (AsA) or lactoferrin (LF). The results of quantitative real-time PCR showed that expression of the Hbß gene in the gills decreased markedly when exposed to heat stress, whereas that in the epidermis exhibited an increase 3h after infection with E. piscicida. Seven days after starting to feed either immunostimulant, epidermal Hbß gene expression in all AsA or LF dose groups was significantly higher than in the control group. The results of in situ hybridization showed that the abundance and intensity of the stained cells in the epidermis and in the gills were consistent with the expression levels of Hbß gene obtained from the infection and immunosuppressant experiments and the heat stress experiment, respectively. Our results suggest that mucosal Hbβ gene expression is closely related to physiological and immunological status and could be a useful indicator for monitoring condition of fish health.

Combining near-infrared radiance of vegetation and fluorescence spectroscopy to detect effects of abiotic changes and stresses

Solar-induced chlorophyll fluorescence (SIF) shows great potential to assess plants physiological state and response to environmental changes. Recently the near-infrared reflectance of vegetation (NIRv) provides a promising way to quantify the confounding effect of canopy structure in SIF, while the difference between SIF and NIRv under varying environmental conditions has not been well explored. Here we developed a simple approach to extract the fluorescence yield (ΦF) by the combined use of SIF and the near-infrared radiance of vegetation (NIRvR). The proposed NIRvR approach was evaluated in multiple ways, including with the seasonal leaf-level steady-state fluorescence yield. Results indicate that NIRvR-derived ΦF well captured the seasonal variation of the fluorescence yield changes, and achieved similar results with the existing approach. Both SIF and NIRvR were derived from the airborne imaging fluorescence spectrometer HyPlant for three case studies to evaluate the impacts of light adaptation, heat stress and water limitation on ΦF. For the light adaptation case study, ΦF over the low-light adapted sugar beet field was about 1.3 times larger compared to an unaffected reference area while the difference in NIRvR was minimal, which clearly shows the short-term photosynthetic light induction effect and the ability of SIF to detect plant physiological responses. For the heat stress experiment, ΦF decreased during a natural heatwave in 2015 in the fields of rapeseed from 0.0150 to 0.0130, barley from 0.0152 to 0.0144, and wheat from 0.0146 to 0.0142 which showed signs of senescence, while slightly increased from 0.0125 to 0.0130 in the corn field which was still in growing. At the water-limited sugar beet field, ΦF first increased towards solar noon and then slightly decreased during the afternoon over the water-limited areas from 0.017 to 0.021 and 0.020, with high temperature and high light at noon. The advantages to use SIF/NIRvR as a proxy of ΦF to detect stress-induced limitations in photosynthesis include that the impacts of canopy structure and sun-sensor geometry on the ΦF estimation are explicitly cancelled, and photosynthetically active radiation (PAR) is not required as input. Finally, our approach is directly applicable to satellite-derived estimates of SIF, enabling the study of variations in ΦF to detect the effects of abiotic changes and stresses at large scale.

Variation in Coral Thermotolerance Across a Pollution Gradient Erodes as Coral Symbionts Shift to More Heat-Tolerant Genera

Phenotypic plasticity is one mechanism whereby species may cope with stressful environmental changes associated with climate change. Reef building corals present a good model for studying phenotypic plasticity because they have experienced rapid climate-driven declines in recent decades (within a single generation of many corals), often with differential survival among individuals during heat stress. Underlying differences in thermotolerance may be driven by differences in baseline levels of environmental stress, including pollution stress. To examine this possibility, acute heat stress experiments were conducted on Acropora hyacinthus from 10 sites around Tutuila, American Samoa with differing nutrient pollution impact. A threshold-based heat stress assay was conducted in 2014 and a ramp-hold based assay was conducted in 2019. Bleaching responses were measured by assessing color paling. Endosymbiont community composition was assessed at each site using quantitative PCR. RNA sequencing was used to compare differences in coral gene expression patterns prior to and during heat stress in 2019. In 2014, thermotolerance varied among sites, with polluted sites holding more thermotolerant corals. These differences in thermotolerance correlated with differences in symbiont communities, with higher proportions of heat-tolerant Durusdinium found in more polluted sites. By 2019, thermotolerance varied less among sites, with no clear trend by pollution level. This coincided with a shift toward Durusdinium across all sites, reducing symbiont community differences seen in 2014. While pollution and symbiont community no longer could explain variation in thermotolerance by 2019, gene expression patterns at baseline levels could be used to predict thermotolerance thresholds. These patterns suggest that the mechanisms underlying thermotolerance shifted between 2014 and 2019, though it is possible trends may have also been affected by methodological differences between heat stress assays. This study documents a shift in symbiont community over time and captures potential implications of that shift, including how it affects variation in thermotolerance among neighboring reefs. This work also highlights how gene expression patterns could help identify heat-tolerant corals in a future where most corals are dominated by Durusdinium and symbiont-driven thermotolerance has reached an upper limit.

Signaling pathways of heat- and hypersalinity-induced polyp bailout in Pocillopora acuta

Polyp bailout is a drastic response to acute stress where coral coloniality breaks down and polyps detach. We induced polyp bailout in Pocillopora acuta with heat stress and tested for differential gene expression using RNAseq and a qPCR assay. Furthermore, we induced polyp bailout with hypersalinity and compared the results to identify stressor-independent signals and pathways active during polyp bailout. Both stressors led to the onset of polyp bailout and the detachment of vital polyps. We observed activation of microbe-associated molecular pattern receptors and downstream signaling pathways of the innate immune system. Further, we detected growth factors and genes active during Wnt-signaling potentially contributing to wound healing, regeneration, and proliferation. Upregulation of several genes encoding for matrix metalloproteinases and the fibroblast growth factor signaling pathway are the most likely involved in the remodeling of the extracellular matrix, as well as in the detachment of polyps from the calcareous skeleton during polyp bailout. Expression of genes of interest in our qPCR assay of vital polyps from our heat-stress experiment, showed a trend for a normalization of gene expression after polyp bailout. Our results provide new insights into the signaling cascades leading to the observed physiological responses during polyp bailout. Comparison between the two stressors showed that certain signaling pathways are independent of the stressor and suggested that polyp bailout is a general response of corals to acute stress. Furthermore, immune system responses during polyp bailout indicate that microbe-associated partners of corals may lead to the polyp bailout response.

The functions of Patchouli and Elsholtzia in the repair of hen follicular granular cells after heat stress

The objective of this experimental study was to examine the effects of the Chinese herbal medicines Patchouli and Elsholtzia on the follicular granulosa cells of hens undergoing heat stress conditions. In the current investigation, hen follicular granulosa cells were isolated from the prehierarchical follicles of layer hens and then cultured in-vitro. The cells were randomly divided into the 6 groups. Following the completion of this study's experiments using different heat stress and medicinal treatments, the cell activities of each group were measured using an MTT method. The levels of the heat shock protein 70 (HSP70) were detected using ELISA. The expressions of the steroidogenic acute regulatory protein (StAR) mRNA; cytochrome P450 family 11, subfamily A, member 1 (CYP11A1) mRNA; proliferating cell nuclear antigen (PCNA) mRNA; and the follicle stimulating hormone receptor (FSHR) were detected using the real-time quantitative polymerase chain reactions. The concentration levels of estrogen and progesterone in the cell supernatant of each group were measured using ELISA. The results showed that cell activity had significantly decreased following the heat stress treatments at 43℃, 44℃, and 45℃ (P < 0.01), respectively. Meanwhile, cell activities observed in Patchouli and Elsholtzia were found to be much better than those of heat stress group (P < 0.05). In addition, the expression levels of HSP70 in the follicular granulosa cells of Patchouli and Elsholtzia groups were lower than those of heat stress group. Patchouli and Elsholtzia can maintain expressions of the receptor at 43℃. This study determined that the estrogen and progesterone in the supernatant fluid of Patchouli and Elsholtzia were higher than those observed in heat stress. Therefore, the results obtained in this study indicated that the Patchouli and Elsholtzia treatments administered prior the heat stress experiments had successfully protected the follicular granulosa cells from heat damages while maintaining the normal secretory functions of the granulosa cells.

Increased accuracy of genomic predictions for growth under chronic thermal stress in rainbow trout by prioritizing variants from GWAS using imputed sequence data.

Through imputation of genotypes, genome‐wide association study (GWAS) and genomic prediction (GP) using whole‐genome sequencing (WGS) data are cost‐efficient and feasible in aquaculture breeding schemes. The objective was to dissect the genetic architecture of growth traits under chronic heat stress in rainbow trout (Oncorhynchus mykiss) and to assess the accuracy of GP based on imputed WGS and different preselected single nucleotide polymorphism (SNP) arrays. A total of 192 and 764 fish challenged to a heat stress experiment for 62 days were genotyped using a customized 1 K and 26 K SNP panels, respectively, and then, genotype imputation was performed from a low‐density chip to WGS using 102 parents (36 males and 66 females) as the reference population. Imputed WGS data were used to perform GWAS and test GP accuracy under different preselected SNP scenarios. Heritability was estimated for body weight (BW), body length (BL) and average daily gain (ADG). Estimates using imputed WGS data ranged from 0.33 ± 0.05 to 0.55 ± 0.05 for growth traits under chronic heat stress. GWAS revealed that the top five cumulatively SNPs explained a maximum of 0.94%, 0.86% and 0.51% of genetic variance for BW, BL and ADG, respectively. Some important functional candidate genes associated with growth‐related traits were found among the most important SNPs, including signal transducer and activator of transcription 5B and 3 (STAT5B and STAT3, respectively) and cytokine‐inducible SH2‐containing protein (CISH). WGS data resulted in a slight increase in prediction accuracy compared with pedigree‐based method, whereas preselected SNPs based on the top GWAS hits improved prediction accuracies, with values ranging from 1.2 to 13.3%. Our results support the evidence of the polygenic nature of growth traits when measured under heat stress. The accuracies of GP can be improved using preselected variants from GWAS, and the use of WGS marginally increases prediction accuracy.

Nutritional Intervention Strategies Using Dietary Antioxidants and Organic Trace Minerals to Reduce the Incidence of Wooden Breast and Other Carcass Quality Defects in Broiler Birds.

Wooden breast (WB) is a degenerative myopathy seen in modern broiler birds resulting in quality downgrade of breast fillets. Affected filets show increased toughness both before as well as after cooking and have decreased water holding capacity and marinade pick up compared to normal fillets. Although the exact etiology is unknown, the circulatory insufficiency and increased oxidative stress in the breast muscles of modern broiler birds could be resulting in damage and degeneration of muscle fibers leading to myopathies. Three independent experiments were conducted to evaluate the effect of various dietary interventions on the incidence of WB when birds are exposed to oxidative stress associated with feeding oxidized fat and mild heat stress. Feed additives such as dietary antioxidant [Ethoxyquin (ETX)], mineral methionine hydroxy analog chelate (MMHAC) of Zn, Cu, and Mn, and organic selenium (Org Se) were tested at recommended levels. In experiment 1, ETX reduced (P < 0.05) the incidence of severe WB induced by oxidized fat diet. The magnitude of improvement in percentage of normal (no WB) filets and reduction in muscle lipid peroxidation was greater (P < 0.05) when ETX and MMHAC were fed together as shown by experiment 2. In birds exposed to mild heat stress (Experiment 3), feeding MMHAC by itself reduced (P < 0.05) tissue damage by reducing incidence of tibial head lesions, skin scratches, breast blisters, in addition to increasing the incidence of normal (no WB) fillets. When MMHAC was combined with ETX and Org Se, further improvement (P < 0.05) in normal (no WB) filets was observed. In summary, under different oxidative stress conditions, dietary intervention programs that contain ETX, MMHA-Zn, -Cu, and -Mn and Org Se can improve performance and increase carcass integrity, reducing problems, such as WB, either independently or with additive effect. This effect is most likely attained by simultaneously improving the exogenous and endogenous antioxidant status, reducing oxidative stress, and improving tissue healing process of the bird.

Correction to: Thirty years of coral heat-stress experiments: a review of methods

Correction to: Thirty years of coral heat-stress experiments: a review of methods

An overview of heat stress in tomato (Solanum lycopersicum L.).

Heat stress has been defined as the rise of temperature for a period of time higher than a threshold level, thereby permanently affecting the plant growth and development. Day or night temperature is considered as the major limiting factor for plant growth. Earlier studies reported that night temperature is an important factor in the heat reaction of the plants. Tomato cultivars capable of setting viable fruits under night temperatures above 21 °C are considered as heat-tolerant cultivars. The development of breeding objectives is generally summarized in four points: (a) cultivars with higher yield, (b) disease resistant varieties in the 1970s, (c) long shelf-life in 1980s, and (d) nutritive and taste quality during 1990s. Some unique varieties like the dwarf “Micro-Tom”, and the first transgenic tomato (FlavrSavr) were developed through breeding; they were distributed late in the 1980s.High temperature significantly affects seed, pollen viability and root expansion. Researchers have employed different parameters to evaluate the tolerance to heat stress, including membrane thermo stability, floral characteristics (Stigma exertion and antheridia cone splitting), flower number, and fruit yield per plant. Reports on pollen viability and fruit set/plant under heat stress by comparing the pollen growth and tube development in heat-treated and non-heat-stressed conditions are available in literature. The electrical conductivity (EC) have been used to evaluate the tolerance of some tomato cultivars in vitro under heat stress conditions as an indication of cell damage due to electrolyte leakage; they classified the cultivars into three groups: (a) heat tolerant, (b) moderately heat tolerant, and (c) heat sensitive.It is important to determine the range in genetic diversity for heat tolerance in tomatoes. Heat stress experiments under field conditions offer breeders information to identify the potentially heat tolerant germplasm.

Heat stress-induced HSP90 expression is dependent on ERK and HSF1 activation in turbot (Scophthalmus maximus) kidney cells.

Mitogen-activated protein kinases (MAPKs) and heat shock proteins (HSPs) are ubiquitous proteins that are functional mediators in both normal and stressed states of the cell. In this study, we performed heat stress (37 °C) experiments on turbot kidney (TK) cells. Heat stress expression patterns of HSP90, as well as the expression and phosphorylation levels of extracellular-regulated signal kinases (ERKs) and the transcription factor HSF1 and c-Fos, were examined. The results show that heat stress activates ERK1/2 and HSF1, and induces HSP90 gene expression in TK cells. Inhibition of ERK activation attenuates heat stress-induced HSP90 gene expression. The double luciferase reporter gene experiment showed that HSF1 is an important transcription factor for heat-induced HSP90 gene expression. Likewise, c-Fos does not directly regulate the heat-induced expression of HSP90 in turbot kidney cells. To our knowledge, this is the first study to report a signaling pathway that regulates the heat shock response in turbot cells. Our results may facilitate an understanding of the underlying molecular mechanisms of the cellular stress response in marine fish.

The Seagrass Methylome Is Associated With Variation in Photosynthetic Performance Among Clonal Shoots.

Evolutionary theory predicts that clonal organisms are more susceptible to extinction than sexually reproducing organisms, due to low genetic variation and slow rates of evolution. In agreement, conservation management considers genetic variation as the ultimate measure of a population’s ability to survive over time. However, clonal plants are among the oldest living organisms on our planet. Here, we test the hypothesis that clonal seagrass meadows display epigenetic variation that complements genetic variation as a source of phenotypic variation. In a clonal meadow of the seagrass Zostera marina, we characterized DNA methylation among 42 shoots. We also sequenced the whole genome of 10 shoots to correlate methylation patterns with photosynthetic performance under exposure to and recovery from 27°C, while controlling for somatic mutations. Here, we show for the first time that clonal seagrass shoots display DNA methylation variation that is independent from underlying genetic variation, and associated with variation in photosynthetic performance under experimental conditions. It remains unknown to what degree this association could be influenced by epigenetic responses to transplantation-related stress, given that the methylomes showed a strong shift under acclimation to laboratory conditions. The lack of untreated control samples in the heat stress experiment did not allow us to distinguish methylome shifts induced by acclimation from such induced by heat stress. Notwithstanding, the co-variation in DNA methylation and photosynthetic performance may be linked via gene expression because methylation patterns varied in functionally relevant genes involved in photosynthesis, and in the repair and prevention of heat-induced protein damage. While genotypic diversity has been shown to enhance stress resilience in seagrass meadows, we suggest that epigenetic variation plays a similar role in meadows dominated by a single genotype. Consequently, conservation management of clonal plants should consider epigenetic variation as indicator of resilience and stability.

Heat stress responses and population genetics of the kelp Laminaria digitata (Phaeophyceae) across latitudes reveal differentiation among North Atlantic populations.

To understand the thermal plasticity of a coastal foundation species across its latitudinal distribution, we assess physiological responses to high temperature stress in the kelp Laminaria digitata in combination with population genetic characteristics and relate heat resilience to genetic features and phylogeography. We hypothesize that populations from Arctic and cold‐temperate locations are less heat resilient than populations from warm distributional edges. Using meristems of natural L. digitata populations from six locations ranging between Kongsfjorden, Spitsbergen (79°N), and Quiberon, France (47°N), we performed a common‐garden heat stress experiment applying 15°C to 23°C over eight days. We assessed growth, photosynthetic quantum yield, carbon and nitrogen storage, and xanthophyll pigment contents as response traits. Population connectivity and genetic diversity were analyzed with microsatellite markers. Results from the heat stress experiment suggest that the upper temperature limit of L. digitata is nearly identical across its distribution range, but subtle differences in growth and stress responses were revealed for three populations from the species’ ecological range margins. Two populations at the species’ warm distribution limit showed higher temperature tolerance compared to other populations in growth at 19°C and recovery from 21°C (Quiberon, France), and photosynthetic quantum yield and xanthophyll pigment responses at 23°C (Helgoland, Germany). In L. digitata from the northernmost population (Spitsbergen, Norway), quantum yield indicated the highest heat sensitivity. Microsatellite genotyping revealed all sampled populations to be genetically distinct, with a strong hierarchical structure between southern and northern clades. Genetic diversity was lowest in the isolated population of the North Sea island of Helgoland and highest in Roscoff in the English Channel. All together, these results support the hypothesis of moderate local differentiation across L. digitata's European distribution, whereas effects are likely too weak to ameliorate the species’ capacity to withstand ocean warming and marine heatwaves at the southern range edge.

Standardized short-term acute heat stress assays resolve historical differences in coral thermotolerance across microhabitat reef sites.

Coral bleaching is one of the main drivers of reef degradation. Most corals bleach and suffer mortality at just 1-2°C above their maximum monthly mean temperatures, but some species and genotypes resist or recover better than others. Here, we conducted a series of 18-hr short-term acute heat stress assays side-by-side with a 21-day long-term heat stress experiment to assess the ability of both approaches to resolve coral thermotolerance differences reflective of in situ reef temperature thresholds. Using a suite of physiological parameters (photosynthetic efficiency, coral whitening, chlorophyll a, host protein, algal symbiont counts, and algal type association), we assessed bleaching susceptibility of Stylophora pistillata colonies from the windward/exposed and leeward/protected sites of a nearshore coral reef in the central Red Sea, which had previously shown differential mortality during a natural bleaching event. Photosynthetic efficiency was most indicative of the expected higher thermal tolerance in corals from the protected reef site, denoted by an increased retention of dark-adapted maximum quantum yields at higher temperatures. These differences were resolved using both experimental setups, as corroborated by a positive linear relationship, not observed for the other parameters. Notably, short-term acute heat stress assays resolved per-colony (genotype) differences that may have been masked by acclimation effects in the long-term experiment. Using our newly developed portable experimental system termed the Coral Bleaching Automated Stress System (CBASS), we thus highlight the potential of mobile, standardized short-term acute heat stress assays to resolve fine-scale differences in coral thermotolerance. Accordingly, such a system may be suitable for large-scale determination and complement existing approaches to identify resilient genotypes/reefs for downstream experimental examination and prioritization of reef sites for conservation/restoration. Development of such a framework is consistent with the recommendations of the National Academy of Sciences and the Reef Restoration and Adaptation Program committees for new intervention and restoration strategies.

Thirty years of coral heat-stress experiments: a review of methods

For over three decades, scientists have conducted heat-stress experiments to predict how coral will respond to ocean warming due to global climate change. However, there are often conflicting results in the literature that are difficult to resolve, which we hypothesize are a result of unintended biases, variation in experimental design, and underreporting of critical methodological information. Here, we reviewed 255 coral heat-stress experiments to (1) document where and when they were conducted and on which species, (2) assess variability in experimental design, and (3) quantify the diversity of response variables measured. First, we found that two-thirds of studies were conducted in only three countries, three coral species were more heavily studied than others, and only 4% of studies focused on earlier life stages. Second, slightly more than half of all heat-stress exposures were less than 8 d in duration, only 17% of experiments fed corals, and experimental conditions varied widely, including the level and rate of temperature increase, light intensity, number of genets used, and the length of acclimation period. In addition, 95%, 55%, and > 35% of studies did not report tank flow conditions, light–dark cycle used, or the date of the experiment, respectively. Finally, we found that 21% of experiments did not measure any bleaching phenotype traits, 77% did not identify the Symbiodiniaceae endosymbiont, and the contribution of the coral host in the physiological response to heat-stress was often not investigated. This review highlights geographic, taxonomic, and heat-stress duration biases in our understanding of coral bleaching, and large variability in the reporting and design of heat-stress experiments that could account for some of the discrepancies in the literature. Development of some best practice recommendations for coral bleaching experiments could improve cross-studies comparisons and increase the efficiency of coral bleaching research at a time when it is needed most.

Thermotolerance of camel (Camelus dromedarius) somatic cells affected by the cell type and the dissociation method.

Researchers dealing with heat stress experiments use different cell kinds and use trypsin that has been reported to affect the cellular proteins of cultured cells. Therefore, we compared the effects of acute and chronic exposures to high temperature (45 °C) on camel skin fibroblast and granulosa cells. Primary culture of fibroblasts and granulosa cells tolerated the acute heat shock for 2 h; however, granulosa cells cultured for long duration (20 h) showed thermotolerance when compared with the fibroblasts. Moreover, the effect of cell dispersion method (trypsin and mechanical dissociation) on the thermotolerance of sub-cultured cells was examined. Trypsin altered the morphology of fibroblasts and granulosa cells exposed to 45 °C for 4 h. Moreover, trypsin significantly reduced the fibroblast and granulosa cell migration in the wound healing assay. The current results demonstrate that cell passaging and cell type can affect the thermotolerance of the cells; it also revealed that trypsin could alter the cellular response to the heat shock. We raise the demand for another alternative method for cell dispersion in experiments dealing with cellular responses to the heat shock.

Characterizing the heat response of Arabidopsis thaliana from the perspective of codon usage bias and translational regulation

mRNA translation is carefully regulated at both the initiation and the elongation step. Under heat stress, it is known that particular genes change their expression and translation levels to respond to the environment. Attention has been paid to the detailed mechanisms of how a few proteins work, and little is done to analyze whether the global evolutionary patterns affect the translational changes. Determinants like codon usage bias and its related evolutionary features are less studied in heat stress experiments, especially for plants.Utilizing the RNA-seq and Ribo-seq data of normal and heat-stressed Arabidopsis thaliana generated from a previous study, we conducted gene-level (global) and codon-resolution (local) translation analyses. We studied how codon usage bias and other evolutionary features could impact the translation patterns in the heat response of the plant.We found that the evolutionary features including codon usage bias, tAI, nitrogen cost, and conservation (identity) could affect the global and local translation efficiency. Under heat stress, the optimal and conserved codons are more likely to alter their local translation elongation speed to modulate the global translation of host genes. Meanwhile, we also verified the widely accepted notions that the secondary structures and proline codons could largely slow down the translation rate.Our results revealed the effect of codon usage bias and other evolutionary patterns on the translation regulation under heat stress. Unveiling the effect of these features on translational regulation of plants might be helpful in understanding the relationship and interaction between plants and the environment.

Comparison of the abilities of vegetation indices and photosynthetic parameters to detect heat stress in wheat

With the changing of the climate, the detection of heat stress as early as possible has become increasingly important for wheat (Triticum aestivum L.) production. Previous studies have demonstrated that photosynthetic parameters can serve as indicators of the stress conditions, and vegetation indices (VIs) provide the ability to non-destructively monitor photosynthetic parameters. However, it remains unclear whether VIs can be used to detect heat stress in a similar manner as the photosynthetic parameters. In addition, the optimal VIs for indicating heat stress and detecting the stress status are also currently unknown. In the present study, a heat stress experiment was designed with four temperature levels [T1, 17 °C/27 °C (Tmin/Tmax), T2 (25 °C/35 °C), T3 (29 °C/39 °C), and T4 (33 °C/43 °C)] and three treatment durations [three days (D1), six days (D2) and nine days (D3)]. Three photosynthetic parameters [leaf chlorophyll content (LCC), net photosynthesis rate (Pn), and maximum efficiency of photosystem II (Fv/Fm)] and 17 published VIs were selected to compare their sensitivity and assess their feasibility for detecting heat stress. The results showed that Fv/Fm was the most sensitive photosynthetic parameter to heat stress and had the ability to indicate the start and end of heat stress at the slight level or the early stage. The chlorophyll index-red edge (CIred-edge), normalized difference red edge index (NDRE) and photochemical reflectance index (PRI) were sensitive to heat stress owing to their close relationships with photosynthetic parameters. Among these three VIs, PRI displayed the highest sensitivity. Nevertheless, the sensitivity of PRI was less than that of Fv/Fm, and it failed to detect the beginning and end of heat stress lasting for three days. The ability of PRI to detect heat stress became similar to that of Fv/Fm when the duration of heat stress was increased to seven days. In conclusion, Fv/Fm is the optimum indicator for detecting early-stage heat stress, in which only the photosynthetic functions change. In contrast, PRI, a non-destructive indicator, works well to indicate relatively late-stage heat stress, in which the chemical and physical characteristics of leaves (e.g., chlorophyll content) are affected.

Effects of corn particle size on nutrient utilization in pigs evaluated under optimal and heat stress conditions

The effects of corn particle size on nutrient digestibility and energy utilization in pigs were determined under optimal (experiment 1, 25 ± 1 °C) or heat stress (experiment 2, 37 ± 1 °C) conditions. In Exp. 1 and 2, five experimental diets were tested using a 5 × 5 Latin square design involving five barrows (Landrace × Yorkshire × Duroc, average initial body weight of 30 ± 1 kg and 45.0 ± 1.8 kg, respectively, in individual metabolic cages). Dietary treatments were as follows: 200-, 300-, 400-, 600-, 800-μm corn particle sizes obtained by mesh screens. Under optimal thermal conditions, digestibility of dry matter (DM) and crude fiber (CF) from 200-μm diet was higher (P < 0.05) compared to that from the 300-μm and 400-μm diets. The digestibility of crude protein (CP) and ether extract (EE) was the highest (P < 0.05) at the 200-μm particle size. The apparent total tract digestibility of energy was significantly higher (P < 0.05) on the 200-μm diet. Under heat stress, digestibility of CF when corn was ground to 600 μm was higher (P < 0.05) compared to 300 and 400 μm. Digestibility of NDF and ADF was the highest (P < 0.05) at 600-μm corn particle size. In conclusion, grinding corn to 200-μm corn particles had a positive effect on DM, CP, EE, and CF under optimal thermal condition, while the 600-μm corn particle size had positive effects on digestibility of CF, NDF, and ADF than 200-μm corn particle size under heat stress.