Livestock-Forest integrated system attenuates deleterious heat stress effects in bovine oocytes.

Global warming poses significant challenges to the fertility of tropical dairy cattle. One promising approach to mitigate heat stress effects on reproductive function and reduce the carbon footprint is the use of integrated livestock-forest (ILF) systems. The aim of this study was to investigate the effects of two different systems, namely Full Sun (FS) and ILF, on maternal hyperthermia and oocyte quality of Holstein and Girolando heifers during the tropical summer season. The temperature-humidity index (THI) data revealed intense heat stress during the experiment. Both the system (P<0.01) and the breed (P<0.01) factors had a significant impact on vaginal temperature, being hyperthermia more pronounced in the FS system and in the Holstein breed. Over the five time points collected at a 33-day interval, we observed distinct patterns for ILF (P=0.65) and FS (P<0.001) systems, suggesting an adaptive response in animals kept in FS systems. Furthermore, oocyte quality assessment revealed an effect of the system for oocyte diameter (P<0.001) and levels of IGFBP2 (P<0.001), and caspase 3 levels showed a decrease in ILF compared to FS for both Holstein (P<0.001) and Girolando (P<0.001) breeds. Collectively, these parameters indicate that oocyte quality during the summer months was superior in animals maintained in the ILF system. In conclusion, the ILF system demonstrated promising results in attenuating maternal hyperthermia and mitigating its effects on oocyte quality. Additionally, our observations suggest that animals in the FS system may exhibit an adaptive response to heat stress.

The lack of plasticity and interspecific variability in thermal limits produce a highly heat-tolerant tropical host-parasitoid system.

Thermal limits are often used as proxies to assess the vulnerability of ectotherms to environmental change. While meta-analyses point out a relatively low plasticity of heat limits and a large interspecific variability, only few studies have compared the heat tolerance of interacting species. The present study focuses on the thermal limits, and their plasticity (heat hardening), of three species co-occurring in Western Africa: two ectoparasitoid species, Dinarmus basalis (Rondani) (Hymenoptera: Pteromalidae) and Eupelmus vuilleti (Crawford) (Hymenoptera: Eupelmidae), and their common host, Callosobruchus maculatus (F.) (Coleoptera: Bruchidae). The investigation delves into the Critical Thermal Maximum (CTmax), representing the upper tolerance limit, to understand how these species may cope with extreme thermal events. The CTmax of all three species appeared similarly high, hovering around 46.5 °C, exceeding the global mean CTmax observed in insects by 3.5 °C. Short-term exposure to moderate heat stress showed no impact on CTmax, suggesting a potential lack of heat hardening in these species. Therefore, we emphasized the similarity of heat tolerance in these interacting species, potentially stemming from both evolutionary adaptations to high temperatures during development and the stable and similar microclimate experienced by the three species over the years. While the high thermal tolerance should allow these species to endure extreme temperature events, the apparent lack of plasticity raises concerns about their ability to adapt to future climate change scenarios. Overall, this research provides valuable insights into the thermal physiology of these interacting species, providing a basis for understanding their responses to climate change and potential implications for the host-parasitoid system.

Association analysis of HSP90AA1 polymorphism with thermotolerance in tropically adapted Indian crossbred cattle.

Raising cattle is a lucrative business that operates globally but is confronted by many obstacles, such as thermal stress, which results in substantial monetary losses. A vital role of heat shock proteins (HSPs) is to protect cells from cellular damage. HSP90 is a highly prevalent, extremely adaptable gene linked to physiological resilience in thermal stress. This study aimed to find genetic polymorphisms of the HSP90AA1 gene in Karan Fries cattle and explore their relationship to thermal tolerance and production traits. One SNP (g.3292 A > C) was found in the Intron 8 and three SNPs loci (g.4776 A > G, g.5218T > C and g.5224 A > C) were found in the exon 11 of 100 multiparous Karan Fries cattle. The association study demonstrated that the SNP1-g.3292 A > C was significantly (P < 0.01) linked to the variables respiratory rate (RR), heat tolerance coefficient (HTC) and total milk yield (TMY (kg)) attributes. There was no significant correlation identified between any of the other SNP sites (SNP2-g.4776 A > G; SNP3-g.5218T > C; SNP4-g.5224 A > C) with the heat tolerance and production attributes in Karan Fries cattle. Haploview 4.2 and SHEsis software programs were used to analyse pair linkage disequilibrium and construct haplotypes for HSP90AA1. Association studies indicated that the Hap3 (CATA) was beneficial for heat tolerance breeding in Karan Fries cattle. In conclusion, genetic polymorphisms and haplotypes in the HSP90AA1 were associated with thermal endurance attributes. This relationship can be utilized as a beneficial SNP or Hap marker for genetic heat resistance selection in cow breeding platforms.

Large potential impacts of marine heatwaves on ecosystem functioning.

Ocean warming is driving significant changes in the structure and functioning of marine ecosystems, shifting species' biogeography and phenology, changing body size and biomass and altering the trophodynamics of the system. Particularly, extreme temperature events such as marine heatwaves (MHWs) have been increasing in intensity, duration and frequency. MHWs are causing large-scale impacts on marine ecosystems, such as coral bleaching, mass mortality of seagrass meadows and declines in fish stocks and other marine organisms in recent decades. In this study, we developed and applied a dynamic version of the EcoTroph trophodynamic modelling approach to study the cascading effects of individual MHW on marine ecosystem functioning. We simulated theoretical user-controlled ecosystems and explored the consequences of various assumptions of marine species mortality along the food web, associated with different MHW intensities. We show that an MHW can lead to a significant biomass reduction of all consumers, with the severity of the declines being dependent on species trophic levels (TLs) and biomes, in addition to the characteristics of MHWs. Biomass of higher TLs declines more than lower TLs under an MHW, leading to changes in ecosystem structure. While tropical ecosystems are projected to be sensitive to low-intensity MHWs, polar and temperate ecosystems are expected to be impacted by more intense MHWs. The estimated time to recover from MHW impacts is twice as long for polar ecosystems and one-third longer for temperate biomes compared with tropical biomes. This study highlights the importance of considering extreme weather events in assessing the effects of climate change on the structures and functions of marine ecosystems.

Laser-Induced Stress-Driven Nanoplate Jumping Visualized by Ultrafast Electron Microscopy.

Understanding laser-induced jumping has attracted great interest in nanomaterial launching and transfer but requires a high spatiotemporal resolution visualization. Here, we report a jumping dynamics of nanoplate driven by ultrafast laser-induced stress using time-resolved transmission electron microscopy. Single-shot imaging reveals a nondestructive launching of gold nanoplates in several nanoseconds after the pulsed femtosecond laser excitation. The temperature rise and acoustic vibration, derived from ultrafast electron crystallography with a picosecond time resolution, confirm the existence of a laser-induced elastic stress wave. The generation, propagation, and reflection of thermal stress waves are further clarified by atomic simulation. The nonequilibrium ultrafast laser heating produces a compressive stress wave within several picoseconds, constrained by the supporting substrate under nanoplate to provide thrust force. This compressive stress is subsequently reflected into tensile stress by the substrate, promoting the nanoplate to jump off the substrate. Furthermore, the uneven interface adhesion results in the jumping flip of nanoplates, as well as, diminished their jumping speed. This study unveils the jumping regime driven by impulsive laser-excited stress and offers understanding of light-matter interaction.

Seasonal Proteome Variations in Orbicella faveolata Reveal Molecular Thermal Stress Adaptations.

Although seasonal water temperatures typically fluctuate by less than 4 °C across most tropical reefs, sustained heat stress with an increase of even 1 °C can alter and destabilize metabolic and physiological coral functions, leading to losses of coral reefs worldwide. The Caribbean region provides a natural experimental design to study how corals respond physiologically throughout the year. While characterized by warm temperatures and precipitation, there is a significant seasonal component with relative cooler and drier conditions during the months of January to February and warmer and wetter conditions during September and October. We conducted a comparative abundance of differentially expressed proteins with two contrasting temperatures during the cold and warm seasons of 2014 and 2015 in Orbicella faveolata, one of the most important and affected reef-building corals of the Caribbean. All presented proteoforms (42) were found to be significant in our proteomics differential expression analysis and classified based on their gene ontology. The results were accomplished by a combination of two-dimensional gel electrophoresis (2DE) to separate and visualize proteins and mass spectrometry (MS) for protein identification. To validate the differentially expressed proteins of Orbicella faveolata at the transcription level, qRT-PCR was performed. Our data indicated that a 3.1 °C increase in temperature in O. faveolata between the cold and warm seasons in San Cristobal and Enrique reefs of southwestern Puerto Rico was enough to affect the expression of a significant number of proteins associated with oxidative and heat stress responses, metabolism, immunity, and apoptosis. This research extends our knowledge into the mechanistic response of O. faveolata to mitigate thermal seasonal temperature variations in coral reefs.

Multiscale Simulation of Laser-Based Direct Energy Deposition (DED-LB/M) Using Powder Feedstock for Surface Repair of Aluminum Alloy.

Laser-based direct energy deposition (DED-LB/M) has been a promising option for the surface repair of structural aluminum alloys due to the advantages it offers, including a small heat-affected zone, high forming accuracy, and adjustable deposition materials. However, the unequal powder particle size during powder-based DED-LB/M can cause unstable flow and an uneven material flow rate per unit of time, resulting in defects such as pores, uneven deposition layers, and cracks. This paper presents a multiscale, multiphysics numerical model to investigate the underlying mechanism during the powder-based DED-LB/M surface repair process. First, the worn surfaces of aluminum alloy components with different flaw shapes and sizes were characterized and modeled. The fluid flow of the molten pool during material deposition on the worn surfaces was then investigated using a model that coupled the mesoscale discrete element method (DEM) and the finite volume method (FVM). The effect of flaw size and powder supply quantity on the evolution of the molten pool temperature, morphology, and dynamics was evaluated. The rapid heat transfer and variation in thermal stress during the multilayer DED-LB/M process were further illustrated using a macroscale thermomechanical model. The maximum stress was observed and compared with the yield stress of the adopted material, and no relative sliding was observed between deposited layers and substrate components.

Alleviating Coral Thermal Stress via Inoculation with Quorum Quenching Bacteria.

In the background of global warming, coral bleaching induced by elevated seawater temperature is the primary cause of coral reef degradation. Coral microbiome engineering using the beneficial microorganisms for corals (BMCs) has become a hot spot in the field of coral reef conservation and restoration. Investigating the potential of alleviating thermal stress by quorum quenching (QQ) bacteria may provide more tools for coral microbial engineering remediation. In this study, QQ bacteria strain Pseudoalteromonas piscicida SCSIO 43740 was screened among 75 coral-derived bacterial strains, and its quorum sensing inhibitor (QSI) compound was isolated and identified as 2,4-di-tert-butylphenol (2,4-DTBP). Then, the thermal stress alleviating potential of QQ bacteria on coral Pocillopora damicornis was tested by a 30-day controlled experiment with three different treatments: control group (Con: 29 °C), high temperature group (HT: 31 °C), and the group of high temperature with QQ bacteria inoculation (HTQQ: 31 °C + QQ bacteria). The results showed that QQ bacteria SCSIO 43740 inoculation can significantly mitigate the loss of symbiotic algae and impairment of photosynthesis efficiency of coral P. damicornis under thermal stress. Significant difference in superoxide dismutase (SOD) and catalase (CAT) enzyme activities between HT and HTQQ was not observed. In addition, QQ bacteria inoculation suppressed the coral microbial community beta-dispersion and improved the stability of microbial co-occurrence network under thermal stress. It was suggested that QQ bacteria inoculation can alleviate coral thermal stress via reshaping microbial interaction and maintain community stability of coral microbiome. This study provided new evidence for the probiotic function of QQ bacteria in corals, which shedding light on the development of new microbiological tools for coral reef conservation.

Influence of Cold Wave Diversities on Thermal Stress and Thermal Fatigue Life of Asphalt Mixture.

Apart from low-temperature cracking, asphalt pavement may also suffer from thermal fatigue cracking. To clarify the impact of cold waves on the thermal fatigue performance of asphalt mixtures, the typical atmospheric temperature characteristics of different regions in China from 2012 to 2019 were analyzed, and the frequency of cold waves in these regions was determined. The viscoelastic parameters of an asphalt mixture are determined through an indirect tensile relaxation test. The thermal stress of the asphalt mixture is simulated and analyzed by using the finite element method. The effect of cold waves on the thermal fatigue life of the asphalt mixture was evaluated. The results show that the frequency of cold waves is different from region to region in China, and the cold waves mainly occurred from October of one year to February of the next year. Northeast China has the most frequency and the largest temperature drop amplitude, followed by North China. The maximum thermal stress increases with the decrease in temperature drop and initial temperature and is unrelated to the duration of cold waves. The thermal stress calculated based on the atmospheric thermal boundary is higher than the value using the road surface temperature. The thermal fatigue lives of asphalt mixtures in North China and Northeast China are very short, while the thermal fatigue life of the mixture in Central China is the longest. To meet the requirement of thermal fatigue damage caused by cold waves during the designed service stage, the recommended threshold for thermal stress is 0.39-0.77 MPa.

Influence of Monochromatic Light during Incubation on the Production and Metabolism of Low-Temperature Broiler Chicks.

The use of artificial lighting during the incubation phase is a tool that has been studied with the aim of increasing the production rates and hatchability. Using this, this study aims to investigate the effects of the luminous incidence of white and red monochromatic light on the production and metabolism of broiler chicks subjected to low temperatures. A total of 315 eggs of Ross 708 heavy breeders were used. The eggs were distributed randomly, with 35 eggs per tray, totaling 105 eggs per incubator. The treatments were the following: incubation without the use of light; the use of white monochromatic light; and the use of red monochromatic light. The lamps used were of the LED type. The samples were distributed in the factorial completely randomized experimental design with position effect on the tray. Candling, egg weighing, calculating the probability of survival and egg weight loss were performed. Temperatures were recorded using a thermographic camera. At birth, three chicks per tray were euthanized for evaluation: weight with and without yolk residue, gastrointestinal tract biometry, and blood and liver biochemistry. Analyses were performed using the R computational program. It was observed that there was a significant effect of the treatments on the levels of calcium, phosphorus, cholesterol, amylase, glucose, urea and glutamate pyruvate transaminase on the biochemical profile of the blood and on the thermographic temperatures of the eggs; the experiment was kept at low temperatures resulting in thermal stress, with an average temperature of 34.5 °C. Therefore, the use of red and white monochromatic light in the artificial incubation process for brown-colored eggs is not recommended, because in the post-hatching phase, it promoted the metabolism dysregulation on the blood biochemical profile to control the differentiation in the wavelength of traditional incubation.

A Practical Approach for Determination of Thermal Stress and Temperature-Dependent Material Properties in Multilayered Thin Films.

Multilayered thin films are essential to most microelectro-mechanical systems (MEMSs). The reliability and predictability of the behavior of such systems, especially when intended for usage at high temperatures or in harsh environments, demand the consideration of thermo-mechanical properties of the individual films of the multilayer arrangement during the design stage. This paper introduces a newly derived analytical model for the convenient indirect determination of the temperature-dependent Young's modulus and the thermally induced stress of individual layers within a multilayered thin film system, i.e., a multilayer-adapted Stoney equation. It is based on sample curvature measurement and requires data from only a single experiment. Experimental and numerical investigations of the new models are carried out using a five-layered sample of a RuAl metallization system developed for wireless high-temperature acoustic sensing. The results highlight the usability of the new model in practical MEMS analysis, enabling insights into complex layer stacks by overcoming current experimental limitations.

Heat stress in symbiotic dinoflagellates: Implications on oxidative stress and cellular changes.

Global warming has been shown to harmfully affect symbiosis between Symbiodiniaceae and other marine invertebrates. When symbiotic dinoflagellates (the genus Breviolum) were in vitro exposed to acute heat stress of +7 °C for a period of 5 days, the results revealed the negative impact on all physiological and other cellular parameters measured. Elevated temperatures resulted in a severe reduction in algal density of up to 9.5-fold, as well as pigment concentrations, indicating the status of the physiological stress and early signs of photo-bleaching. Reactive oxygen species (ROS) were increased in all heated dinoflagellate cells, while the antioxidant-reduced glutathione levels initially dropped on day one but increased under prolonged temperature stress. The cell viability parameters were reduced by 97 % over the heating period, with an increased proportion of apoptotic and necrotic cells. Autofluorescence (AF) for Cy5-PE 660-20 was reduced from 1.7-fold at day 1 to up to 50-fold drop at the end of heating time, indicating that the AF changes were highly sensitive to heat stress and that it could be an extremely sensitive tool for assessing the functionality of algal photosynthetic machinery. The addition of the drug 5-AZA-2'-deoxycytidine (5-AZA), which inhibits DNA methylation processes, was assessed in parallel and contributed to some alterations in algal cellular stress response. The presence of drug 5-AZA combined with the temperature stress had an additional impact on Symbiodiniaceae density and cell complexity, including the AF levels. These variations in cellular stress response under heat stress and compromised DNA methylation conditions may indicate the importance of this epigenetic mechanism for symbiotic dinoflagellate thermal tolerance adaptability over a longer period, which needs further exploration. Consequently, the increased ROS levels and changes in AF signals reported during ongoing heat stress in dinoflagellate cells could be used as early stress biomarkers in these microalgae and potentially other photosynthetic species.

Improved Research on Two-Step Thermal Stress Calculation Method for Asphalt Mixture: Extended Creep Compliance Test.

The two-step thermal stress calculation method (TTSCM) is commonly used to predict the cracking temperature of asphalt mixture. The aim of this study is to improve TTSCM's mathematical model so as to enhance its prediction accuracy. First, this study evaluated the errors of predicted cracking temperatures of original TTSCM for AC-16 and AC-25 asphalt mixtures by thermal stress-restrained specimen test (TSRST). Then, an improved method called the extended creep compliance test (ECCT) was developed to modify the TTSCM. The test results show that the cracking predictions of the original TTSCM are not always accurate. Particularly for AC-16 asphalt mixture, the predicted cracking temperature is 2.9 °C (-10.6%) higher than the measured value by the TSRST. The ECCT method has been proven to be an effective way to enhance the prediction accuracy of the TTSCM. The predicted cracking temperatures modified by the ECCT method for both asphalt mixtures are relatively accurate, having an error within ±2%. The ECCT method changed the calculated thermal stress values at different temperatures of the TTSCM; however, they still conformed to a basic changing trend with respect to the initial temperature and cooling rate. Finally, a recommendation regarding the ECCT method was presented.

Thermal Comfort of Nelore Cattle (Bos indicus) Managed in Silvopastoral and Traditional Systems Associated with Rumination in a Humid Tropical Environment in the Eastern Amazon, Brazil.

The objective of this study was to evaluate the thermal comfort of Nelore cattle (Bos indicus) managed in silvopastoral and traditional systems associated with rumination behavior in a humid tropical environment in the Eastern Amazon, Brazil. The study was carried out on a rural property in Mojuí dos Campos, Pará, Brazil, during the transition period of the year, from June to July 2023. Over these two months, six consecutive data collection days were held. We selected 20 clinically healthy non-castrated male Nelore cattle, aged between 18 and 20 months, with an average weight of 250 kg and body condition score of 3.5 (1-5). These animals were randomly divided into two groups: traditional system (TS) and silvopastoral system (SS). The physiological variables evaluated included RR, RT and BST. The variables were analyzed using the linear mixed model. For agrometeorological variables, higher values were observed between 10:00 a.m. (33 °C) and 6:00 p.m. (30 °C), with the highest temperature observed at 4:00 p.m. (40 °C). The RR showed interactions (p = 0.0214) between systems and times; in general, higher RR were obtained in the Traditional. The animals' RT showed no significant difference (p < 0.05) between the production systems, but there was a statistically significant difference in relation to the time of collection (p < 0.0001). In the BGHI, it was possible to observe that there was mild stress in the period from 22:00 at night to 6:00 in the morning and moderate stress in the period of greatest increase in temperature, from 10:00 in the morning to 18:00 at night. BST showed no statistical difference between the regions studied or between the SP (35.6 °C) and TS (36.25 °C) systems. RT in the TS showed a positive correlation with AT (r = 0.31507; p = 0.0477). RT in the SP showed a positive correlation with THI (r = 35583; p = 0.0242). On the other hand, RT in the SP (r = 0.42873; p = 0.0058) and ST (r = 0.51015; p = 0.0008) showed a positive correlation with BGHI. RR in the TS showed a positive correlation with BGHI (r = 0.44908; p = 0.0037). The greatest amounts of rumination were carried out by animals in the SP system, generally ruminating lying down (p < 0.05). With regard to rumination behavior in the morning and afternoon, there were higher numbers of WS and LD in the TS (p > 0.05). Most of the time, the cattle were LD during the morning and afternoon shifts, and at night and dawn they were WS in the TS. Therefore, the SP offers more thermal comfort advantages compared to the TS system.

Effects of red-light irradiation and melatonininjection on the antioxidant capacity and occurrence of apoptosis in abalones (Haliotis discus hannai) subjected to thermal stress.

High ocean temperatures caused by global warming induce oxidative stress in aquatic organisms. Melatonin treatment and irradiation using red light-emitting diodes (LEDs) have been reported to reduce oxidative stress in a few aquatic organisms. However, the effects of red LED irradiation and melatonin injection on the antioxidant capacity and degree of apoptosis in abalones, which are nocturnal organisms, have not yet been reported. In this study, we compared the expression levels of antioxidant enzymes, total antioxidant capacity, and the degree of apoptosis in abalones subjected to red LED irradiation and melatonin treatment. The results revealed that at high water temperatures (25 °C), the mRNA expression levels of the superoxide dismutase (SOD) and glutathione peroxidase (GPx) genes and the antioxidant activity of SOD decreased in abalones in the red-LED irradiated and melatonin-treated groups compared with those in abalones in the control group. Although high water temperatures induced DNA damage in the abalone samples, the degree of apoptosis was lower in the red-LED irradiated and melatonin-treated groups than in the control group. Overall, the abalones in the melatonin-treated and red-LED irradiated groups showed reduced oxidative stress and increased antioxidant enzyme levels under thermal stress compared with those in the control group. Therefore, red LED irradiation is a promising alternative to melatonin treatment, which is difficult to administer continuously for a long time, for protecting abalones from oxidative stress.

Screening and Identification of the Biomarkers Applied for the Evaluation of Acute and Chronic Thermal Tolerance Ability in Largemouth Bass (Micropterus salmoides).

Affected by the continuously rising temperature, thermal stress leads to a delinked growth rate and resistance to stress in cultured largemouth bass (Micropterus salmoides, LMB) in China. Identification of LMB with better thermal resistance will benefit the breeding of new varieties. However, there has been limited reporting on the evaluation to identify LMB with better thermal resistance. LMB consists of the northern LMB (Micropterus salmoides salmoides, NLMB) and the Florida LMB (Micropterus salmoides floridanus, FLMB). Due to their different geographical distributions, it has been suggested that FLMB exhibit better thermal resistance compared to NLMB. In this study, NLMB and FLMB were subjected to thermal stress for 3 h (acute) and 60 d (chronic) at 33 °C, respectively. Subsequently, the variations of 12 candidate biomarkers between NLMB and FLMB were analyzed. Exposure to acute thermal stress significantly increased plasma cortisol, blood glucose, and lactate levels; activities of superoxide dismutase (SOD), glutathione peroxidase (GPX), catalase (CAT), glucose kinase (GK), pyruvate kinase (PK), lactate dehydrogenase (LDH), and glucose 6 phosphatase (G6Pase); and the expressions of hsp70 and hsp90 in both NLMB and FLMB (p < 0.05). Compared to NLMB, FLMB exhibited a lower plasma cortisol level and a higher expression of hsp90 under acute thermal stress (p < 0.05). Exposure to chronic thermal stress significantly increased plasma cortisol and blood glucose levels, as well as activities of GK, PK, LDH, and G6Pase, as well as expressions of hsp70 and hsp90 in both NLMB and FLMB (p < 0.05). Additionally, FLMB showed a lower expression of hsp70 compared to NLMB (p < 0.05). In conclusion, our results showed that LMB with lower plasma cortisol level and higher expression of hsp90 under acute thermal stress, as well as lower expression of hsp70 under chronic thermal stress were suggested to have better thermal resistance. Our study provides valuable information for identifying and breeding LMB varieties with better thermal resistance in the future.

Experimental heatwaves reduce the effectiveness of ejaculates at occupying female reproductive tracts in a model insect.

Globally, heatwaves have become more common with hazardous consequences on biological processes. Research using a model insect (Tribolium castaneum) found that 5-day experimental heatwave conditions damaged several aspects of male reproductive biology, while females remained unaffected. However, females' reproductive fitness may still be impacted, as insects typically store sperm from multiple males in specialized organs for prolonged periods. Consequently, using males which produce sperm with green fluorescent protein (GFP)-tagged sperm nuclei, we visualized in vivo whether thermal stress affects the ejaculate occupancy across female storage sites under two scenarios; (i) increasing time since insemination and (ii) in the presence of defending competitor sperm. We reconfirmed that sperm from heatwave-exposed males sired fewer offspring with previously mated females and provided new scenarios for in vivo distributions of heat-stress-exposed males' sperm. Sperm from heatwave-exposed males occupied a smaller area and were at lower densities across the females' storage sites. Generally, sperm occupancy decreased with time since insemination, and sperm from the first male to mate dominated the long-term storage site. Reassuringly, although heated males' ejaculate was less successful in occupying female tracts, they were not lost from female storage at a faster rate and were no worse than control males in their offensive ability to enter storage sites occupied by competitor sperm. Future work should consider the potential site-specificity of factors influencing sperm storage where amenable.

Molecular characterization and function of hif1a and fih1 in response to acute thermal stress in American shad (Alosa sapidissima).

In order to evaluate the function of hypoxia-inducible factor-1 alpha (hif1α) and factor inhibiting hif1α (fih1) in response to thermal stress, we first conducted a functional analysis of A. sapidissima hif1α and fih1, and determined hif1α and fih1 expressions in different tissues in response to thermal stress based on identified housekeeping genes (HKGs). The results showed that hif1α and fih1 were mainly located in the nucleus and cytoplasm. The full length cDNA sequence of hif1α and fih1 was 4073 bp and 2759 bp, respectively. The cDNA sequence of hif1α includes 15 exons encoding 750 amino acid residues, and the full length cDNA sequence of fih1 contains 9 exons encoding 354 amino acid residues. During the acute thermal stress transferring from 16 ± 0.5 °C (control) to 20 ± 0.5 °C, 25 ± 0.5 °C, and 30 ± 0.5 °C for 15 min, it was found that the expression trends of hif1α and fih1 showed an inhibitory regulation in the heart, while they consistently expressed in brain, intestine, muscle, gill, kidney and liver. In conclusion, this is the first study to identify the tissue-specific HKGs in A. sapidissima and found that ef1α and ß-actin are the most suitable HKGs. Hif1α and Fih1 are mainly the nuclear and cytoplasmic proteins, respectively, having high levels in the heart and brain. Alosa sapidissima countered a temperature increase from 16 to 25 ℃ by regulating the expressions of hif1α and fih1, but their physiological regulatory functions were unable to cope with acute thermal stress when the temperature difference was 14 ℃ (from 16 to 30 ℃).

Heat exposure limits pentose phosphate pathway activity in bumblebees.

Bumblebee populations across the globe are experiencing substantial declines due to climate change, with major consequences for pollination services in both natural and agricultural settings. Using an economically important species, Bombus impatiens, we explored the physiological mechanisms that may cause susceptibility to extreme heat events. We tested the hypothesis that heat exposure limits the activity of the pentose phosphate pathway (PPP)-a parallel pathway to glycolysis that can use nectar sugar to generate antioxidant potential and combat oxidative stress. Using isotopically labelled glucose, we tracked PPP activity in B. impatiens at rest, during exercise and during a post-exercise recovery period under two different temperature regimes (22°C and 32°C). We found that the PPP is routinely used by B. impatiens at moderate temperatures, but that its activity is markedly reduced when ATP demands are high, such as during periods of exercise and heat exposure. We also exposed B. impatiens to either 22°C or 32°C for 5 hours and assessed levels of oxidative damage (lipid peroxidation, protein carbonyls) and antioxidant potential [reduced (GSH) and oxidized (GSSG) glutathione concentrations]. Interestingly, bees exhibited little oxidative damage after the thermal exposure, but we found a lower GSH:GSSG ratio in 32°C-exposed bees, reflecting lower antioxidant potential. Overall, our study demonstrates that acute heat stress severely limits PPP activity and may constrain antioxidant potential in B. impatiens. The repeated attenuation of this pathway in a warming climate may have more severe physiological consequences for this species, with potential implications for pollination services across North America.

Hot climate, hot koalas: the role of weather, behaviour and disease on thermoregulation.

Thermoregulation is critical for endotherms living in hot, dry conditions, and maintaining optimal core body temperature (Tb) in a changing climate is an increasingly challenging task for mammals. Koalas (Phascolarctos cinereus) have evolved physiological and behavioural strategies to maintain homeostasis and regulate their Tb but are thought to be vulnerable to prolonged heat. We investigated how weather, behaviour and disease influence Tb for wild, free-living koalas during summer in north-west New South Wales. We matched Tb with daily behavioural observations in an ageing population where chlamydial disease is prevalent. Each individual koala had similar Tb rhythms (average Tb = 36.4 ± 0.05°C), but male koalas had higher Tb amplitude and more pronounced daily rhythm than females. Disease disrupted the 24-hr circadian pattern of Tb. Koala Tb increased with ambient temperature (Ta). On the hottest day of the study (maximum Ta = 40.8°C), we recorded the highest (Tb = 40.8°C) but also the lowest (Tb = 32.4°C) Tb ever documented for wild koalas, suggesting that they are more heterothermic than previously recognized. This requires individuals to predict days of extreme Ta from overnight and early morning conditions, adjusting Tb regulation accordingly, and it has never been reported before for koalas. The large diel amplitude and low minimum Tb observed suggest that koalas at our study site are energetically and nutritionally compromised, likely due to their age. Behaviour (i.e. tree hugging and drinking water) was not effective in moderating Tb. These results indicate that Ta and koala Tb are strongly interconnected and reinforce the importance of climate projections for predicting the future persistence of koalas throughout their current distribution. Global climate models forecast that dry, hot weather will continue to escalate and drought events will increase in frequency, duration and severity. This is likely to push koalas and other arboreal folivores towards their thermal limit.