Effects of drinking water, sanitation, handwashing and nutritional interventions on stress physiology, oxidative stress, and epigenetic programming in young children living in rural Bangladesh: A randomized clinical trial

Importance: A regulated stress response is essential for healthy trajectories, but the integrated effects of early childhood environmental and nutritional interventions on stress physiology are unknown. Objective: To assess the effects of a combined nutritional, water, sanitation, and handwashing intervention on physiological stress response, oxidative stress, and DNA methylation. Design, Setting, and Participants: In a trial in rural Bangladesh, we randomized geographical clusters of pregnant women and their in-utero children into either the combined nutritional, water, sanitation, and handwashing intervention or the control group. Physiological stress response, oxidative stress, and methylation levels of 757 children were measured at ages one and two years. Analysis was intention-to-treat. Interventions: The intervention group received combined nutritional counseling and lipid-based nutrient supplements, chlorinated drinking water, upgraded sanitation, and handwashing with soap (N+WSH). The control group did not receive interventions. Main Outcomes and Measures: We measured four isomers of urinary F2-isoprostanes [iPF(2α)-III; 2,3-dinor-iPF(2α)-III; iPF(2α)-VI; 8,12-iso-iPF(2α)-VI] at year one. At year two, we measured pre- and post-stressor concentrations of salivary alpha-amylase and cortisol, overall methylation of the glucocorticoid receptor (NR3C1) exon 1F promoter including methylation levels at the nerve growth factor-inducible protein A (NGFI-A) binding site, mean arterial pressure, and resting heart rate. Results: Children in the N+WSH group had lower levels of F2-isoprostanes compared to controls (difference -0.16 to -0.19 log ng/mg of creatinine, P<0.01). Compared to the control group, post-stressor cortisol levels were elevated (0.24 log μg/dl; 95% CI, 0.07 to 0.4; P<0.01) and the residualized gain score for cortisol was higher (0.06 μg/dl; 95% CI, 0.01 to 0.12; P=0.023) in the N+WSH group. Children in the N+WSH group exhibited decreased methylation of the NGFI-A transcription factor binding site (-0.04 logit-transformed %; 95% CI, -0.08 to 0; P=0.037). Conclusions and Relevance: A nutritional, water, sanitation, and handwashing intervention reduced oxidative stress, enhanced hypothalamic-pituitary-adrenocortical axis activity, and reduced methylation levels in a transcription factor binding site of the glucocorticoid receptor gene. A targeted environmental and nutritional intervention affected the set point, reactivity, and regulation of the physiological stress system in early childhood, which may have implications for long-term health and developmental trajectories. Trial Registration: ClinicalTrials.gov NCT01590095

Stress and Pain Before, During and After the First Wave of the COVID-19 Pandemic: An Exploratory Longitudinal Mixed Methods Study

Aims: This study explores the association between subjective feeling of stress and pain experience in the context of the COVID-19 pandemic with a focus on characteristics known to trigger a physiological stress response [sense of low control, threat to ego, unpredictability and novelty (STUN)].Methods: This exploratory longitudinal convergent mixed methods design consisted of online questionnaires over three time points (before, during and after the 1st wave of the COVID-19 pandemic) (N = 49) and qualitative interviews (N = 27) during the 1st wave of the pandemic on distinct samples of individuals living with chronic pain (CP). Both types of data sources were mixed upon integration using joint display.Results: Mean pain intensity scores remained stable across time points, while pain unpleasantness and pain interference scores significantly improved. Global impression of change scores measured during the first wave of the pandemic do not entirely concord with pain scores evolution. Two thirds of participants reported a global deterioration of their pain condition at the beginning of the pandemic. Stress and pain catastrophizing before the pandemic were associated with pain scores throughout the pandemic; while most specific measures of stress due to the novel, uncontrollable, unpredictable and threatening nature of the pandemic were not. Qualitative data demonstrated that the deterioration reported in pain status reflected additional dimensions, including spatial expansion of the painful area, reduced access to treatments and challenges in adapting pain management strategies.Conclusions: Helping individuals to negotiate stressful aspects of the pandemic might help offset the negative impacts of stress on pain status in this context or other important life events.

Physiological response of weaned piglets to two transport durations observed in a Canadian commercial setting

Observational studies describing the impact of transport duration on weaned piglet welfare are limited. Current Canadian transport regulations are heavily informed by studies involving market hogs. Due to physiological differences between weaned piglets and market hogs, additional data on their response to transport are needed for age-specific evidence-based recommendations. A cohort study was conducted to describe and compare mortality, injury, weight change, hematological or biochemical changes in hydration, muscle injury and stress response observed in weaned piglets undergoing short duration (SD, &lt;3h), or long duration (LD, &gt;30 h) commercial summertime transport events. Data collection on 440 of 11,434 transported piglets occurred the morning of the day before transport (T0), at arrival (T1) and approximately three to four days (78-93h) after arrival at the nursery barn (T2). Low mortality occurred over all transport events (0.06%) with no association observed between transport duration and odds of death during transport (P=0.62). The incidence of lameness between T0 and T1 was low (1.84% of the 435 focal piglets scored) with all lameness cases identified as mild in severity. Lesions on ears and skin were more prevalent than other injury types after transport (T1) and may have been related to mixing aggression associated with weaning rather than transport alone. LD piglets weighed 0.39 kg less than SD piglets at T1 (P&lt;0.01), but no difference in group weight was observed at T2 (P=0.17). Hematological and biochemical differences were present between groups at T1. LD piglets had increased hematocrit levels compared to SD piglets (P=0.01) suggesting increased body water losses. SD piglets showed greater levels of muscle injury compared to LD piglets including elevated aspartate aminotransferase (P&lt;0.01) and creatine kinase (P&lt;0.01). However, these parameters were within normal reference ranges for piglets of this age group. Indicators of physiological stress response including cortisol and neutrophil to lymphocyte ratios were elevated in SD piglets compared to LD piglets (P=0.02 and P&lt;0.01, respectively). The results of this study demonstrate that both short and long transport durations can result in detectable physiological changes in weaned piglets. The overall impact of these durations on piglet welfare should be further explored by analyzing behavioural time budgets during and after transport.

Long-Tailed Pygmy Rice Rats Modify Their Behavioural Response and Faecal Corticosterone Metabolites in Response to Culpeo Fox but Not to Lesser Grison

Even though behavioural and physiological reactions to predation risk exhibited by prey species have received considerable attention in scientific journals, there are still many questions still unsolved. Our aim was to broaden the knowledge on one specific question: do long-tailed pygmy rice rats adapt their behavioural and physiological antipredator strategies depending on the predator species? For this question, we live-trapped in a temperate forest in Southern Chile long-tailed pygmy rice rats (Oligoryzomys longicaudatus), which were exposed to three predator odour phases (Phase 0: preliminary, no predator cues; Phase 1: one plot with culpeo fox faeces (Lycalopex culpaeus), one plot with lesser grison (Galictis cuja) faeces and one plot acting as a control with no odour; Phase 2: post treatment, no predator cues). We measured the behavioural response by the capture ratio. To assess the physiological stress response, we collected fresh faecal samples to quantify faecal corticosterone metabolites (FCM). Our results showed that O. longicaudatus increased both the capture ratio and FCM levels in the presence of culpeo cues. Culpeo foxes have higher densities in the study area than G. cuja and exhibit a higher activity pattern overlap with O. longicaudatus. Moreover, it has been also been reported in other regions that L. culpaeus consumption of O. longicaudatus is more frequent compared to G. cuja diet. The increase in capturability could be because traps can be regarded as a shelter in high-risk settings, but it can also be explained by the predator inspection behaviour. The increase in FCM concentrations during culpeo treatment can be linked to the adaptive mobilisation of energy to execute antipredator responses to increase survival chances.

Cortisol, Temperament and Serotonin in Karate Combats: An Evolutionary Psychobiological Perspective

Objectives There is evidence suggesting that in martial arts competitions athletes characterized by higher anxiety and harm avoidance may be more likely to lose a fight. This psychological profile has been hypothesized to explain in part the observation that cortisol is higher in losers before and in response to a competition. An important research target that needs further exploration is the identification of phenotypic traits that can be helpful in predicting athletes’ performance. Here we present a brief description of the theoretical bases that drives our research in the evolutionary psychobiology of sports and illustrate preliminary data on the relationship between the 5HTTLPR genotype, salivary cortisol, temperament and competition. Methods Sixty-five healthy male non-professional athletes provided saliva samples 10 min before and after a kumite session and filled out the Tridimensional Personality Questionnaire. Results Salivary cortisol levels 10 min before the competition were higher in losers and in athletes with the S allele. Temperament was associated with competition outcome and cortisol: losers were characterized by higher scores of harm avoidance and harm avoidance was positively correlated with cortisol levels. Conclusions The results confirm previous findings linking temperamental traits, pre-and post- competition physiological stress response with competition outcome in kumite fight. Moreover, they indicate an association between the 5HTTLPR polymorphism and pre-competition salivary cortisol, thus providing a preliminary but non-conclusive evidence on the role played by the 5HTTLPR genotype as a vulnerability factor in sport competition.

Connecting post-release mortality to the physiological stress response of large coastal sharks in a commercial longline fishery

Bycatch mortality is a major factor contributing to shark population declines. Post-release mortality (PRM) is particularly difficult to quantify, limiting the accuracy of stock assessments. We paired blood-stress physiology with animal-borne accelerometers to quantify PRM rates of sharks caught in a commercial bottom longline fishery. Blood was sampled from the same individuals that were tagged, providing direct correlation between stress physiology and animal fate for sandbar (Carcharhinus plumbeus, N = 130), blacktip (C. limbatus, N = 105), tiger (Galeocerdo cuvier, N = 52), spinner (C. brevipinna, N = 14), and bull sharks (C. leucas, N = 14). PRM rates ranged from 2% and 3% PRM in tiger and sandbar sharks to 42% and 71% PRM in blacktip and spinner sharks, respectively. Decision trees based on blood values predicted mortality with >67% accuracy in blacktip and spinner sharks, and >99% accuracy in sandbar sharks. Ninety percent of PRM occurred within 5 h after release and 59% within 2 h. Blood physiology indicated that PRM was primarily associated with acidosis and increases in plasma potassium levels. Total fishing mortality reached 62% for blacktip and 89% for spinner sharks, which may be under-estimates given that some soak times were shortened to focus on PRM. Our findings suggest that no-take regulations may be beneficial for sandbar, tiger, and bull sharks, but less effective for more susceptible species such as blacktip and spinner sharks.

Quantifying biological responses of catch-and-release angling in understudied fish species and practices

Many catch-and-release angling events involve air exposure and exhaustive exercise that elicit a physiological stress response, and depending on a variety of factors, delayed mortality is a possible outcome. There have been ample studies in this area, however, significant gaps exist in the literature for species that are targeted by more specialized anglers, such as freshwater drum (Aplodinotus grunniens). I quantified physiological and reflex responses in freshwater drum following angling, across seasons. Once a fish was on the line, the fight duration and time exposed to air were varied to account for differences in angler skill level (fight time range: 5 s to 2 min; air exposure range: 20 s to 3 min). Location and severity of injury were determined, blood biopsies were taken to quantify physiological stress, and reflex impairment was assessed. Thirtyone percent of fish captured were deeply hooked in the esophagus tissue. Freshwater drum experienced a disruption in homeostasis as blood glucose, plasma cortisol and plasma lactate increased significantly from baseline values following angling. Additionally, seasonal differences were observed for blood glucose and plasma cortisol as higher values were observed in the summer when compared to the spring. The ‘orientation’ reflex was the most frequently impaired (29 % of fish lacked this reflex), but impairment did not differ seasonally. Because freshwater drum have the largest latitudinal range of any North American freshwater fish and are being targeted more frequently by anglers as of late, it is important to fill this knowledge gap regarding their responses to angling events to develop best practices for anglers to promote conservation. The wide distribution of freshwater drum may also make them a candidate model species for addressing the convergence between assessing the impacts of catch-and-release angling and other environmental issues facing freshwater fishes, such as climate change.

Multi-Transcript Level Profiling Revealed Distinct mRNA, miRNA, and tRNA-Derived Fragment Bio-Signatures for Coping Behavior Linked Haplotypes in HPA Axis and Limbic System

The molecular basis of porcine coping behavior (CB) relies on a sophisticated interplay of genetic and epigenetic features. Deep sequencing technologies allowed the identification of a plethora of new regulatory small non-coding RNA (sncRNA). We characterized mRNA and sncRNA profiles of central parts of the physiological stress response system including amygdala, hippocampus, hypothalamus and adrenal gland using systems biology for integration. Therefore, ten each of high- (HR) and low- (LR) reactive pigs (n = 20) carrying a CB associated haplotype in a prominent QTL-region on SSC12 were selected for mRNA and sncRNA expression profiling. The molecular markers related to the LR group included ATP1B2, MPDU1, miR-19b-5p, let-7g-5p, and 5′-tiRNALeu in the adrenal gland, miR-194a-5p, miR-125a-5p, miR-7-1-5p, and miR-107-5p in the hippocampus and CBL and PVRL1 in the hypothalamus. Interestingly, amygdalae of the LR group showed 5′-tiRNA and 5′-tRF (5′-tRFLys, 5′-tiRNALys, 5′-tiRNACys, and 5′-tiRNAGln) enrichment. Contrarily, molecular markers associated with the HR group encompassed miR-26b-5p, tRNAArg, tRNAGlyiF in the adrenal gland, IGF1 and APOD in the amygdala and PBX1, TOB1, and C18orf1 in the hippocampus and miR-24 in the hypothalamus. In addition, hypothalami of the HR group were characterized by 3′-tiRNA enrichment (3′-tiRNAGln, 3′-tiRNAAsn, 3′-tiRNAVal, 3′-tRFPro, 3′-tiRNACys, and 3′-tiRNAAla) and 3′-tRFs enrichment (3′-tRFAsn, 3′-tRFGlu, and 3′-tRFVal). These evidence suggest that tRNA-derived fragments and their cleavage activity are a specific marker for coping behavior. Data integration revealed new bio-signatures of important molecular interactions on a multi-transcript level in HPA axis and limbic system of pigs carrying a CB-associated haplotype.