Primary Stress Response Research Articles

Deep phenotyping reveals CRH and FKBP51-dependent behavioral profiles following chronic social stress exposure in male mice.

The co-chaperone FKBP51, encoded by FKBP5 gene, is recognized as a psychiatric risk factor for anxiety and depressive disorders due to its crucial role in the stress response. Another key modulator in stress response regulation is the corticotropin releasing hormone (CRH), which is co-expressed with FKBP51 in many stress-relevant brain-regions and cell-types. Together, they intricately influence the balance of the hypothalamic-pituitary-adrenal (HPA) axis, one of the primary stress response systems. Previous research underscores the potential moderating effects these genes have on the regulation of the stressful life events towards the vulnerability of major depressive disorder (MDD). However, the specific function of FKBP51 in CRH-expressing neurons remains largely unexplored. Here, through deep behavioral phenotyping, we reveal heightened stress effects in mice lacking FKBP51 in CRH co-expressing neurons (CRHFKBP5-/-), particularly evident in social contexts. Our findings highlight the importance of considering cell-type specificity and context in comprehending stress responses and advocate for the utilization of machine-learning-driven phenotyping of mouse models. By elucidating these intricacies, we lay down the groundwork for personalized interventions aimed at enhancing stress resilience and individual well-being.

ATR inhibition activates cancer cell cGAS/STING-interferon signaling and promotes antitumor immunity in small-cell lung cancer.

Patients with small-cell lung cancer (SCLC) have poor prognosis and typically experience only transient benefits from combined immune checkpoint blockade (ICB) and chemotherapy. Here, we show that inhibition of ataxia telangiectasia and rad3 related (ATR), the primary replication stress response activator, induces DNA damage-mediated micronuclei formation in SCLC models. ATR inhibition in SCLC activates the stimulator of interferon genes (STING)-mediated interferon signaling, recruits T cells, and augments the antitumor immune response of programmed death-ligand 1 (PD-L1) blockade in mouse models. We demonstrate that combined ATR and PD-L1 inhibition causes improved antitumor response than PD-L1 alone as the second-line treatment in SCLC. This study shows that targeting ATR up-regulates major histocompatibility class I expression in preclinical models and SCLC clinical samples collected from a first-in-class clinical trial of ATR inhibitor, berzosertib, with topotecan in patients with relapsed SCLC. Targeting ATR represents a transformative vulnerability of SCLC and is a complementary strategy to induce STING-interferon signaling-mediated immunogenicity in SCLC.

Physiological assessment of the effects of experimental capture and transport in Pacific spiny dogfish

Sharks are collected from the wild as live specimens for display in aquariums and use in research. The methods by which live sharks are captured and transported can be associated with sub-lethal physiological outcomes or even mortality, which requires evaluation to determine optimal collection and transport methods for a given species. The present study evaluated the primary (i.e., endocrine) and secondary (i.e., biochemical) stress responses and mortality in Pacific spiny dogfish (Squalus suckleyi) following capture and subsequent transport between experimental rod-and-reel angling and longline fishing. Dogfish were resilient to transport following capture by either method, exhibiting no at-vessel mortality and 0.03% delayed mortality. Longline captured dogfish exhibited signs of primary (i.e., elevated corticosterone) and secondary (i.e., elevated plasma potassium, osmolality, glucose, and lactate; decreased blood pH) stress responses, whereas dogfish that experienced rod-and-reel capture and transport only exhibited a secondary stress response (i.e., elevated plasma potassium, osmolality, and lactate; decreased blood pH). Together, these data demonstrate that Pacific spiny dogfish are robust to capture and transport using experimental hook-and-line fishing techniques.

Multifunctional role of dietary copper to regulate stress-responsive gene for mitigation of multiple stresses in Pangasianodon hypophthalmus

It is an urgent needs to address climate change and pollution in aquatic systems using suitable mitigation measures to avoid the aquatic animals' extinction. The vulnerability and extinction of the aquatic animals in the current scenario must be addressed to enhance safe fish food production. Taking into consideration of such issues in fisheries and aquaculture, an experiment was designed to mitigate high temperature (T) and low pH stress, as well as arsenic (As) pollution in fish using copper (Cu) containing diets. In the present investigation, the Cu-containing diets graded with 0, 4, 8, and 12 mg kg-1 were prepared and fed to Pangasianodon hypophthalmus reared under As, low pH, and high-temperature stress. The gene expression was highly affected in terms of the primary, secondary, and tertiary stress response, whereas supplementation of Cu-containing diet mitigates the stress response. Oxidative stress genes such as catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) were significantly upregulated by stressors (As, As + T, and As + pH + T). Whereas, heat shock protein (HSP 70), inducible nitric oxide synthase (iNOS), metallothionine (MT), caspase 3a (Cas 3a), and cytochrome P450 (CYP 450) were highly upregulated by stressors, while dietary Cu at 8 mg kg-1 diet significantly downregulated these gene expressions. Indeed, the immunity-related genes viz. TNFα, Ig, TLR, and immune-related attributes viz. albumin, globulin, total protein, A:G ratio, blood glucose, NBT, and myeloperoxidase (MPO) were also improved with Cu-containing diets. Cu containing diets substantially improved neurotransmitter enzyme (AChE) and vitamin C (Vit C). DNA damage was also reduced with supplementation of Cu at 8 mg kg-1 diet. The growth index viz. final body weight gain (%), specific growth rate, protein efficiency ratio, food conversion ratio, relative feed intake, and daily growth index were noticeably enhanced by Cu diets (4 and 8 mg kg-1 diet). The growth-related genes expressions viz. growth hormone (GH), growth hormone regulator 1 (Ghr1), growth hormone regulator β (Ghrβ,) myostatin (MYST), and somatostatin (SMT) supported the growth enhancement with Cu at 8 mg kg-1 diet. The bioaccumulation of As was reduced with Cu-containing diets. The fish were infected with Aeromonas hydrophila at the end of the 105 days experimental trial. Cu at 8 mg kg-1 diet improved immunity, reduced the cumulative mortality, and enhanced the relative percentage survival of the fish. The results revealed that the innovative Cu diets could reduce the extinction of the fish against climate change and pollution era and produce the safest production that is safe to humans for consumption.

Secondary Stress Response of Tilapia (Oreochromis Niloticus) in Different Doses of Hydrogen Peroxide (H2O2) During the Transport Period

Fish transportation is a crucial step in maintaining fish quality during the shipping process from one region to another. The closed-wet system, which is commonly used for fish transportation, has several obstacles in the form of fish stress and mortality. Stressed fish generate primary, secondary, and tertiary stress responses. The purpose of this study is to assess the secondary stress response of tilapia during transportation by giving different doses of hydrogen peroxide. 3% hydrogen peroxide applied to water resulting in three different doses i.e. 0.3 ml.L-1 , 0.1 ml.L-1 , 0.08 ml.L-1 and without hydrogen peroxide used as control groups. Data was obtained by measuring blood glucose levels and the ventilation rate of tilapia. The results showed an alteration in blood glucose levels and ventilation rate of tilapia during transportation. Treatment with 0.1 ml.L-1 hydrogen peroxide gave the lowest glucose levels during 10 hours of transportation. Changes in ventilation rate of tilapia were observed in the treatment with 0.1 ml.L-1 of hydrogen peroxide which indicated miled stress conditions. It can be concluded that the optimal dose of hydrogen peroxide in the transportation process is 0.1 ml.L-1 of hydrogen peroxide.

Evaluation of the effects of exogenous cortisol manipulation and the glucocorticoid antagonist, RU486, on the exploratory tendency of bluegill sunfish (Lepomis macrochirus)

In teleost fishes, activation of the hypothalamic-pituitary-interrenal axis leads to an elevation of circulating cortisol levels as a primary stress response. While acute elevation of cortisol is generally beneficial, long-term elevation, a common characteristic of chronic stress, may lead to detrimental effects on health and physiological performance in fishes. Some stress-mediated behavioural shifts, such as variation along the shy-boldness axis in fish, may influence individual fitness. The present study evaluated the role of cortisol and its mechanisms of action in the exploratory behaviour of the bluegill sunfish (Lepomis macrochirus). Fish were implanted with cocoa butter alone (sham treatment), or cocoa butter containing cortisol, or cortisol and the glucocorticoid receptor antagonist, RU486. A control (untreated) group was also used. Animals were held for 48 h following treatment and then were subjected to a Z-maze trial to characterize the exploratory behaviour. Cortisol treatment had no measurable effect on the exploratory behaviour of bluegill sunfish. Despite presenting a higher probability of refuge emergence, fish treated with cortisol combined with RU486 behaved similarly to cortisol-treated and control groups. While these results suggest that cortisol may not be involved in the mechanisms controlling boldness, the influence of cortisol elevation across longer time periods plus validation in different contexts will be necessary to confirm this conclusion.

Short-term cortisol adaption to discrimination and Mexican-origin adolescents' mental and sleep health.

Discrimination experiences are a salient contributor to the health disparities facing Latina/x/o youth. The biopsychosocial model of minority health posits that discrimination influences health through wear and tear on the biological stress responses, including the hypothalamic-pituitary-adrenal (HPA) axis, which is a primary stress response system in the body. Emerging evidence suggests that discrimination alters the secretion of cortisol, the end product of the HPA axis, yet, whether the daily processes between discrimination and diurnal cortisol response influence mental and sleep health remains unanswered. This study integrated daily diary and post-diary survey data to examine whether daily diurnal cortisol responses to discrimination influence adolescents' mental (depressive symptoms, anxiety) and sleep (sleep quality, duration) health in a sample of Mexican-origin youth (N = 282; M age = 17.10; 55% female). Results showed that adolescents who experienced more discrimination across the four-day diary period exhibited steeper diurnal cortisol slopes and lower evening cortisol; however, such physiological responses tended to be associated with poorer adolescents' mental and sleep health. The current study underscores the potential adaptation cost associated with short-term cortisol adaptation in the face of discrimination.

Physiological stress responses of European seabass Dicentrarchus labrax related to changing operations of floating net cages in intensive farming conditions at the Moroccan M'diq bay.

The present study purposed to quantify and compare in-situ the primary and secondary physiological stress responses, related to the operation of floating net-cages changing, in both sub-adults (523 days post hatching) and adults (916 dph) European sea bass Dicentrarchus labrax, under intensive farming conditions in the Moroccan M'diq Bay. Levels of cortisol, glucose, total cholesterol, total protein and lactate, as well as the percentage of hematocrit were measured before and after stress. The results showed significantly elevated levels of cortisol and blood glucose in both age groups, while total cholesterol and protein levels were unaffected. In fact, blood lactate was significantly reduced in sub-adults, while in adults this parameter was not affected by the operation. However, the hematocrit percentages measured after the operation were significantly higher than those found before the operation in both groups of fish, which is linked to the increased rate of oxygen renewal by the new net cages and the lower water temperature inside the cages. With regard to the age-specific response during this essential operation - before and after the operation - plasma cortisol, blood glucose and lactate concentrations, as well as plasma total protein levels, were significantly higher in sub-adults than in adults. It is concluded that aquaculture practices such as changing the aquaculture net-cage could have repercussions in terms of the classic physiological responses to stress in D. labrax. This article is protected by copyright. All rights reserved.

Differential Aluminum tolerance in Arabidopsis thaliana ecotypes is seemingly related to metabolite changes

Plants are negatively affected by aluminum (Al) in acidic soils (pH ≤ 5.0), which impairs root growth and ultimately plant yield. Organic acids are closely related to Al neutralization improving the metal tolerance with an expansive metabolic cost. Here, we investigated phenotypic, metabolic, and genetic responses of three Arabidopsis thaliana ecotypes, Columbia (Col-0), Wassilewskija (Ws) and Landsberg erecta (Ler) in response to Al. By comparing the respective control plants with Al-exposed plants, the ecotypes Col-0 and Ws displayed stronger reductions in root growth and reproductive yield than Ler. All ecotypes presented high expression of ALMT1, an Al-resistance associated gene. Further analyses revealed a large accumulation of diverse amino acids, carbohydrates and organic acids following Al stress in both shoot and root tissues of Col-0 and Ws plants, but not in Ler. Altogether, our results suggest that a higher capacity for using and translocating reduced carbon molecules seems crucial to overcoming Al stress in the Ler ecotype. In addition, lower expenses in carbon molecules might be linked to a higher capacity to deal with Al stress, supporting a tight relationship between primary metabolism and Al stress responses. Finally, novel insights on the influence of Al over plant growth and primary metabolism in both shoots and roots are described.

Chronic stress negatively impacts wound healing, welfare, and stress regulation in internally tagged Atlantic salmon (Salmo salar).

The desire to understand fish welfare better has led to the development of live monitoring sensor tags embedded within individuals for long periods. Improving and understanding welfare must not come at the cost of impaired welfare due to a tag's presence and implantation process. When welfare is compromised, the individual will experience negative emotions such as fear, pain, and distress, impacting the stress response. In this study, Atlantic salmon (Salmo salar) underwent surgical implantation of a dummy tag. Additionally, half of this group was introduced to daily crowding stress. Both groups and an untagged group were followed for 8weeks using triplicate tanks per group. Sampling took place once a week, and where stress was given, it was conducted 24h before sampling. Stress-related measurements were taken to understand if tagging caused chronic stress and explore the chronic stress response and its impact on wound healing. Primary stress response hormones measured included CRH, dopamine, adrenocorticotropic hormone, and cortisol. Secondary stress response parameters measured included glucose, lactate, magnesium, calcium, chloride, and osmolality. Tertiary stress response parameters measured included weight, length, and five fins for fin erosion. Wound healing was calculated by taking the incision length and width, the inflammation length and width, and the inside wound length and width. The wound healing process showed that stressed fish have a larger and longer-lasting inflammation period and a slower wound healing process, as seen from the inside wound. The tagging of Atlantic salmon did not cause chronic stress. In contrast, daily stress led to an allostatic overload type two response. ACTH was elevated in the plasma after 4weeks, and cortisol followed elevation after 6weeks, highlighting a breakdown of the stress regulation. Fin erosion was elevated alongside cortisol increase in the stressed group. This data suggests that tagging previously unstressed fish in a controlled environment does not negatively affect welfare regarding stress responses. It also indicates that stress delays wound healing and increases the inflammatory response, highlighting how continued stress causes a breakdown in some stress responses. Ultimately, the tagging of Atlantic salmon can be successful under certain conditions where proper healing is observed, tag retention is high, and chronic stress is not present, which could allow for the possible measurement of welfare indicators via smart-tags.

Effect of Handling Stress on Primary and Secondary Stress Responses of the Catfish, Clarias batrachus

Cortisol is a major hormone directly associated with stress in fish and is a reliable physiological indicator of primary stress response in fish, whereas glucose and osmolality are the indicators of secondary stress response in fish. This study explored the stress levels in the catfish, Clarias batrachus (Magur) by measuring the cortisol, glucose, and osmolality levels in plasma by exposing the fish to three different kinds of interventions namely, non-anaesthetized, anaesthetized, and stressed. No statistically significant changes were reported in the plasma cortisol, plasma glucose, and plasma osmolality levels when the blood samples were collected after the three interventions. These results indicated that Clarias batrachus is a sturdy fish, which can withstand routine laboratory handling, and that the blood samples can be collected without anaesthetization.

Impact of heat priming on heat shock responses in Origanum vulgare: Enhanced foliage photosynthetic tolerance and biphasic emissions of volatiles

Climate change enhances the frequency of heatwaves that negatively affect photosynthesis and can alter constitutive volatile emissions and elicit emissions of stress volatiles, but how pre-exposure to mildly warmer temperatures affects plant physiological responses to subsequent severe heat episodes remains unclear, especially for aromatic plants with high and complex volatile defenses. We studied the impact of heat shock (45 °C/5 min) applied alone and after exposure to moderate heat stress (35 °C/1 h, priming) on foliage photosynthesis and volatile emissions in the aromatic plant Origanum vulgare through 72 h recovery period. Heat stress decreased photosynthesis rates and stomatal conductance, whereas the reductions in photosynthesis were primarily due to non-stomatal factors. In non-primed plants, heat shock-induced reductions in photosynthetic activity were the greatest, but photosynthetic activity completely recovered by the end of the experiment. In primed plants, a certain inhibition of photosynthetic activity remained, suggesting a sustained priming effect. Heat shock enhanced the emissions of volatiles including lipoxygenase pathway volatiles, long-chained fatty acid-derived compounds, mono- and sesquiterpenes, geranylgeranyl diphosphate pathway volatiles, and benzenoids, whereas different heat treatments resulted in unique emission blends. In non-primed plants, stress-elicited emissions recovered at 72 h. In primed plants, volatile emissions were multiphasic, the first phase, between 0.5 and 10 h, reflected the primary stress response, whereas the secondary rise, between 24 and 72 h, indicated activations of different defense metabolic pathways. Our results demonstrate that exposure to mild heat leads to a sustained physiological stress memory that enhances plant resistance to subsequent severe heat stress episodes.

Comparative assessment of plasma cortisol and fecal corticoid metabolites (FCM) of Atlantic salmon (Salmo salar L.) subjected to acute- and long-term stress

Farmed Atlantic salmon (Salmo salar L.) are subject to a variety of stressors throughout production. Elevations of cortisol level in the blood are one of the major endocrine primary stress responses in vertebrates and are widely used as stress indicator. However, blood sampling is invasive and stressful procedures. Cortisol instantly released into the blood at sampling can easily interfere with the initial stress response of interest. Fecal corticoid metabolites (FCM) have been suggested for a less invasive assessment of stress in fish. In the present study, we evaluated stress responses by using plasma cortisol and FCM as stress indicators in Atlantic salmon. Atlantic salmon (with average weight 42 g) were exposed to 3 different stressors: 1) parr-smolt transformation; 2) infestation with salmon lice (Lepeophtheirus salmonis) for two-weeks; 3) infection with infectious salmon anaemia virus (ISAV) for four weeks. The results demonstrated that FCM levels correlated well with the plasma cortisol levels at single-point sampling during long-term stress (p < 0.05). Significant increases of both plasma cortisol and FCM were found two weeks after initiated 24 h light. Each tank was sampled twice, with around 40-min interval. The chasing during the first sampling was used as an acute stressor. The effects of sampling procedures on plasma cortisol and FCM levels are compared. Blood and feces were analyzed by an Enzyme-Linked Immunosorbent Assay (ELISA). Plasma cortisol and FCM levels increased within 40 min after fish perceived an acute stressor when the average weight of fish was below 100 g. Attenuated cortisol stress responses to acute stress were found after fish experiencing long-term stress. Collectively, FCM reflects changes in plasma cortisol and can be an alternative to analysis of plasma cortisol in Atlantic salmon, complementing evidence of using FCM as stress indicators for monitoring and improving fish welfare. The results highlight that standard sampling procedures and good experimental designs should be established to ensure robust and repeatable results in the study of stress based on the FCM measurement. Future study will investigate representative timepoints for FCM sampling, with particular attention to the types of stress and sizes of the fish.

Primary and Secondary Physiological Stress Responses of European Sea Bass (Dicentrarchus labrax) Due to Rearing Practices under Aquaculture Farming Conditions in M'diq Bay, Moroccan Mediterranean: The Case of Sampling Operation for Size and Weight Measurement.

Globally, aquaculture contributes to sustainable food and nutritional stability. However, stress conditions constitute a major threat affecting farmed-fish welfare and fish farms’ performances. In this regard, the present study was aimed at measuring and comparing in-situ (offshore) the physiological stress responses caused by recurrent sampling operations for length−weight measurement. Studied fish were European sea bass Dicentrarchus labrax sub-adults and adults reared in intensive farming conditions in M’diq Bay, on the Moroccan Mediterranean coast. The physiological stress response was evaluated by measuring blood biomarkers, including cortisol, glucose, lactate, total cholesterol and total proteins levels, and hematocrit percentage. The hypercortisolemia, hyperglycemia, hyperlactatemia and hypoproteinemia detected in the post-sampling state in both age groups of D. labrax indicated stress tendencies and a high sensitivity to aquaculture practice-related stress factors, with major and minor responses detected in the same age and same stress conditions. It is also interesting to note that the fish age and the time-course of the sampling operation had a statistically significant effect in terms of the physiological response (with p < 0.01 and p < 0.001, respectively). In conclusion, the present study showed that sea bass subjected to various stressful situations in intensive farming conditions displayed a physiological stress response specific to its age, to the individual status, as well as to the duration of stressor.

Economic Improvement of Artisanal Fishing by Studying the Survival of Discarded Plectorhinchus mediterraneus.

Europe calls for the end to fisheries discards, which means bringing all caught fish (subject to minimum sizes or quotas) to land. This decision is beneficial to the ecosystem, since it forces the selectivity of the fishing gears to improve. However, artisanal fishermen find themselves in a vulnerable situation where their subsistence depends on catches with small profit margins. An exemption to this landing obligation exists, as it is also ruled that those animals whose survival is scientifically guaranteed may be returned to the sea. Here we study the survival of Plectorhinchus mediterraneus captured by hookline and gillnet, as well as their physiological recovery. Survival exceeds 93% in both cases. The physiological assessment of primary (cortisol) and secondary (energy mobilization, acid-base and hydromineral balance, and immune system) stress responses indicates that surviving animals are able to recover after fishing. Thus, we propose the optimal size of capture of this species to achieve greater economic benefit. For this, we rely on the prices according to size in recent years, as well as on the growth curves of the species. In this way, by releasing fish of less than 1 kg, the current benefits could be multiplied between 2.3 and 9.6 times. This pilot study lays the groundwork for regulating artisanal fisheries through scientific data related to survival of discards along with information on the sale prices.

Toxicity of the disinfectant benzalkonium chloride (C14) towards cyanobacterium Microcystis results from its impact on the photosynthetic apparatus and cell metabolism

Quaternary ammonium compounds (QACs) are commonly used in a variety of consumer and commercial products, typically as a component of disinfectants. During the COVID-19 pandemic, QACs became one of the primary agents utilized to inactivate the SARS-CoV-2 virus on surfaces. However, the ecotoxicological effects of QACs upon aquatic organisms have not been fully assessed. In this study, we examined the effects of a widely used QAC (benzalkonium chloride-C14, BAC-14) on two toxigenic Microcystis strains and one non-toxigenic freshwater Microcystis strain and carried out an analysis focused on primary, adaptive and compensatory stress responses at apical (growth and photosynthesis) and metabolic levels. This analysis revealed that the two toxic Microcystis strains were more tolerant than the non-toxic strain, with 96 hr-EC50 values of 0.70, 0.76, and 0.38 mg/L BAC-14 for toxigenic M. aeruginosa FACHB-905, toxigenic M. aeruginosa FACHB-469, and non-toxigenic M. wesenbergii FACHB-908, respectively. The photosynthetic activities of the Microcystis, assessed via Fv/Fm values, were significantly suppressed under 0.4 mg/L BAC-14. Furthermore, this analysis revealed that BAC-14 altered 14, 12, and 8 metabolic pathways in M. aeruginosa FACHB-905, M. aeruginosa FACHB-469, and M. wesenbergii FACHB-908, respectively. It is noteworthy that BAC-14 enhanced the level of extracellular microcystin production in the toxigenic Microcystis strains, although cell growth was not significantly affected. Collectively, these data show that BAC-14 disrupted the physiological and metabolic status of Microcystis cells and stimulated the production and release of microcystin, which could result in damage to aquatic systems.

Physiological Effects of Recapture and Transport from Net-Cages in Lumpfish

Lumpfish are widely used for removing sea lice in salmonid sea-based aquaculture. If these fish are to be harvested and used for human consumption, it is necessary to know how the physical strain associated with removing the lumpfish from the net-cages affects the fish in the short-term, and if live-storage in tanks, well-boats, or nets awaiting slaughter, will result in stress and mortalities. In this study, we investigated the effect of physical stress and mortality in a group of lumpfish recaptured from commercial net-cages, transported to holding tanks, and stored for one week. In addition to cortisol (primary stress response), we analyzed ions directly related to osmoregulation (Na+ and Cl−), osmotic stress (Ca2+), and blood plasma pH as an indicator of a secondary stress response. The aim of the study was to increase the basic physiological understanding of the physiological effects of handling procedures and transport in lumpfish. Only minor, and temporary, effects on primary stress response and secondary stress response were seen in lumpfish recaptured from net-cages and transported to holding facilities, indicating that lumpfish cope well with short transport (here 5 h). These findings are important in a context where lumpfish are harvested for reuse, e.g., human consumption or processing, following their lice-eating stage in net-cages.

Characterizing the stress response in juvenile Delta smelt exposed to multiple stressors

The Delta Smelt (Hypomesus transpacificus), once an abundant fish endemic to the Sacramento-San Joaquin Estuary, is now on the brink of extinction. Due to the high sensitivity of this species, knowledge of their stress response will be vital to their future survival and sustainability. Understanding the magnitude and kinetics of cortisol induction in Delta Smelt will provide valuable information when interpreting the degree of environmentally relevant stressors, such as warming and predator exposure. As little is known about the primary stress response and cortisol dynamics in Delta Smelt, the first aim of this study was to measure basal and maximal whole-body cortisol prior to and following exposure to a sublethal and significant netting stress at 17 and 21 °C. Our findings reveal that juvenile Delta Smelt held at 21 °C display an exacerbated stress response and a reduction in available energy compared to fish held at 17 °C. There was no evidence of the secondary stress response to the netting stress as whole-body glucose and lactate levels in treatment groups remained similar to basal values. The second aim of this study was to investigate the effect of a largemouth bass (Micropterus salmoides) predator cue, which was found to induce a significant increase in cortisol relative to control levels in juvenile Delta Smelt. Indices such as cortisol can be used as bioindicators of stress in the field and results from this study suggest that moderate temperatures and reduced predation are optimal release conditions during hatchery-based supplementation to minimize stress to this highly sensitive species.

Acoustic Delicing of Atlantic Salmon (Salmo salar): Fish Welfare and Salmon Lice (Lepeophtheirus salmonis) Dynamics

Acoustic lice treatment (AcuLice) is a newly developed system, which uses a composite acoustic sound image with low-frequency sound to remove salmon lice (Lepeophtheirus salmonis) from Atlantic salmon (Salmo salar). This field study documents the stress effects on Atlantic salmon and the effect on salmon lice dynamics during large-scale use of the AcuLice system. The effect of the AcuLice treatment on salmon lice dynamics was measured by weekly salmon lice counting at the facilities from mid-summer 2019 to late-spring 2020. The number of salmon lice treatments in the same period was also compared to a reference group. In addition, the number of weeks until the first salmon lice treatment (mechanical treatment) was compared between the two groups. Apart from a slight increase in plasma glucose, no significant differences were observed in the primary, secondary, or tertiary stress responses measured. For the mature female salmon lice, a significantly lower number (mean ± SEM) was shown for the AcuLice group (0.24 ± 0.03) compared to the reference group (0.44 ± 0.04). In addition, a lower number (mean ± SEM) of salmon lice treatments and a longer production period before the first salmon lice treatment occurred was observed at the AcuLice facilities (33.2 ± 3 weeks) compared to the reference facilities (20.3 ± 2 weeks). These data suggest that the use of the AcuLice system reduces the need for traditional salmon lice treatments with no added stress to the fish.

The retrograde signaling regulator ANAC017 recruits the MKK9-MPK3/6, ethylene, and auxin signaling pathways to balance mitochondrial dysfunction with growth.

In plant cells, mitochondria are ideally positioned to sense and balance changes in energy metabolism in response to changing environmental conditions. Retrograde signaling from mitochondria to the nucleus is crucial for adjusting the required transcriptional responses. We show that ANAC017, the master regulator of mitochondrial stress, directly recruits a signaling cascade involving the plant hormones ethylene and auxin as well as the MAP KINASE KINASE (MKK) 9–MAP KINASE (MPK) 3/6 pathway in Arabidopsis thaliana. Chromatin immunoprecipitation followed by sequencing and overexpression demonstrated that ANAC017 directly regulates several genes of the ethylene and auxin pathways, including MKK9, 1-AMINO-CYCLOPROPANE-1-CARBOXYLATE SYNTHASE 2, and YUCCA 5, in addition to genes encoding transcription factors regulating plant growth and stress responses such as BASIC REGION/LEUCINE ZIPPER MOTIF (bZIP) 60, bZIP53, ANAC081/ATAF2, and RADICAL-INDUCED CELL DEATH1. A time-resolved RNA-seq experiment established that ethylene signaling precedes the stimulation of auxin signaling in the mitochondrial stress response, with a large part of the transcriptional regulation dependent on ETHYLENE-INSENSITIVE 3. These results were confirmed by mutant analyses. Our findings identify the molecular components controlled by ANAC017, which integrates the primary stress responses to mitochondrial dysfunction with whole plant growth via the activation of regulatory and partly antagonistic feedback loops.