This paper describes a 4-month pilot study that tested the suitability of a physical activity intervention for first graders (children aged 6 and 7 years) in a public school in Santiago, Chile. Teachers were trained to deliver the programme in the classroom during the school day. Teachers were surveyed to determine if this intervention fit within their curriculum and classroom routines and they reported in a focus group that it was suitable for them. All children actively participated in the programme and positive changes in their attitudes towards physical activity were observed by their teachers. Anthropometrics, blood pressure and hand grip strength were measured in the students. A significant reduction was observed in children with high waist circumference ≥ 90th percentile, and in mean systolic blood pressure. However, statistical power values for those comparisons were rather low. Anthropometry and hand grip strength were not modified. The latter calculations and the lack of a control group are showing the weaknesses of this pilot study and that further research with a larger sample size and an experimental design is strongly needed.

Studies in many Western countries have consistently shown that monetary diet cost is positively associated with diet quality, but this may not necessarily be the case in Japan. This cross-sectional study examined the nutritional correlates of monetary diet cost among 3963 young (all 18 years old), 3800 middle-aged (mean age 48 years) and 2211 older (mean age 74 years) Japanese women. Dietary intakes were assessed using a comprehensive self-administered diet history questionnaire for young and middle-aged women and a brief self-administered diet history questionnaire for older women. Monetary diet cost was estimated using retail food prices. Total vegetables, fish and shellfish, green and black tea, white rice, meat, fruit and alcoholic beverages contributed most (79–89 %) to inter-individual variation in monetary diet cost. Multiple regression analyses showed that monetary diet cost was negatively associated with carbohydrate intake, but positively with intakes of all other nutrients examined (including not only dietary fibre and key vitamins and minerals but also saturated fat and Na) in all generations. For food group intakes, irrespective of age, monetary diet cost was associated inversely with white rice and bread but positively with pulses, potatoes, fruit, total vegetables, fruit and vegetable juice, green and black tea, fish and shellfish, and meat. In conclusion, in all three generations of Japanese women and contrary to Western populations, monetary diet cost was positively associated with not only healthy dietary components (including fruits, vegetables, fish and shellfish, dietary fibre, and key vitamins and minerals), but also less healthy components (including saturated fat and Na).

The effects of different temperature and time conditions during retorting of canned cat food on physicochemical characteristics and palatability were examined. For this purpose, lacquer cans containing an unprocessed loaf-type commercial cat food were heated in a pressurised retorting system at three specified temperature–time profiles (113°C/232 min, 120°C/103 min and 127°C/60 min) to equal a similar lethality (F0 value = 30). Physicochemical properties (viscosity, texture, particle size, pH) were determined, and a 10 d three-bowl palatability test was performed with ten European shorthair cats. Retorting at 113°C/232 min resulted in differences in all the physical parameters examined (<viscosity, firmness, adhesiveness, and > particle size). Significant pH differences were observed (6·53, 6·63 and 6·66 for T113/232, 120 and 127°C, respectively). Preference ratios were 0·38, 0·31 and 0·31 for T113/232, 120 and 127°C, respectively (P = 0·067). It can be concluded that different retorting temperature–time profiles with equal F0 value significantly affect physical characteristics and tended to affect palatability of moist cat food.

The present study aimed to evaluate the inter-individual variability in fermentation of standard fibrous substrates by faecal inocula from ten healthy adult female cats. Substrates were citrus pectin (CP), fructo-oligosaccharides (FOS), guar gum (GG), sugar beet pulp (SBP) and wheat middlings (WM). Each substrate was incubated with faecal inoculum from each cat. Gas production was measured continuously during the 48 h incubation and SCFA and organic matter disappearance (only SBP and WM) after incubation. Out of ten cats, nine produced faeces on the days of inoculum preparation. The substrates contrasted in terms of fermentation parameters measured. The inter-individual variability was in general lower for the more simple and pure substrates (CP, FOS, GG) than for the more complex substrates containing mixtures of fibres (SBP, WM). Furthermore, for total SCFA and gas produced, inter-individual variability was lower than for proportions of butyrate and of branched-chain fatty acids and for the parameters of gas production kinetics. It is concluded that the variability in in vitro fermentation parameters is associated with the complexity of fibrous substrates. The presented data are instrumental for the calculation of number of faecal donors required for precise in vitro characterisation of the fermentability of dietary fibres. In addition, the number of faecal donors should be adjusted to the specific fermentation parameter(s) of interest.

To gain knowledge on the precision of an in vitro method for characterisation of the fermentability of dietary fibres, this study aimed to evaluate the repeatability and reproducibility of such a method. Substrates used were citrus pectin (CP), fructo-oligosaccharides (FOS), guar gum (GG), sugar beet pulp (SBP) and wheat middlings (WM). Each substrate was incubated with faecal inoculum from five cats with three replicates for each substrate–cat combination. Gas production was measured continuously during the 48 h incubation and SCFA and organic matter disappearance (only SBP and WM) were determined after incubation. Four consecutive runs were performed. The within-run variability (repeatability) was generally lower for the more simple and pure substrates (CP, FOS, GG) than for the more complex substrates containing mixtures of fibres (SBP, WM). Replicates showed high variability, in particular for SCFA profiles and parameters of gas production kinetics. The between-run CV (reproducibility) for the measured parameters were, in general, below 10 % for CP, FOS and GG and higher values were obtained for SBP and WM. It is concluded that for precise dietary fibre characterisation, the number of replicates should be multiple and adjusted according to the variability of the parameters of interest and the complexity of fibres. The method yielded reproducible results with some variation in absolute values obtained, which may have an impact on the significance level of the differences among substrates.

The 2006 National Research Council (NRC) equation calculating puppy energy requirements does not account for reported breed differences in growth pattern. Energy requirements of toy breed puppies are unknown and it is unclear whether feeding guidelines should differ between breeds. Energy requirements of Yorkshire terrier (YT) puppies were observed over their first year of life and compared with those predicted by the NRC and those previously observed in large (Labrador retriever) and medium (miniature Schnauzer; MS) breed puppies. Twenty-two puppies (from eight litters) were offered complete and balanced diets to maintain ideal body condition score (BCS). Energy intake, body weight and BCS were recorded from 10 to 52 weeks of age. Every 12 weeks, health was monitored by veterinary examination, routine haematology and plasma biochemistry. Puppies remained clinically healthy with normal skeletal development throughout. After analysis by linear mixed models it was observed that the NRC equation overestimates YT energy requirements between 10 and 20 weeks of age by up to 324·3 (95 % CI 390·4, 258·2) kJ/kg0·75. Energy intake was lower (P < 0·05) in YT than Labradors until 29 weeks by up to 376·6 (95 % CI 477·4, 275·3) kJ/kg0·75 and lower than MS between 16 and 25 weeks by up to 216·3 (95 % CI 313·0, 119·7) kJ/kg0·75 (P < 0·05). Data indicate differences in toy, medium and large breed energy requirements for growth. The NRC equation for puppy energy requirements overestimated the requirements of this YT population, suggesting the need for breed-specific feeding guides for growth to avoid overfeeding.

The physiological demands of weight-pulling dogs have yet to be investigated. Two groups of competitive weight-pulling dogs both underwent two identical pull series 3 h apart. The control group (n 8) was compared with a group fed a rapidly digestible carbohydrate and protein supplement after the first pull series (n 9). Blood was drawn before and after each pull series as well as at 15 and 30 min after the first pull series finished. Biochemistry values remained unremarkable throughout the study in both groups regardless of supplementation or exercise over time. Lactic acid showed mild significant increases post-exercise (2·1 (sd 1·2) mmol/l) compared with baseline (1·4 (sd 0·3) mmol/l; P = 0·03) after the initial pull series. When examining the effects of time there was a significant increase in insulin from baseline (median of 10·8 (range 6·8–17·4) μIU/ml) compared with 30 min after supplementation (17·0 (range 8·1–33·0) μIU/ml) and at 3 h after supplementation (19·2 (range 9·7–53·4) μIU/ml). In the treatment group there was also a time effect, with glucagon being elevated from baseline (median of 100 (range 79–115) pg/ml) compared with 30 min after supplementation (114 (range 90–183) pg/ml) and after the second pull series (131 (range 107–152) pg/ml). Evaluation of each dog's ability to pull the same or greater amount of weight on the second pull series revealed no significant differences. In conclusion, weight-pulling dogs have mild elevations in lactate reflecting little anaerobic metabolism compared with other canine sprinting athletes; hormonal changes associated with carbohydrate absorption are reflected within the treatment group, and supplementation had no effect on performance.

Working canines are deployed by the Federal Emergency Management Agency (FEMA), as part of a National Disaster Response Plan. Stress associated with helicopter flight and the resulting physical effects on the dog are unknown. Our objective was to test the hypotheses that (1) helicopter travel affects the physiology and faecal microbiota of working canines, but that (2) physiological consequences of helicopter travel will not negatively affect their work performance. A total of nine FEMA canines were loaded onto helicopters and flown for 30 min in July 2015. Rectal temperature, behavioural stress indicators and saliva swabs (for cortisol) were collected at baseline, loading, mid-flight and post-flight. After flight, canines completed a standardised search exercise to monitor work performance. Faecal samples were collected for microbial DNA extraction and Illumina sequencing. All canines were on a standardised diet (CANIDAE® Grain Free PURE Land®) for 3 weeks prior to the study. Visible indicators of stress were observed at loading and at mid-flight and corresponded with an increase (P < 0·05) in salivary cortisol from 5·4 µg/l (baseline) to 6·4 µg/l (loading). Additionally, rectal temperature increased (P < 0·05) from 38·61°C (baseline) to 39·33°C (mid-flight) and 39·72°C (post-flight). Helicopter travel did not affect search performance (P > 0·05). We found that α- and β-diversity measures of faecal microbiota were not affected (P > 0·05). Our data suggest that although helicopter travel may cause physiological changes that have been associated with stress in working dogs, it does not make an impact on their search performance or the stability of faecal microbiota.

The dog chew studied here is a starch-based, twin-screw cooker extruded dog care and treat (C&T) product with oral health benefits. The manufacturing process and nutrient profile of such products are markedly different from those of main meal pet foods. Predicted metabolisable energy (PME) in pet food is calculated using equations derived from main meal feeding studies so it is unclear whether these equations can be applied to C&T products. The present study aimed to directly measure metabolisable energy (ME) content of the dog chew in dogs and compare with calculated PME. A batch of dog chews was manufactured and the product rendered micronutrient complete to allow solus feeding. Following a 3 d standard diet pre-feed phase, the test product was fed solus to a panel of seven adult dogs for a period of 8 d. Dietary intake was recorded daily and faecal matter collected for the last 5 d. Test product and pooled faecal samples were analysed for proximate nutrients, and digestibility coefficients were calculated as the difference in intake and faecal excretion (7–11 d). Digestible energy was converted to ME by correcting for energy losses in urine. PME was calculated using proximate analysis and modified Atwater factors according to National Research Council 2006. The results showed close agreement between actual ME (1272 (sd 12·1) kJ/100 g) and calculated PME (1268 (sd 12·6) kJ/100 g), indicating transferability of the NRC 2006 PME equations to the dog chew tested here.

We previously found a weak response in serum 25-hydroxyvitamin D3 (25(OH)D3) concentrations when dogs were supplemented with oral vitamin D3 (D3). In the present study, we determined the relative potency of oral 25(OH)D3 compared with D3 for increasing vitamin D status in dogs with low serum 25(OH)D concentrations. Four male and three female, 4-year-old, intact, lean, genetically related, Chinese-crested/beagle dogs were studied in a randomised, single cross-over trial. After feeding a low-vitamin D diet (<4 IU/100 g) for 30 d, four dogs received daily D3 supplementation at 2·3 µg/kg body weight0·75, while three dogs received a molar equivalency as 25(OH)D3. The supplements, dissolved in ethanol, were applied to a commercial treat for consumption. Serum 25(OH)D3 and 24R,25-dihydroxyvitamin D3 (24R,25(OH)2D3) were analysed weekly using a validated HPLC method. Both supplementations increased (P ≤ 0·01) serum 25(OH)D3 concentrations. However, oral 25(OH)D3 resulted in greater (P < 0·0001) concentrations than D3 by week 1, with a difference of 173 % (P < 0·0001) by week 2. The supplementation period was limited to 14 d after serum 25(OH)D3 concentrations were not appearing to plateau. Thereafter, a washout period of 1 month separated the cross-over. Following 25(OH)D3, but not D3 supplementation, serum 24R,25(OH)2D3 concentrations increased (P ≤ 0·02), 3 to 5 weeks after initiating supplementation. Vitamin D status, as indicated by serum 25(OH)D3 and 24R,25(OH)2D3 concentrations, is more rapidly and efficiently increased in adult dogs by oral supplementation of 25(OH)D3 than D3.

The present study aimed to determine the minimum requirements of the essential n-3 fatty acids EPA and DHA in Atlantic salmon (Salmo salar) that can secure their health under challenging conditions in sea cages. Individually tagged Atlantic salmon were fed 2, 10 and 17 g/kg of EPA + DHA from 400 g until slaughter size (about 3·5 kg). The experimental fish reared in sea cages were subjected to the challenging conditions typically experienced under commercial production. Salmon receiving the lowest EPA + DHA levels showed lower growth rates in the earlier life stages, but no significant difference in final weights at slaughter. The fatty acid composition of various tissues and organs had remarkably changed. The decreased EPA + DHA in the different tissue membrane phospholipids were typically replaced by pro-inflammatory n-6 fatty acids, most markedly in the skin. The EPA + DHA levels were maintained at a higher level in the liver and erythrocytes than in the muscle, intestine and skin. After delousing at high water temperatures, the mortality rates were 63, 52 and 16 % in the salmon fed 2, 10 and 17 g/kg EPA + DHA. Low EPA + DHA levels also increased the liver, intestinal and visceral fat amount, reduced intervertebral space and caused mid-intestinal hyper-vacuolisation. Thus, 10 g/kg EPA + DHA in the Atlantic salmon diet, a level previously regarded as sufficient, was found to be too low to maintain fish health under demanding environmental conditions in sea cages.

The present study evaluated recipes of home-prepared diets for dogs and cats published in Portuguese. A total of 106 diets were evaluated: eighty for dogs, twenty-four for cats and two intended for both species. A commercial software package was used to analyse the diets, and an ingredient chemical composition database was built based on the Brazilian Tables of Food Composition and United States Department of Agriculture Nutrient Database. The estimated chemical composition of each recipe was compared with the Nutritional Guidelines for Complete and Complementary Pet Food for Cats and Dogs (Fédération Européenne de L'industrie des Aliments Pour Animaux Familiers; FEDIAF, 2014) recommendations for maintenance (as units/MJ). Most recipes (48 %) had no precise determination of ingredients and quantities. All diets had at least one nutrient below the recommendations, and all investigated nutrients were deficient in at least one diet. The most frequent nutrients below recommendation were: Fe (68·3 % of the recipes for dogs; 100 % of the recipes for cats); vitamin E (82·9 % of the dog recipes; 84·6 % of the cat recipes); Zn (75·6 % for dogs; 88·4 % for cats); Ca (73·2 % for dogs; 73 % for cats); Cu (85·4 % for dogs; 69·2 % for cats); choline (85·4 % for dogs; 69·2 % for cats); riboflavin (65·8 % for dogs; 11·5 % for cats); thiamine (39 % for dogs; 80·7 % for cats); and vitamin B12 (61 % for dogs; 34·6 % for cats). These recipes may potentially expose animals to nutritional deficiencies, and it is important to inform the owners of the risks of providing home-prepared diets. Better training of professionals that intend to prescribe home-prepared diets is advisable.

High flavonoid consumption can improve vascular health. Exploring flavonoid–metabolome relationships in population-based settings is challenging, as: (i) there are numerous confounders of the flavonoid–metabolome relationship; and (ii) the set of dependent metabolite variables are inter-related, highly variable and multidimensional. The Metabolite Fingerprint Score has been developed as a means of approaching such data. This study aims to compare its performance with that of more traditional methods, in identifying the metabolomic fingerprint of high and low flavonoid consumers. This study did not aim to identify biomarkers of intake, but rather to explore how systemic metabolism differs in high and low flavonoid consumers. Using liquid chromatography–tandem MS, 174 circulating plasma metabolites were profiled in 584 men and women who had complete flavonoid intake assessment. Participants were randomised to one of two datasets: (a) training dataset, to determine the models for the discrimination variables (n 399); and (b) validation dataset, to test the capacity of the variables to differentiate higher from lower total flavonoid consumers (n 185). The stepwise and full canonical variables did not discriminate in the validation dataset. The Metabolite Fingerprint Score successfully identified a unique pattern of metabolites that discriminated high from low flavonoid consumers in the validation dataset in a multivariate-adjusted setting, and provides insight into the relationship of flavonoids with systemic lipid metabolism. Given increasing use of metabolomics data in dietary association studies, and the difficulty in validating findings using untargeted metabolomics, this paper is of timely importance to the field of nutrition. However, further validation studies are required.

Cellular oxidative damage is thought to be one of the key mechanisms underlying age-related cognitive impairment in dogs. Several nutritional interventions to limit cognitive decline are reported in the literature. To our knowledge, the association of grape and blueberry extracts has never been tested in aged dogs. Our objective was to evaluate the effect of a polyphenol-rich extract from grape and blueberry (PEGB) on oxidative status and cognitive performances in aged dogs. A total of thirty-five beagle dogs (aged 8·0–14·5 years) were fed a basal diet with PEGB at either 0 parts per million (ppm) (n 11; control), 240 ppm (n 12; PEGB1) or 480 ppm (n 12; PEGB2) for 75 d. To investigate the effects of PEGB supplementation on cognition and oxidative status, a delayed non-matching to position (DNMP) test and RT-PCR on genes involved in oxidative stress were evaluated. The dogs fed PEGB1 showed a higher superoxide dismutase mRNA expression compared with dogs fed PEGB2 (P = 0·042) and with the control group (P = 0·014). Moreover, the dogs fed PEGB2 showed higher nuclear factor-like 2 (Nrf2) mRNA expression compared with the dogs fed PEGB1 (P = 0·027). Concerning the DNMP test, the proportion of dogs showing cognitive improvements relative to their baseline level was significantly higher in dogs fed the PEGB, regardless of the dosage, than in dogs receiving no supplementation (P = 0·030). The results obtained in the DNMP test suggested a potential benefit of the PEGB on working memory. However, this hypothesis should be further investigated to confirm this cognitive effect.

The present study assessed the effect of dietary supplementation of pomegranate peel (methanolic) extract (PPE) on the nutrient digestibility, faecal fermentative metabolites and antioxidant status of dogs. Six medium-sized dogs of mixed breeds were allocated to two groups in a replicated cross-over design and fed a basal diet with and without supplementation of PPE at 50 mg/kg body weight. Each of the experimental periods consisted of 30 d, with a 21 d washout period in between. Results indicated that PPE supplementation led to no significant changes in nutrient digestibility except a trend of improvement (P = 0·070) in crude fibre digestibility. PPE supplementation significantly (P < 0·001) influenced faecal pH, ammonia and lactate production, indicative of positive effects of PPE polyphenols. PPE supplementation further increased faecal SCFA concentration accompanied by a reduction (P < 0·01) in branched-chain fatty acids. PPE supplementation also improved (P < 0·05) indices of erythrocytic antioxidants, namely reduced glutathione, catalase, glutathione peroxidase and glutathione S-transferase together with an improvement in lipid peroxidation. Overall, it can be concluded that dietary supplementation with PPE at 50 mg/kg body weight had a positive impact on hindgut fermentation and antioxidant status in dogs, thereby demonstrating its potential as a gut health-promoting nutraceutical.

EPA and DHA are often used in veterinary medicine due to their beneficial effects for several medical conditions such as osteoarthritis. EPA and DHA are administered to dogs through different matrices. The aim of the present study was to determine the effects on the plasma levels in dogs caused by various matrices for EPA and DHA administration. In this study, three different n-3 PUFA formulations were used: soft chew tablet (CCx); liquid fish oil (LFO); and enriched kibbles (EK). The formulations were administered single-dose and compared in a randomised, cross-over designed study with a 1-week wash-out period. Several variables were observed after the administration of these formulations in thirteen dogs: the NEFA plasma concentration, the AUC for 1 d (AUC0–24 h), and maximum plasma concentration for both EPA and DHA. All plasma fatty acid levels reached baseline levels within 72 h. CCx (median = 2·987) had a significantly lower AUC0–24 h for EPA compared with LFO (median = 5·647, P = 0·043) and EK (median = 5·119, P = 0·032) (F2,22 = 4·637, P = 0·021). CCx (median = 2·471) AUC0–24 h for DHA was significantly lower compared with LFO (median = 4·837, Z = −2·56, P = 0·011) and EK (median = 4·413, Z = −2·59, P = 0·01). EPA and DHA plasma levels were affected by matrix, as with the CCx, the AUC0–24 h of EPA and DHA were both lower compared with LFO and EK. The effect of matrix on bioavailability is important for product development as well as for clinical trials studying effects of EPA and DHA.

The present experiment was undertaken to validate a probiotic of canine origin for its potential use in dogs. A total of fifteen adult female Labrador dogs were allocated to three equal groups and fed a basal diet without probiotic (control) or with probiotic of either canine (Lactobacillus johnsonii CPN23; cPRO) or dairy (L. acidophilus NCDC 15; dPRO) origin for 9 weeks. The digestibility of most macronutrients remained similar among the groups; however, fibre digestibility was improved (P = 0·034) in dogs receiving cPRO. The faecal fermentative metabolites ammonia (P < 0·05) and lactate (P = 0·094) were altered favourably, indicating a positive influence of both probiotics. Faecal concentrations of acetate, propionate and butyrate were increased (P < 0·01) in both probiotic groups. However, improvements were higher in cPRO v. dPRO. The delayed-type hypersensitivity reaction to intradermal inoculation of phytohaemagglutinin-P was higher (P = 0·053) in cPRO as compared with control. The antibody response to sheep erythrocytes was, however, similar across the three groups. Overall, in dogs, the canine-origin probiotic was superior when compared with the dairy-origin probiotic.

Pomegranate (Punica granatum), a polyphenol-rich fruit, has been suggested to reduce cardiovascular risk due to its antioxidant properties. Hypertension and obesity are the most preventable cardiovascular risk factors. Few studies on blood pressure and/or body-weight status have been conducted in human subjects. Previous investigations have tended to focus on pomegranate juice. The aim of the present study was to investigate the effect of pomegranate extract (PE) on blood pressure and anthropometric measures in adults with no symptomatic disease. A total of fifty-five participants enrolled in a randomised double-blinded placebo-controlled clinical trial where they were assigned to either PE capsules or placebo capsules for 8 weeks. Blood pressure, body weight, waist circumference, waist:hip ratio (WHR) and body composition (lean body mass, body fat) were measured at baseline, week 4 and week 8. Results showed a significant decrease in diastolic blood pressure after 8 weeks (by 2·79 (sd 5·32) mmHg; P < 0·05), while the decrease in systolic blood pressure did not reach statistical significance (2·57 (sd 7·4) mmHg; P > 0·05). Body fat percentage, lean body mass, waist circumference and WHR did not significantly differ between groups at the end of the intervention. Results suggest that PE may reduce blood pressure and possibly prevent hypertension in the normotensive population. Further large trials are required to elucidate this effect.

Neutering is a common veterinary recommendation and is often associated with obesity development. Thus, the aim of the present study was to evaluate the effects of two different amounts of protein intake by neutered dogs regarding maintenance energy requirement (MER), body composition, and biochemical and hormonal parameters. A total of fourteen healthy adult dogs were fed either a diet containing 59·7 g protein/1000 kcal (4184 kJ) (P60) or a diet with 94·0 g protein/1000 kcal (4184 kJ) (P94) for 26 weeks after neutering to maintain their body weight prior to neutering. A mixed model was fitted to verify diet, time and diet × time interaction effects on biochemical parameters, serum concentrations of insulin, glucagon, leptin and insulin-like growth factor-1 (IGF-1). MER and the body composition data were evaluated within diets (paired t test) and within times (unpaired t test). A time effect was found for fructosamine, TAG, total lipids and IGF-1 serum concentrations. The diet × time interaction was significant for glucagon (P < 0·05). No differences between diets in the MER within each time were found. However, there was a reduction in the MER of dogs fed the P60 diet 26 weeks after neutering (P = 0·042). The fat body mass of dogs fed the P60 diet increased (P < 0·05) after neutering, even without a body-weight change. Some of the biochemical parameters changed over time, but all remained within the normal range. For the period evaluated in the present study, a diet with 94·0 g of protein/1000 kcal (4184 kJ) metabolisable energy seems to be a beneficial nutritional strategy to maintain the MER and the body composition of dogs after neutering.