Differences in the range of thermoneutral zone between mouse strains: potential effects on translational research.

Laboratory mice are commonly used for studies emulating human metabolism. To render human energetics, their ratio of daily (DEE) to basal (BMR) energy expenditure of 1.7-1.8 should be maintained. However, the DEE/BMR ratio strongly depends on whether a given study using a mouse model is carried out above, or below the lower critical temperature (LCT) of the thermoneutral zone, which is rarely considered in translational research. Here, we used mice artificially selected for high or low rates of BMR along with literature data to analyze the effect of ambient temperature on possible systematic bias in DEE/BMR. We demonstrated that the estimated LCTs of mice from the high and low BMR lines differ by more than 7°C. Furthermore, the range of variation of LCTs of mouse strains used in translational research spans from 23 to 33°C. Differences between LCTs in our selected mice and other mouse strains are mirrored by differences in their DEE-to-BMR ratio, on average increasing it at the rate of 0.172°C-1 at temperatures below LCT. Given the wide range of LCTs in different mouse strains, we conclude that the energetic cost of thermoregulation may differ greatly for different mouse strains with a potentially large impact on translational outcomes. Thus, the LCT of a given mouse strain is an important factor that must be considered in designing translational studies.

Decay rates of arboreal and terrestrial nests of Eastern chimpanzees (Pan troglodytes schweinfurthii) in the Bugoma Central Forest Reserve, Uganda: Implications for population size estimates.

Chimpanzees were once thought to sleep primarily in the trees, but recent studies indicate that some populations also construct terrestrial night nests. This behavior has relevance not only to understanding the behavioral diversity of Pan troglodytes, but also to the conservation of the species, given that nest encounter rates are often used to estimate great ape population densities. A proper estimate of decay rates for ground nests is necessary for converting the encounter rate of nests to the density of weaned chimpanzees. Here we present the results of the first systematic comparative study between the decay rates of arboreal and terrestrial chimpanzee nests, from the Bugoma Central Forest Reserve in western Uganda. We followed the decay of 56 ground and 51 tree nests in eight nest groups between April 2020 and October 2021. For 15 of the ground and 19 of the tree nests, we collected detailed information on the condition of the nests every two weeks; we checked the remaining 73 nests only twice. On average, ground nests lasted 238 days versus 276 days for tree nests (p = 0.05). Of the 107 total nests surveyed, 51% of tree and 64% of ground nests had disappeared after six months. Based on our results, we propose a modification of the formula used to convert nest density into chimpanzee density. Our results highlight the importance of taking into account potential differences in decay rates between ground versus tree nests, which will likely influence our understanding of the distribution of ground nesting behavior in chimpanzee across tropical Africa, as well as our estimations of the densities of ground nesting populations.

Factors affecting nest height and ground nesting behaviour in Eastern chimpanzees (Pan troglodytes schweinfurthii) of the northern Democratic Republic of the Congo.

In order to achieve a better understanding of the factors that might have led our hominin ancestors to transition to a more terrestrial niche, including sleeping on the ground, we have conducted a study on the ground nesting behavior of chimpanzees (Pan troglodytes). Chimpanzees, like all other species of great apes, build nests in which to sleep each night, but little is known about regional differences in their nesting habits. Previously, nesting on the ground was considered typical of gorillas, but rare in most populations of chimpanzees. Using data acquired during our extensive chimpanzee nesting survey conducted between 2004 and 2013 across a > 50 000 km2 region in northern Democratic Republic of the Congo, we report a distinctive ground nesting behaviour of eastern chimpanzees (P. t. schweinfurthii). We have mapped the geographical distribution of ground nesting and compared its frequency at 20 survey areas on both sides of a large river, the Uele. We found that ground nests made up more than 1% of total nests at 15 of the 20 survey regions. For a subset of 16 of these regions, we utilized statistical models to investigate whether forest type and structure, as well as the abundance of carnivores and large herbivores, and the activities of humans impacted the frequency of ground nesting and nest height. We predicted that higher encounter rates of human and dangerous animal signs would be associated with lower rates of ground nesting as well as increased nest height. Overall, 10.4% of the Bili-Uéré chimpanzee nests were terrestrial, but the frequency of ground nesting varied extensively between the survey areas (0-29% of nests). The occurrence of ground nests was positively associated with denser forests (p = 0.004), herb patches (p < 0.001), and light gaps (p < 0.001). Light gaps (p < 0.001), herb patches (p = 0.044), and vine tangles (p = 0.016) also had a strong negative effect on nest height. Hunting by humans had a negative effect on the probability of the occurrence of ground nests (p = 0.001) and a positive one on nest height (p = 0.013), with a similar but likely marginal effect of large herbivores on nest height (p = 0.023). In addition, the chimpanzees nested at significantly lower heights with increasing distance from roads and settlements (p < 0.001). Carnivore encounter rates, however, had no significant impact on ground nest frequency or nest height. Our results indicate that ground nesting can no longer be considered a rare and patchily-occurring phenomenon in Pan troglodytes, but is instead a major component of the chimpanzee behavioural repertoire across a considerable fraction of the range of the Eastern subspecies. Our study highlights that neither the large body size of gorillas nor the taming of fire are necessary conditions for hominids to sleep overnight on the ground, even in areas inhabited by multiple species of large carnivore. Human hunting, however, appears to reduce the probability of ground nesting, or eliminate the behavior altogether.

Mice selected for a high basal metabolic rate evolved larger guts but not more efficient mitochondria.

Intra-specific variation in both the basal metabolic rate (BMR) and mitochondrial efficiency (the amount of ATP produced per unit of oxygen consumed) has profound evolutionary and ecological consequences. However, the functional mechanisms responsible for this variation are not fully understood. Mitochondrial efficiency is negatively correlated with BMR at the interspecific level but it is positively correlated with performance capacity at the intra-specific level. This discrepancy is surprising, as theories explaining the evolution of endothermy assume a positive correlation between BMR and performance capacity. Here, we quantified mitochondrial oxidative phosphorylation activity and efficiency in two lines of laboratory mice divergently selected for either high (H-BMR) or low (L-BMR) levels of BMR. H-BMR mice had larger livers and kidneys (organs that are important predictors of BMR). H-BMR mice also showed higher oxidative phosphorylation activity in liver mitochondria but this difference can be hypothesized to be a direct effect of selection only if the heritability of this trait is low. However, mitochondrial efficiency in all studied organs did not differ between the two lines. We conclude that the rapid evolution of BMR can reflect changes in organ size rather than mitochondrial properties, and does not need to be accompanied obligatorily by changes in mitochondrial efficiency.

Brain size, gut size and cognitive abilities: the energy trade-offs tested in artificial selection experiment.

The enlarged brains of homeotherms bring behavioural advantages, but also incur high energy expenditures. The 'expensive brain' (EB) hypothesis posits that the energetic costs of the enlarged brain and the resulting increased cognitive abilities (CA) were met by either increased energy turnover or reduced allocation to other expensive organs, such as the gut. We tested the EB hypothesis by analysing correlated responses to selection in an experimental evolution model system, which comprises line types of laboratory mice selected for high or low basal metabolic rate (BMR), maximum (VO2max) metabolic rates and random-bred (unselected) lines. The traits are implicated in the evolution of homeothermy, having been pre-requisites for the encephalization and exceptional CA of mammals, including humans. High-BMR mice had bigger guts, but not brains, than mice of other line types. Yet, they were superior in the cognitive tasks carried out in both reward and avoidance learning contexts and had higher neuronal plasticity (indexed as the long-term potentiation) than their counterparts. Our data indicate that the evolutionary increase of CA in mammals was initially associated with increased BMR and brain plasticity. It was also fuelled by an enlarged gut, which was not traded off for brain size.

Significance of variation in basal metabolic rate in laboratory mice for translational experiments.

The basal metabolic rate (BMR) accounts for 60-70% of the daily energy expenditure (DEE) in sedentary humans and at least 50% of the DEE in laboratory mice in the thermoneutral zone. Surprisingly, however, the significance of the variation in the BMR is largely overlooked in translational research using such indices as physical activity level (PAL), i.e., the ratio of DEE/BMR. In particular, it is unclear whether emulation of human PAL in mouse models should be carried out within or below the thermoneutral zone. It is also unclear whether physical activity within the thermoneutral zone is limited by the capacity to dissipate heat generated by exercise and obligatory metabolic processes contributing to BMR. We measured PAL and spontaneous physical activity (SPA) in laboratory mice from two lines, divergently selected towards either high or low level of BMR, and acclimated to 30 °C (i.e., the thermoneutral zone), 23 or 4 °C. The mean PAL did not differ between both lines in the mice acclimated to 30 °C but became significantly higher in the low BMR mouse line at the lower ambient temperatures. Acclimation to 30 °C reduced the mean locomotor activity but did not affect the significant difference observed between the selected lines. We conclude that carrying out experiments within the thermoneutral zone can increase the consistency of translational studies aimed at the emulation of human energetics, without affecting the variation in physical activity correlated with BMR.

Plant-herbivore interactions: Combined effect of groundwater level, root vole grazing, and sedge silicification.

Accumulation of silica (Si) by plants can be driven by (1) herbivory pressure (and therefore plant-herbivore interactions), (2) geohydrological cycles, or (3) a combination of (1) and (2), with (1-3) possibly affecting Si concentration with a 1-year delay.To identify the relative significance of (1-3), we analyzed the concentration of Si in fibrous tussock sedge (Carex appropinquata), the population density of the root vole (Microtus oeconomus), and the groundwater level, over 11 years.The largest influence of autumn Si concentration in leaves (Sileaf) was on the level of the current-year groundwater table, which was positive and accounted for 13.3% of its variance. The previous year's vole population density was weakly positively correlated with Sileaf, and it alone explained 9.5% of its variance.The only variable found to have a positive, significant effect on autumn Si concentration in rhizomes (Sirhiz) was the current-year spring water level, explaining as much as 60.9% of its variance.We conclude that the changes in Si concentration in fibrous tussock sedge are predominantly driven by hydrology, with vole population dynamics being secondary.Our results provide only partial support for the existence of plant-herbivore interactions, as we did not detect the significant effects of Si tussock concentration on the vole density dynamics. This was mainly due to the low level of silicification of sedges, which was insufficient to impinge herbivores.Future studies on plant-herbivore interactions should therefore aim at disentangling whether anti-herbivore protection is dependent on threshold values of herbivore population dynamics. Furthermore, studies on Si accumulation should focus on the effect of water-mediated Si availability.

Larger guts and faster growth in mice selected for high basal metabolic rate.

Postnatal growth in birds and mammals is the time of highest vulnerability and relatively high energy demands and therefore shapes the organisms' future outcomes. Several different factors might impose limitations on growth in juveniles, one of them being the efficiency of the digestive process and size of the gastrointestinal tract. We tested the gut size-growth rate relationship using a unique experimental model-mice from a selection experiment designed to produce two lines with divergent levels of basal metabolic rate (BMR): the high BMR (H-BMR) and low BMR (L-BMR) line types. These lines differ with respect to not only BMR, but also correlated traits-internal organ size and food intake. Applying a cross-fostering design and a thermoregulatory burden imposed by shaving the mothers, we demonstrated that the mass of intestine strongly affected the growth rate, with the H-BMR pups having larger intestines and growing faster, and with reduced growth rate of pups of both lines nursed by shaved L-BMR mothers. Our study also provides a functional link between high growth rate of neonates and high BMR of adults, partly reflecting metabolic costs of maintenance of their guts.

Low basal metabolic rate as a risk factor for development of insulin resistance and type 2 diabetes.

INTRODUCTION: Identification of physiological factors influencing susceptibility to insulin resistance and type 2 diabetes (T2D) remains an important challenge for biology and medicine. Numerous studies reported energy expenditures as one of those components directly linked to T2D, with noticeable increase of basal metabolic rate (BMR) associated with the progression of insulin resistance. Conversely, the putative link between genetic, rather than phenotypic, determination of BMR and predisposition to development of T2D remains little studied. In particular, low BMR may constitute a considerable risk factor predisposing to development of T2D. RESEARCH DESIGN AND METHODS: We analyzed the development of insulin resistance and T2D in 20-week-old male laboratory mice originating from three independent genetic line types. Two of those lines were subjected to divergent, non-replicated selection towards high or low body mass-corrected BMR. The third line type was non-selected and consisted of randomly bred animals serving as an outgroup (reference) to the selected line types. To induce insulin resistance, mice were fed for 8 weeks with a high fat diet; the T2D was induced by injection with a single dose of streptozotocin and further promotion with high fat diet. As markers for insulin resistance and T2D advancement, we followed the changes in body mass, fasting blood glucose, insulin level, lipid profile and mTOR expression. RESULTS: We found BMR-associated differentiation in standard diabetic indexes between studied metabolic lines. In particular, mice with low BMR were characterized by faster body mass gain, blood glucose gain and deterioration in lipid profile. In contrast, high BMR mice were characterized by markedly higher expression of the mTOR, which may be associated with much slower development of T2D. CONCLUSIONS: Our study suggests that genetically determined low BMR makeup involves metabolism-specific pathways increasing the risk of development of insulin resistance and T2D.

Coevolution of body size and metabolic rate in vertebrates: a life-history perspective.

Despite many decades of research, the allometric scaling of metabolic rates (MRs) remains poorly understood. Here, we argue that scaling exponents of these allometries do not themselves mirror one universal law of nature but instead statistically approximate the non-linearity of the relationship between MR and body mass. This 'statistical' view must be replaced with the life-history perspective that 'allows' organisms to evolve myriad different life strategies with distinct physiological features. We posit that the hypoallometric allometry of MRs (mass scaling with an exponent smaller than 1) is an indirect outcome of the selective pressure of ecological mortality on allocation 'decisions' that divide resources among growth, reproduction, and the basic metabolic costs of repair and maintenance reflected in the standard or basal metabolic rate (SMR or BMR), which are customarily subjected to allometric analyses. Those 'decisions' form a wealth of life-history variation that can be defined based on the axis dictated by ecological mortality and the axis governed by the efficiency of energy use. We link this variation as well as hypoallometric scaling to the mechanistic determinants of MR, such as metabolically inert component proportions, internal organ relative size and activity, cell size and cell membrane composition, and muscle contributions to dramatic metabolic shifts between the resting and active states. The multitude of mechanisms determining MR leads us to conclude that the quest for a single-cause explanation of the mass scaling of MRs is futile. We argue that an explanation based on the theory of life-history evolution is the best way forward.

Functional and structural changes in aorta of mice divergently selected for basal metabolic rate.

Cardiovascular diseases (CVD) are one of the most common causes of mortality likely genetically linked to the variation in basal metabolic rate (BMR). A robust test of the significance of such association may be provided by artificial selection experiments on animals selected for diversification of BMR. Here we asked whether genetically determined differences in BMR correlate with anatomical shift in endothelium structure and if so, the relaxation and contraction responses of the aorta in mice from two lines of Swiss-Webster laboratory mice (Mus musculus) divergently selected for high or low BMR (HBMR and LBMR lines, respectively). Functional and structural study of aorta showed that a selection for divergent BMR resulted in the between-line difference in diastolic aortic capacity. The relaxation was stronger in aorta of the HBMR mice, which may stem from greater flexibility of aorta mediated by higher activity of Ca2+-activated K+ channels. Structural examination also indicated that HBMR mice had significantly thicker aorta's middle layer compared to LBMR animals. Such changes may promote arterial stiffness predisposing to cardiovascular diseases. BMR-related differences in the structure and relaxation ability of aortas in studied animals may be reminiscent of potential risk factors in the development of CVD in humans.

Not that hot after all: no limits to heat dissipation in lactating mice selected for high or low BMR.

Heat dissipation has been suggested as a limit to sustained metabolic effort, e.g. during lactation, when overheating is a possible risk. We tested this hypothesis using mice artificially selected for either high or low BMR that also differed with respect to parental effort. We used fixed size cross-fostered families and recorded litter mass daily until the 14th day of lactation. Midway through the experiment (day 8) half of the mothers from each group had fur from the dorsal body surface removed to increase their thermal conductance and facilitate heat dissipation. Our results showed that neither high nor low BMR mouse lines benefited from increasing their thermal conductance at peak lactation. On the contrary, growth of the litters reared by the low BMR females was compromised. Thus, our results do not support the heat dissipation limitation hypothesis.

Long-Term Trait Consistency in Mice Selected for Swim-Induced High Aerobic Capacity.

The majority of studies show that metabolic rates are usually repeatable at the individual level, although their repeatabilities tend to decline with time and to be strongly affected by physiological changes. Changes in individual repeatabilities may therefore affect putative differences between experimental groups or populations. This problem is particularly relevant to artificial selection experiments that apply the selection protocol at early life stages, running the risk of a poor correlation of the trait with itself throughout the life cycle of individuals. Moreover, significant physiological changes (e.g., induced by reproduction) may affect traits under selection and therefore their postreproductive differentiation between selected lines. Here, using a unique animal model-mice from four lines selected for [Formula: see text] during swimming in 25°C water and four random-bred control (reference) lines-we analyzed the long-term consistency of aerobic capacity as well as postswim hypothermia in primiparous and nonreproducing females at 12, 25, and 29 wk of age. Our results show that significant between-line type divergence in [Formula: see text] and hypothermia persists over time and is only weakly affected by past reproduction. Furthermore, both traits are also repeatable within lines at the individual level. More generally, our results suggest that past reproduction events are unlikely to significantly affect between-population and between-individual differences in [Formula: see text] and related traits.

The nexus of hair corticosterone level, immunocompetence, metabolic rates and overwinter survival in the root vole, Microtus oeconomus.

Although corticosterone (CORT) regulates many physiological mechanisms, the associations between CORT levels, immunocompetence, energy expenditures and overwinter survival have not been examined. Here, we studied individual variation in CORT level extracted from hair, immunocompetence quantified as the neutrophil-to-lymphocyte (N/L) ratio, total white blood cells (WBC) and natural antibody levels (NAbs), along with the resting (RMR) and peak metabolic rates (PMR) and mortality during three consecutive winter seasons in a natural population of the root vole, Microtus oeconomus. In early winter, hair CORT level was strongly positively associated with body mass and inversely related to voles' ability to survive. We suggest that the observed association between hair CORT level and body mass may be the key component of the physiological nexus driving the survivorship of individual rodents. Additionally, hair CORT was a significant predictor of variation of the whole body RMR, which in turn enhanced overwinter survival in the studied population. On the other hand, hair CORT was not significantly associated with changes in the blood indices. Interestingly, the analysis carried out only during the first year of study (2008), which was characterized by a high population density and prevalence of infestation with a blood protozoan, Babesia spp., showed that the intensity of the infestation was negatively correlated with both the hair CORT level and the N/L ratio. Because CORT is often considered immunosuppressive, we expected a positive association between its level and the N/L ratio. However, hair CORT did not significantly correlate with the N/L ratio. We suggest that the lack of an association between hair CORT and the N/L ratio resulted from a small inter-individual variation in the N/L ratio in 2008, which was much higher and less variable than in the other years of our study.

Selection for high aerobic capacity has no protective effect against obesity in laboratory mice.

Aerobic capacity (VO2max measured during intensive physical exercise) both trained and intrinsic (i.e. genetically determined) has recently been deemed a good predictor of cardiometabolic risks. However, the underlying mechanisms linking VO2max and health risk factors are not entirely clear, as it seems that not VO2max per se, but rather some correlated traits, like spontaneous physical activity (SPA) are responsible for sustaining the lean phenotype. Here we investigated the link between genetically determined aerobic capacity, SPA and resistance to diet-induced health risks using replicated lines of mice selected for high aerobic capacity during swimming in mid-cold water (25°C) and Randomly Bred control mice. After four months of consumption of the western type HFat and HCarb diets and no forced nor voluntary training, we found no evidence of protective effects of intrinsic high VO2max. The Selected mice displayed similar levels of blood glucose, cholesterol, triglycerides and body fat as the Random Bred control animals. Most notably we found no correlation between VO2max and SPA levels. Our results therefore call into question the ubiquity of VO2max as a predictor of metabolic health and leanness, at least in animal models.

Metabolic risk factors in mice divergently selected for BMR fed high fat and high carb diets.

Factors affecting contribution of spontaneous physical activity (SPA; activity associated with everyday tasks) to energy balance of humans are not well understood, as it is not clear whether low activity is related to dietary habits, precedes obesity or is a result of thereof. In particular, human studies on SPA and basal metabolic rates (BMR, accounting for >50% of human energy budget) and their associations with diet composition, metabolic thrift and obesity are equivocal. To clarify these ambiguities we used a unique animal model-mice selected for divergent BMR rates (the H-BMR and L-BMR line type) presenting a 50% between-line type difference in the primary selected trait. Males of each line type were divided into three groups and fed either a high fat, high carb or a control diet. They then spent 4 months in individual cages under conditions emulating human "sedentary lifestyle", with SPA followed every month and measurements of metabolic risk indicators (body fat mass %, blood lipid profile, fasting blood glucose levels and oxidative damage in the livers, kidneys and hearts) taken at the end of study. Mice with genetically determined high BMR assimilated more energy and had higher SPA irrespective of type of diet. H-BMR individuals were characterized by lower dry body fat mass %, better lipid profile and lower fasting blood glucose levels, but higher oxidative damage in the livers and hearts. Genetically determined high BMR may be a protective factor against diet-induced obesity and most of the metabolic syndrome indicators. Elevated spontaneous activity is correlated with high BMR, and constitutes an important factor affecting individual capability to sustain energy balance even under energy dense diets.

Effect of calorie restriction on spontaneous physical activity and body mass in mice divergently selected for basal metabolic rate (BMR).

Spontaneous physical activity (SPA) represents an important component of daily energy expenditures in animals and humans. Intra-specific variation in SPA may be related to the susceptibility to metabolic disease or obesity. In particular, reduced SPA under conditions of limited food availability may conserve energy and prevent loss of body and fat mass ('thrifty genotype hypothesis'). However, both SPA and its changes during food restriction show wide inter-individual variations. We studied the effect of 30% caloric restriction (CR) on SPA in laboratory mice divergently selected for high (H-BMR) and low (L-BMR) basal metabolic rate. Selection increased SPA in the H-BMR line but did not change it in the L-BMR mice. This effect reflected changes in SPA intensity but not SPA duration. CR increased SPA intensity more strongly in the L-BMR line than in the H-BMR line and significantly modified the temporal variation of SPA. However, the initial between-line differences in SPA were not affected by CR. Loss of body mass during CR did not differ between both lines. Our results show that the H-BMR mice can maintain their genetically determined high SPA under conditions of reduced food intake without sacrificing their body mass. We hypothesize that this pattern may reflect the higher flexibility in the energy budget in the H-BMR line, as we showed previously that mice from this line reduced their BMR during CR. These energy savings may allow for the maintenance of elevated SPA in spite of reduced food intake. We conclude that the effect of CR on SPA is in large part determined by the initial level of BMR, whose variation may account for the lack of universal pattern of behavioural responses to CR.

Heat dissipation does not suppress an immune response in laboratory mice divergently selected for basal metabolic rate (BMR).

The capacity for heat dissipation is considered to be one of the most important constraints on rates of energy expenditure in mammals. To date, the significance of this constraint has been tested exclusively under peak metabolic demands, such as during lactation. Here, we used a different set of metabolic stressors, which do not induce maximum energy expenditures and yet are likely to expose the potential constraining effect of heat dissipation. We compared the physiological responses of mice divergently selected for high (H-BMR) and low basal metabolic rate (L-BMR) to simultaneous exposure to the keyhole limpet haemocyanin (KLH) antigen and high ambient temperature (Ta). At 34°C (and at 23°C, used as a control), KLH challenge resulted in a transient increase in core body temperature (Tb) in mice of both line types (by approximately 0.4°C). Warm exposure did not produce line-type-dependent differences in Tb (which was consistently higher by ca. 0.6°C in H-BMR mice across both Ta values), nor did it result in the suppression of antibody synthesis. These findings were also supported by the lack of between-line-type differences in the mass of the thymus, spleen or lymph nodes. Warm exposure induced the downsizing of heat-generating internal organs (small intestine, liver and kidneys) and an increase in intrascapular brown adipose tissue mass. However, these changes were similar in scope in both line types. Mounting a humoral immune response in selected mice was therefore not affected by ambient temperature. Thus, a combined metabolic challenge of high Ta and an immune response did not appreciably compromise the capacity to dissipate heat, even in the H-BMR mice.

Effect of reproduction on the consistency of the between-line type divergence in laboratory mice selected on Basal metabolic rate.

Artificial selection experiments are an effective tool for testing evolutionary hypotheses, because they allow one to separate genetic and environmental variances of the phenotype. However, it is unclear whether trait divergence typically selected early in life persists over an animal's life and altered physiological states, such as reproduction. Here we analyzed the long-term consistency of the between-line type divergence in basal metabolic rate (BMR) selected at 12 wk of age in laboratory mice. We measured BMR in nonreproducing and reproducing females at the age of 22 wk and then at 27 wk of age. Our results show that within both the reproducing group and the control group, the between-line type separation in BMR is consistently retained over time and reproductive status. Metabolically active internal organs (heart, liver, kidneys, and small intestine) also consistently differed in size between the two line types with no significant long-term effect of reproduction. The observed consistency of the between-line type divergence in BMR suggests the existence of the persistent effect of the selection on metabolic traits applied early in life. Moreover, BMR variation achieved by means of artificial selection is considerably higher than that found in natural/unmanipulated populations. The latter may therefore be characterized by insufficient variance to statistically resolve correlations involving BMR.