Relationships Between Gut Microbiota, Metabolome, Body Weight, and Glucose Homeostasis of Obese Dogs Fed with Diets Differing in Prebiotic and Protein Content.

Obesity is a major issue in pets and nutritional strategies need to be developed, like promoting greater protein and fiber intake. This study aimed to evaluate the effects of dietary protein levels and prebiotic supplementation on the glucose metabolism and relationships between the gut, microbiota, metabolome, and phenotype of obese dogs. Six obese Beagle dogs received a diet containing 25.6% or 36.9% crude protein, with or without 1% short-chain fructo-oligosaccharide (scFOS) or oligofructose (OF), in a Latin-square study design. Fecal and blood samples were collected for metabolite analysis, untargeted metabolomics, and 16S rRNA amplicon sequencing. A multi-block analysis was performed to build a correlation network to identify relationships between fecal microbiota, metabolome, and phenotypic variables. Diets did not affect energy homeostasis, but scFOS supplementation modulated fecal microbiota composition and induced significant changes of the fecal metabolome. Bile acids and several amino acids were related to glucose homeostasis while specific bacteria gathered in metavariables had a high number of links with phenotypic and metabolomic parameters. It also suggested that fecal aminoadipate and hippurate act as potential markers of glucose homeostasis. This preliminary study provides new insights into the relationships between the gut microbiota, the metabolome, and several phenotypic markers involved in obesity and associated metabolic dysfunctions.

Study of ingredients and nutrient composition of commercially available treats for dogs.

Forty-one dog treats were selected from the market with the aim of providing more insight into supplemental pet food composition. Thirty-two products (four biscuits, nine tender treats, two meat-based strips, five rawhides, eight chewable sticks, four dental care sticks) were analysed for proximate nutrient composition and quantification of minerals, hydroxyproline (Hyp), starch, glucose, fructose and sucrose. Labelled ingredients were often expressed as non-specific categories. A treat supplied a mean of 332.0±39.2 kcal metabolisable energy (ME)/100 g, and the most energy-dense product was a tender treat (475.0 kcal ME/100 g). Small dogs receive the highest percentage of maintenance energy requirement when producers' feeding instructions are followed. Treat categories revealed variability in dry matter, crude protein, ash, Hyp and starch. Rawhides showed the highest Hyp content. Simple sugars were identified in most treats, and sucrose was the most prevalent. Results of the study suggest treat labelling should include more information on the ingredients used, and the varying nutrient and caloric density of treats should be considered. Specific attention should be given to the use of treats in dogs with specific ingredient sensitivities or nutrient considerations.

The Genome of a Tortoise Herpesvirus (Testudinid Herpesvirus 3) Has a Novel Structure and Contains a Large Region That Is Not Required for Replication In Vitro or Virulence In Vivo.

UNLABELLED: Testudinid herpesvirus 3 (TeHV-3) is the causative agent of a lethal disease affecting several tortoise species. The threat that this virus poses to endangered animals is focusing efforts on characterizing its properties, in order to enable the development of prophylactic methods. We have sequenced the genomes of the two most studied TeHV-3 strains (1976 and 4295). TeHV-3 strain 1976 has a novel genome structure and is most closely related to a turtle herpesvirus, thus supporting its classification into genus Scutavirus, subfamily Alphaherpesvirinae, family Herpesviridae. The sequence of strain 1976 also revealed viral counterparts of cellular interleukin-10 and semaphorin, which have not been described previously in members of subfamily Alphaherpesvirinae. TeHV-3 strain 4295 is a mixture of three forms (m1, m2, and M), in which, in comparison to strain 1976, the genomes exhibit large, partially overlapping deletions of 12.5 to 22.4 kb. Viral subclones representing these forms were isolated by limiting dilution assays, and each replicated in cell culture comparably to strain 1976. With the goal of testing the potential of the three forms as attenuated vaccine candidates, strain 4295 was inoculated intranasally into Hermann's tortoises (Testudo hermanni). All inoculated subjects died, and PCR analyses demonstrated the ability of the m2 and M forms to spread and invade the brain. In contrast, the m1 form was detected in none of the organs tested, suggesting its potential as the basis of an attenuated vaccine candidate. Our findings represent a major step toward characterizing TeHV-3 and developing prophylactic methods against it. IMPORTANCE: Testudinid herpesvirus 3 (TeHV-3) causes a lethal disease in tortoises, several species of which are endangered. We have characterized the viral genome and used this information to take steps toward developing an attenuated vaccine. We have sequenced the genomes of two strains (1976 and 4295), compared their growth in vitro, and investigated the pathogenesis of strain 4295, which consists of three deletion mutants. The major findings are that (i) TeHV-3 has a novel genome structure, (ii) its closest relative is a turtle herpesvirus, (iii) it contains interleukin-10 and semaphorin genes (the first time these have been reported in an alphaherpesvirus), (iv) a sizeable region of the genome is not required for viral replication in vitro or virulence in vivo, and (v) one of the components of strain 4295, which has a deletion of 22.4 kb, exhibits properties indicating that it may serve as the starting point for an attenuated vaccine.

Effect of short-chain fructooligosaccharide-enriched energy-restricted diet on weight loss and serum haptoglobin concentration in Beagle dogs.

The effects of the dietary inclusion of two levels of short-chain fructooligosaccharides (sc-FOS) on weight loss, biochemical parameters and serum haptoglobin concentration were investigated in twelve experimental obese Beagle dogs. Dogs were randomised into two groups and submitted to a weight loss program (WLP): the control group (C) received a commercial energy-restricted high-protein diet containing 1 % DM sc-FOS, whereas the test group (T) received the same diet enriched with sc-FOS to attain a 3 % DM content. Body weight (BW) and body condition score were weekly assessed in each dog and blood was collected before and after WLP to measure total plasma cholesterol (CHOL), TAG, NEFA, glucose (GLUC), insulin, serum leptin and haptoglobin. Groups showed similar BW and blood parameters before treatment. When values before and after treatment of the dogs were compared, significant reductions were observed for all parameters, with the exception of NEFA and GLUC. However, when these reductions were compared between C and T groups, significant differences were detected only for haptoglobin (T before v. T after: 1545 v. 605 mg/l, P = 0.03; C before v. C after: 1635 v. 1400 mg/l, P = NS). Positive correlations between haptoglobin and CHOL and between haptoglobin and TAG were observed before but not after WLP. In conclusion, feeding obese dogs with the energy-restricted diet caused significant weight loss and reduction of blood parameters, irrespective of the sc-FOS content included. However, serum haptoglobin level, and the subclinical inflammatory condition associated with it, was significantly lowered in the T but not in the C group.

Short-chain fructooligosaccharides influence insulin sensitivity and gene expression of fat tissue in obese dogs.

Dietary fibers may modulate insulin resistance and glucose homeostasis in dogs. Their efficacy is, however, dependent on their origin, physical properties, and fermentability in the large bowel. Eight healthy Beagle dogs were fed a commercial diet at twice their maintenance requirements until they became obese. They were then maintained in the obese state and used in a cross-over design study to evaluate the effects of short-chain fructooligosaccharide (scFOS) supplementation (1% wt:wt dry matter in the diet). The euglycemic hyperinsulinemic clamp technique was performed before and after fattening and at the end of each 6-wk cross-over period. Fat tissue biopsies were taken in food-deprived and postprandial phases to measure mRNA abundance of genes involved with fatty acid, glucose metabolism, or inflammation. Insulin resistance appeared progressively with fattening and the rate of glucose infusion during euglycemic clamp was lower (P < 0.05) at the end of the fattening period (7.39 mg.kg(-1).min(-1)) than at baseline (21.21 mg.kg(-1).min(-1)). In stable obese dogs, scFOS increased (P < 0.05) the rate of glucose infusion compared with control (7.77 vs. 4.72 mg.kg(-1).min(-1)). Plasma insulin and triglyceride concentrations were greater in obese than in lean dogs but were not altered by scFOS. Whereas mRNA was not affected in food-deprived dogs, scFOS increased uncoupling protein 2 (P = 0.05) and tended to increase carnitine palmitoyl transferase 1 adipose mRNA levels during the postprandial period (P = 0.09). Adding 1% scFOS to the diet of obese dogs decreases insulin resistance and appears to modulate the transcription of genes involved in fatty acid or glucose metabolism.

Effects of chronic obesity and weight loss on plasma ghrelin and leptin concentrations in dogs.

The objective of this study was to evaluate, in dogs, the effects of obesity and weight loss on plasma total ghrelin and leptin concentrations. Twenty-four Beagle dogs, 12 control lean and 12 obese dogs of both genders and aged between 1 and 9 years, were used for the experiments. Mean body weight was 12.7+/-0.7 kg for the lean group and 21.9+/-0.8 kg for the obese group. The trial was divided into three phases. During phase 1, all 24 Beagle dogs were fed a maintenance diet. During phase 2, the obese dogs were submitted to a weight loss protocol with a high protein-low energy diet. The weight loss protocol ended once dogs reached optimal body weight. During phase 3, the dogs that were submitted to the weight loss protocol were maintained at their optimal body weight for 6 months. Plasma total ghrelin, leptin, insulin and glucose concentrations were measured to evaluate the effects of obesity and weight loss on these parameters in dogs. Body weight, body condition score, thoracic and pelvic perimeters, and ingested food amounts were also recorded during the study. Obese dogs demonstrated a significant decrease in plasma ghrelin and a significant increase in plasma leptin and insulin concentrations when compared with control dogs. During weight loss, significant increases in plasma total ghrelin and glucose and significant decreases in plasma leptin and insulin were observed. The increase in plasma ghrelin concentrations seemed to be transient. Body weight and the morphometric parameters correlated positively with leptin concentrations and negatively with total ghrelin concentrations. These results suggest that ghrelin and leptin could play a role in dogs in the adaptation to a positive or negative energy balance, as observed in humans.

Influence of obesity on plasma lipid and lipoprotein concentrations in dogs.

OBJECTIVE: To determine effects of obesity and diet in dogs on plasma lipid and lipoprotein concentrations by assaying plasma leptin and ghrelin concentrations and determining total plasma cholesterol and triglyceride concentrations as well as the concentrations of cholesterol and triglycerides in various lipoprotein classes (ie, very-low-density, low-density, and high-density lipoproteins). ANIMALS: 24 Beagles; 12 lean (mean [+/- SEM] body weight, 12.7 +/- 0.7 kg) and 12 chronically obese (21.9 +/- 0.8 kg) dogs of both sexes, between 1 and 9 years old. PROCEDURES: Total plasma cholesterol and triglyceride concentrations; lipoprotein cholesterol and triglyceride concentrations; and plasma ghrelin, leptin, free fatty acids, insulin, and glucose concentrations were measured and compared between lean and obese dogs, both of which were fed a complete and balanced maintenance diet. Chronically obese dogs were subsequently fed a high-protein low-energy diet to evaluate effects of diet composition on plasma lipid and lipoprotein measurements. RESULTS: Chronic obesity resulted in a significant decrease in plasma ghrelin concentration and a significant increase in plasma leptin, cholesterol, and triglyceride concentrations in dogs. High total plasma cholesterol and triglyceride concentrations resulted from increased cholesterol and triglyceride concentrations in all lipoprotein fractions. In obese dogs, modification of diet composition resulted in beneficial effects on plasma lipid and leptin concentrations, even before weight loss was observed. CONCLUSIONS AND CLINICAL RELEVANCE: Correlations exist between obesity and plasma measurements (ie, lipoproteins, leptin, insulin, and ghrelin) commonly associated with obesity. Modification of diet composition to control energy intake improves plasma lipid and leptin concentrations in obese dogs.