Clinical haematology and blood plasma chemistry can be used as a valuable tool to provide substantial diagnostic information for fish. A wide range of parameters can be used to assess nutritional status, digestive function, disease identification, routine metabolic levels, general physiological status and even the assessment and management of wild fish populations. However to evaluate such data accurately, baseline reference intervals for each measurable parameter must be established for the species of fish in question. Baseline data for ornamental fish species are limited, as research is more commonly conducted using commercially cultured fish. Blood samples were collected from sixteen red top ice blue cichlids (Metriaclima greshakei), an ornamental freshwater fish, to describe a range of haematology and plasma chemistry parameters. Since this cichlid is fairly large in comparison with most tropical ornamental fish, two independent blood samples were taken to assess a large range of parameters. No significant differences were noted between sample periods for any parameter. Values obtained for a large number of parameters were similar to those established for other closely related fish species such as tilapia (Oreochromis spp.). In addition to reporting the first set of blood values for M. Greshakei, to our knowledge, this study highlights the possibility of using previously established data for cultured cichlid species in studies with ornamental cichlid fish.

Hunting with dogs in winter conditions is practised in the Nordic countries. The present study aimed at determining daily energy expenditure (DEE) and body water turnover (BWT) by the doubly labelled water technique in eight hunting dogs (body-weight (BW) range 14–27 kg) working 3 h/d for 3 d ( − 6°C) on ground covered with 20–40 cm of loose snow, to provide information on energy and water requirements. The mean distance run during the hunting period was recorded by the global positioning system and averaged 19·4 km/d. DEE increased with increasing BW (P < 0·001) and varied between 7·20 and 16·6 MJ/d (mean 11·0 MJ/d) corresponding to 950–1350 kJ/kg BW0·75 per d (mean 1170 kJ/kg BW0·75 per d). The larger dogs tended to run longer than the smaller dogs and therefore spent more energy per kg BW0·75 but not significantly (P>0·05). DEE values determined were close to the values measured for hunting dogs running for 3 h/d in hot climates, suggesting that climate within the range of the two studies has little impact on energy expenditure per h running activity. Compared with the work of sled dogs per km travelled running on a track, the work performed by the hunting dogs was suggested to be higher when running in a loose snow layer. However, DEE was much lower because sled dogs ran for a longer distance each day. Mean BWT was 217 ml/kg BW0·75 or 19 ml/kJ metabolisable energy.

Tryptophan, a precursor of important molecules such as serotonin, melatonin and niacin, is an essential amino acid for dogs. In pigs, tryptophan supplementation has been shown to induce a significant increase in food intake. The aim of the present study was to assess whether long-term tryptophan supplementation increases voluntary food intake in dogs and to observe whether this was accompanied by a change in serum ghrelin. In the present study, sixteen adult Beagle dogs were used, with four male and four female dogs fed diets supplemented with tryptophan (1 g/dog per d) during 81 d (Trp) and four male and four female dogs that were not supplemented (control). A voluntary food intake test was performed during 5 d following the supplementation period. The Trp group tended to show a higher food intake during the voluntary food intake test (58·0 (se 5·37) v. 77·5 (se 3·65) g/kg metabolic weight per d; P = 0·074). No significant differences were found for serum ghrelin concentrations.

The prevalence of feline obesity is influenced by numerous factors, including inactivity and overconsumption of food. The objective of the present study was to evaluate the effect of feeding frequency on physical activity in adult cats. A total of twelve healthy adult cats were used in a cross-over study consisting of 32 d. In each of the two periods, six cats were fed either two meals or four meals daily. Throughout the study, cats were fed the same diet at amounts to maintain body weight and body condition score. Cats were individually housed 4 h/d at each scheduled feeding time, while for the other 20 h, cats were group-housed to allow for voluntary physical activity in the room with a 16 h light–8 h dark cycle. Voluntary activity levels were evaluated using Actical activity collars for seven consecutive days in each period. Daily average activity level for two-meal-fed cats (20·04 (sem 2·19), activity counts/epoch (15 s)) was not different from four-meal-fed cats (20·14 (sem 2·15), activity counts/epoch (15 s); P>0·05). In conclusion, when group-housed cats are fed to maintain their body weight and body condition score, varied feeding frequency between twice and four times daily may not affect activity levels.

Although horses and donkeys belong to the same genus, their genetic characteristics probably result in specific proteomes and post-translational modifications (PTM) of proteins. Since PTM can alter protein properties, specific PTM may contribute to species-specific characteristics. Therefore, the aim of the present study was to analyse differences in serum protein profiles of horses and donkeys as well as mules, which combine the genetic backgrounds of both species. Additionally, changes in PTM of the protein transthyretin (TTR) were analysed. Serum protein profiles of each species (five animals per species) were determined using strong anion exchanger ProteinChips® (Bio-Rad, Munich, Germany) in combination with surface-enhanced laser desorption ionisation-time of flight MS. The PTM of TTR were analysed subsequently by immunoprecipitation in combination with matrix-assisted laser desorption ionisation-time of flight MS. Protein profiling revealed species-specific differences in the proteome, with some protein peaks present in all three species as well as protein peaks that were unique for donkeys and mules, horses and mules or for horses alone. The molecular weight of TTR of horses and donkeys differed by 30 Da, and both species revealed several modified forms of TTR besides the native form. The mass spectra of mules represented a merging of TTR spectra of horses and donkeys. In summary, the present study indicated that there are substantial differences in the proteome of horses and donkeys. Additionally, the results probably indicate that the proteome of mules reveal a higher similarity to donkeys than to horses.

Little nutritional information has been collected from domestic cats fed raw meat diets. The objective of the present study was to evaluate differences in N metabolism of domestic cats fed raw beef-based diet (66 % crude protein (CP) and 20 % fat), bison-based diet (49 % CP and 39 % fat), elk-based diet (79 % CP and 6 % fat) and horse-based diet (60 % CP and 26 % fat). A total of eight intact adult female cats were fed to maintain body weight in a cross-over design. Daily food intake, faecal and urinary outputs, and N metabolism were measured. Dietary N was highly digestible (96·8 (sem 0·7)) for all treatments. Urinary N accounted for a majority of total N excretion, and differences in total N excretion reflect differences in urinary N. Differences in N intake and N absorption were due to differences in CP levels among diets. N retention was similar to values reported in the literature for domestic cats fed purified and traditional extruded diets. Despite differences in protein concentrations and N intake, all raw meats tested maintained N metabolism.

Dietary restriction for the weight-loss management of obese horses limits the natural trickle-feeding behaviour. During feed restriction, wood shavings are often advised as bedding to prevent dietary supplementation from non-feed sources. Data from twelve overweight/obese horses and ponies of mixed breed and sex, bedded on wood shavings during 16 weeks of feed restriction, were retrospectively evaluated. DM intake (DMI) was restricted to 1·25 % of body mass (BM) daily. Animals were randomly assigned to one of two diets (hay/chaff, n 6; hay/balancer meal, n 6). BM was recorded weekly. Feeding behaviour was recorded by continual observation over 24 h during week 15. The apparent digestibility (gross energy (GE), acid-detergent fibre (ADF) and DM) of feed was determined for all animals by total faecal collection (72 h, week 16). Rates of weight loss were independent of diet type, DM (R2 0·15), GE (R2 0·20) and ADF digestibilities (R2 0·18). Despite similar DMI, faecal DM ranged between 0·52 and 1·16 % of BM daily and was associated with wide ranges in apparent digestibility (GE − 11·34 to 53·08 %; ADF − 50·37 to 42·83 % and DM 2·14 to 57·32 %), which were improbably low for some animals. Apparent digestibilities were associated with DM output (GE R2 0·96; ADF R2 0·99 and DM R2 0·99) and time spent feeding (GE R2 0·62; DM R2 0·61 and ADF R2 0·59), indicating that feed intake was supplemented with wood shavings in at least five of the twelve animals. Quantities of wood shavings ingested (negligible to >3·0 kg/d) were back-calculated from predicted feed digestibilities. All animals remained healthy. Implications of ‘feed-bulking/energy dilution’ for feed-restricted animals need further consideration.

Data comparing age-related alterations in faecal IgA concentrations of dogs are not available in the literature. The present study aimed to compare the faecal concentrations of IgA in puppies, mature and senior dogs. A total of twenty-four beagle dogs were used, including eight puppies (5 months old, four females and four males), eight mature (4·6 years old, eight males) and eight senior dogs (10·6 years old, three males and five females). Fresh faecal samples were collected from each dog for three consecutive days and pooled by animal. After saline extraction, IgA content was measured by ELISA. Data were analysed by one-way ANOVA, and means were compared with Tukey's test (P < 0·05). Results showed that puppies have lower faecal IgA concentrations than mature dogs (P < 0·05); senior animals presented intermediary results. The reduced faecal IgA concentration in puppies is consistent with the reduced serum and salivary IgA concentrations reported previously, suggesting a reduced mucosal immunity in this age group. Although some studies have found an increased serum IgA concentration in older dogs, this may differ from the intestinal secretion of IgA, which appears to be lower in some senior animals (four of the eight dogs studied).

The effects of age on microbiota composition, gut fermentation end-product formation and peripheral lymphocyte numbers were compared between old and young adult Beagle dogs fed four kibble diets differing in yeast cell wall contents. The experiment had a double 4 × 4 Latin square design, one with four mature dogs (4 years old) and the other with four old dogs (10 years old), with four replicates (diets) per dog. In each period a 15 d adaptation period preceded a 5 d total collection of faeces for the digestibility trial. On day 21, fresh faecal samples were collected for the determination of bacterial enumeration, pH, biogenic amine and short-chain fatty acid. Flow cytometry was used for immunophenotypic evaluation. Dogs were fed four kibble diets with similar composition with 0, 0·15, 0·30 and 0·45 % of yeast cell wall (as-fed), respectively. Data were evaluated using general linear models of Statistical Analysis Systems statistical software (P < 0·05). No evidence of a difference in faecal bacteria counts between ages was found (total aerobes, total anaerobes, Bifidobacterium, Lactobacillus, Clostridium and Escherichia coli: P>0·15). Faecal concentrations of butyrate, histamine, agmatine and spermine were lower (P ≤ 0·05) and faecal pH was higher (P = 0·03) in older dogs than in mature adult dogs, suggesting an alteration in bacterial metabolic activity, or in the rate of intestinal absorption of these compounds. Concentrations of T-lymphocytes, T-cytotoxic lymphocytes and B-lymphocytes were also lower (P ≤ 0·01) in older dogs than in mature adult dogs. The study confirmed alterations in peripheral lymphocytes and revealed a reduced concentration of some fermentation end products in the colon of old dogs.

Urolithiasis is a common clinical problem in dogs. Struvite and calcium oxalate are the predominant mineral types in dog urolithiasis. The aim of the present study was to compare the effect of two commercial dry foods formulated for the management of struvite urolithiasis with different anion–cation balance on urinary pH. For the trial, twelve privately owned adult dogs showing struvite urolithiasis were studied. The dogs were randomly divided into two groups (A and B) and fed two dissolving diets for 3 months. The analyses of urine were repeated six times. In both diets, the anion–cation balance was negative ( − 203 and − 192 for diets A and B, respectively). At the first urine analysis, pH values of all the dogs were close to 8·0, and bacteria were present in about 70 % of the samples and thus an antimicrobial was administered for 1 week. Both groups showed a progressive decrease in pH values, and after 2 months, in both cases, the recommended pH values for stone dissolution were achieved. From the sampling at 30 d, group A showed pH values significantly (P < 0·05) lower than group B, probably due to the lower anion–cation balance of diet A. The combination of antimicrobial and dietary therapy allowed the dissolution of struvite uroliths in both groups, even if the utilisation of the diet characterised by the lower anion–cation balance seems to decrease the urinary pH more rapidly. In this case, it seems necessary to interrupt the dietary treatment in order to avoid the risk of other diseases.

The present study compared the intravenous glucose tolerance test (IVGTT) and the glucose postprandial response (GPPR) test for the evaluation of glucose metabolism in obese dogs. A total of ten owned obese dogs (body condition score (BCS) of 9; fat mass, 45·7 (sem 1·51) %) were used. These dogs had their weight reduced by 20 % (BCS, 8; fat mass, 33·5 (sem 1·92) %; P < 0·001), designated as weight-reduced (WR) group. A control group of ten Beagle dogs was also included (BCS, 4·5; fat mass, 18·3 (sem 1·38) %; P < 0·01). Glucose tolerance was measured by two methods: IVGTT (infusion of 0·5 g of glucose/kg body weight) and GPPR (consumption of cooked rice to achieve 6 g of starch/kg body weight). When using the IVGTT, the area under the curve (AUC) for glucose and maximum glycaemia were higher for obese dogs compared with controls (P < 0·05), with intermediate results for the WR group (P>0·05). Basal insulin, insulin response peak, insulinogenic index and the AUC for insulin increment from 0 to 15 min and from 60 to 120 min were higher for the obese group (P < 0·05), while the WR group and control dogs showed similar results (P>0·05). When using the GPPR test, the AUC for insulin increment from 0 to 120 min was higher for the obese group compared with the control group (P < 0·05) and intermediate for the WR group (P>0·05). However, the AUC for insulin increment from 120 to 360 min was similar between the obese and WR groups (P>0·05), while it was lower for the control group (P < 0·05). The IVGTT showed that the loss of 20 % body weight resulted in an improvement of glucose control with reduced insulin secretion, and these same WR dogs showed higher insulin secretion with values similar to those of obese dogs when the GPPR test was used.

Among obesity-associated disorders, low-grade inflammation has been described. The putative therapeutic properties of citrus and curcumin polyphenols could be associated with their anti-inflammatory properties. Two diets supplemented either with hesperidin (0·05 %) and naringin (0·1 %) from citrus extract or with highly bioavailable curcumin from Curcuma longa extract (0·09 %) were fed to eight obese cats for two 8-week periods (cross-over study design) while maintaining animals in an obese state. Plasma acute-phase protein (APP; α1-acid glycoprotein (AGP), serum amyloid A and haptoglobin) levels were assessed before and at the end of each test period. TNF-α, IL-1β, IL-2, IL-4, IL-5, IL-10, IL-12, IL-18, transforming growth factor-β, interferon (IFN)-γ mRNA levels were determined in peripheral blood mononuclear cells (PBMC) by real-time PCR. Compared with pre-study values, supplementation with citrus polyphenols resulted in lower plasma AGP and haptoglobin concentrations, while that with curcumin resulted in lower plasma AGP concentration. There were no differences between the supplementations. TNF-α, IL-1β, IL-4, IL-5, IL-10, IL-12, IL-18, transforming growth factor-β, mRNA levels remained unaffected by either dietary supplementation. In contrast, IFN-γ and IL-2 mRNA levels were lower at the end of the citrus and the curcumin supplementation, respectively. There were no differences between the supplementations. The present study results show a slight effect of citrus and curcumin supplementation on inflammatory markers expressed by PBMC, and a decreased concentration of APP, which are mainly expressed by the liver. This would confirm that hesperidin and naringin or highly bioavailable curcumin extract have beneficial effects, targeted in the liver and could improve the obesity-related inflammatory state.

Faecal moisture content can determine whether faeces appear soft or firm, and faecal character can influence whether owners are satisfied with a dog food. In a previous study, dogs appeared to produce softer faeces after noon. The purpose of the present study was to determine whether time of defecation affected canine faecal water content. A total of eight hound dogs were fed one of four canned diets as a single meal each morning for 1 week per diet in a Latin square design. All four diets contained approximately 77 % moisture and, on a DM basis, 24 MJ/kg gross energy, 23 % crude protein, 32 % crude fat, 31 % N-free extract and 1 % crude fibre. The proportion of dietary protein from soya-derived texturised vegetable protein (TVP):beef was 0:100, 14:86, 29:71 and 57:43, respectively. Soya carbohydrate partially replaced maize starch as TVP increased. Faeces were collected by direct catch during the sixth morning and afternoon of each diet period. Mean faecal moisture content was greater in the afternoon than in the morning (79 v. 71 %; P = 0·01) and increased with dietary TVP (P ≤ 0·0001), and there was an interaction between time of day and percentage TVP (P = 0·003). Faecal moisture content differed from morning to afternoon only with TVP in the diet. Faecal wet weight was similar from morning to afternoon. This suggests that time of day and presence of TVP from soya should be taken into account when evaluating the effect of a diet on faecal form and moisture content in dogs fed once daily.

The occurrence and severity of obesity- and insulin resistance-related disorders vary according to the diet. The aim of the present longitudinal study was to examine the effects of a high-fat or a high-fructose diet on body weight (BW), body fat mass, insulin sensitivity (IS) and lipid profiles in a rat model of dietary-induced obesity and low IS. A total of eighteen, 12-week-old male Wistar rats were divided into three groups, and were fed with a control, a high-fat (65 % lipid energy) or a high-fructose diet (65 % fructose energy) for 10 weeks. BW, body fat mass (2H2O dilution method), IS (euglycaemic–hyperinsulinaemic clamp technique), plasma glucose, insulin, NEFA, TAG and total cholesterol were assessed before and at the end of 10-week period. Cholesterol was measured in plasma lipoproteins separated from pooled samples of each group and each time period by using fast-protein liquid chromatography. All rats had similar BW at the end of the 10-week period. Body fat mass was higher in the high-fat group compared to the control group. There was no change in basal glycaemia and insulinaemia. The IS was lower in the high-fat group and was unchanged in the high-fructose group, compared to the control group. Plasma TAG concentration and cholesterol distribution in lipoproteins did not change over time in any group. Plasma NEFA concentration decreased, whereas plasma TAG concentration increased over time, regardless of the diet in both cases. The 10-week high-fat diet led to obesity and low IS, whereas rats fed with the high-fructose diet exhibited no change in IS and lipidaemia. The high-fat diet had more deleterious response than high-fructose diet to induce obesity and low IS in rats.

In dry-extruded canine diets, starch ileal digestibility varies with the starch source, amount and processing parameters. Starch that escapes duodeno-ileal digestion can affect faecal quality by stimulating colonic bacterial fermentation. The aim of the present study was to assess the effect of various resistant starch (RS) sources and levels on the faecal score of dogs of different breeds and sizes. A total of twenty-one healthy adult female dogs (body weight ranging 5·0–30·6 kg) were used. The maintenance diet for the dogs was supplemented with increasing amounts of RS from two sources: high-amylose starch from maize (to 2·5, 4·3 and 7·4 % RS) and raw potato starch (to 7·4 and 11·4 % RS). Each level of RS was tested over a 7 d period followed by a 7 d washout period. Faecal scores were evaluated by one person using a scale ranging from 1 (for hard and dry faeces) to 5 (for liquid stools). Faeces were considered ‘optimal’ at scores of 2·5–3·0, ‘acceptable’ at scores of 3·0–3·75 and ‘unacceptable’ at scores >3·75. Small dogs showed very little sensitivity to RS based on the faecal score, while large dogs were quite responsive to RS supplementation. These results suggest that small dogs are poor models for assessing the effect of starch sources on ileal digestibility. They also indicate that a low RS content (strongly affected by source and processing) is an important factor for ensuring an optimal faecal score in large breed dogs.