Quantifying energy expenditure in Göttingen Minipigs with the 13C-bicarbonate method under basal and drug-treated conditions.

Effective treatments of obesity focusing on energy expenditure (EE) are needed. To evaluate future EE-modulating drug candidates, appropriate animal models and methods to assess EE are needed. This study aimed to evaluate the stable isotope 13C-bicarbonate method (13C-BM) for estimating EE in Göttingen minipigs under basal and drug-treated conditions. Four experiments (Expt.1-4) were conducted to assess: 1) the 13C-BM reproducibility using breath sample collection (n = 8), on two consecutive days, 2) the effect of two dose levels (5 and 10 mg/kg body weight (BW)) of the mitochondrial uncoupler dinitrophenol (DNP) in a crossover design (n = 8), 3) sampling method agreement; blood vs. exhaled air (n = 6) and 4) 13C-BM using constant isotope infusion compared with indirect calorimetry (IC) (n = 3). Results correlated significantly (p < 0.001) between days (Expt.1), with an average coefficient of variance of 5.4 ± 2.3%. Administration of 10 mg DNP/kg BW increased (p < 0.01) EE by 33.2 ± 6.4% (Expt.2). Results based on different sampling methods correlated significantly (p < 0.001) and EE increased after 10 mg DNP/kg BW (p < 0.05) in Expt.3. However, results based on blood sampling were significantly higher (p < 0.01) than those of exhaled air. No effect of DNP and significantly different EE results (p < 0.05) was observed in a limited number of animals, when constant isotope infusion and blood sampling was compared with IC (Expt.4). In conclusion, the 13C-BM is useful for investigating treatment effects on EE in minipigs. However, further validation under standardized conditions is needed to provide accurate estimates of the 13C recovery factor and respiratory quotient, both of decisive importance when using the 13C-BM.

Effect of Cyberlindnera jadinii yeast on growth performance, nutrient digestibility, and gut health of broiler chickens from 1 to 34 d of age.

The effect of dietary graded levels of Cyberlindnera jadinii yeast (C. jadinii) on growth performance, nutrient digestibility, and gut health of broilers was evaluated from 1 to 34 d of age. A total of 360 male broiler chicks were randomly allocated to 1 of 4 dietary treatments (6 replicate pens each) consisting of a wheat-soybean meal-based pelleted diet (Control or CJ0), and 3 diets in which 10% (CJ10), 20% (CJ20), and 30% (CJ30) of the crude protein were supplied by C. jadinii, by gradually replacing protein-rich ingredients. Body weight and feed intake were measured at d 1, 11, 22, and 32. Pellet temperature, durability, and hardness increased linearly (P < 0.05) with C. jadinii inclusion, with highest (P < 0.05) values for CJ30. Up until d 22, feed conversion ratio (FCR) was similar between treatments (P = 0.169). Overall, increasing C. jadinii inclusion linearly increased (P = 0.047) feed intake but had no effect on weight gain or mortality. FCR increased (P < 0.05) linearly with increasing C. jadinii inclusion but only birds fed CJ30 had a significantly poorer FCR compared to the Control. Ileal digestibility was not affected by C. jadinii inclusion, however, there was a significant linear decrease in crude protein and phosphorus, and a tendency for a decrease in fat digestibility. Apparent metabolizable energy (AME) decreased (P < 0.001) quadratically with increasing C. jadinii and was significantly lower in CJ30 compared to the Control. Ileal concentrations of volatile fatty acids (VFAs) were not affected by C. jadinii inclusion, but butyric acid and total VFAs were linearly and quadratically increased and were significantly higher in cecal digesta of birds fed CJ20 and CJ30. Increasing C. jadinii inclusion was associated with an increase (P < 0.05) in the relative abundance of lactobacillus in the ileum and cecum. In conclusion, C. jadinii yeast can supply up to 20% of the total dietary protein without negatively affecting performance, digestibility, or gut health of broilers. The potential confounding role of feed processing and C. jadinii cell wall components on broiler performance is discussed.

Development of In Vitro Assays with the Canine Hookworm Uncinaria stenocephala and Assessment of Natural Plant Products for Anti-Parasitic Activity.

Enteric helminth infection is an increasing concern in companion animals due to reports of resistance to commonly used anthelmintic drugs. Thus, the assessment of new therapeutic options such as bioactive dietary additives is of high importance. Here, we adapted egg hatch, larval migration, and larval motility assays to screen extracts of several natural ingredients against the canine hookworm Uncinaria stenocephala, a prevalent parasite of dogs in northern Europe. Egg hatch and larval migration assays were established showing that the anthelmintic drugs levamisole and albendazole had strong anti-parasitic activity against U. stenocephala, validating the use of these assays for the assessment of novel anti-parasitic substances. Subsequently, we identified that extracts from the seaweed Saccharina latissima, but not extracts from grape seed or chicory, significantly inhibited both hatching and larval migration. Finally, we showed that α-linolenic acid, a putative anti-parasitic compound from S. latissima, also exhibited anti-parasitic activity. Collectively, our results established a platform for the screening for anthelmintic resistance or novel drug candidates against U. stenocephala and highlighted the potential use of seaweed extracts as a functional food component to help control hookworm infection in dogs.

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.