Methods for estimating and modelling spruce budworm development rates at constant temperatures

We describe an experimental protocol for measuring the response of spruce budworm postdiapause larval development to temperature. This protocol is specifically designed to include measurements of development near their upper and lower thermal thresholds. The application of this protocol to a laboratory colony allowed for the first experimental evidence that spruce budworm larval development occurs at temperatures as low as 5 °C and as high as 35 °C, and it provides data to fit stage-specific development models. Our protocol is also designed to minimise mortality near the thermal development thresholds, thus allowing for multigenerational studies. We observed developmental plasticity in larvae reared at constant temperatures, particularly the occurrence of up to 42% of some individuals requiring only five instars to complete development compared to the expected six instars. The occurrence exhibited no clear relation to temperature. Although this protocol is specifically designed for spruce budworm, it provides a template for the study of other species’ developmental responses to temperature.

Ocular Surface Pathology in Patients Suffering from Mercury Intoxication

Purpose: To report the ocular surface pathology of patients suffering from acute/subacute mercury vapor intoxication. Design: Cross-sectional study. Participants: Male workers intoxicated with inorganic mercury referred for ophthalmic involvement and healthy control subjects. Methods: The following tests were performed: dry eye (DE)-related symptoms indicated by the ocular surface disease (OSDI) index questionnaire; tear osmolarity; analysis of 23 tear cytokine concentrations and principal component and hierarchical agglomerative cluster analyses; tear break-up time (T-BUT); corneal fluorescein and conjunctival lissamine green staining; tear production by Schirmer and tear lysozyme tests; mechanical and thermal corneal sensitivity (non-contact esthesiometry); and corneal nerve analysis and dendritic cell density by in vivo confocal microscopy (IVCM). Results: Twenty-two out of 29 evaluated patients entered the study. Most had DE-related symptoms (OSDI values > 12), that were severe in 63.6% of them. Tear osmolarity was elevated (>308 mOsms/L) in 83.4% of patients (mean 336.23 (28.71) mOsm/L). Corneal and conjunctival staining were unremarkable. T-BUT was low (<7 s) in 22.7% of patients. Schirmer test and tear lysozyme concentration were low in 13.6% and 27.3% of cases, respectively. Corneal esthesiometry showed patient mechanical (mean 147.81 (53.36) mL/min) and thermal thresholds to heat (+2.35 (+1.10) °C) and cold (−2.57 (−1.24) °C) to be significantly higher than controls. Corneal IVCM revealed lower values for nerve density ((6.4 (2.94) n/mm2), nerve branching density (2 (2.50) n/mm2), and dendritic cell density (9.1 (8.84) n/mm2) in patients. Tear levels of IL-12p70, IL-6, RANTES, and VEGF were increased, whereas EGF and IP-10/CXCL10 were decreased compared to controls. Based on cytokine levels, two clusters of patients were identified. Compared to Cluster 1, Cluster 2 patients had significantly increased tear levels of 18 cytokines, decreased tear lysozyme, lower nerve branching density, fewer dendritic cells, and higher urine mercury levels. Conclusions: Patients suffering from systemic mercury intoxication showed symptoms and signs of ocular surface pathology, mainly by targeting the trigeminal nerve, as shown by alterations in corneal sensitivity and sub-basal nerve morphology.

Lengthening of Warm Periods Increased the Intensity of Warm-season Marine Heatwaves Over the Past Four Decades

Marine heatwaves (MHWs), periods of anomalously warm sea surface temperature (SST) which can have significant impacts on marine ecosystems, have increased in frequency and severity over recent decades. Many coastal systems (e.g. coral reefs) are particularly vulnerable to warm-season heat stress when temperature can exceed organisms’ thermal thresholds and lead to mass mortality. While many studies have examined the change of the warm-season heat stress occurrence over time, e.g., for coral reefs, there has been less analysis of the thermal structure of heat stress events. Here we examined the trend in the characteristics of warm-season heat stress (referred to as warm-season MHWs) at the global-scale from 1985 to 2019, using multiple metrics for each of duration, peak intensity, accumulated heat stress, heating rates and level of intensity. The results showed that warm-season MHWs have become more frequent, longer-lasting, featured higher peak intensity and accumulated heat stress across most of the ocean over the past four decades. Furthermore, decomposition of the trends in warm-season MHWs structure showed that the increased accumulated heat stress was predominantly driven by the increased duration rather than the increased intensity, especially in the western and central equatorial Pacific. The results contribute to improving the understanding of warm-season MHWs, which may help inform the prediction of their impacts on marine ecosystems as well as marine conservation and management under climate change.

Marine Heatwaves Exceed Cardiac Thermal Limits of Adult Sparid Fish (Diplodus capensis, Smith 1884)

Climate change not only drives increases in global mean ocean temperatures, but also in the intensity and duration of marine heatwaves (MHWs), with potentially deleterious effects on local fishes. A first step to assess the vulnerability of fishes to MHWs is to quantify their upper thermal thresholds and contrast these limits against current and future ocean temperatures during such heating events. Heart failure is considered a primary mechanism governing the upper thermal limits of fishes and begins to occur at temperatures where heart rate fails to keep pace with thermal dependency of reaction rates. This point is identified by estimating the Arrhenius breakpoint temperature (TAB), which is the temperature where maximum heart rate (fHmax) first deviates from its exponential increase with temperature and the incremental Q10 breakpoint temperature (TQB), which is where the Q10 temperature coefficient (relative change in heart rate for a 10°C increase in temperature) for fHmax abruptly decreases during acute warming. Here we determined TAB, TQB and the temperature that causes cardiac arrhythmia (TARR) in adults of the marine sparid, Diplodus capensis, using an established technique. Using these thermal indices results, we further estimated adult D. capensis vulnerability to contemporary MHWs and increases in ocean temperatures along the warm-temperate south-east coast of South Africa. For the established technique, we stimulated fHmax with atropine and isoproterenol and used internal heart rate loggers to measure fHmax under conditions of acute warming in the laboratory. We estimated average TAB, TQB, and TARR values of 20.8°C, 21.0°C, and 28.3°C. These findings indicate that the physiology of D. capensis will be progressively compromised when temperatures exceed 21.0°C up to a thermal end-point of 28.3°C. Recent MHWs along the warm-temperate south-east coast, furthermore, are already occurring within the TARR threshold (26.6–30.0°C) for cardiac function in adult D. capensis, suggesting that this species may already be physiologically compromised by MHWs. Predicted increases in mean ocean temperatures of a conservative 2.0°C, may further result in adult D. capensis experiencing more frequent MHWs as well as a contraction of the northern range limit of this species as mean summer temperatures exceed the average TARR of 28.3°C.

Comprehensive Phenotyping of Cutaneous Afferents Reveals Rapid-Onset Alterations in Nociceptor Response Properties Following Spinal Cord Injury

Spinal cord injury (SCI) is a complex syndrome that has profound effects on patient well-being, including the development of medically-resistant chronic pain. The mechanisms underlying SCI pain have been the subject of thorough investigation but still remain poorly understood. While the majority of the research has focused on changes occurring within and surrounding the site of injury in the spinal cord, there is now a consensus that alterations within the peripheral nervous system, namely sensitization of nociceptors, contribute to the development and maintenance of chronic SCI pain. Here we demonstrate that thoracic spinal contusion injury results in the emergence of autotomy and spasticity, both indicators of spontaneous pain, in areas below the level of the injury within 24 hr of SCI. These behaviors were associated with hindpaw edema and elevated cutaneous calcitonin gene-related peptide (CGRP) concentration. Electrophysiological recordings using an ex vivo skin/nerve/DRG/spinal cord preparation demonstrated that SCI increased mechanical and thermal sensitivity, as well as the incidence of spontaneous activity (SA) and afterdischarge (AD), in below-level C-fiber nociceptors 24 hr following injury. Interestingly, the distribution of nociceptors that exhibit SA and AD are not identical, and the development of SA was observed more frequently in nociceptors with low thermal thresholds, while AD was found more frequently in nociceptors with high thermal thresholds. These results demonstrate that SCI causes rapid-onset of peripheral inflammation-like processes that sensitize nociceptors, which may contribute to the early emergence and persistence of chronic SCI pain.

Plasticity in parental effects confers rapid larval thermal tolerance in the estuarine anemone Nematostella vectensis

ABSTRACTParental effects can prepare offspring for different environments and facilitate survival across generations. We exposed parental populations of the estuarine anemone, Nematostella vectensis, from Massachusetts to elevated temperatures and quantified larval mortality across a temperature gradient. We found that parental exposure to elevated temperatures resulted in a consistent increase in larval thermal tolerance, as measured by the temperature at which 50% of larvae die (LT50), with a mean increase in LT50 of 0.3°C. Larvae from subsequent spawns returned to baseline thermal thresholds when parents were returned to normal temperatures, indicating plasticity in these parental effects. Histological analyses of gametogenesis in females suggested that these dynamic shifts in larval thermal tolerance may be facilitated by maternal effects in non-overlapping gametic cohorts. We also compared larvae from North Carolina (a genetically distinct population with higher baseline thermal tolerance) and Massachusetts parents, and observed that larvae from heat-exposed Massachusetts parents had thermal thresholds comparable to those of larvae from unexposed North Carolina parents. North Carolina parents also increased larval thermal tolerance under the same high-temperature regime, suggesting that plasticity in parental effects is an inherent trait for N. vectensis. Overall, we find that larval thermal tolerance in N. vectensis shows a strong genetic basis and can be modulated by parental effects. Further understanding of the mechanisms behind these shifts can elucidate the fate of thermally sensitive ectotherms in a rapidly changing thermal environment.

Plasma levels of a methadone constant rate infusion and their corresponding effects on thermal and mechanical nociceptive thresholds in dogs

Background The present study aimed to collect pharmacokinetic data of a methadone continuous rate infusion (CRI) and to investigate its effect on mechanical and thermal nociceptive thresholds. Seven, 47 to 54 months old beagle dogs, weighing 9.8 to 21.2 kg, were used in this experimental, randomized, blinded, placebo-controlled crossover study. Each dog was treated twice with either a methadone bolus of 0.2 mg kg− 1 followed by a 0.1 mg kg− 1 h− 1 methadone CRI (group M) or an equivalent volume of isotonic saline solution (group P) for 72 h. Mechanical and thermal thresholds, as well as vital parameters and sedation were measured during CRI and for further 24 h. Blood samples for methadone plasma concentrations were collected during this 96 h period. Results Percentage thermal excursion (%TE) increased significantly from baseline (BL) until 3 h after discontinuation of CRI in M. Within P and between treatment groups differences were not significant. Mechanical threshold (MT) increased in M until 2 h after CRI discontinuation. Bradycardia and hypothermia occurred in M during drug administration and dogs were mildly sedated for the first 47 h. Decreased food intake and regurgitation were observed in M in five and four dogs, respectively. For methadone a volume of distribution of 10.26 l kg− 1 and a terminal half-life of 2.4 h were detected and a clearance of 51.44 ml kg− 1 min− 1 was calculated. Effective methadone plasma concentrations for thermal and mechanical antinociception were above 17 ng ml− 1. Conclusion A methadone CRI of 0.1 mg kg− 1 h− 1 for 3 days after a loading dose results in steady anti-nociceptive effects in an acute pain model in healthy dogs. Main side effects were related to gastrointestinal tract, hypothermia, bradycardia and sedation.

Contrasting heat stress response patterns of coral holobionts across the Red Sea suggest distinct mechanisms of thermal tolerance

Corals from the northern Red Sea, in particular the Gulf of Aqaba (GoA), have exceptionally high bleaching thresholds approaching >5°C above their maximum monthly mean (MMM) temperatures. These elevated thresholds are thought to be due to historical selection, as corals passed through the warmer Southern Red Sea during re-colonization from the Arabian Sea. To test this hypothesis, we determined thermal tolerance thresholds of GoA versus Central Red Sea (CRS) Stylophora pistillata corals using the Coral Bleaching Automated Stress System (CBASS) to run a series of standardized acute thermal stress assays. Relative thermal thresholds of GoA and CRS corals were indeed similar and exceptionally high (~7°C above MMM). However, absolute thermal thresholds of CRS corals were on average 3°C above those of GoA corals. To explore the mechanistic underpinnings, we determined gene expression response and microbiome dynamics of coral holobiont compartments. Transcriptomic responses differed markedly, with a strong response to the thermal stress in GoA corals versus a remarkably muted response in corals from the CRS. This pattern was recapitulated in the algal symbionts that showed site-specific genetic differentiation. Concomitant to this, a subset of coral and algal genes showed temperature-induced expression in GoA corals, while exhibiting fixed high expression, i.e. front-loading, in CRS corals. Bacterial community composition of GoA corals changed dramatically under heat stress, whereas CRS corals displayed consistent assemblages, indicating distinct microbial response patterns. Our work demonstrates distinct patterns underlying thermal tolerance across spatial scales, even for the same species and ocean basin. We interpret the response of GoA corals as that of a resilient population approaching a tipping point in contrast to a pattern of consistently elevated thermal resistance in CRS corals that cannot further attune. Such response differences suggest distinct thermal tolerance mechanisms that affect the response of coral populations to ocean warming.