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Utilizing metabolomics, a tool for measuring and characterizing low-molecular-weight substances (LMWs), to identify eventual changes in response to dietary intervention is novel in cats with chronic kidney disease (CKD), a condition characterized by retention of uremic solutes. This study aims to assess the serum metabolomic profile of cats in early stages of CKD and to compare the serum metabolomic of CKD cats after 60 days of a renal diet to evaluate the effect of dietary intervention on these metabolites. Twenty-five domestic cats were included in the study. Fifteen cats with CKD stages 1 ( n = 6) and 2 ( n = 9) according to the International Renal Interest Society (IRIS) were included in the renal groups, and a control group consisting of 10 cats was included. All animals were enrolled on a maintenance diet for 30 days before the experimental period. The metabolomics analysis was performed by gas chromatography-mass spectrometry (GC-MS). Partial least squares discriminant analysis (PLS-DA) was performed on Metaboanalyst 4.0 software. Forty-three metabolites were identified. Citric acid and monostearin were altered in the CKD2 group when compared to CKD1 and the control group at T0. A total of seven serum metabolites differed after 60 days of the renal diet: glycine, fructose, glutamic acid, arachidonic acid, stearic acid, creatinine, and urea. Changes were seen in the serum metabolomic profile after 60 days of the renal diet, and some of the metabolites that changed in response to the diet have beneficial effects on health. Overall, metabolomics markers have the potential to identify early stages of CKD, providing insights into the possible pathophysiologic processes that contribute to the development and progression of CKD.

Background: Among the undesirable changes associated with obesity, one possibility recently raised is dysbiosis of the intestinal microbiota. Studies have shown changes in microbiota in obese rats and humans, but there are still few studies that characterize and compare the fecal microbiota of lean, obese and dogs after weight loss. Thus, this study aimed to evaluate the effects of a weight loss program (WLP) in fecal microbiota of dogs in addition to comparing them with those of lean dogs. Twenty female dogs of different breeds, aged between 1 and 9 years were selected. They were equally divided into two groups: Obese group (OG), with body condition score (BCS) 8 or 9/9, and body fat percentage greater than 30%, determined by the deuterium isotope dilution method, and lean group (LG) with BCS 5/9, and maximum body fat of 15%. Weight loss group (WLG) was composed by OG after loss of 20% of their current body weight. Fecal samples were collected from the three experimental groups. Total DNA was extracted from the feces and these were sequenced by the Illumina methodology. The observed abundances were evaluated using a generalized linear model, considering binomial distribution and using the logit link function in SAS (p < 0.05)., Results: The WLP modulated the microorganisms of the gastrointestinal tract, so that, WLG and LG had microbial composition with greater biodiversity than OG, and intestinal uniformity of the microbiota (Pielou's evenness index) was higher in OG than WLG dogs (P = 0.0493) and LG (P = 0.0101). In addition, WLG had values of relative frequency more similar to LG than to OG., Conclusion: The fecal microbiota of the studied groups differs from each other. The weight loss program can help to reverse the changes observed in obese dogs., (© 2022. The Author(s).)

This observational study aimed to evaluate serum and urinary amino acid (AA) concentrations in healthy dogs and dogs with chronic kidney disease (CKD) fed a commercial therapeutic renal diet with reduced protein and phosphorus levels. Ten dogs with CKD stages 3 or 4 composed the study group and received the renal diet for 180 days (RG T180). A control group (CG T30) composed of seven healthy dogs was fed a renal diet for 30 days. When comparing serum AA between RG T180 and CG T30, histidine, isoleucine, leucine, lysine, phenylalanine, tryptophan, cysteine, citrulline, ornithine, taurine, branched-chain amino acids (BCAA), and total essential amino acids (EAA) were higher in RG T180. Meanwhile, arginine, asparagine, aspartate, glutamine, serine, and tyrosine were higher in CG T30. Serum phenylalanine, tryptophan, and hydroxyproline were higher in RG T0 (dogs with CKD before consuming a renal diet) when compared to RG T180. In addition, the serum ratios of arginine/citrulline, tyrosine/phenylalanine, and serine/glycine were higher in CG T30 than in RG T180. Concerning urinary AA concentrations in CKD dogs, isoleucine, phenylalanine, tryptophan, aspartate, cysteine, and BCAA were higher in RG T180. In urine, the total EAA/total non-essential AA ratio in RG T180 was higher than in CG T30 as well as tyrosine/phenylalanine ratio higher in CG T30. In conclusion, the combination of renal diet and conservative treatment over 6 months in dogs with CKD stages 3 or 4 affected the AAs metabolism when compared to healthy adult dogs.

Chronic kidney disease (CKD) is highly prevalent in dogs, and metabolomics investigation has been recently introduced for a better understanding of the role of diet in CKD. This study aimed to compare the serum metabolomic profile of healthy dogs (CG) and dogs with CKD (CKD-T0 and CKD-T6) to evaluate whether the diet would affect metabolites. Six dogs (5 females; 1 male; 7.47 ± 2.31 years old) with CKD stage 3 or 4 (IRIS) were included. CG consisted of 10 healthy female dogs (5.89 ± 2.57 years old) fed a maintenance diet. Serum metabolites were analyzed by 1 H nuclear magnetic resonance ( 1 H NMR) spectra. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were performed to assess differences in metabolomic profiles between groups and before (CKD-T0) and after renal diet (CKD-T6). Data analysis was performed on SIMCA-P software. Dogs with CKD showed an altered metabolic profile with increased urea, creatinine, creatine, citrate, and lipids. Lactate, branched-chain amino acids (BCAAs), and glutamine were decreased in the CKD group. However, after 6 months of diet, the metabolite profiles of CKD-T0 and CKD-T6 were similar. Metabolomics profile may be useful to evaluate and recognize metabolic dysfunction and progression of CKD, and the diet may have helped maintain and retard the progression of CKD.

The post-operative period can generate immunological stress and can be modulated through supplementation with the omega-3 series of polyunsaturated fatty acids. This study aimed to evaluate the effects of diets enriched with high doses of eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids and glutamine on inflammatory mediators in dogs before and after ovariohysterectomy (OVH). Twelve female dogs were divided into two groups: group A was fed a commercial diet without the addition of EPA and DHA, and group B was fed an experimental diet enriched with EPA and DHA (0.2 g/100 kcal). Experimental diet intake initiated 21 days before surgery and continued until 30 days after OVH. Parameters measured were serum cytokines (TNF-α, IL-6 and IL-10), C-reactive protein (CRP), IGF-1, lymphoproliferation and body composition before and after surgery. Statistical analyses were performed with SAS software considering the effects of age and diet and their interactions, and means were compared by the Tukey test. There was no difference between groups in body weight (p = .682), lean mass (p = .101) and body fat (p = .103). There were no group differences in serum concentrations of TNF-α, IL-6, IL-10, IGF-1, CRP and the percentage of lymphocyte proliferation. However, a time effect for TNF-α was observed (p < .001), in which T 0P (10 days after the surgical procedure) presented lower values of this cytokine when compared to the other evaluation time points; and interaction effects between group and time were observed for serum concentrations of IL-6 (p < .001) and IL-10 (p = .002). OVH procedure was not considered invasive enough to increase inflammatory cytokines after 30 days of surgery, as well as the dosage of the EPA and DHA used before and after the surgery did not modulate the inflammatory markers., (© 2019 Blackwell Verlag GmbH.)

Myxomatous mitral valve disease (MMVD) is characterized by thickening of the valve leaflets and omega-3 (ω-3) supplementation has been associated with modulation of blood pressure (BP) and heart rate, improvement of doppler echocardiographic indices, antiarrhythmic, anti-inflammatory and anti-dislipidemic effects in dogs and humans, although prospective studies of it single use are still absent in the veterinary literature. The objective of this study was to evaluate the influence of ω-3 supplementation in dogs with MMVD. Twenty-nine dogs were followed quarterly for 12 months by clinical evaluation, arterial blood pressure, electrocardiography, doppler echocardiography, thoracic radiography and laboratory tests including inflammatory mediators and cardiac biomarker blood concentrations. The dogs were classified in stages B2 and C, according to the classification proposed by ACVIM 2019. They were randomly assigned to either ω-3 group (ω-3G) or control group (CG). The ingestion of ω-3 reduced the chance of developing arrhythmias by 2.96 times (p = 0.003). The vertebral heart size (VHS) measurements were higher in the control group (p = 0.033). In conclusion, at the dosages used in this study, ω-3 dietary supplementation reduces the volumetric overload, has antiarrhythmic effect and keeps dogs with B2 and C stages of MMVD in milder stages of the disease., Competing Interests: J.T.J. and C.F.F.P. are employed by a commercial company (Grandfood Indústria e Comércio, São Paulo, Brazil), however, this does not alter our adherence to PLOS ONE policies on sharing data and materials.

Purpose: Sperm DNA fragmentation is a major cellular mechanism underlying varicocele-related male infertility. However, the type of DNA fragmentation - whether oxidative or of another nature - remains unknown. Thus, the aim of this study was to evaluate single- and double-stranded sperm DNA fragmentation, and oxidative-induced sperm DNA damage in men with varicocele., Materials and Methods: A cross-sectional study was performed, including 94 normozoospermic adults, of which 39 men without varicocele (controls) and 55 men with varicocele grades II or III, uni- or bilaterally. All men collected semen by masturbation. After semen analysis, the remaining volume was used for evaluation of three types of sperm DNA damage: (i) total DNA fragmentation, using an alkaline comet assay, (ii) double-stranded DNA fragmentation, using a neutral comet assay, and (iii) oxidative DNA damage, using an alkaline comet assay associated with the DNA glycosylase formamidopyrimidine enzyme. In each assay, percentage of sperm with any degree of DNA fragmentation, and with high DNA fragmentation were compared between the groups using an unpaired Student's t test or a Mann-Whitney test., Results: The varicocele group presented a higher rate of sperm with fragmented DNA (both any and high DNA fragmentation), considering single-stranded DNA fragmentation, double-stranded DNA fragmentation, or a combination of both, as well as oxidative-induced DNA fragmentation., Conclusions: Patients with varicocele have an increase in sperm DNA fragmentation levels, particularly in oxidative stress-induced sperm DNA damage., Competing Interests: None declared., (Copyright® by the International Brazilian Journal of Urology.)

Introduction: The study of metabolic profile can be an important tool to better understand, at a systemic level, metabolic alterations caused by different pathological conditions, such as obesity. Furthermore, it allows the discovery of metabolic biomarkers, which may help to diagnose alterations caused by obesity., Objective: To investigate the metabolic profile of blood serum of obese dogs, control dogs, and dogs that were subjected to a weight loss program., Methods: Ten obese adult spayed female dogs were included, and their body composition was determined by the deuterium isotope dilution method. The dogs were subjected to a weight loss program and formed a new experimental group after losing 20% of the initial body weight. A third experimental group was composed of ten lean adult spayed female dogs. The metabolic profile of blood serum was evaluated through nuclear magnetic resonance (NMR). Principal Component Analyses (PCA) and Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA) models were constructed using Pareto scaling pre-processing. Pathway analysis was also performed using the MetaboAnalist online tool., Results: The PCA shows that the control and after weight loss groups presented a trend to negative PC1, indicating similarities between these two groups. In contrast, obese animals presented a tendency to appear on negative PC2 indicating a different metabolic profile. The OPLS-DA analysis of the serum indicated that healthy groups presented higher content of glucose, while animals that lost weight had higher levels of cholesterol and lactate than the control group. On the other hand, the analysis showed that lipid content, cholesterol, and branched-chain amino acids were highest in obese animals. Variable Influence on Projection (VIP) analysis demonstrated that Lactate is the most important metabolite for the OPLS-DA model and Hierarchical Cluster Analysis (HCA) corroborated the similarity between the control group and the obese after weight loss groups. Moreover, the pathway analysis indicated the most important metabolic pathways related to this dataset., Conclusions: The metabolomic assessment based on NMR of blood serum differed between obese dogs and animals in optimal body condition. Moreover, the weight loss resulted in metabolic profiles similar to those observed in lean animals.

Background: Hyperlipidaemia is considered a cause of other diseases that are clinically important and potentially life threatening. Combination of pea and barley as exclusive starch sources is known to interfere with glycemic control in diabetic dogs, but their effect on lipid profile of hiperlipidaemic dogs is yet to be evaluated. Twelve adult diabetic dogs were fed three dry extruded diets with different starch sources and different fat levels: peas and barley (PB), maize (Mi), and peas, barley and rice (Ba) with 15.7, 15.6 and 9.0% of their dry matter as fat, respectively. Plasmatic cholesterol and triglycerides concentration curves over 10 h were obtained after 60 days on each diet and with the same NPH insulin dose. ANOVA test or Friedman test were used to compare the dietary effects on triglycerides and cholesterol variables among the diets., Results: Dogs presented lower mean (p = 0.05), fasting (p = 0.03), and time 8-h postprandial (p = 0.05) triglyceridemia after PB diet period than Ba diet period and time 4-h postprandial (p = 0.02) lower after PB than Mi diet. Cholesterolemia mean, minimum, maximum, area under the cholesterol curve and times points: 2, 4, 6, 8 and 10-h postprandial, had lower values after PB ingestion in comparison to Mi, without difference to Ba diet., Conclusion: Inclusion of pea and barley, as exclusive starch sources, in therapeutic diets for diabetic dogs can minimize plasmatic triglycerides and cholesterol concentration at fasting and at different postprandial time, compared to the maize diet or diet with lesser fat content.

An integrated study on the effect of renal diet on mineral metabolism, fibroblast growth factor 23 (FGF-23), total antioxidant capacity, and inflammatory markers has not been performed previously. In this study, we evaluated the effects of renal diet on mineral metabolism, oxidative stress and inflammation in dogs with stage 3 or 4 of chronic kidney disease (CKD). Body condition score (BCS), muscle condition score (MCS), serum biochemical profile, ionized calcium (i-Ca), total calcium (t-Ca), phosphorus (P), urea, creatinine, parathyroid hormone (PTH), FGF-23, interleukin 6 (IL-6), interleukin 10 (IL-10), tumor necrosis factor alpha (TNF-α) and total antioxidant capacity (TAC) were measured at baseline (T0) and after 6 months of dietary treatment (T6). Serum urea, P, t-Ca, i-Ca, PTH, FGF-23, IL-6, IL-10, TNF-α and TAC measurements did not differ between T0 and T6. Serum creatinine (SCr) was increased at T6 and serum PTH concentrations were positively correlated with serum SCr and urea. i-Ca was negatively correlated with urea and serum phosphorus was positively correlated with FGF-23. Urea and creatinine were positively correlated. The combination of renal diet and support treatment over 6 months in dogs with CKD stage 3 or 4 was effective in controlling uremia, acid-base balance, blood pressure, total antioxidant capacity, and inflammatory cytokine levels and in maintaining BCS and MCS.

The source of starch may interfere with glycaemic control in dogs, but few studies have evaluated these aspects in diabetic dogs. This study compared the effects of two isonutrient diets with different starch sources, peas and barley (PB) v. maize (Mi), on diabetic dogs. The Mi diet was processed in order to generate a lower starch gelatinisation index. In all, fifteen adult diabetic dogs without other conditions were included. The animals were fed two dry extruded rations with moderate levels of fat and starch and high levels of protein and fibre using a random, double-blind cross-over design. Glycaemic curves over 48 h were developed via continuous glucose monitoring after 60 d on each diet and with the same neutral protamine Hagedorn (NPH) insulin dosage. The following were compared: fasting, mean, maximum and minimum blood glucose, maximum and minimum glycaemia difference, glycaemic increment, area under the glycaemic curve, area under the glycaemic increment curve and serum fructosamine concentration. Paired t tests or Wilcoxon signed-rank tests were used to compare the amount of food and nutrients ingested and the dietary effects on glycaemic variables between the diets. Dogs fed the PB diet presented a lower average mean interstitial glucose (P=0·01), longer mean hypoglycaemic time (P<0·01), shorter mean hyperglycaemic time (P<0·01) and smaller difference between maximum and minimum blood glucose levels (P=0·03). Thus, the processing applied to the Mi diet was not sufficient to achieve the same effects of PB on glycaemic control in diabetic dogs.

The aim of this study was to evaluate the antioxidant effect of in vitro supplementation with vitamin E in human spermatozoon incubated with an oxidative stress inducer. In this study, semen samples from 30 patients were collected and with one aliquot we performed semen analysis according to WHO. The remaining volume was divided into four aliquots: group C: incubated with BWW medium; group I: incubated with 5 mmol 1-1 hydrogen peroxide; group A: incubated with 40 μmol 1-1 vitamin E; and group AI: incubated with both them. After incubations, sperm functional analyses were performed and included: evaluation of oxidative stress, acrosome integrity, mitochondrial activity and DNA fragmentation. Groups were compared using a Friedman test with Bonferroni post hoc (α = 5%). In this study, we observed that in group I there was a decrease in acrosome integrity and mitochondrial activity, and an increase in DNA fragmentation, when compared to group C. Group AI showed an increase in acrosome integrity and mitochondrial activity when compared with group I. Based on our findings, we conclude that the vitamin E supplementation had a positive effect in protecting human spermatozoon from induced oxidative stress., (© 2018 Blackwell Verlag GmbH.)

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.

Many dog owners see homemade diets as a way of increasing the bond with their pets, even though they may not have the convenience of commercial diets. Modifications of ingredients, quality and proportion might change the nutritional composition of the diet, generating nutritional imbalances. The present study evaluated how dog owners use and adhere to homemade diets prescribed by veterinary nutritionists over an extended period of time. Forty-six owners of dogs fed a homemade diet for at least 6 months were selected for the present study. Owners were invited to answer questions by first reading all possible answers and then selecting the one that best indicated their opinion. The results were evaluated through descriptive statistics. Thirty-five owners (76·1 %) found that the diets are easy to prepare. Fourteen owners (30·4 %) admitted to modifying the diets, 40 % did not adequately control the amount of provided ingredients, 73·9 % did not use the recommended amounts of soyabean oil and salt, and 34·8 % did not correctly use the vitamin, mineral or amino acid supplements. Twenty-six owners (56·5 %) reported that their dogs refused to eat at least one food item. All of these alterations make the nutritional composition of the diets unpredictable and likely nutritionally imbalanced. Although homemade diets could be a useful tool for the nutritional management of dogs with certain diseases, not all owners are able to appropriately use this type of diet and adhere to it for an extended period of time and this limitation needs to be considered when recommending the use of homemade diets.

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.

The effects of six extruded diets with different starch sources (cassava flour, brewer's rice, corn, sorghum, peas or lentils) on dog total tract apparent digestibility and glycemic and insulinemic response were investigated. The experiment was carried out on thirty-six dogs with six dogs per diet in a completely randomized design. The diets containing brewer's rice and cassava flour presented the greatest digestibility of dry matter, organic matter and gross energy (p < 0.05), followed by corn and sorghum; pea and lentil diets had the lowest. Starch digestibility was greater than 98% in all diets and was greater for brewer's rice and cassava flour than for lentils and peas diets (p < 0.05). Dogs' immediate post-prandial glucose and insulin responses (AUC < or = 30 min) were greater for brewer's rice, corn, and cassava flour diets (p < 0.05), and later meal responses (AUC > or = 30 min) were greater for sorghum, lentil and pea diets (p < 0.05). Variations in diet digestibility and post-prandial response can be explained by differences in chemical composition of each starch source including fibre content and starch granule structure. The nutritional particularities of each starch ingredient can be explored through diet formulations designed to modulate glycemic response. However, more studies are required to support these.