The 13C-bicarbonate technique as a tool for measurement of energy expenditure in overweight dogs undergoing body weight reduction and the effect of different dietary composition.

Changes in body size and composition, i.e., body weight (BW) gain or loss, affect the daily energy expenditure (EE). To ensure an appropriate BW reduction and to find an efficient strategy to reduce and maintain a target BW, regular evaluations and adjustments of energy allowance are important. This study aimed to provide a detailed knowledge about the possible changes in resting EE using the oral 13C-bicarbonate technique (o13CBT) as a research tool in 16 overweight pet dogs undergoing BW reduction. Dietary composition (i.e., in % of dry matter [DM] being a high protein [33.3], low fat [9.6], and high crude fiber [18.0] diet [LFHFibre], and a high protein [37.9], high fat [52.0], carbohydrate-free diet [HFat]) during 16 wk of energy restriction were evaluated regarding effects on resting EE, rate of BW reduction, body composition, and plasma concentrations of metabolic hormones involved in energy metabolism and appetite regulation. The mean BW loss was higher (P < 0.05) for the dogs fed the LFHFibre diet (1.1%/wk) than that for dogs fed the HFat diet (0.8%/wk), but the total BW reduction of 14.6% and 12.0% of initial BW did not differ significantly (P > 0.05). Resting EE was lower (P < 0.02) after the BW reduction; 414 kJ (99 kcal)/kg BW0.75/d at the start (week 0) and 326 kJ (78 kcal)/kg BW0.75/d at the end (week 16) of the study. The BW reduction in both groups (P > 0.05) consisted of both fat mass (FM) and fat-free mass (FFM). Energy expenditure, calculated in relation to amount of FFM, was not significantly (P > 0.05) affected by BW reduction. Dietary composition did not significantly affect (P > 0.05) plasma concentrations of insulin, leptin, and ghrelin, and no effect (P > 0.05) of BW reduction was observed on hormone concentrations. In conclusion, the o13CBT proved to be a useful research method for studying short-term EE in overweight dogs. Even though all dogs lost BW, most dogs were still overweight at the end of the study. Due to a high individual variation among dogs, a longer experimental period with a larger sample size would be desirable.

The most common nutritional disorder in dogs is overweight, and knowledge about dogs' energy requirement is therefore important to adjust daily feed allowance. Changes in body weight may affect energy expenditure (EE) and, thereby, energy requirement. This study aimed to measure such potential changes under resting conditions in overweight dogs. It was found that the minimally invasive 13C-bicarbonate technique was a useful research method for studies regarding EE during weight loss (WL) in dogs. EE decreased when the dogs lost weight, and energy allowance needed to be reduced to maintain WL. The second objective of this study was to evaluate the effects of feeding diets with different macronutrient compositions on EE, rate of WL, body composition, and plasma concentrations of hormones involved in energy metabolism and appetite regulation. The mean WL rate was slightly higher for dogs fed a diet with high protein, low fat, and high crude fiber contents than those fed a carbohydrate-free diet with a high protein and fat contents. However, diet did not affect the resting EE, measured plasma hormone concentrations, or the total WL at the end of the study.

Estimation of energy expenditure using the oral 13 C-bicarbonate technique in privately owned adult and senior dogs of three different body sizes.

Knowledge about energy requirements (ER) of dogs is important in order to provide dogs with an appropriate energy supply. In this study, the oral 13 C-bicarbonate technique (o13 CBT) was used to estimate energy expenditure (EE) in 50 privately owned dogs of different body sizes (small: Danish-Swedish Farmdog (DSF, n = 16), medium: Beagle (n = 15), large: Labrador Retriever (n = 19)), of different ages (adult: 2-7 years (n = 33), senior: ≥8 years (n = 17)). The aim was to evaluate the feasibility of the method in the field and to get more information about ER of dogs. All dogs were measured twice, on two separate days under standardized, resting conditions. The dogs were fasted overnight prior to measurements, which were carried out in home environment. Only measurements of dogs resting calmly were included in the statistical analyses. No significant difference (p > 0.05) in EE was found between days of measurements. The EE measured in Labrador Retrievers (405 kJ (97 kcal)/kg BW0.75 /day) was significantly lower (p < 0.001) than of Beagles (530 kJ (127 kcal)/kg BW0.75 /day) and DSF (497 kJ (119 kcal)/kg BW0.75 /day), the difference between Beagles and DSF being non-significant. Senior dogs had significantly lower (p < 0.001) EE values than adult dogs with least square means (LSM) of 441 kJ (105 kcal)/kg BW0.75 /day and 513 kJ (123 kcal)/kg BW0.75 /day respectively. Analysed within breeds, EE of senior Labrador Retrievers (370 kJ (88 kcal)/kg BW0.75 /day)) was significantly lower (p < 0.01) than adults (439 kJ (105 kcal)/kg BW0.75 /day), as well as in DSF (453 kJ (108 kcal)/kg BW0.75 /day and 541 kJ (129 kcal)/kg BW0.75 /day for senior and adult dogs respectively). However, the EE was not significantly different (p > 0.05) between adult (548 kJ (131 kcal)/kg BW0.75 /day) and senior (499 kJ (119 kcal)/kg BW0.75 /day) Beagles. This study suggests that when measured under standardized resting conditions, the o13 CBT can provide reliable results of EE and be a helpful tool to get more knowledge about ER of dogs of different sizes, breeds and ages.

The oral 13C-bicarbonate technique for determination of energy expenditure in dogs: dietary and environmental factors affecting the respiratory quotient and 13C recovery factor.

The oral 13C-bicarbonate technique (o13CBT) can be used for short-term measurements of CO2 production (RCO2) and energy expenditure (EEx). The method relies on appropriate estimates for the respiratory quotient (RQ) and recovery factor (RF) of 13C. Four Retriever dogs were included in four experiments to validate the o13CBT against indirect calorimetry (IC), and determine RQ and RF; Expt. 1: feeding different protein:fat:carbohydrate ratios [in % of metabolisable energy]: 25:33:42 in a maintenance (Mnt.) diet; 38:26:36 in a high-protein high-fibre (HFibre) diet and 27:56:17 in a high-fat (HFat) diet, simultaneously with start of measurements (T0); Expt. 2: the Mnt. diet at T0 or 4 h postprandial (T4); Expt. 3: T4 at different ambient temperatures, 22°C and 15°C; Expt. 4: T4 after 1 h physical activity. The RCO2 and EEx were determined from the respiration chamber measurements made simultaneously with IC and the o13CBT (o13CBTonline), and in Expts. 1 and 2, also on two consecutive days using o13CBT with collection of breath into breath bags (o13CBTbreathbags). The RQ values obtained at T0 reflected dietary compositions, with the highest least square mean (LSM) of 0.954  for the Mnt. diet, 0.905 for the HFibre and 0.877 for the HFat diet (p < 0.05). An increased interval between meal and measurement period decreased RQ significantly (p < 0.05) in Expt. 2, LSM being 0.954 at T0 and 0.909 at T4. Ambient temperature (Expt. 3) and physical activity (Expt. 4) did not influence postprandial RQ. The RF values were not significantly affected by diet (Expt. 1). Measurements starting at T0 (Expt. 2) resulted in higher (p < 0.05) RF values than at T4 (LSM = 0.971 and 0.836, respectively). The ambient temperatures (Expt. 3) did not influence postprandial RF. However, when dogs were physically active prior to measurements (Expt. 4), RF values (LSM = 1.019) were higher (p < 0.05) than when resting only (LSM = 0.836). Calculations based on RQ and RF determined in each experiment resulted in RCO2 and EEx values which were not different regardless of method used, except for Expt. 1 where EEx-values [kJ · kg BW-0.75 · d-1] were higher (p < 0.05) when measured with o13CBTbreathbags (460) than by IC (421) and o13CBTonline (420). Provided accurate RQ and RF values, the o13CBTbreathbags can be used as an independent and minimally invasive research tool to determine EEx in dogs under carefully standardised conditions.

Effects of long-term feeding of rapeseed meal on skeletal muscle transcriptome, production efficiency and meat quality traits in Norwegian Landrace growing-finishing pigs.

This study was performed to investigate the effects of dietary inclusion of 20% rapeseed meal (RSM) as an alternative to soybean meal (SBM) in a three-month feeding experiment with growing finishing pigs. Dietary alteration affected growth performance, several carcass traits and transcriptional responses in the skeletal muscle, but did not affect measured meat quality traits. In general, pigs fed the RSM test diet exhibited reduced growth performance compared to pigs on SBM control diet. Significant transcriptional changes in the skeletal muscle of growing pigs fed RSM diet were likely the consequence of an increased amount of fiber and higher polyunsaturated fatty acids, and presence of bioactive phytochemicals, such as glucosinolates. RNAseq pipeline using Tophat2-Cuffdiff identified 57 upregulated and 63 downregulated genes in RSM compared to SBM pigs. Significantly enriched among downregulated pathways was p53-mediated signalling involved in cellular proliferation, while activation of negative growth regulators (IER5, KLF10, BTG2, KLF11, RETREG1, PRUNE2) in RSM fed pigs provided further evidence for reduced proliferation and increased cellular death, in accordance with the observed reduction in performance traits. Upregulation of well-known metabolic controllers (PDK4, UCP3, ESRRG and ESRRB), involved in energy homeostasis (glucose and lipid metabolism, and mitochondrial function), suggested less available energy and nutrients in RSM pigs. Furthermore, several genes supported more pronounced proteolysis (ABTB1, OTUD1, PADI2, SPP1) and reduced protein synthesis (THBS1, HSF4, AP1S2) in RSM muscle tissue. In parallel, higher levels of NR4A3, PDK4 and FGF21, and a drop in adropin, ELOVL6 and CIDEC/FSP27 indicated increased lipolysis and fatty acid oxidation, reflective of lower dressing percentage. Finally, pigs exposed to RSM showed greater expression level of genes responsive to oxidative stress, indicated by upregulation of GPX1, GPX2, and TXNIP.

The ¹³C bicarbonate method: an inverse end product method for measuring CO2 production and energy expenditure.

We reconsider the principle of the (13)C bicarbonate (NaH(13)CO3) method ((13)C-BM) for the determination of the CO2 production to obtain an estimate of energy expenditure (EE). Its mathematical concept based on a three-compartmental model is related to the [(15)N]glycine end product method. The CO2 production calculated by the (13)C-BM, RaCO2((13)C) is compared to the result from the indirect calorimetry, RCO2(IC). In an interspecies comparison (dog, goat, horse, cattle, children, adult human; body mass ranging from 15 to 350 kg, resting and fasting conditions) we found an excellent correlation between the results of (13)C-BM and IC with RCO2(IC) = 0.703 × RaCO2((13)C), (R(2) = 0.99). The slope of this correlation corresponds to the fractional (13)C recovery (RF((13)C)) of (13)C in breath CO2 after administration of NaH(13)CO3. Significant increase in RF((13)C) was found in physically active dogs (0.95 ± 0.14; n = 5) vs. resting dogs (0.71 ± 0.10, n = 17; p = .015). The (13)C recovery in young bulls was greater in blood CO2 (0.81 ± 0.05) vs. breath CO2 (0.73 ± 0.05, n = 12, p < .001) and in ponies with oral (0.76 ± 0.03, n = 8) vs. intravenous administration of NaH(13)CO3 (0.69 ± 0.07; n = 8; p = .026). We suggest considering the (13)C-BM as a 'stand-alone' method to provide information on the total CO2 production as an index of EE.

The oral [(13)C]bicarbonate technique for measurement of short-term energy expenditure of sled dogs and their physiological response to diets with different fat:carbohydrate ratios.

The oral [(13)C]bicarbonate technique (o(13)CBT) was assessed for the determination of short-term energy expenditure (EE) under field conditions. A total of eight Alaskan huskies were fed two experimental diets in a cross-over experiment including two periods of 3 weeks. Effects of diets on EE, apparent total tract digestibility (ATTD) and on plasma hormones, blood lactate and glucose were furthermore investigated. The percentages of metabolisable energy derived from protein (P), fat (F) and carbohydrates (C) were 26:58:16 in the PFC diet and 24:75:1 in the PF diet. Measurements of EE were performed in the post-absorptive state during rest. Blood samples were collected during rest and exercise and ATTD was determined after days with rest and with exercise. EE was higher (P < 0·01) in period 2 than in period 1 (68 v. 48 kJ/kg body weight(0·75) per h). The ATTD of organic matter, crude protein and crude fat was higher (P < 0·01) in the PF diet compared with the PFC diet, and lower (P < 0·01) for total carbohydrates. Exercise did not affect ATTD. Higher (P < 0·01) insulin-like growth factor 1 and leptin concentrations were measured when fed the PF diet compared with the PFC diet. Concentrations of insulin decreased (P < 0·01), whereas cortisol and ghrelin increased (P < 0·05), after exercise. There was no effect of diet on blood lactate and glucose, but higher (P < 0·001) lactate concentrations were measured in period 1 than in period 2. The results suggest that the o(13)CBT can be used in the field to estimate short-term EE in dogs during resting conditions. Higher ATTD and energy density of the PF diet may be beneficial when energy requirements are high.

Evaluation of the oral (13)C-bicarbonate technique for measurements of energy expenditure in dogs before and after body weight reduction.

BACKGROUND: Overweight and obesity are the most common nutritional disorders in dogs and may lead to various secondary diseases and decreased lifespan. In obesity research, measurement of energy expenditure (EE) and determination of the energy requirements are essential. The objective with this study was to validate and evaluate the suitability of the oral (13)C-bicarbonate technique (o(13)CBT) for measuring EE in dog obesity studies. A further objective was to investigate the impact of body weight (BW) reduction and changes in body composition on the EE when measured under conditions corresponding to the basal metabolic rate (BMR). RESULTS: The EE in five privately owned, overweight dogs was measured simultaneously with the o(13)CBT and indirect calorimetry (IC) for comparison of the results. Two measurements per dog were performed under the same standardised conditions (i.e. fasted and resting state) at the start, and after completing a 12-week BW reduction program. Additionally, measurements of body composition by Dual-energy X-ray absorptiometry (DEXA) were conducted at the beginning and at the end of the BW reduction program. There were no differences in EE results obtained by the o(13)CBT and IC. Overweight and the BW reduction did not affect the estimates for the respiratory quotient (RQ) or the recovery factor for the (13)C-tracer (RF), both needed when using the o(13)CBT. The dogs lost 16% (SD ± 2.0) of their initial BW in reduced fat mass (P < 0.001), whereas fat free mass (FFM) remained unchanged. There was no effect of the BW reduction on the determined EE expressed in kJ/kg BW/d, or in kJ/kg BW(0.75)/d. However, EE was lower (P < 0.001) after the BW reduction program when expressed in relation to FFM (kJ/kg FFM/d). CONCLUSIONS: Results from the present study show that the o(13)CBT can be a used in obesity research to determine EE in fasted dogs and under resting conditions. Furthermore, the results suggest that the BMR does not change with reduced BW in overweight dogs as long as the FFM remains unchanged. This indicates that the BMR to maintain one gram of fat is equal to maintaining one gram of FFM in overweight dogs.

The oral (13)C-bicarbonate technique for estimation of energy expenditure in dogs: validation against indirect calorimetry.

To get more knowledge about the energy requirements of dogs and to formulate appropriate feeding guidelines, it is essential to determine their energy expenditure (EE) in a reliable and feasible way. In this study, the non-invasive oral stable isotope (13)C-bicarbonate technique (o(13)CBT) was validated against indirect calorimetry (IC) for the determination of CO2-production and EE in dogs. Eleven privately owned dogs were simultaneously measured with IC and the o(13)CBT after being fasted overnight. All dogs were measured twice on two separate days. For calculation, measurements were divided into two groups depending on dogs' behaviour during the measurement. Dogs of Group 1 (n = 17) were resting calmly in the chamber and dogs of Group 2 (n = 5) were more active. Mean heart rate was significantly higher in Group 2 (102 beats per minute [bpm]) than in Group 1 (77 bpm) (p < 0.001). Within groups, the CO2-production and EE [kJ d(-1) kg BW(-0.75)] estimated by the o(13)CBT or IC did not differ significantly (Group 1: [Formula: see text] = 368; EEIC = 363; Group 2: [Formula: see text] = 701; EEIC = 718). However, the estimated (13)C recovery factor (RF) for the estimation of CO2-production was significantly different between Groups 1 and 2 (0.72 and 0.94, respectively, p < 0.001). The respiratory quotient (RQ), which is needed for the estimation of EE, did not differ between groups. This study shows that the non-invasive o(13)CBT can be used for accurate estimation of the CO2-production rate and EE in resting dogs. A value of 0.77 can be applied as an estimate of the RQ in fasted dogs and 0.72 as an appropriate estimate for RF when dogs are resting calmly during the measurements.

Evaluation of the oral ¹³C-bicarbonate tracer technique for the estimation of CO2 production and energy expenditure in dogs during rest and physical activity.

For feeding of working dogs during their daily life, illness, routine jobs or sporting activities, an accurate determination of their nutritional requirements is essential to ensure their optimal health and performance. To predict the appropriate guidelines about how to feed dogs, it appears essential to determine the energy expenditure (EE) in a reliable and feasible way. In the present experiment, the non-invasive oral ¹³C-bicarbonate tracer technique (o¹³CT), i.e. collection of breath samples after oral administration of NaH¹³CO3, was used for the estimation of CO2 production and EE in dogs. Measurements were conducted during two days of rest, and during three days with 3 h of exercise per day. Average EE was 483 and 876 kJ kg⁻°·75 d⁻¹ during rest and exercise, respectively. The o¹³CT seems appropriate to use as a minimal restrictive and non-invasive method to obtain reliable estimates of EE in dogs at different activity levels under near natural conditions.

Partitioning of late gestation energy expenditure in ewes using indirect calorimetry and a linear regression approach.

Late gestation energy expenditure (EE(gest)) originates from energy expenditure (EE) of development of conceptus (EE(conceptus)) and EE of homeorhetic adaptation of metabolism (EE(homeorhetic)). Even though EE(gest) is relatively easy to quantify, its partitioning is problematic. In the present study metabolizable energy (ME) intake ranges for twin-bearing ewes were 220-440, 350- 700, 350-900 kJ per metabolic body weight (W0.75) at week seven, five, two pre-partum respectively. Indirect calorimetry and a linear regression approach were used to quantify EE(gest) and then partition to EE(conceptus) and EE(homeorhetic). Energy expenditure of basal metabolism of the non-gravid tissues (EE(bmng)), derived from the intercept of the linear regression equation of retained energy [kJ/W0.75] and ME intake [kJ/W(0.75)], was 298 [kJ/ W0.75]. Values of the intercepts of the regression equations at week seven, five, and two pre-partum were 311, 398, and 451 [kJ/W0.75], respectively. The difference between the intercepts for different weeks was used to calculate EE(homeorhetic). The remaining part of EE(gest) was considered to be EE(conceptus). In conclusion, the good agreement between our values of EE(conceptus) and those in the literature indicates the method's validity.