Elżbieta Król

There has been debate over the appropriate equations for calculation of daily energy expenditure (DEE) in doubly labelled water (DLW) studies. Previous studies that have used tritium as the hydrogen label have generally not corrected for fractionation effects, while studies using deuterium generally have. Some validation data are available to support both positions, but as yet very little work has compared the different approaches in animals simultaneously injected with both isotopes. We compared the metabolizable energy intake (MEI) of mice ( Mus musculus , outbred MF1, n = 15) averaged over 5 days, to the calculated DEE measured on day 6 using the DLW method, for mice injected simultaneously with tritium, deuterium and ¹⁸ oxygen. This allowed us to compare in the same animals the validity of the contrasting equations. Using the deuterium data and the equations that correct for deuterium fractionation, the average difference between DEE and MEI was 0.6 ± 11.4%. Using tritium data and equations that account for tritium fractionation, the average difference was 1.2 ± 11.6%. The discrepancy was greater if fractionation effects were ignored. Using the deuterium data, the discrepancy averaged 2.5 ± 11.4% and using the tritium data it averaged 3.7 ± 11.7%. Applying the tritium data in the equations that correct for deuterium fractionation resulted in an intermediate discrepancy of 1.9 ± 11.6%. Using the difference between deuterium and tritium washouts to estimate the fractionated water loss produced physiologically impossible individual estimates in 13 of 15 mice. Mice ate less food and lost body mass on the day when the DLW method was applied compared to the previous 5 days, but energy expenditure was apparently unaffected.

Gill health has become a significant global challenge for Atlantic salmon (Salmo salar) aquaculture, particularly during the marine phase of farming. The increasing prevalence of gill pathologies has been linked to rising seawater temperatures, underscoring the need to evaluate existing tools for monitoring gill health and to develop novel approaches for early detection. In this study, we investigated the gill responses of commercially farmed Atlantic salmon to natural infection with Neoparamoeba perurans during an outbreak of amoebic gill disease (AGD) in Tasmania. Our focus spanned the low AGD prevalence, high AGD prevalence, and post-freshwater treatment stages of the outbreak. Evaluations of gill tissue included assessments of the gross AGD score, histopathological score, abundance of N. perurans (measured by 18S rRNA gene expression), and expression levels of inflammation-related transcripts. We demonstrated a strong correlation between different measures of AGD-related gill pathology and significant differences between distinct stages of the N. perurans outbreak. Post-treatment, fish exhibited considerable variability in their responses to the freshwater bath, highlighting the necessity for personalized management strategies that consider genetic, environmental, and health status factors. The expression patterns of angiogenin-1 (ANG1) and complement C1q tumour necrosis factor-related protein 3-like (C1QTNF3) emphasize their potential as biomarkers for early detection of gill damage in salmon aquaculture worldwide.

Peak lactation occurs when milk production is at its highest. The factors limiting peak lactation performance have been subject of intense debate. Milk production at peak lactation appears limited by the capacity of lactating females to dissipate body heat generated as a by-product of processing food and producing milk. As a result, manipulations that enhance capacity to dissipate body heat (such as fur removal) increase peak milk production. We investigated the potential correlates of shaving-induced increases in peak milk production in laboratory mice. By transcriptomic profiling of the mammary gland, we searched for the mechanisms underlying experimentally increased milk production and its consequences for mother–young conflict over weaning, manifested by advanced or delayed involution of mammary gland. We demonstrated that shaving-induced increases in milk production were paradoxically linked to reduced expression of some milk synthesis-related genes. Moreover, the mammary glands of shaved mice had a gene expression profile indicative of earlier involution relative to unshaved mice. Once provided with enhanced capacity to dissipate body heat, shaved mice were likely to rear their young to independence faster than unshaved mothers.

Seasonal energetic challenges may constrain an animal's ability to respond to changing individual and environmental conditions. Here, we investigated variation in heart rate, a well-established proxy for metabolic rate, in Svalbard reindeer (Rangifer tarandus platyrhynchus), a species with strong seasonal changes in foraging and metabolic activity. In 19 adult females, we recorded heart rate, subcutaneous temperature and activity using biologgers. Mean heart rate more than doubled from winter to summer. Typical drivers of energy expenditure, such as reproduction and activity, explained a relatively limited amount of variation (2–6% in winter and 16–24% in summer) compared to seasonality, which explained 75% of annual variation in heart rate. The relationship between heart rate and subcutaneous temperature depended on individual state via body mass, age and reproductive status, and the results suggested that peripheral heterothermy is an important pathway of energy management in both winter and summer. While the seasonal plasticity in energetics makes Svalbard reindeer well-adapted to their highly seasonal environment, intraseasonal constraints on modulation of their heart rate may limit their ability to respond to severe environmental change. This study emphasizes the importance of encompassing individual state and seasonal context when studying energetics in free-living animals. This article is part of the theme issue ‘Measuring physiology in free-living animals (Part II)’.

The gill of teleost fish is a multifunctional organ involved in many physiological processes such as gas exchange, osmotic and ionic regulation, acid-base balance and excretion of nitrogenous waste. Due to its extensive interface with the environment, the gill plays a key role as a primary mucosal defense tissue against pathogens, as manifested by the presence of the gill-associated lymphoid tissue (GIALT). In recent years, the prevalence of multifactorial gill pathologies has increased significantly, causing substantial losses in Atlantic salmon aquaculture. The transition from healthy to unhealthy gill phenotypes and the progression of multifactorial gill pathologies, such as proliferative gill disease (PGD), proliferative gill inflammation (PGI) and complex gill disorder (CGD), are commonly characterized by epithelial hyperplasia, lamellar fusion and inflammation. Routine monitoring for PGD relies on visual inspection and non-invasive scoring of the gill tissue (gross morphology), coupled with histopathological examination of gill sections. To explore the underlying molecular events that are associated with the progression of PGD, we sampled Atlantic salmon from three different marine production sites in Scotland and examined the gill tissue at three different levels of organization: gross morphology with the use of PGD scores (macroscopic examination), whole transcriptome (gene expression by RNA-seq) and histopathology (microscopic examination). Our results strongly suggested that the changes in PGD scores of the gill tissue were not associated with the changes in gene expression or histopathology. In contrast, integration of the gill RNA-seq data with the gill histopathology enabled us to identify common gene expression patterns associated with multifactorial gill disease, independently from the origin of samples. We demonstrated that the gene expression patterns associated with multifactorial gill disease were dominated by two processes: a range of immune responses driven by pro-inflammatory cytokines and the events associated with tissue damage and repair, driven by caspases and angiogenin.

Mus musculus molossinus (MSM) is a wild-derived mouse strain which maintains the ability to synthesize melatonin in patterns reflecting the ambient photoperiod. The objective of this study was to characterize the effects of photoperiodic variation on metabolic and reproductive traits, and the related changes in pituitary-hypothalamic gene expression in MSM mice. MSM mice were kept in long (LP) or short photoperiod (SP) for 6 weeks. Our results demonstrate that MSM mice kept in LP, as compared to mice kept in SP, display higher expression of genes encoding thyrotropin (TSH) in the pars tuberalis, thyroid hormone deiodinase 2 (dio2) in the tanycytes, RFamide-related peptide (RFRP3) in the hypothalamus and lower expression of dio3 in the tanycytes, along with larger body and reproductive organ mass. Additionally, to assess the effects of the gestational photoperiodic environment on the expression of these genes, we kept MSM mice in LP or SP from gestation and studied offspring. We show that the gestational photoperiod affects the TSH/dio pathway in newborn MSM mice in a similar way to adults. This result indicates a transgenerational effect of photoperiod from the mother to the fetus in utero. Overall, these results indicate that photoperiod can influence neuroendocrine regulation in a melatonin-proficient mouse strain, in a manner similar that documented in other seasonal rodent species. MSM mice may therefore become a useful model for research into the molecular basis of photoperiodic regulation of seasonal biology.

Maximal animal performance may be limited by the ability of animals to dissipate heat; the heat dissipation limitation (HDL) theory. Since diets vary in the incidental heat produced during digestion (specific dynamic action, SDA), the HDL theory predicts lactating female mice consuming diets with lower SDA should have increased reproductive performance. Dietary fat has a lower SDA than dietary carbohydrate. Female mice were fed low (LF), medium (MF) or high fat (HF) diets (10%, 45% and 60% energy from fat respectively) from days 4-18 of lactation. HF and MF-fed mice weaned significantly heavier litters than LF mice. This was because they not only consumed more energy (metabolisable energy intake, E_mei; HF:306.5±25.0, MF:340.5±13.5 kJ d⁻¹) at peak lactation, but also delivered more milk energy to their pups (milk energy output, E_milk: 203.2±49.9, 229.3±42.2 kJ d⁻¹ respectively) than the LF-fed mice (E_mei =266.7±4.5, E_milk =164.60±30.59 kJ d⁻¹). This effect was greater than predicted from the SDA of the different diets combined with a mathematical model based on the HDL theory. Fatty acid profiles of the diets, milk and pups, showed significant correlations between the profiles. Besides reduced SDA, HF and MF-fed mice were probably able to directly transfer absorbed dietary fat into milk, reducing the heat production of lactogenesis, and enabling them to perform better than expected from the HDL model. In summary, HF and MF diets had beneficial effects on reproductive performance compared to the LF diet because they enabled mice to generate milk more efficiently with less incidental heat production.