Open-circuit Indirect Calorimetry System Research Articles

Description and Validation of Flow-Through Chambers of Respirometry for Measuring Gas Exchange in Animal Trials.

Indirect calorimetry (IC) is a widely used method to study animal energy metabolism by measuring gas exchange. The accuracy of IC depends on detecting variations in signals reflecting the metabolic response, which can be challenging due to measurement noise and external factors. This study proposes a methodology to validate IC systems, including an easy-to-use spreadsheet for data computing, to verify accuracy and detect whole-system leaks. We conducted a recovery test using a simulation of CO2 dynamics in MS Excel and injecting a known CO2 concentration into four respirometry chambers. The thought flow rate of CO2 was observed and compared to the expected rate from the simulation. Data filtering and computing, including a detailed calculation of signals calibration, Bartholomew transformation, and noise reduction, was developed to obtain the gas exchange and heat production parameters using an open-circuit IC system. The results from the recovery test in our system show that the proposed methodology is accurate and precise. The proposed methodology and recovery test can be used to standardize the validation of IC systems together with adequate data computing, providing accurate measurements of animal energy metabolism in different environmental conditions and energy utilization from feeds.

Energy, nitrogen partitioning, and methane emissions in dairy goats differ when an isoenergetic and isoproteic diet contained orange leaves and rice straw crop residues

The aim of this study was to evaluate the effects of incorporating rice straw and orange leaves into the diets for goats. Ten Murciano-Granadina goats at mid lactation weighing 45 ± 0.3 kg were used in a crossover design. Two isoproteic and isoenergetic diets (180 g/kg DM and 17 MJ/kg DM, respectively) with alfalfa hay as forage source (33% of DM) were fed. A control diet (CON) incorporated barley as energy source and soy hulls as fiber component. The experimental diet (ORG) replaced barley and soy hulls with orange leaves (19% on DM basis), rice straw (12%, on DM basis) and soya oil (2%). Peas and horsebeans were the protein source in both diets. Each goat received the 2 treatments in 2 periods. Goats were fed the experimental diets and after 14 d on their respective treatments moved to individual metabolism cages for another 7 d. Subsequently, feed intake, total fecal and urine output and milk yield were recorded daily over the first 5 d. During the next 2 d ruminal fluid and blood samples were collected, and then individual gas-exchange measurements were recorded by a mobile open-circuit indirect calorimetry system using a head box. No differences in dry matter intake were detected, and apparent total-tract digestibility was greater in CON than ORG. Efficiency of metabolizable energy intake for milk and maintenance also was lower in response to ORG (0.65 vs. 0.63), with energy balance being negative (-12 kJ/kg of BW0.75) due to mobilization of fat (-16 g/animal vs. 68 g/animal for ORG and CON, respectively). Although actual milk yield was lower in goats fed ORG (2.32 vs. 2.06 kg/d, respectively), energy-corrected milk did not differ (2.81 kg/d on average). In terms of milk quality, milk fat content, and concentrations of monounsaturated (18.54 vs. 11.55 g/100 g milk fat) and polyunsaturated fatty acids (5.75 vs. 3.99 g/100 g milk fat) were greater in goats fed ORG. Based on various indices, the milk produced by ORG would be less atherogenic and thrombogenic than CON milk. Compared with CON, enteric CH4 emission was lower due to feeding ORG (reduction of 38 g CH4/kg milk fat). Data suggest that greater fat mobilization in goats fed ORG might have been due to the apparent lack of synchrony between degradable protein and carbohydrate and the lipogenic nutrients associated with the lower cereal content of the ORG diet. Thus, goats fed ORG seemed to rely more on fat depots to help meet energy requirements and reach optimal performance. As such, the lower content of glucogenic nutrients in ORG did not favor body fat deposition and partitioning of ME into body tissue. Overall, responses in terms of CH4 emissions and milk quality suggest that inclusion of rice straw and orange leaves in diets for small ruminants could be a valuable alternative to reuse, recycle and revalue agricultural by-products.

Inclusion of lemon leaves and rice straw into compound feed and its effect on nutrient balance, milk yield, and methane emissions in dairy goats

The objective of this experiment was to study the effects of incorporating lemon leaves and rice straw into the compound feed of diets for dairy goats. Ten Murciano-Granadina dairy goats (n = 5 per group) in mid-lactation were used in a crossover design experiment (2 treatments across 2 periods). Goats were fed a mixed ration with barley grain (control, CON) or CON plus lemon leaves [189 g/kg of dry matter (DM)] and rice straw (120 g/kg of DM) in place of barley grain (LRS). Soybean oil (19 g/kg of DM) was added to the LRS diet to make it isoenergetic (17 MJ of gross energy/kg of DM) relative to CON. After 14 d on their respective treatments, goats were allocated to individual metabolism cages for another 7 d. Subsequently, feed intake, total fecal and urine output, and milk yield were recorded daily over the first 5 d. During the last 2 d, ruminal fluid and blood samples were collected, along with individual gas exchange measurements recorded by a mobile open-circuit indirect calorimetry system using a head box. No differences in DM intake were detected, and ME intake in LRS was lower than in CON (1,095 vs. 1,180 kJ/kg of metabolic body weight). No differences were observed in milk production, but milk fat content was greater in LRS (6.4%) than in CON (5.6%). Greater concentrations of monounsaturated (14.94 vs. 11.96 g/100 g of milk fat) and polyunsaturated fatty acids (4.53 vs. 4.03 g/100 g of milk fat) were detected in the milk of goats fed LRS compared with CON. Atherogenicity (2.68 vs.1.91) and thrombogenic (4.58 vs. 2.81) indices were lower with LRS compared with CON. Enteric CH4 emission was lower in LRS (24.3 g/d) compared with CON (31.1 g/d), probably due to the greater lipid content and unsaturated fatty acid profile of lemon leaves and the soybean oil added in the LRS diet. Overall, data suggest that incorporating lemon leaves and rice straw into lactating goat diets is effective in reducing CH4 emissions while allowing improvements in milk fat production and milk thrombogenic index without affecting production performance. Thus, their inclusion in compound feeds fed to small ruminants appears warranted and would have multiple positive effects, as on efficiency of nutrient use, human health, and the environment.

An Open-Circuit Indirect Calorimetry Head Hood System for Measuring Methane Emission and Energy Metabolism in Small Ruminants.

Simple SummaryAn open-circuit indirect calorimetry system for small ruminants was updated. Calibration factors for CH4, CO2 and O2 close to 1 confirmed the absence of leaks in the indirect calorimetry system and the accurate performance of this device. An experimental test quantified the gas exchange and the repeatability for CH4 and heat production measurements, which were 79% and 61%, respectively. The heat production obtained by indirect calorimetry was close to the heat production obtained by carbon and nitrogen balance. Discrepancies between the two methods averaged 1.92% when expressed as a percentage of the intake of metabolizable energy, a rather satisfactory value considering the substantial amount of technical and analytical work involved. The close agreement found between them can be considered as an indicative of the absence of systematic error. When diets with different forages were compared, the daily CH4 production was 1.54 and 1.25 L/h for diets-based in alfalfa hay and alfalfa silage, respectively.Methane (CH4) is a natural by-product of microbial fermentation in the rumen and is a powerful greenhouse gas. An open-circuit indirect calorimetry system for continuous determination of CH4 and CO2 production and O2 consumption and, thereafter, heat production (HP) calculation for small ruminants was described and validated. The system consisted of a computerized control, data acquisition and recording system for gases and air flux. The average value ± standard deviation for the calibration factors in the system were 1.005 ± 0.0007 (n = 6), 1.013 ± 0.0012 (n = 6) and 0.988 ± 0.0035 (n = 6) for O2, CO2 and CH4, respectively. Calibration factors close to 1 confirmed the absence of leaks in the indirect calorimetry system. In addition, an experimental test with 8 goats at mid lactation was conducted to validate the system. The repeatability for CH4 and heat production measured with the open-circuit indirect calorimetry system was 79% and 61%, respectively. Daily average HP measured by indirect calorimetry (Respiration Quotient method) was close to the average HP determined from Carbon-Nitrogen balance (CN method), accounting for 685 and 667 kJ per kg metabolic body weight, respectively. Therefore, discrepancies averaged 1.92%, a rather satisfactory value considering the substantial amount of technical and analytical work involved. The close agreement found between both methods can be considered as being indicative of the absence of systematic error. Two diets with different forage were tested: 40% was either alfalfa hay (HAY) or alfalfa silage (SIL), and the proportion of concentrate was the same in both groups (60%). The experimental trial shown that HP and CH4 were higher in HAY than SIL diet (differences between treatments of 28 kJ of HP per kg of metabolic body weight and 7.1 L CH4/day were found). The data acquisition and recording device developed improved the accuracy of the indirect calorimetry system by reducing the work involved in managing output data and refining the functionality for measuring gas exchange and energy metabolism in small ruminants.

The natural compound, formononetin, extracted from Astragalus membranaceus increases adipocyte thermogenesis by modulating PPARγ activity.

Increasing energy expenditure through adipocyte thermogenesis is generally accepted as a promising strategy to mitigate obesity and its related diseases. However, few clinically effective and safe agents are known to promote adipocyte thermogenesis. In this study, 20 traditional Chinese herbal medicines were screened to examine whether they induced adipocyte thermogenesis. The effects of Chinese herbal medicines or components isolated from extracts of A. membranaceus, on adipocyte thermogenesis were analysed by assessing expression of uncoupling protein 1 (UCP1) by qPCR. Eight-week-old C57BL6/J male mice were fed a high-fat diet for 8weeks and then randomized to two groups treated with vehicle or formononetin for another 8weeks. Glucose tolerance tests and staining of adipose tissue with haematoxylin and eosin were carried out. Whole-body oxygen consumption was measured with an open-circuit indirect calorimetry system. Extracts of A. membranaceus increased expression of Ucp1 in primary cultures of mouse adipocytes. Formononetin was the only known component of A. membranaceus extracts to increase adipocyte Ucp1 expression. Diet-induced obese mice treated with formononetin gained less weight and showed higher energy expenditure than untreated mice. In addition, formononetin binds directly with PPARγ. Taken together, our study demonstrates that the Chinese herbal medicine from A. membranaceus and its constituent formononetin have the potential to reduce obesity and associated metabolic disorders. Our results suggest that formononetin regulates adipocyte thermogenesis as a non-classical PPARγ agonist.

Effect of lemon leaves on energy and C-N balances, methane emission, and milk performance in Murciano-Granadina dairy goats.

The objective of this experiment was to find out the effect of lemon leaves on energy and C-N balances, methane emission, and milk performance in dairy goats. Lemon leaves were used to replace alfalfa as forage in a diet for Murciano-Granadina goats. Ten Murciano-Granadina dairy goats (44.1 ± 4.47 kg of BW) in late lactation (185 ± 7.2 d) were selected in a crossover design experiment, where each goat received 2 treatments in 2 periods. One group was fed a mixed ration with 450 g of pelleted alfalfa per kilogram of DM (ALF diet) and, the other group replaced alfalfa with 450 g of pelleted lemon leaves per kilogram DM (LEM diet). The concentrate was pelleted, being the same for the two groups (forage to concentrate ratio was 45/55). The goats were allocated to individual metabolism cages. After 14 d of adaptation, feed intake, total fecal and urine output, and milk yield were recorded daily over a 5-d period. Then, gas exchange measurements were recorded individually by an open-circuit indirect calorimetry system using a head box. Higher dietary lipids in LEM diet reduced DMI (200 g/d) and energy intake (251 kJ/kg of BW0.75), although no differences between treatments were observed for ME intake (998 kJ/kg of BW0.75, on average) and oxidation of nutrients (64% and 25% for carbohydrates and fat oxidation, respectively, on heat production from oxidation basis). Greater (P < 0.05) milk fat values for C18:2n6t and CLA 9c11t + 9t11c were found in LEM compared with ALF diet. Goats fed LEM diet produced significantly fewer CH4 emissions than ALF diet (18%). Likewise, the use of lemon leaves as forage reduced the amount of CH4 in 2.7 g/kg of milk. Results suggest that lemon leaves are effective in reducing CH4 emission without detrimental effect on milk yield.

Long-term intake of saccharin decreases post-absortive energy expenditure at rest and is associated to greater weight gain relative to sucrose in wistar rats

BackgroundNon-nutritive sweeteners (NNS) have been associated with increased prevalence of obesity. In previous studies, we demonstrated that saccharin could induce an increase in weight gain either when compared to sucrose or to a non-sweetened control at a similar total caloric intake. These data raised the hypothesis that reduced energy expenditure (EE) could be a potential mechanism explaining greater weight gain with saccharin use in rats. The aim of the present study was to compare long-term energy expenditure at rest between rats using saccharin or sucrose and correlate it with weight gain. MethodsIn the present study, we examine the potential impact of saccharin compared to sucrose in the EE of Wistar rats. In a controlled experiment of 17 weeks, 24 Wistar rats were divided into 2 groups: saccharin-sweetened yogurt (SAC) or sucrose-sweetened yogurt (SUC), plus a free chow diet. Only rats that consumed at least 70% of the offered yogurt were included. EE (kcal/day) was determined at rest through open circuit indirect calorimetry system in the early post-absorptive period with determinations of both VO2 consumption and CO2 production. Measurements were evaluated at baseline, 5 and 12 weeks of dietary intervention. Weight gain, caloric intake (from yogurt, from chow and total) were determined weekly.ResultsBody weight and EE were similar between groups at baseline: (p = .35) and (p = .67) respectively. At the end of the study, SAC increased total weight gain significantly more in relation to SUC (p = .03). Cumulative total caloric intake (yogurt plus chow) was similar between groups during the whole period (p = .54). At 12 weeks, the EE was smaller in SAC compared to SUC (p = .009). Considering both groups, there was a strong negative correlation between total weight gain and change in EE observed [r(20) = −.61, p = .003]. However, when analyzing the groups separately we found that SUC maintained this inverse correlation [r(8) = −.68, p = .03], while SAC did not [r(10) = −.33, p = .29].ConclusionThese data support the hypothesis that long-term use of saccharin may blunt post-absorptive EE at rest in Wistar rats, which is related to weight gain. On the other hand, long-term sucrose intake can increase energy expenditure in rats. This effect combined can explain, at least partially, the weight gain increases associated to saccharin in relation to sucrose in these animals.

Replacement of alfalfa hay (Medicago sativa) with maralfalfa hay (Pennisetum sp.) in diets of lactating dairy goats

The objective of this experiment was to study the effects of substituting alfalfa (Medicago sativa) with maralfalfa (Pennisetum sp.) on energy, nitrogen and carbon balance, methane emission, and milk performance in dairy goats. Ten Murciano-Granadina dairy goats in late lactation (45.7±2.96kg of body weight [BW]) were selected in a 2-treatment and crossover design experiment where each goat received both treatments in 2 periods. One group of five goats was fed a mixed ration with alfalfa as forage (A diet) and the other diet replaced alfalfa with maralfalfa (M diet) in a forage concentrate ratio of 40/60. Diets were isoenergetic and isoproteic. The goats were allocated to individual metabolism cages. After 17 d of adaptation, feed intake, total faecal and urine output and milk yield were recorded daily over a 5 d period. Then, gas exchange measurements were recorded individually by a mobile open-circuit indirect calorimetry system using a head box. Daily dry matter intake was higher on A than on M diets (1.8 vs. 1.6, for A and M respectively), but no differences was observed in metabolizable energy intake (1089kJ/kg of BW0.75, on average). Greater values in M than A were found in rumen propionic acid (17.5 vs. 15.6mol/100mol, respectively) and milk C15:0 (0.81 vs. 0.62g/100g), C17:0 (0.33 vs. 0.24g/100g) fatty acids, indicating predominance of amylolytic fermentation. Milk uric acid, a potential biomarker of rumen nitrogen flow and feed efficiency, was higher in A than M (133 vs. 102μmol/L). Higher milk yield (1.8 vs. 1.7kg/d) and CH4 emissions (28.5 vs. 25.9g/d) in A compared to M diet were observed, respectively. Use of tropical grasses such as maralfalfa in temperate climates could be a strategy for farmers to incorporate forage into feed for flocks, as milk chemical composition did not change markedly and maralfalfa inclusion reduced CH4 emissions by goats (although this reduction did not occur in CH4 emission compared to dry matter intake and milk yield).

Murciano-Granadina Goat Performance and Methane Emission after Replacing Barley Grain with Fibrous By-Products.

The aim of this experiment was to study the effects of substituting dietary barley grain with orange pulp or soybean hulls on energy, nitrogen and carbon balance, methane emission and milk performance in dairy goats. Twelve Murciano-Granadina dairy goats in midlactation were selected and divided into three groups based on similar body weight (42.1 ± 1.2 kg) and milk yield (2.16 ± 0.060 kg/goat/day). The experiment was conducted in an incomplete crossover design where one group of four goats was fed a mixed ration of barley grain (BRL), another group of four goats replaced barley grain with orange pulp (OP) and the last group of four goats with soybean hulls (SH). After adaptation to diets, the goats were allocated to individual metabolism cages and intake, faeces, urine and milk were recorded and analysed. Then, gas exchange measurements were recorded by a mobile open-circuit indirect calorimetry system using a head box. Dry matter intake was similar for all three groups (2.03 kg/d, on average). No influence of the diet was observed for energy balance and the efficiency of use of metabolizable energy for milk production was 0.61. The OP and SH diets showed greater (P < 0.05) fat mobilization (-42.8 kJ/kg of BW0.75, on average) than BRL (19.2 kJ/kg of BW0.75). Pentadecanoic acid (15:0) and heptadecanoic acid (17:0) were potential biomarkers of rumen function because the higher contents found in the milk of OP and SH goats than BRL suggest a negative impact of these diets on rumen bacterial metabolism; probably linked to the lower nitrogen supply of diet OP to synthesize microbial protein and greater content of fat in diet SH. Replacement of cereal grain with fibrous by-products did not increased enteric methane emissions (54.7 L/goat per day, on average). Therefore, lactating goats could utilize dry orange pulp and soybean hulls diets with no detrimental effect on milk performance.

Effect of rice bran as a replacement for oat grain in energy and nitrogen balance, methane emissions, and milk performance of Murciano-Granadina goats

The objective of this experiment was to study the effects of substituting oat grain with rice bran on energy, nitrogen and carbon balance, methane emissions, and milk performance in dairy goats. Ten Murciano-Granadina dairy goats in late lactation (46.1±3.07kg) were assigned to 2 treatments in a crossover design, where each goat received both treatments in 2 periods. One group of 5goats was fed a mixed ration with 379g of oat grain/kg of dry matter (O diet) and the other group of 5 goats was fed a diet that replaced oat grain with 379g/kg dry matter of rice bran (RB diet). Diets were formulated to be isoenergetic and isoproteic, so bypass fat was added to reach the same amount of energy in both diets. The goats were allocated to individual metabolism cages. After 14 d of adaptation, feed intake, total fecal and urine outputs, and milk yield were recorded daily over a 5-d period. Then, gas exchange measurements were recorded individually by a mobile open-circuit indirect calorimetry system using a head box. Dry matter intake was different for both diets [1.83±0.11 vs. 1.61±0.08 (means ± SD), for O and RB, respectively]. Metabolizable energy intake and heat production were not significantly different between diets, with average values of 1,254 [standard error of the mean (SEM)=110.0] and 640 (SEM=21.0) kJ/kg of BW0.75, respectively. Significant differences were found in milk fat content (5.3 and 6.9%, SEM=0.36; for O and RB, respectively) and milk fatty acids: medium-chain fatty acids (17.17 vs. 12.90g/100g, SEM=0.969; for O and RB, respectively) and monounsaturated fatty acids (20.63 vs. 28.29g/100g, SEM=1.973; for O and RB, respectively). Enteric CH4 emission was lower for the RB diet (23.2 vs. 30.1g/d, SEM=2.14; for O and RB, respectively), probably because of the higher lipid content in RB diets than O diets (11.7 vs. 4.1%, respectively). Lactating goats utilized RB without detrimental effects on energy metabolism. Higher milk fat and lower CH4 emissions were observed with the RB diet compared with the O diet.

Use of dry citrus pulp or soybean hulls as a replacement for corn grain in energy and nitrogen partitioning, methane emissions, and milk performance in lactating Murciano-Granadina goats

The aim of this study was to assess the effect of substitution of dietary corn grain by dry citrus pulp or soybean hulls on energy and nitrogen partitioning, substrate oxidation, methane emission, and milk performance in dairy goats during midlactation. Twelve multiparous Murciano-Granadina goats of similar body weight (41.7±2.8kg) were split in 3 groups in an incomplete crossover design. One group of 4 goats was fed a mixed ration with 605g/kg of dry matter of corn grain (CRG), another group replaced corn grain with dry citrus pulp (CTP), and the last with soybean hulls (SYH). The goats were allocated to individual metabolism cages. After 14 d of adaptation, feed intake, total fecal and urine output, and milk yield were recorded daily over a 5-d period. Then, gas exchange measurements were recorded by a mobile open-circuit indirect calorimetry system using a head box. Dry matter intake was similar for all 3 groups (1.53kg/d, on average). Total replacement of the concentrate with fibrous by-products increased fiber apparent digestibility. The metabolizable energy intake was significantly greater for diet CRG than SYH (1,193 vs. 1,079kJ/kg of BW0.75, respectively), CTP showed an intermediate value. The heat production was higher for the fiber diet than starchy diet (908 vs. 843kJ/kg of BW0.75 for SYH and CRG, respectively). The efficiency of use of metabolizable energy for milk production obtained by regression was 0.59. Goats fed CTP and SYH diets produced similar CH4 emissions (34.8g/d, on average), significantly higher compared with goats fed the CRG diet (24.7g/d). Goats of the 3 treatments were in negative energy balance, so the oxidation of fat was greater than for carbohydrates. No significant differences were observed for milk production (1.72kg/d), and milk fat was significantly greater for a more fibrous diet compared with a starchy diet (6.57 vs. 4.95% in SYH and CRG, respectively).

Novel Indirect Calorimetry Technology to Analyze Metabolism in Individual Neonatal Rodent Pups

BackgroundThe ability to characterize the development of metabolic function in neonatal rodents has been limited due to technological constraints. Low respiratory volumes and flows at rest pose unique problems, making it difficult to reliably measure O2 consumption, CO2 production, respiratory quotient (RQ), and energy expenditure (EE). Our aim was to develop and validate a commercial-grade indirect calorimetry system capable of characterizing the metabolic phenotype of individual neonatal rodents.Methodology/Principal FindingsTo address this research need, we developed a novel, highly sensitive open-circuit indirect calorimetry system capable of analyzing respiratory gas exchange in a single neonatal rodent pup. Additionally, we derived an equation from known metabolic relationships to estimate inlet flow rates, improving the efficiency of data collection. To validate the neonatal rodent indirect calorimetry system and evaluate the applicability of the derived equation for predicting appropriate flow rates, we conducted a series of experiments evaluating the impact of sex, litter size, time of day (during the light phase), and ambient temperature on neonatal rat metabolic parameters. Data revealed that the only metabolic parameter influenced by litter size is a neonatal rat's RQ, with rat pups reared in a small litter (5 pups) having lower RQ's than rat pups reared in either medium (8 pups) or large (11 pups) litters. Furthermore, data showed that ambient temperature affected all metabolic parameters measured, with colder temperatures being associated with higher CO2 production, higher O2 consumption, and higher energy expenditure.Conclusion/SignificanceThe results of this study demonstrate that the modified Panlab Oxylet system reliably assesses early postnatal metabolism in individual neonatal rodents. This system will be of paramount importance to further our understanding of processes associated with the developmental origins of adult metabolic disease.

Technical note: A new facility for continuous respiration measurements in lactating cows

An open-circuit indirect calorimetry system consisting of 4 climate-controlled respiration chambers for cattle has been constructed and validated. The system allows for the continuous monitoring of O2, CO2, and CH4 concentrations in chamber air, and the simultaneous determination of feed and water intake, overall physical activity, position changes, standing and lying times, and animal behavior. For complete balance trials, feces, urine, and milk can be collected quantitatively. Most importantly, lactating cows can be milked in the chamber, and blood samples can be drawn from permanent catheters without disruption of the measurements. The investigator, on entering the chamber, wears a facemask connected to the ambient air during the whole milking process. Data are routed to a data acquisition system with appropriate data evaluation software developed in our research unit. Thus, dynamic changes of the above-named parameters during the course of the day or of longer time periods can be monitored. Such data are critical for understanding the complex regulation and interplay of feed intake, energy metabolism, climatic conditions, and milk production.

Long-Term Effects of Feeding Monensin on Methane Production in Lactating Dairy Cows

The objective of this study was to determine the long-term effects of feeding monensin on methane (CH4) production in lactating dairy cows. Twenty-four lactating Holstein dairy cows (1.46±0.17 parity; 620±5.9kg of live weight; 92.5±2.62 d in milk) housed in a tie-stall facility were used in the study. The study was conducted as paired comparisons in a completely randomized design with repeated measurements in a color-coded, double-blind experiment. The cows were paired by parity and days in milk and allocated to 1 of 2 treatments: 1) the regular milking cow total mixed ration (TMR) with a forage-to-concentrate ratio of 60:40 (control TMR; placebo premix) vs. a medicated TMR (monensin TMR; regular TMR + 24mg of Rumensin Premix/kg of dry matter) fed ad libitum. The animals were fed and milked twice daily (feeding at 0830 and 1300h; milking at 0500 and 1500h) and CH4 production was measured prior to introducing the treatments and monthly thereafter for 6 mo using an open-circuit indirect calorimetry system. Monensin reduced CH4 production by 7% (expressed as grams per day) and by 9% (expressed as grams per kilogram of body weight), which were sustained for 6 mo (mean, 458.7 vs. 428.7±7.75 g/d and 0.738 vs. 0.675±0.0141, control vs. monensin, respectively). Monensin reduced milk fat percentage by 9% (3.90 vs. 3.53±0.098%, control vs. monensin, respectively) and reduced milk protein by 4% (3.37 vs. 3.23±0.031%, control vs. monensin, respectively). Monensin did not affect the dry matter intake or milk yield of the cows. These results suggest that medicating a 60:40 forage-to-concentrate TMR with 24mg of Rumensin Premix/kg of dry matter is a viable strategy for reducing CH4 production in lactating Holstein dairy cows.

CCAAT/Enhancer-binding Protein β Deletion Reduces Adiposity, Hepatic Steatosis, and Diabetes in Lepr Mice

CCAAT/enhancer-binding protein beta (C/EBPbeta) plays a key role in initiation of adipogenesis in adipose tissue and gluconeogenesis in liver; however, the role of C/EBPbeta in hepatic lipogenesis remains undefined. Here we show that C/EBPbeta inactivation in Lepr(db/db) mice attenuates obesity, fatty liver, and diabetes. In addition to impaired adipogenesis, livers from C/EBPbeta(-/-) x Lepr(db/db) mice had dramatically decreased triglyceride content and reduced lipogenic enzyme activity. C/EBPbeta deletion in Lepr(db/db) mice down-regulated peroxisome proliferator-activated receptor gamma2 (PPARgamma2) and stearoyl-CoA desaturase-1 and up-regulated PPARalpha independent of SREBP1c. Conversely, C/EBPbeta overexpression in wild-type mice increased PPARgamma2 and stearoyl-CoA desaturase-1 mRNA and hepatic triglyceride content. In FAO cells, overexpression of the liver inhibiting form of C/EBPbeta or C/EBPbeta RNA interference attenuated palmitate-induced triglyceride accumulation and reduced PPARgamma2 and triglyceride levels in the liver in vivo. Leptin and the anti-diabetic drug metformin acutely down-regulated C/EBPbeta expression in hepatocytes, whereas fatty acids up-regulate C/EBPbeta expression. These data provide novel evidence linking C/EBPbeta expression to lipogenesis and energy balance with important implications for the treatment of obesity and fatty liver disease.

Development of a mobile, open-circuit indirect calorimetry system

A system for the rapid determination of O2 consumption, CO2 production and subsequent indirect estimation of energy metabolism of ruminants has been updated and validated. The accurate determination of gas temperature, relative humidity, absolute pressure and flow is important to the overall use of the system. Analogue outputs from the specific analyzers were multiplexed and then routed to a data acquisition system with appropriate software control. Subsequent data processing included modification to fit LOTUS 1-2-3 files. The analyzers were calibrated with standard test gases for the gas meters. In an experimental confirmation trial, eight sheep were fed at maintenance (M) or 2× maintenance (2M) or fasted 48 h after each level of intake (MF and 2MF, respectively) and subsequent energy metabolism was determined. Oxygen consumption (mL min−1), metabolic rate (W) and CO2 production (mL min−1) were greater (P &lt; 0.05) for animals fed 2M (111.2, 379.5 and 101.9, respectively) than for those fed M (85.4, 291.2 and 86.1, respectively). Maintenance values were, in turn, also higher (P &lt; 0.05) than those for either MF (56.7, 193.4 and 48.0, respectively) or 2MF (54.6, 186.4 and 46.3, respectively). M sheep had a higher respiratory quotient (1.03) (P &lt; 0.05) than either MF sheep (0.86) or 2MF sheep (0.89), with 2M sheep being intermediate (0.95). There were no differences (P &gt; 0.05) between the two fasted states for all parameters. Results from the confirmation experiment agree with previously published work and were consistent with those from similar systems. Key words: Indirect calorimetry, oxygen consumption, sheep

Effect of age on body composition and resting metabolic rate

The relationship between fat-free mass (FFM) and resting metabolic rate (RMR) was compared in young men (n = 24; age 18-33 yr), old men (n = 24; 69-89 yr), and old women (n = 20; 67-75 yr). Body composition was assessed using anthropometry, bioelectrical impedance analysis (BIA), and isotope dilution with 18O-labeled water. RMR was measured at least twice using an open-circuit indirect calorimetry system with a ventilated hood. The results indicate that the different methods for assessing body composition vary substantially and should not be used interchangeably. Anthropometry was not adequate to assess group differences in body fatness, although skinfold measures may be appropriate for within-group comparisons. BIA correlated well with the isotope-dilution technique and may be a useful measure of FFM. Finally, RMR was lower in the old men than the young (1.04 +/- 0.02 vs. 1.24 +/- 0.03 kcal/min, P less than 0.001) and remained lower even when adjusted for FFM estimated by isotope dilution (P less than 0.001). RMR in the women was also lower (0.84 +/- 0.02 kcal/min), but in contrast to the difference between young and old men, RMR adjusted for FFM did not differ (P = 0.16) between old men and women. Therefore, it is clear that differences in FFM cannot fully account for the lower RMR in the old, suggesting that aging is associated with an alteration in tissue energy metabolism.

Construction and operation of an open-circuit indirect calorimetry system for small ruminants.

An open-circuit indirect respiration calorimetry system for the collection of long-term metabolic heat production data for medium and small animals is described. The system is designed for use with small ruminants, but it has the capacity for measuring heat production in nonruminant species up to 150 kg. The system consists of an internal climatic control system, air flow control and volume measuring system, and gas analyzers for O2, CO2 and CH4. Carbon dioxide recovery checks of the four chambers produced recovery values ranging from 96.73 to 100.89%. The actual mean fasting heat production measured for 12 wether sheep was 63.13 kcal/kg.75 +/- .83. The total estimated cost of four chambers and the associated instrumentation was less than $17,000 in 1984.

268. Induction of the “estrous FSH-surge” following exogenous LH and ovulation in the golden hamster and its suppression by estradiol and progesterone

The objective of this study was to determine the long-term effects of feeding monensin on methane (CH4) production in lactating dairy cows. Twenty-four lactating Holstein dairy cows (1.46 ± 0.17 parity; 620 ± 5.9 kg of live weight; 92.5 ± 2.62 d in milk) housed in a tie-stall facility were used in the study. The study was conducted as paired comparisons in a completely randomized design with repeated measurements in a color-coded, double-blind experiment. The cows were paired by parity and days in milk and allocated to 1 of 2 treatments: 1) the regular milking cow total mixed ration (TMR) with a forage-to-concentrate ratio of 60:40 (control TMR; placebo premix) vs. a medicated TMR (monensin TMR; regular TMR + 24 mg of Rumensin Premix/kg of dry matter) fed ad libitum. The animals were fed and milked twice daily (feeding at 0830 and 1300 h; milking at 0500 and 1500 h) and CH4 production was measured prior to introducing the treatments and monthly thereafter for 6 mo using an open-circuit indirect calorimetry system. Monensin reduced CH4 production by 7% (expressed as grams per day) and by 9% (expressed as grams per kilogram of body weight), which were sustained for 6 mo (mean, 458.7 vs. 428.7 ± 7.75 g/d and 0.738 vs. 0.675 ± 0.0141, control vs. monensin, respectively). Monensin reduced milk fat percentage by 9% (3.90 vs. 3.53 ± 0.098%, control vs. monensin, respectively) and reduced milk protein by 4% (3.37 vs. 3.23 ± 0.031%, control vs. monensin, respectively). Monensin did not affect the dry matter intake or milk yield of the cows. These results suggest that medicating a 60:40 forage-to-concentrate TMR with 24 mg of Rumensin Premix/kg of dry matter is a viable strategy for reducing CH4 production in lactating Holstein dairy cows.