Comparison of the precursor, amino acid oxidation, and end-product methods for the evaluation of protein turnover in senior dogs.

Stable isotope methods have been used to study protein metabolism in humans; however, there application in dogs has not been frequently explored. The present study compared the methods of precursor (13C-Leucine), end-products (15N-Glycine), and amino acid oxidation (13C-Phenylalanine) to determine the whole-body protein turnover rate in senior dogs. Six dogs (12.7 ± 2.6 years age, 13.6 ± 0.6 kg bodyweight) received a dry food diet for maintenance and were subjected to all the above-mentioned methods in succession. To establish 13C and 15N kinetics, according to different methodologies blood plasma, urine, and expired air were collected using a specifically designed mask. The volume of CO2 was determined using respirometry. The study included four methods viz. 13C-Leucine, 13C-Phenylalanine evaluated with expired air, 13C-Phenylalanine evaluated with urine, and 15N-Glycine, with six dogs (repetitions) per method. Data was subjected to variance analysis and means were compared using the Tukey test (P<0.05). In addition, the agreement between the methods was evaluated using Pearson correlation and Bland-Altman statistics. Protein synthesis (3.39 ± 0.33 g.kg-0,75. d-1), breakdown (3.26 ± 0.18 g.kg-0.75.d-1), and flux estimations were similar among the four methods of study (P>0.05). However, only 13C-Leucine and 13C-Phenylalanine (expired air) presented an elevated Pearson correlation and concordance. This suggested that caution should be applied while comparing the results with the other methodologies.

Energy expenditure of red-footed tortoises (Chelonoidis carbonaria) fed kibble diets with high levels of fibre, starch, or fat.

Knowledge of energy requirements is necessary to optimise the nutritional management of animals. For tortoises, very little is known about their nutrient and energy requirements. Data on substrate oxidation and the implications of starch or fat intake on the energy metabolism are lacking. The present study compared the daily energy expenditures (DEE) of red-footed tortoises (Chelonoidis carbonaria) that were fed three extruded diets: a control diet high in fibre and two test diets, one with high starch content and another with high fat content. A total of 18 tortoises (5.5 ± 1.18 kg) were used in a completely randomised design, with 6 animals per diet. After 14 months of experimental diet intake and 48 h of preliminary fasting, the animals were kept for 12 h in 70-l respiratory chambers. An open "push mode" system was used to determine the carbon dioxide production and oxygen consumption levels for the subsequent calculations of DEE. The data were analysed with ANOVA, and the means were compared by using Tukey's test (p < 0.05). The body weights, chamber temperatures and food intakes of the tortoises were similar among the treatments (p > 0.05). There were no significant differences in oxygen consumption (21.7 ± 3.16 ml · kg-1 · h-1), carbon dioxide production (18.1 ± 1.96 ml · kg-1 · h-1), or DEE (9.7 ± 1.04 kJ · kg-1 d-1) between diets or sex (p > 0.05). The respiratory quotients (0.84 ± 0.11) were also similar among the diets (p > 0.05). The DEE of red footed tortoises did not differ after a long-term starch or fat intake.

Effect of dietary fiber on fecal androgens levels: An experimental analysis in brown brocket deer (Mazama gouazoubira).

Non-invasive endocrinology is an important tool for animal conservation, but its success depends on many factors (e.g. adequate hormonal extraction, diet, antibody used in the assay). Dietary fiber is one of the main sources that can lead to erroneous interpretation of the endocrine status provided by EIA analysis. This study aimed to evaluate the dietary fiber effect on the fecal androgen metabolites (FAM) detection, on the daily defecation rate and fecal production, as well as to analyze the gastrointestinal passage and retention time of the experimental diets. Eight brown brocket deer (Mazama gouazoubira) males were randomly assigned to two groups and submitted to both isocaloric and isoproteic experimental diets for 10 days, in a crossover system: low fiber percentage feed (LF, 7% fiber) and high fiber percentage feed (HF, 19% fiber). Such groups were alternated in the middle of this period, with an interval of 10 days between them. In addition, there was a five-day adaptation phase at the beginning of each diet. Fecal collection for FAM measurement was performed during 10 days of treatment, whereas,the defecation rate and fecal production were performed every two hours, for 6 days. The mean FAM level in the HF group was 5038.0 ± 1529.1 ng/g, while for LF, 2178.7 ± 824.9 ng/g (p < 0.05). The mean HF fecal production was 182.6 ± 36.2 g DM/day and 117.5 ± 12.6 g DM/day for LF (p < 0.05). There were no differences in terms of mean defecation rate, passage, and retention times between groups. The results suggest that dietary fiber affects the FAM detection, and this should be taken into consideration before conducting experiments using fecal samples as a source of reproductive hormones profiling.

Influence of incubation temperature on embryo development, hatchling morphology and early growth rate in red-footed tortoise (Chelonoidis carbonaria).

Reptile embryos respond to temperature changes with metabolic and physiological adjustments that influence hatchling success, phenotype, behaviour, and growth rate. Climate change and global warming can affect the reptile population by altering the frequencies of hatchling survival and phenotypes. Therefore, previous studies proposed artificial incubation as a potential strategy for mitigating these effects. Red-footed tortoise (Chelonoidis carbonaria) eggs were collected and incubated at constant temperatures of 27.5 °C and 29.5 °C to investigate the physiological effects of temperature on embryo development, hatchling morphology, and early post-hatch growth rate. The direct effect of temperature on the incubation period, egg mass loss, hatching success, hatchling size, and mass was evaluated at hatching and three months of age. Hatchlings from 29.5 °C presented a shorter incubation period (141 days) than those from 27.5 °C (201 days; p < 0.05). Egg mass loss, hatchling mass, and size at hatching were not different between the incubation temperatures (p > 0.05). However, the hatching success (survival rate) was lower (64.5% versus 100%) in eggs incubated at 29.5 °C, but the hatchling mass and straight plastron width were higher at three months of age than those from eggs incubated at 27.5 °C (p < 0.05). These results indicate that incubation temperature influences hatching success and hatchling size and mass in the first months by influencing the early growth rate.

Effects of crude protein and sodium intake on water turnover in cats fed extruded diets.

The comprehension of strategies to increase urine production may be important, especially in kibble diets to prevent urolithiasis in cats. The effects of increasing amounts of crude protein (CP) and sodium on the water turnover of cats were evaluated using the water balance (WB) method and the deuterium dilution technique. The study followed a randomized block design, with three blocks of eight cats, two cats per food type in each block, and six cats per food. Four extruded diets with different amounts of CP and sodium were evaluated (on DM basis): 28% CP and 0.58% sodium; 39% CP and 0.64% sodium; 52% CP and 0.76% sodium; and 64% CP and 0.87% sodium. Cats were individually housed in cages for 8 days to measure WB, urea excretion, and faecal and urine characteristics. Deuterium oxide was used to evaluate water turnover, and during the period cats were housed in a collective cattery. The data were analysed by an F test, and the means were compared by polynomial contrasts. The ɑ level of significance was set at 0.05. The methods were compared by Pearson correlation, and Bland and Altman analysis. The increase in the CP content elevated linearly the renal excretion of urea (p < .001), and, together with the higher sodium intake, elevated the renal solute load, which resulted in a linear increase in urine production and water intake (p < .01). The urine density, metabolic water, and faecal and insensible water losses did not differ (p > .05). The water flux increased linearly when using the deuterium method (p < .001), but the obtained values were 20.85 ± 11.11 ml/cat/day higher than those verified using the WB method (p = .001). Higher CP and sodium amounts in dry diets increased the urine production and water consumption of cats, and this can be explored as a possible option to increase urination.

Validation of saliva and urine use and sampling time on the doubly labelled water method to measure energy expenditure, body composition and water turnover in male and female cats.

Less invasive protocols are necessary to study energy expenditure (EE) of cats living in homes for expressing their normal living conditions. The present study compared sampling times and the use of saliva, urine and blood to measure 2H and 18O to apply the doubly labelled water method. In the first study, four cats were used to evaluate the enrichment (2, 4, 6, 7 and 8 h) and elimination (2, 4, 6, 8, 10, 12, 14, 16, 18 and 20 d) of 2H and 18O (subcutaneously injected). The maximum enrichment was after 5 h (R2 0·82) of injection, with an Ln linear elimination of both isotopes (P < 0·001; R2 0·99). The results of EE were similar, regardless of the sampling time used (P = 0·999). In the second study, seven male cats and seven female cats were used. Before and after isotope injection (5 h, 7 d, 10 d and 14 d), blood, saliva and urine were collected. Isotope enrichment was lower in urine (P < 0·05) and at the similar level in blood and saliva. Isotope elimination was similar for all fluids (P < 0·473). The EE calculated with blood and saliva was similar but higher for urine (P = 0·015). According to Bland-Altman statistics, blood and saliva presented low bias and high correlation (P < 0·001), but this was not observed for urine (P = 0·096). Higher EE was observed for male cats (384 (se 39) kJ/kg0·67 per d) than for female cats (337 (se 34) kJ/kg0·67 per d; P < 0·05). The sampling time for the method is flexible, and saliva can be used as a substitute for blood.

Feed processing effects on digestibility, palatability, excreta fermentation products and blood parameters in blue-fronted amazon parrots (Amazona aestiva)†.

Captive parrots show a high incidence of obesity and other metabolic disorders due to the consumption of unbalanced diets. Therefore, this study evaluated the digestibility and metabolic effects of transitioning blue-fronted amazons from a high fat diet (sunflower seeds) to processed diets with three degrees of starch gelatinization (SG). The same feed formulation was processed to obtain pelletized feed (PEL) at 27.1% SG; low-cooked extruded feed (EXTL ) at 81.6% SG; and high-cooked extruded feed (EXTH ) at 98.5% SG. Thirty adult parrots were fed sunflower seeds for 90 days, then were distributed in a completely randomized design with 10 repetitions per treatment, and fed one of the three prepared diets for 160 days. Feed palatability, apparent digestibility, excreta concentrations of volatile fatty acids, lactate and ammonia, initial and final radiographic examinations, blood cell counts and glucose, triglycerides, cholesterol, total protein, albumin, aspartate aminotransferase (AST) and uric acid levels were evaluated. The data were analysed by an analysis of variance and compared by Tukey's test (p < 0.05). Sunflower seed was more digestible than processed feeds (p < 0.05). Diet processing interfered with fat and starch digestibility (p < 0.001), being higher in the PEL than in the EXTH and EXTL respectively. Transitioning from sunflower seeds to balanced diets reduced serum glucose, triglycerides, cholesterol and AST (p < 0.05) and increased red blood cell, haemoglobin, lymphocyte, monocyte and leucocyte counts (p < 0.01). Radiographs indicated a decreased hourglass (p = 0.015) and a reduced heart-liver ratio after ingesting the processed feeds (p < 0.05). Feed processing did not affect blood cell counts, serum biochemistry or radiographic examinations. In conclusion, parrots preferred the extruded diet and did not require an extensive SG to properly digest the feed. Consuming the processed diets improved the birds' metabolism and health.