Evaluation of the oral 13C-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 ¹³C-bicarbonate technique (o¹³CBT) 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 o¹³CBT 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 o¹³CBT and IC. Overweight and the BW reduction did not affect the estimates for the respiratory quotient (RQ) or the recovery factor for the ¹³C-tracer (RF), both needed when using the o¹³CBT. 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 o¹³CBT 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.     

Response to lysine in a wheat gluten diet in adult minipigs after short-and long-term dietary adaptation as assessed with an indicator amino acid oxidation and balance technique

An experiment was conducted to examine the response to wheat gluten (WG)-based diets at two lysine levels in adult minipigs (23 kg BW) using the indicator AA oxidation (IAAO) approach and N balance. Twenty minipigs (n = five per group), fitted with reentrant ileoileal cannulas allowing collection of ileal effluents, were fed restrictively two WG-based diets (WG and WG + Lys; 2.7 and 6.6 g of lysine/kg, respectively) for adaptation periods of 10 and 100 d. On d 7 and 9, for pigs fed the diets for 10 d, and on d 97 and 99, for pigs fed the diets for 100 d, primed i.v. fasted/fed tracer protocols with [(13)C]bicarbonate, and [(13)C]leucine were performed. With the WG diet, [(13)C]bicarbonate recoveries (%) were lower irrespective of the adaptation period, and higher during the fed period (fasted: WG + Lys = 82.5, and WG = 69.1; fed: WG + Lys = 90.6, and WG = 85.9; P < 0.05). Leucine oxidation rate was higher with the lower lysine intake (WG = 194.6 vs. 109.5 mg/[kg BW x d]; P < 0.05). Wheat gluten feeding resulted in a negative leucine balance independent of the adaptation period (WG = -29.1, and WG + Lys = 48.2 mg/[kg BW x d]; P < 0.05). In contrast with the IAAO method, N balance did not differ between the two lysine intakes, possibly because of an underestimation of N losses. The finding of a lower (13)C bicarbonate recovery with the lower dietary lysine intake suggests that caution should be taken in using a single recovery factor for all AA oxidation studies.     

Utilization of glucose and long-chain fatty acids in lactating dairy cows fed a fat-enriched diet

The fate of carbon from long-chain fatty acids and glucose in dairy cows which were fed with protected fat was studied using stable isotope technique. The experiment was carried out on two groups of dairy cows (n=16 in each group) during the first 15 weeks of the lactation period. The cows were fed isoenergetic and isoproteinogenous diets based on corn silage. About 1.8 kg of tapioca starch in the diet of the starch group was substituted by about 0.7 kg of rumen protected fat (Ca salts of palm oil and soybean oil) in the diet of the fat group. The carbon atoms of dietary fat were naturally depleted in 13C as compared to carbon atoms of starch. Daily milk performance and lactose output were significantly (P < 0.05) higher among the cows fed with fat diet. In comparison to the starch group, the enrichment of milk fat with 13C was significantly lower, while that of breath CO2 was significantly higher in the fat group (P < 0.05). This means the fatty acids were incorporated into milk fat in preference to metabolic oxidation. Further studies showed that blood glucose is oxidized to a lower extent and is used for the synthesis of lactose to a higher proportion if the cows were fed with the fat diet. The glucose entry rate into the body glucose pool was not different between the diets. In conclusion, the dietary fatty acids perform a glucose sparing effect and improve the glucose supply for the mammary gland.     

Early selection for extreme resistance to potato virus Y and tobacco etch virus in potato using a β-glucuronidase-tagged virus

The Ry_sto gene from Solanum stoloniferum introduced into potato cultivars (Solanum tuberosum L. ssp. tuberosum) confers resistance to potato virus A, potato virus V and potato virus Y (PVY). In addition to PVY, tobacco etch virus (TEV) and a TEV construct that encodes β‐glucuronidase (TEV‐GUS) were inoculated to determine the inheritance of resistance to these viruses in progenies obtained from potato cultivars containing the Ry_sto gene. While cultivars ‘Karlena’ and ‘Delikat’ were susceptible, ‘Bettina’ and clone 927eY were resistant to PVY, TEV and TEV‐GUS, as determined by enzyme linked immunosorbent assay, biotest and GUS assay, respectively. The segregation ratios obtained from the progenies of ‘Bettina’בDelikat’ and 816eY בKarlena’ indicate that resistances to PVY and TEV are governed by one dominant gene or two genes tightly linked in coupling phase. Evidently, Ry_sto confers broad spectrum resistance to potyviruses. TEV resistance could be reliably detected 4 days after inoculation with the TEV‐GUS construct by GUS assay. Therefore, the GUS‐tagged TEV construct can be used for early selection for resistances based on the gene Ry_sto or closely linked genes.     

Diversity studies in genetic resources of <Emphasis Type="Italic">Solanum</Emphasis> spp. (section Petota) by comparative application of ISAP markers

The Genebank at the Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) harbours three different potato collections comprising 2846 wild potato accessions, 2757 accessions of cultivars, landraces and breeding material of Solanum tuberosum L. and 579 Solanum clones originating mainly from South and Central America (as of July 2016). The three collections are maintained via seeds, in vitro culture, cryopreservation or in the field. All four processes are labour and cost intensive. Therefore, methods facilitating the efficient management and maintenance of the collections are needed. We describe the experimental application of the retrotransposon-based Inter-SINE amplified polymorphism (ISAP) marker system for supporting the management of potato collections. Three ISAP primer combinations were evaluated on a set of 259 cultivated and 89 wild potato accessions focussing on three different aspects: diversity analysis, duplicate identification and phylogenetic studies. Compared to up to 29 microsatellites, ISAPs were similarly efficient in monitoring genetic diversity and detecting duplicated accessions. In addition, the three ISAP primer combinations could be applied to wild potato species, although their use in phylogenetic studies is limited due to their highly polymorphic nature. Generally, ISAPs are a very valuable tool in the maintenance of clonally maintained potato collections and for the verification of the identity of accessions.     

The C bicarbonate dilution technique to determine energy expenditure in young bulls validated by indirect calorimetry

We explored the applicability of the ¹³C bicarbonate dilution technique for determination of energy expenditure (EE) in young bulls in comparison to whole body indirect calorimetry (IC). Twelve bulls of a F2 German Holstein x Charolais cross (4.5 months, 332 ± 16 kg BM) received a diet providing 1000 kJ ME d^− 1 kg BM^− 0.75 and 4.3 g crude protein d^− 1 kg BM^− 0.75. Bulls were housed in respiration chambers and received an intravenous bolus of NaH¹³CO₃ (A: 3 μmol kg BM^− 1 (n = 2), B: 7 μmol kg BM^− 1 (n = 4), C: 17.5 μmol kg BM^− 1 (n = 6), 99 at.% ¹³C) into the jugular vein to measure EE. Simultaneously, EE was determined by IC. After the ¹³C administration blood samples and breath gas were collected from the animals in the respiration chamber during a 24-h period (7.00–7.00 h). The recovery of ¹³C in breath CO₂ (% of ¹³C dose) was irrespective of NaH¹³CO₃ dose (A: 69.7 ± 2.7%, B: 70.5 ± 4.5%, C: 75.0 ± 4.9%; P > 0.05). Only small amounts of ¹³C were excreted in urine (3.4 ± 2.6%) and feces (2.0 ± 1.3%). The EE determined by the ¹³C bicarbonate method using breath and blood ¹³C recovery rates as correction factors was not different from that measured by IC (816 ± 81 [blood] or 827 ± 101 [breath] vs. 820 ± 90 kJ d^− 1 kg BM^− 0.75). Bland–Altman analysis showed a 95% confidence interval for EE of ± 99 and ± 109 kJ d^− 1 kg BM^− 0.75 based on blood and breath ¹³C recovery, respectively. In conclusion, the ¹³C bicarbonate dilution method is appropriate to obtain reliable estimates of EE in young bulls using blood CO₂ or breath CO₂ under standardized experimental conditions, i.e. in the fasting state.     

Optimizing multi-stage shrimp production systems

Multi-stage penaeid shrimp grow-out systems have considerable advantage over conventional single-stage grow-out systems. A multi-stage shrimp grow-out system has more than one production stage wherein the shrimp stocking density changes as the shrimp grow in size and are moved from one production stage to the next. The shrimp stocking density (shrimp/m²) is at its highest when the shrimp post larvae enter the first production stage and is reduced each time the shrimp are transferred to subsequent stages. The use of multi-stage instead of single-stage production systems has been considered by numerous authors. This paper presents a methodology by which to select the optimum number of stages for a production system and, using available data, demonstrates that optimum efficiency can be achieved, in most cases, by using a two-stage production system consisting of a prolonged nursery stage followed by a grow-out stage.