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ObjectiveTo validate a recently developed commercially available leptin radioimmunoassay (RIA) for use with feline serum and evaluate the relationship between serum leptin concentrations and body fat mass in domestic cats.Animals19 sexually intact male specific-pathogen-free domestic cats that weighed 3.8 to 7.1 kg and were 1.1 to 3.5 years old.ProcedureSpecificity for feline leptin was evaluated by use of gel filtration chromatography and reverse-phase high-performance liquid chromatography fractionation of serum. Body fat mass was determined by use of the deuterium oxide (D2O) dilution method. Serum water D2O enrichment was measured by use of gas-phase Fourier transform infrared spectroscopy.ResultsBody fat mass and percentage body fat ranged from 0.3 to 2.3 kg and 7.5 to 34.9%, respectively. Serum leptin concentrations were lower in the unfed versus the fed state and ranged between 1.6 and 4.9 ng/ml human equivalent (HE); mean +/- SD value was 2.9 +/- 0.2 ng/ml HE. Leptin concentrations increased with increasing body fat mass and percentage of body fat.ConclusionsLeptin is in the serum of domestic cats in free (> 78%) and apparently bound forms. The relationship between body fat and serum leptin concentration was similar to that observed in humans and rodents and indicative of a lipostatic role for leptin in cats. Cats that have an overabundance of body fat appear to be less sensitive to the weight-normalizing action of leptin than cats of ideal body condition.

Naturally occurring diabetes mellitus (NODM) is one of the most common endocrine disorders in dogs and its etiology closely resembles type 1 diabetes mellitus (T1DM) in people. Human patients with T1DM commonly have cellular derangements consistent with inflammation, impaired immune function, and hypovitaminosis D. There is little information available regarding inflammatory biomarkers, immune function, and vitamin D status in diabetic dogs. Therefore, our objectives were to assess inflammatory biomarkers, vitamin D metabolites, and phagocytic capacity in diabetic dogs and determine whether associations exist with these variables and the level of clinical control or vitamin D metabolites. This was a prospective case-control study that included 20 otherwise healthy diabetic dogs (clinically controlled, n  = 10; uncontrolled, n  = 10) and 20 non-diabetic, healthy, age (± 2 years), breed, and sex matched controls. Complete blood count, biochemical panel, urinalysis, and fructosamine were performed at a single commercial reference laboratory. Basal plasma tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-8, and IL-10 were measured using a canine-specific multiplex bead-based assay. Serum C-reactive protein (CRP) was measured using a commercially available ELISA kit. Serum 25-hydroxyvitamin (OH)D 3 and 24,25-dihydroxyvitamin (OH) 2 D 3 were measured with HPLC. Phagocytosis of opsonized- Escherichia coli ( E. coli ) was evaluated with flow cytometry. Diabetic dogs had higher serum CRP concentrations than controls ( p  = 0.02). Plasma IL-8 concentrations were higher in diabetic dogs with uncontrolled clinical disease compared to controls ( p  = 0.02). Diabetic dogs had a lower percentage of leukocytes that phagocytized opsonized- E. coli ( p  = 0.02), but an increased number of bacteria phagocytized per cell ( p  < 0.001) compared to controls. No between-group differences were identified in vitamin D metabolites, nor were associations found between vitamin D and any variables. Fructosamine had a positive association with serum CRP concentration (rho = 0.35, p  = 0.03) and number of bacteria phagocytized per cell (rho = 0.45, p  = 0.004) in our cohort ( n  = 40). Like people with T1DM, diabetic dogs have a proinflammatory phenotype and phagocytic dysregulation that may be correlated with glycemic control., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Jaffey, Backus, Kreisler, Graves, Al-Nakkash and Allison.)

Background: Vitamin D deficiency and excess in clinically presented cats conventionally is diagnosed by comparison of patient plasma 25-hydroxyvitamin D (25 (OH)D) concentration with plasma reference intervals determined in healthy adult cats. For immature cats, validity of this vitamin D status assessment method is uncertain., Objective: The overall objective was determination of whether plasma concentration of 25 (OH) D and other vitamin D metabolites in immature cats markedly change with developmental age as has been reported in other species., Methods: Four male and 4 female domestic short-hair kittens from weaning were continuously presented a single nutritionally adequate growth-diet. Concentrations of 25 (OH) D and 24,25-dihydroxyvitamin D (24,25 (OH) 2 D), and calcitriol were quantified in plasma of jugular venous blood collected at 12, 15, 18, and 21 weeks and 1 year of age. Plasma was liquid and solid-phase extracted and fractionation by normal-phase HPLC, and 25 (OH) D and 24,25 OH) 2 D quantified by reverse-phase HPLC-UV and calcitriol by RIA., Results: Plasma 3-epi-25 (OH) D and 25 (OH) D concentrations increased ( p  < 0.001) with age so that by study end the concentrations rose by 1-and 2-fold, respectively. Concentrations of 3-epi-25 (OH) D relative to 25 (OH) D were 30% at 12 weeks and 20% at 1 year. Between ages 12 and 21 weeks, rises in 25 (OH) D concentration were positively correlated with body weight gains ( ρ  = 0.952, p  < 0.001) and 24,25 (OH) 2 D concentrations were consistently greater than 25 (OH) D concentrations ( p  < 0.001). At 1 year of age, concentrations of 24,25 (OH) 2 D declined below those of 25 (OH) D and 3-epi-24,25 (OH) 2 D consistency occurred in low concentrations. Vitamin D 2 metabolites and sex differences in metabolite concentrations were not observed., Conclusion: Reliance on quantification of plasma 25 (OH) D concentration for vitamin D status assessment in kittens may be confounded by developmental changes in 25 (OH) D independent of vitamin D intake. High 24,25 (OH) 2 D concentration and occurrence of 3-epi-25 (OH) D in plasma additionally may interfere with the quantification., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Backus and Ueda.)

Human patients with type 1 diabetes mellitus (T1DM) are susceptible to several long-term complications that are related to glycemic control and immune dysregulation. Immune function remains relatively unexplored in dogs with naturally occurring diabetes mellitus (NODM). Calcitriol improves various aspects of immune function in a variety of species, but its effect in diabetic dogs remains unexplored. Therefore, the objectives of this study were to (i) evaluate immune function in dogs with NODM and determine if differences exist based on the level of clinical control and (ii) assess the immunomodulatory effects of calcitriol. Twenty diabetic dogs (clinically controlled, n = ten, not controlled, n = ten) and 20 non-diabetic, healthy control dogs were included in this prospective, case-control study. Whole blood was incubated with calcitriol (10 -7 M) or negative control, after which the samples were divided for phagocytosis and leukocyte cytokine response experiments. The phagocytosis of opsonized Escherichia coli ( E. coli ) was evaluated with flow cytometry. The samples for leukocyte cytokine response evaluations were stimulated with lipopolysaccharide (LPS), lipoteichoic acid (LTA), or phosphate buffer solution (PBS; negative control), and tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-8, and IL-10 were measured in supernatant using a canine-specific multiplex bead-based assay. The leukocytes from diabetic dogs produced higher concentrations of IL-10 ( p = 0.01), IL-6 ( p < 0.0001), and IL-8 ( p < 0.0001) than the control dogs while controlling for the intervention and stimulant. Calcitriol decreased the supernatant concentrations of TNF-α ( p < 0.001) and IL-8 ( p = 0.04) with concomitant increases in IL-6 ( p = 0.005). Diabetic dogs had a lower percentage of leukocytes undergoing phagocytosis ( p < 0.0001) but a higher number of bacteria phagocytized per cell ( p = 0.001) when compared to the control dogs. Calcitriol had no effect on phagocytic capacity. Lastly, the status of clinical control in diabetic dogs did not yield differences in immune function. These results support that dogs with NODM exhibit immune dysregulation and warrant additional investigation.

Background: Serum 25-hydroxyvitamin (OH)D, C-reactive protein (CRP), and haptoglobin are useful biomarkers in various infectious diseases and inflammatory disorders in dogs, but their utility in histoplasmosis is unknown., Objective: Determine if serum 25(OH)D, CRP, and haptoglobin concentrations are different in dogs with histoplasmosis compared to healthy controls and whether serum globulin, albumin, CRP, or haptoglobin are associated with 25(OH)D concentration., Animals: Twenty-two client-owned dogs (histoplasmosis, n = 12; controls, n = 10)., Methods: Prospective case-control study. Dogs with histoplasmosis were categorized as pulmonary, disseminated, or gastrointestinal (GI) tract. Serum 25(OH)D was measured using modified high-performance liquid chromatography (HPLC). Serum CRP and haptoglobin were measured with ELISA assays., Results: Dogs with histoplasmosis were grouped as disseminated (n = 8) and GI tract (n = 4). No dogs had pulmonary tract involvement alone. Dogs with histoplasmosis (median, interquartile range [IQR]; 11.6 ng/mL, 16.8) had lower serum 25(OH)D concentrations than controls (35.7 ng/mL, 17.6; P < .001). Serum CRP and haptoglobin concentrations were higher in dogs with histoplasmosis (CRP: median, IQR; 63.5 mg/L, 37.1 and haptoglobin: 459.7 mg/dL, 419.6) than controls (CRP: 1.9 mg/L, 2; P < .001 and haptoglobin: 85.5 mg/dL, 106.7; P = .003). Serum 25(OH)D concentration was positively associated with fold change in serum albumin concentration (ρ = 0.77; P < .001), and negatively associated with fold change in serum globulin (ρ = -0.61; P = .003) and CRP concentrations (ρ = -0.56; P = .01)., Conclusion and Clinical Importance: Assay of serum 25(OH)D, CRP, and haptoglobin could have clinical value in dogs with histoplasmosis., (© 2023 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC on behalf of American College of Veterinary Internal Medicine.)

Background: Clinicopathologic variables predictive of disseminated coccidioidomycosis are known in humans but have not been explored in dogs. Serum 25-hydroxyvitamin (OH)D correlates with severity of disease of various etiologies in dogs but its role in coccidioidomycosis is unknown., Objective: Determine whether serum 25(OH)D concentrations are different in dogs with coccidioidomycosis compared with healthy controls and if clinicopathologic variables are associated with extent of disease., Animals: Thirty-five dogs with coccidioidomycosis (pulmonary, n = 13; disseminated, n = 15; uncharacterized, n = 7), and 25 healthy control dogs., Methods: Prospective cohort study. Serum 25(OH)D and C-reactive protein (CRP) concentrations were measured with modified-HPLC and a commercial ELISA kit, respectively., Results: There was no difference in 25(OH)D concentrations between dogs with coccidioidomycosis (median, interquartile range [IQR]; 31.9 ng/mL, 23.3-49.2) and controls (29.5 ng/mL, 25.6-40.8, P = .73). Serum 25(OH)D concentration was lower in dogs with coccidioidomycosis and IgG titers ≥1:32 than dogs with titers below this cut-off (P = .02). Dogs with IgG titers ≥1:32 were more likely to have disseminated disease (OR, 7.5; 95% CI: 1.1-68; P = .03). Serum CRP concentrations were higher in dogs with IgG titers ≥1:16 (median, IQR; 4474.8 ng/mL, 2885.8-8236.1) than in those below this cut-off (151.2 ng/mL, 30.4-2907.3; P = .02). There was a significant inverse association between serum 25(OH)D and CRP at 25(OH)D concentrations ≤33 ng/mL., Conclusion and Clinical Importance: Serum 25(OH)D concentration was lower for dogs with IgG titers ≥1:32, indicating a potential association between semi-quantitative titers and 25(OH)D concentrations in dogs with coccidioidomycosis. IgG titers ≥1:32 yielded higher odds of disseminated disease, but was inadequate as a standalone test to determine form of disease., (© 2023 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC on behalf of American College of Veterinary Internal Medicine.)

Objectives: The study aimed to evaluate the safety and effectiveness of vitamin D supplementation with dietary 25-hydroxyvitamin D 3 (25[OH]D 3 ) in adult cats., Methods: Three levels of dietary 25(OH)D 3 concentrations (4.9, 8.4, 11.8 µg/kg as fed) were received by five adult cats for 9 weeks, each in a randomized complete block design. Effects were determined on plasma or serum concentrations of 25(OH)D 3 , 24,25-dihydroxyvitamin D 3 , calcitriol, parathyroid hormone, ionized calcium, urinary excretions of phosphorus, calcium and magnesium, and clinical hematology and chemistry panels., Results: The lowest concentration of dietary 25(OH)D 3 supported elevation of vitamin D status, with no adverse effects. Supplementation of 8.4 µg/kg 25(OH)D 3 had significant effects on the urinary magnesium: creatinine ratio. Increasing supplementation up to 11.8 µg/kg 25(OH)D 3 had significant effects on plasma concentrations of calcium and magnesium, and vitamin D metabolites., Conclusions and Relevance: Dietary supplementation with approximately 5.0 µg/kg of 25(OH)D 3 or the ingested equivalence of 0.09 µg of 25(OH)D 3 per metabolic body weight (kg 0.67 ) is a safe, potent and effective means for raising vitamin D status in cats. A higher dose with approximately 11.8 µg/kg of 25(OH)D 3 resulted in elevation in C-3 epimers of 25(OH)D 3 and slight elevation in plasma magnesium and calcium concentrations above their respective reference intervals.

Background: Taurine status is impacted by dietary supply of methionine and cysteine (SAA) and possibly intestinal microbial activity, where plasma and whole blood taurine concentrations are currently used to evaluate taurine status., Objective: We determined effects of dietary SAA restriction on rate and extent of taurine depletion of blood and skeletal muscle in dogs of two body sizes, and whether oral antibiotic administration affected the taurine depletion and fecal bile acid excretion of the dogs., Methods: Adult, male, Beagles ( n = 6; 10.1-13.1 kg) and larger mixed-breed dogs ( n = 6; 28.5-41.1 kg) were given four dry-expanded diets, whereby each successive diet contained lower protein and/or SAA concentration. After receiving the final diet for 44 weeks, all dogs were orally administered a mixture of ampicillin, neomycin sulfate, and metronidazole for 12 weeks. Taurine concentrations were determined every 2-4 weeks in venous blood and voided urine and every 4 to 16 weeks in biopsied semimembranosus muscle. Fecal bile acid excretion before and after antibiotics administration were quantified., Results: When given for 36 weeks the lowest SAA diet, 3.4% methionine and 2.9% cystine, taurine concentrations in whole blood were not different between groups, while taurine in plasma declined ( P < 0.05) in large but not in small dogs, and taurine in biopsied muscle decreased ( P < 0.05) by 50% in large and by 37% in small dogs. Concentrations of taurine in muscle were lower ( P < 0.01) and fecal bile acids greater ( P = 0.001) in large than small dogs. Antibiotic administration restored plasma and muscle taurine to initial concentrations and halved fecal bile acid excretion by dogs of both groups., Conclusions: Blood taurine concentration may not be a sensitive indictor of taurine depletion caused by low intake of bioavailable SAA in dogs, especially in large dogs. Taurine status and dietary SAA requirements of dogs may substantively depend on taurine loss mediated by intestinal microbiota., Competing Interests: This study received funding from Royal Canin, SAS, France. The funder had the following involvement with the study: production and analysis of diets used and intellectual contributions to design of experimental trials, interpretation of results, and writing of the report. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Tôrres, Biourge and Backus.)

Objective: To compare the effects of short-term dietary supplementation with vitamin D 3 and 25-hydroxyvitamin D 3 (25[OH]D 3 ) on indicators of vitamin D status in healthy dogs., Animals: 13 purpose-bred adult dogs., Procedures: 20 extruded commercial dog foods were assayed for 25(OH)D 3 content. Six dogs received a custom diet containing low vitamin D concentrations and consumed a treat with vitamin D 2 (0.33 μg/kg 0.75 ) plus 1 of 3 doses of 25(OH)D 3 (0, 0.23, or 0.46 μg/kg 0.75 ) once daily for 8 weeks followed by the alternate treatments in a crossover-design trial. In another crossover-design trial, 7 dogs received a custom diet supplemented with vitamin D 3 or 25(OH)D 3 (targeted content, 3,250 U/kg [equivalent to 81.3 μg/kg] and 16 μg/kg, respectively, as fed) for 10 weeks followed by the alternate treatment. In washout periods before each trial and between dietary treatments in the second trial, dogs received the trial diet without D-vitamer supplements. Dietary intake was monitored. Serum or plasma concentrations of vitamin D metabolites and biochemical variables were analyzed at predetermined times., Results: 25(OH)D 3 concentrations were low or undetected in evaluated commercial diets. In the first trial, vitamin D 2 intake resulted in quantifiable circulating concentrations of 25-hydroxyvitamin D 2 but not 24R,25-dihydroxyvitamin D 2 . Circulating 25(OH)D 3 concentration appeared to increase linearly with 25(OH)D 3 dose. In the second trial, circulating 25(OH)D 3 concentration increased with both D vitamer-supplemented diets and did not differ significantly between treatments. No evidence of vitamin D excess was detected in either trial., Conclusions and Clinical Relevance: Potency of the dietary 25(OH)D 3 supplement estimated on the basis of targeted content was 5 times that of vitamin D 3 to increase indicators of vitamin D status in the study sample. No adverse effects attributed to treatment were observed in short-term feeding trials.

Feline vitamin D status is based on dietary consumption but metabolism of this essential nutrient and the efficacy of supplementation forms are poorly described in cats. The aim of this study was to further elucidate the metabolites of vitamin D 2 in cats and to compare the effectiveness of vitamin D 2 and 25(OH)D 2 for increasing feline vitamin D status. Eight adult male castrated domestic shorthair cats received vitamin D 2 or 25(OH)D 2 in a single crossover design. Vitamin D 2 was dosed daily in a molar equivalent dosage to vitamin D 3 ingested in the diet while 25(OH)D 2 was provided at a daily dose of 20% molar equivalent intake of dietary vitamin D 3 based on its expected higher potency. Plasma concentrations of 25-hydroxyvitamin D epimers were evaluated at baseline then every 2 weeks for a total of 10 weeks. Analysis of multiple vitamin D metabolite concentrations was completed at the end of each supplementation period, followed by a washout period preceding the second phase of the crossover trial. Results showed that supplementation with 25(OH)D 2 more effectively and rapidly raised circulating 25(OH)D 2 levels in cat plasma compared to vitamin D 2 . Formation of C-3 epimers of 25(OH)D 3 , 25(OH)D 2 , and 24,25 R (OH) 2 D 3 , but not 24,25(OH) 2 D 2 , were observed in feline plasma. The abundant concentrations of epimeric forms of vitamin D metabolites found in circulation suggest that these metabolites should be considered during vitamin D analyses in cats. Further studies using 25(OH)D and vitamin D 2 forms are needed to conclude safety and efficacy of these vitamers for supplementation in this species., Competing Interests: CR was employed by the University of Missouri when this research was conducted. CR was employed by the company Hill's Pet Nutrition at the time of manuscript submission. Hill's Pet Nutrition was not involved in the conception of the study, study design, data analysis, or writing of the paper. The remaining author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Ruggiero and Backus.)

Vitamin D insufficiency is associated with various disease processes. We determined whether consumption of a diet supplemented with HyD ® , a 25-hydroxycholecalciferol (25(OH)D 3 ) source, would safely increase plasma 25(OH)D 3 concentrations in Golden Retrievers with low vitamin D status. We hypothesised that dietary supplementation with HyD ® would rapidly increase and sustain plasma 25(OH)D 3 levels in healthy Golden Retrievers with low vitamin D status compared with supplementation with vitamin D 3 . Of fifty-seven privately owned dogs recruited with written owner consent, eighteen dogs with low vitamin D status were identified and sorted between two groups to have similar initial plasma 25(OH)D 3 concentrations, sex distributions, ages and body weights. Dogs of each group were fed a dry dog food supplemented with either 16 μg/kg of 25(OH)D 3 as HyD ® ( n 10) or 81 μg/kg of cholecalciferol (D 3 ) ( n 8) for 4 months. Plasma 25(OH)D 3 concentrations were determined monthly. A significant time effect ( P < 0⋅001) and time by group interaction ( P = 0⋅0045) were found for monthly determined plasma 25(OH)D 3 concentrations. Dogs fed the HyD ® -supplemented diet experienced a 40⋅5 % rise in plasma 25(OH)D 3 values after 1 month ( P < 0⋅001) and no change thereafter. Plasma 25(OH)D 3 values of dogs supplemented with vitamin D 3 did not increase ( P > 0⋅05) and were less than values of dogs supplemented with HyD ® ( P = 0⋅044). With few exceptions, average haematologic, biochemical and urinalyses results remained within the reference range for both groups. Dietary supplementation with HyD ® is sufficient to safely increase and sustain plasma 25(OH)D 3 levels in healthy dogs., (© The Author(s) 2021.)

This study sought to establish a baseline understanding of immune function and its association with serum vitamin D in shelter dogs. Ten apparently healthy shelter dogs housed in the Arizona Humane Society for ≥7 days and 10 apparently healthy, age, breed, and sex-matched control dogs were included. Serum 25-hydroxyvitamin D (25[OH]D), the major circulating vitamin D metabolite, was measured using high performance liquid chromatography. Whole blood samples were stimulated with lipopolysaccharide (LPS), lipoteichoic acid, or phosphate buffer solution, and tumor necrosis factor (TNF)-ɑ, interleukin (IL)-6, and IL-10 were measured using a canine-specific multiplex bead-based assay. Phagocytosis of opsonized-Escherichia coli and E. coli-induced oxidative burst were evaluated with flow cytometry. Shelter dogs had decreased percentages of granulocytes and monocytes (GM) that had phagocytized opsonized-E coli (P = 0.019) and performed E. coli-induced oxidative burst (P = 0.011). There were no significant differences in TNF-α, IL-6, IL-10, or 25(OH)D concentrations between shelter and control dogs. Serum 25(OH)D concentrations had a weak positive association with the intensity of GM E. coli-induced oxidative burst (r 2 = 0.23, P = 0.03). There was a moderate inverse association between serum 25(OH)D concentration and LPS-stimulated TNF-ɑ production in shelter dogs (r 2 = 0.40, P = 0.04). These results demonstrate immune dysregulation in vitro in shelter dogs housed for ≥7 days when compared to age, breed, and sex-matched control dogs. While serum 25(OH)D concentrations did not differ between shelter and control dogs, significant associations between 25(OH)D concentration and immune function parameters in vitro were identified., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

More than one-third of humans and companion dogs in Western societies are overweight or obese. In people, vitamin D deficiency is widespread and associated with obesity, a now recognised inflammatory state. Low vitamin D status occurs in dogs with inflammatory conditions, but its relationship with obesity has not been investigated. In otherwise healthy privately owned adult dogs of ideal body condition (control, n 7) and dogs with overweight to obese body condition (treatment, n 8), serum 25-hydroxyvitamin D (25(OH)D) concentration and body composition as inferred from 2 H-labelled water dilution space were evaluated. Subsequently, the dogs were transitioned to a commercial canine therapeutic weight-loss diet; control dogs were fed to maintain body weight and treatment dogs were energy-restricted to achieve a safe weight-loss rate. Thereafter, serum 25(OH)D concentration was re-evaluated 8 weeks after diet transition, and at the study end, which was 6 months or when ideal body condition was achieved. At study end, body composition analysis was repeated. Initial body condition scores and percentage body fat were positively correlated (ρ = 0·891; P < 0·001). However, percentage body fat and serum 25(OH)D concentration were not significantly correlated. Final serum 25(OH)D concentrations were greater ( P < 0·05) than initial concentrations for control and treatment groups, indicating a diet but not weight-loss effect on vitamin D status. These findings suggest that vitamin D status of dogs is not affected by obesity or loss of body fat with therapeutic weight reduction.

Hypovitaminosis D has been extensively documented in critically ill humans. However, whether or not critically ill dogs have alterations in vitamin D concentrations remains unconfirmed. The primary aims of our study were to compare serum 25-hydroxycholecalciferol [25(OH)D] concentrations in critically ill dogs with healthy control dogs, determine the prognostic utility of serum 25(OH)D concentration as a biomarker in critically ill dogs, and to assess if serum 25(OH)D concentrations in critically ill dogs are associated with length of stay in the intensive care unit or illness severity. Serum concentrations of 25(OH)D together with a range of other clinical, biochemical, and hematological parameters, were measured in 99 dogs within 24 hours of admission to the Intensive Care Unit (ICU). Critically ill dogs (P = 0.001) and dogs with sepsis (P = 0.002) had significantly lower serum 25(OH)D concentrations compared to healthy control dogs. In addition, serum 25(OH)D concentration was an independent predictor of in-hospital and 30 day survival. Using a cut-off of 33 ng/mL, serum 25(OH)D concentrations had excellent sensitivity (0.94; 95% CI, 0.71-1.00), but poor specificity (0.41; 95% CI, 0.31-0.53) for detection of survival. Serum 25(OH)D concentrations were inversely associated with acute patient physiologic and laboratory evaluation (APPLE) fast score but were not associated with ICU length of stay. Hospitalized dogs with critical illness have decreased serum 25(OH)D concentrations compared to healthy dogs and can be used to predict survival in this cohort.

The giant anteater (Mymercophaga tridactyla) is a highly specialized insectivore for which nutrient requirements are not clearly established, making diet formulation challenging for this species. Multiple clinical reports suggest anteaters have an obligate dietary taurine (TAU) requirement. Sulphur amino acid (SAA) metabolism in adult anteaters was evaluated using noninvasive methods to measure TAU synthesis potential from dietary methionine (MET) and a basal diet containing on a dry matter (DM) basis 1.7 mg TAU/kg DM and 6.9 g MET/kg DM. Urinary equilibrium times for TAU excretion were determined by feeding the basal diet with or without 1.5 g/kg DM supplemental TAU (crossover design; n = 4). Effects of supplemental dietary TAU (1.7, 2.0, 2.4, 2.7, 3.0, 3.3 g/kg DM) or MET (6.9, 9.0, 11.2 g/kg DM) on urinary TAU were evaluated (randomized block trials; n = 5 or 4 respectively). All urinary values (TAU, MET, unbound inorganic sulphate) were normalized to creatinine (CRT). Results indicate urinary TAU equilibrium in anteaters requires at least 2 weeks of feeding. Urinary ratio of TAU to CRT (TAU:CRT) increased as dietary TAU content increased from 1.7 to 3.0 g/kg DM, consistent with renal homoeostatic modulation of TAU excretion. Our data indicate that TAU needs were met by TAU in the basal diet or by de novo synthesis. Supplemental MET resulted in ~five- to eightfold increases in urinary TAU:CRT excretion, further supporting existence of mechanisms for TAU synthesis from dietary SAA in anteaters. Adult anteaters appear able to synthesize TAU when diets contain adequate SAA, but dietary TAU may be critical if protein intakes are low or of poor quality. This study may provide guidance on choice of domestic canids vs. felids as suitable physiologic models for improved nutrition in giant anteaters, and also outlines a noninvasive method for assessing TAU status/metabolism that may be useful across species., (© 2017 Blackwell Verlag GmbH.)

Objectives The aim of this report is to describe the identification of a novel vitamin D metabolite, a C-3, alpha-epimer of 25-hydroxycholecalciferol (3-epi-25(OH)D 3 ), in serum and plasma extracts of cat blood and compare its abundance in cat, dog and rat serum to 25-hydroxycholecalciferol (25(OH)D 3 ), a conventional marker of vitamin D status. Methods Serum 25(OH)D 3 and 3-epi-25(OH)D 3 concentrations were measured in healthy cohorts of cats (n = 8), dogs (n = 8) and rats (n = 17) using validated reverse and normal-phase high-performance liquid chromatography methods. The methods were verified using liquid chromatography tandem mass spectrophotometry. Dietary intake and dietary concentrations of vitamin D were also measured for evaluation of species differences and effect of dietary change on vitamin D metabolite concentrations. Differences between cat serum and plasma metabolite concentrations were determined. Results Detectable concentrations of 3-epi-25(OH)D 3 were observed in all cats and rats. No 3-epi-25(OH)D 3 was detected in dogs, where our limit of detection was 5 ng/ml. There were significant differences ( P <0.05) in serum concentrations of 25(OH)D 3 and 3-epi-25(OH)D 3 among species, with cats having the greatest concentrations of both metabolites. Serum and plasma results were not significantly different. A diet change, which resulted in an increase in vitamin D intake among the cats, affected serum concentration with an increase ( P = 0.004) in 3-epi-25(OH)D 3 but no significant change in 25(OH)D 3 . Conclusions and relevance Serum and plasma of cats contain 3-epi-25(OH)D 3 in varied and extraordinary concentrations, much greater than in rats and certainly than that of dogs, a species for which the metabolite was not detected. Importantly, this finding indicates a C-3 epimerization pathway is quantitatively significant for vitamin D metabolism in domestic cats, making 3-epi-25(OH)D 3 assays essential for the evaluation of vitamin D status in cats and positioning the cat as a novel model for study of this pathway.

Objective: To evaluate the effects of intravenous (IV) infusion of fish oil (FO) emulsion following ovariohysterectomy (OVH) on inflammatory mediators and plasma omega-3 nonesterified fatty acids (NEFA) concentrations in dogs., Design: Prospective clinical study., Setting: University teaching hospital., Animals: Twenty-nine privately owned dogs undergoing routine OVH., Interventions: Postoperative 3-hour IV infusion of saline (n = 9), FO (Omegaven, n = 10), or soybean oil (SO, intralipid, n = 10) emulsion and blood collected before, 5 and 24 hours following OVH for plasma NEFA and RBC membrane fatty acids (FAs) concentrations, leukocyte cytokine production capacity, and C-reactive protein (CRP) measurement., Measurements and Main Results: Plasma omega-3 NEFA, eicosapentaenoic acid, docosahexaenoic acid, and total long-chain omega-3 FA significantly increased shortly after FO infusion (8.8 ± 3.3 μM, 13.6 ± 5.6 μM, and 25.1 ± 9.6 μM, respectively) compared to SO (0.7 ± 0.9, 2.3 ± 1.8, and 4.2 ± 3.0 μM, respectively) and saline infusion (1.6 ± 2.5, 2.6 ± 3.1, and 5.9 ± 6.4 μM, respectively). Plasma CRP concentration significantly increased after OVH, but with no significant group differences. A weak negative correlation occurred between post-OVH CRP and postinfusion total long-chain omega-3 FA concentrations (r 2 = 0.21, P = 0.014). Stimulated leukocyte interleukin (IL) 6 production capacity increased (P = 0.001) after OVH in all groups; SO infusion resulted in reduced leukocyte IL-6 production capacity (1048.1 ± 277.7 pg/mL) compared to FO (1299.9 ± 302.1 pg/mL, P = 0.048) and saline infusions (1499.0 ± 363.1 pg/mL, P = 0.01). No significant group difference was observed in leukocyte IL-10 and tumor necrosis factor α production capacities., Conclusions: Postoperative administration of FO emulsion increases plasma omega-3 NEFA concentrations promptly, but does not significantly attenuate CRP production or leukocyte cytokine production capacity. FO infusion at the dosage used in the present study can be safely used in dogs, but it was not clearly beneficial in decreasing post-OVH indices of inflammation., (© Veterinary Emergency and Critical Care Society 2017.)

We previously found a weak response in serum 25-hydroxyvitamin D 3 (25(OH)D 3 ) concentrations when dogs were supplemented with oral vitamin D 3 (D 3 ). In the present study, we determined the relative potency of oral 25(OH)D 3 compared with D 3 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 D 3 supplementation at 2·3 µg/kg body weight 0·75 , while three dogs received a molar equivalency as 25(OH)D 3 . The supplements, dissolved in ethanol, were applied to a commercial treat for consumption. Serum 25(OH)D 3 and 24 R ,25-dihydroxyvitamin D 3 (24 R ,25(OH) 2 D 3 ) were analysed weekly using a validated HPLC method. Both supplementations increased ( P  ≤ 0·01) serum 25(OH)D 3 concentrations. However, oral 25(OH)D 3 resulted in greater ( P  < 0·0001) concentrations than D 3 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)D 3 concentrations were not appearing to plateau. Thereafter, a washout period of 1 month separated the cross-over. Following 25(OH)D 3 , but not D 3 supplementation, serum 24 R ,25(OH) 2 D 3 concentrations increased ( P  ≤ 0·02), 3 to 5 weeks after initiating supplementation. Vitamin D status, as indicated by serum 25(OH)D 3 and 24 R ,25(OH) 2 D 3 concentrations, is more rapidly and efficiently increased in adult dogs by oral supplementation of 25(OH)D 3 than D 3 .

Bisphenol A (BPA) is a widely present endocrine disruptor chemical found in many household items. Moreover, this chemical can bioaccumulate in various terrestrial and aquatic sources; thereby ensuring continual exposure of animals and humans. For most species, including humans, diet is considered the primary route of exposure. However, there has been little investigation whether commercial-brands of dog foods contain BPA and potential health ramifications of BPA-dietary exposure in dogs. We sought to determine BPA content within dog food, whether short-term consumption of these diets increases serum concentrations of BPA, and potential health consequences, as assessed by potential hematological, serum chemistry, cortisol, DNA methylation, and gut microbiome changes, in dogs associated with short-term dietary exposure to BPA. Fourteen healthy privately-owned dogs were used in this study. Blood and fecal samples were collected prior to dogs being placed for two-weeks on one of two diets (with one considered to be BPA-free), and blood and fecal samples were collected again. Serum/plasma samples were analyzed for chemistry and hematology profiles, cortisol concentrations, 5-methylcytosine in lymphocytes, and total BPA concentrations. Fecal samples were used for microbiome assessments. Both diets contained BPA, and after two-weeks of being on either diet, dogs had a significant increase in circulating BPA concentrations (pre-samples=0.7±0.15ng/mL, post-samples=2.2±0.15ng/mL, p<0.0001). Elevated BPA concentrations positively correlated with increased plasma bicarbonate concentrations and associated with fecal microbiome alterations. Short-term feeding of canned dog food increased circulating BPA concentrations in dogs comparable to amounts detected in humans, and greater BPA concentrations were associated with serum chemistry and microbiome changes. Dogs, who share our internal and external environments with us, are likely excellent indicators of potential human health concerns to BPA and other environmental chemicals. These findings may also have relevance to aquatic and terrestrial wildlife., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Little is known regarding optimal vitamin D status in adult dogs. To date no studies on vitamin D supplementation for improving vitamin D status have been reported for adult dogs. The aims of this study were to identify dogs with low vitamin D status and evaluate an oral dosage of cholecalciferol (D3) for effectiveness in increasing vitamin D status. For this, forty-six privately owned dogs were evaluated. Of the dogs, thirty-three (or 71·7 %) had serum 25-hydroxyvitamin D (25(OH)D) concentrations less than 100 ng/ml, a minimum previously suggested for vitamin D sufficiency in dogs. Subsequently, thirteen dogs were enrolled in a supplementation trial. Dogs were given either a D3 supplement (n 7; 2·3 µg/kg(0·75)) or olive oil placebo (n 6) daily with food. Serum concentrations of 25(OH)D were determined at weeks 1, 3 and 6, and at the trial end. Only at the trial end (weeks 9-10) was 25(OH)D significantly greater (P = 0·05) in supplemented v. placebo dogs. Serum concentrations of 24R,25-dihydroxycholecalciferol determined at the trial end were about 40 % of that of 25(OH)D3 and not significantly different between the groups. Concentrations of parathyroid hormone, ionised Ca, P and creatinine measured in initial and final serum samples indicated supplementation caused no toxicity. We conclude that vitamin D3 supplementation at a dosage near the National Research Council recommended safe-upper limit was not effective for rapidly raising serum 25(OH)D concentrations in healthy, adult dogs. Further work is needed in evaluating the metabolism of orally administered D3 in dogs before dosing recommendations can be made.

Objective: To describe the use of postoperative intrajejunal feeding and to evaluate the association of preoperative plasma albumin concentrations with intrajejunal feeding-related complications and clinical outcome., Design: Prospective, observational study., Setting: University veterinary teaching hospital., Animals: Sixty-four dogs., Interventions: Jejunostomy tube placement during abdominal surgery., Measurements and Main Results: Most dogs (81%) survived. The median intrajejunal feeding period was 2.1 days (range: 1-16 days; n = 64). Only 3 (5%) dogs received their estimated resting energy requirement by intrajejunal feeding. Of dogs that were fed intrajejunally (58 out of 64), most (55 out of 58) received intrajejunal feeding within 24 hours after surgery. Energy provision via the jejunal feeding tube did not differ between dogs with and without complications (P = 0.592), or between nonsurvivors and survivors (P = 0.298). Thirty-five dogs ate voluntarily concurrently with intrajejunal feeding. Of dogs that ate voluntarily concurrently with intrajejunal feeding for ≤50% of the postoperative period, most (74%) survived to discharge. Complications were seen in 22% of dogs, and none were life-threatening; gastrointestinal signs were most common. There was no difference in preoperative plasma albumin concentration between dogs with and without complications (P = 0.432) and between nonsurvivors and survivors (P = 0.727). Fecal score was not significantly different between the 2 liquid diets studied (FormulaV Enteral Care HLP and CliniCare Canine/Feline; P = 0.927)., Conclusions: A jejunostomy tube placed during abdominal surgery was likely to be used at the study institution. Few complications were seen and none were life-threatening. Intrajejunal feeding was initiated early after surgery and did not interfere with the initiation of voluntary oral intake. Fecal scores were high and were useful for an objective assessment of fecal consistency in dogs with intrajejunal feeding., (© Veterinary Emergency and Critical Care Society 2016.)

Background: Doxorubicin is a common antineoplastic agent with dose-dependent cardiotoxic adverse effects, and pre-existing myocardial dysfunction is a contraindication to its use., Objectives: To systematically define the hemodynamic and biochemical alterations in dogs undergoing chemotherapy for newly diagnosed lymphoma and assess the reversibility of these alterations with fluid administration., Animals: Twenty-one client-owned dogs with newly diagnosed lymphoma were evaluated 1 week after induction of chemotherapy. Underlying degenerative valve disease was exclusionary. Eighteen healthy age- and weight-matched dogs were used as controls., Methods: Physical examination, blood pressure by Doppler, echocardiography, and biochemical evaluation (routine serum biochemistry, plasma renin activity and aldosterone concentrations, plasma and urine osmolalities, and urine electrolyte concentrations) were measured in dogs with lymphoma and compared to controls. Dogs with lymphoma received crystalloids IV at 6 mL/kg/h for 24 hours. All variables were reassessed at 4 and 24 hours. Deuterium oxide dilution and bromide dilution were used to determine total body water and extracellular water space, respectively., Results: Baseline echocardiograms showed significantly smaller chamber dimensions in dogs with lymphoma compared to controls. These changes were reversed by fluid administration. Systolic blood pressure and urine sodium concentration were significantly increased, and bromide dilution space, PCV, urine specific gravity, and urine potassium concentration were significantly decreased compared to controls., Conclusion and Clinical Importance: Echocardiographic and biochemical abnormalities in dogs with lymphoma appear consistent with volume depletion, and may be the result of systemic hypertension and subsequent pressure natriuresis., (Copyright © 2014 by the American College of Veterinary Internal Medicine.)

Objective: To compare presumed fatty acid content in natural diets of feral domestic cats (inferred from body fat polyunsatrated fatty acids content) with polyunsaturated fatty acid content of commercial feline extruded diets., Sample: Subcutaneous and intra-abdominal adipose tissue samples (approx 1 g) from previously frozen cadavers of 7 adult feral domestic cats trapped in habitats remote from human activity and triplicate samples (200 g each) of 7 commercial extruded diets representing 68% of market share obtained from retail stores., Procedures: Lipid, triacylglycerol, and phospholipid fractions in adipose tissue samples and ether extracts of diet samples were determined by gas chromatography of methyl esters. Triacylglycerol and phospholipid fractions in the adipose tissue were isolated by thin-layer chromatography. Diet samples were also analyzed for proximate contents., Results: For the adipose tissue samples, with few exceptions, fatty acids fractions varied only moderately with lipid fraction and site from which tissue samples were obtained. Linoleic, α-linolenic, arachidonic, eicosapentaenoic, and docosahexaenoic acid fractions were 15.0% to 28.2%, 4.5% to 18.7%, 0.9% to 5.0%, < 0.1% to 0.2%, and 0.6% to 1.7%, respectively. As inferred from the adipose findings, dietary fractions of docosahexaenoic and α-linolenic acid were significantly greater than those in the commercial feline diets, but those for linoleic and eicosapentaenoic acids were not significantly different., Conclusions and Clinical Relevance: The fatty acid content of commercial extruded feline diets differed from the inferred content of natural feral cat diets, in which dietary n-3 and possibly n-6 polyunsaturated fatty acids were more abundant. The impact of this difference on the health of pet cats is not known.

High dietary carbohydrate is suggested to promote development of diabetes mellitus in cats. Glucose tolerance, insulin sensitivity, and insulin secretion were assessed in young [0.8-2.3 (median = 1.1) years, n = 13] and mature [4.0-7.0 (median 5.8) years, n = 12] sexually intact females of a large (n ≅ 700) feline colony in which only dry-type diets (35% metabolizable energy as carbohydrate) were fed from weaning. Insulin sensitivity was assessed from the 'late-phase' (60-120 min) plasma insulin response of intravenous glucose tolerance tests (IVGTTs) and from fractional change in glycaemia from baseline 15 min after an insulin bolus (0.1 U/kg, i.v.). Insulin secretion was assessed from the 'early-phase' (0-15 min) plasma insulin response of IVGTTs. Compared to the young cats, the mature cats had greater body weights [2.3-3.8 (median = 2.9) vs. 3.0-6.3 (median = 4.0) kg, p < 0.01], greater late-phase insulin responses (p < 0.05), lower insulin-induced glycaemic changes (p = 0.06), lower early-phase insulin responses (p < 0.05), and non-significantly different rates of glucose disposal. The late-phase insulin response was correlated with body weight and age (p < 0.05). When group assignments were balanced for body weight, the age-group differences and correlations became non-significant. The findings indicate that body weight gain is more likely than dry-type diets to induce the pre-diabetic conditions of insulin resistance and secretion dysfunction., (© 2010 The Authors. Journal of Animal Physiology and Animal Nutrition © 2010 Blackwell Verlag GmbH.)

Objective: To determine whether lipid particle coalescence develops in veterinary parenteral nutrition (PN) admixture preparations that are kept at room temperature (23 degrees C) for > 48 hours and whether that coalescence is prevented by admixture filtration, refrigeration, or agitation., Sample Population: 15 bags of veterinary PN solutions., Procedures: Bags of a PN admixture preparation containing a lipid emulsion were suspended and maintained under different experimental conditions (3 bags/group) for 96 hours while admixtures were dispensed to simulate IV fluid administration (rate, 16 mL/h). Bags were kept static at 4 degrees C (refrigeration); kept at 23 degrees C (room temperature) and continuously agitated; kept at room temperature and agitated for 5 minutes every 4 hours; kept static at room temperature and filtered during delivery; or kept static at room temperature (control conditions). Admixture samples were collected at 0, 24, 48, 72, and 96 hours and examined via transmission electron microscopy to determine lipid particle diameters. At 96 hours, 2 samples were collected at a location distal to the filter from each bag in that group for bacterial culture., Results: Distribution of lipid particle size in the control preparations and experimentally treated preparations did not differ significantly. A visible oil layer developed in continuously agitated preparations by 72 hours. Bacterial cultures of filtered samples yielded no growth., Conclusions and Clinical Relevance: Data indicated that the veterinary PN admixtures kept static at 23 degrees C are suitable for use for at least 48 hours. Manipulations of PN admixtures appear unnecessary to prolong lipid particle stability, and continuous agitation may hasten lipid breakdown.

The prevalence of obesity in domestic cats is increasing worldwide, and is strongly associated with gonadectomy. We have previously demonstrated the effectiveness of oestradiol in reducing food intake in both male and female neutered cats. This experiment was designed to test the hypothesis that oestradiol or genistein would prevent the increase in food intake following gonadectomy of male and female cats, and would prevent an increase in body fat mass. Three groups of eight cats each were surgically neutered then treated daily with either 0.5 mug oestradiol subcutaneously, 100 mg/kg genistein orally, or vehicle only. Effect of treatment on food intake, vaginal cytology and body weight were recorded, and body composition was assayed using the D(2)O isotopic dilution method. Neutering was followed by an increase in food intake, bodyweight and body fat mass in the control group, which were almost completely prevented by treatment with oestradiol (p < 0.001). Treatment with genistein had no effect on food intake or bodyweight increase, but was associated with a significant increase in lean body mass (p = 0.018), and significantly less body fat accumulation than the control group (p = 0.01). There were no significant differences in responses to treatment between sexes. These findings demonstrate the importance of gonadal oestrogen for the control of food intake in male and female cats, and suggest the provision of an oestrogenic compound could help prevent obesity following neutering. In addition, the findings of this study are consistent with observations in rodents of the efficacy of genistein in inhibiting adipogenesis and promoting lean body tissue development.

A high concentration of dietary carbohydrate is suggested to increase the risk of obesity and diabetes mellitus in domestic cats. To evaluate this, food intake, body weight, fat mass and circulating adiposity-related factors were determined in twenty-four sexually mature (9-12 months) cats assigned to four six-cat dietary groups balanced for body weight and sex. The effect of dietary fat in exchange for carbohydrate at 9, 25, 44 and 64 % of metabolisable energy (ME) in a purified diet of constant protein:ME ratio was studied 13 weeks before and 17 weeks after gonadectomy (GX). Body weight did not significantly change among the cats before GX except for an increase of 17 (sem 5) % in cats given the highest-fat diet. Following GX, all groups gained body weight, and body fat mass was positively correlated (r 0.50; P < 0.04) with dietary fat percentage. Post-GX weight gains were much greater for females (+39 (sem 5) %) than males (+10 (sem 4) %). Plasma ghrelin concentration negatively correlated (P < 0.02) with dietary fat percentage and, before GX, was greater (P < 0.05) in females than males. Plasma insulin concentration increased with weight gain induced by high dietary fat. Plasma glucose, TAG and leptin concentrations were not affected by dietary fat percentage, GX or weight gain. These data provide evidence that in cats, high dietary fat, but not carbohydrate, induces weight gain and a congruent increase in insulin, while GX increases sensitivity to weight gain induced by dietary fat.

The absorption and disposition kinetics of the soy isoflavone genistein were determined in cats (n = 6). An oral dose of 100 mg/kg was administered, which has previously been demonstrated to be the minimum oral estrogenic dose, and was administered intravenously at a dose of 20 mg/kg, being the largest practical dose that could be safely administered. Plasma free, and total (conjugated + free) genistein concentrations were determined by HPLC following organic extraction. Noncompartmental analysis revealed a half-life of 21.67 +/- 7.9 h (free) and 9.95 +/- 2.7 h (conjugated), volume of distribution 31.94 +/- 10.38 L/kg (free) and 11.82 +/- 3.96 L/kg (conjugated) following intravenous administration. Following oral administration the half-lives were determined to be 17 +/- 4.8 h (free) and 8.56 +/- 4.65 h (conjugated), with tmax = 4.4 +/- 0.6 h (free) and 4.42 +/- 0.99 h (conjugated), and Cmax = 0.276 +/- 0.1 microg/mL (free) and 6.24 +/- 6.58 microg/mL (conjugated). Oral bioavailabilities were 1.379 +/- 0.9% (free) and 29.85 +/- 22.61% (conjugated). The ratio of total:free genistein ranged from 25.9 to 5.5. Poor oral absorption and efficient conjugation explain the low bioavailability of free genistein. Accumulation of genistein in peripheral lipophilic compartments may occur.

Dietary genistein has potential as a surrogate estrogen to restore normal control of food intake in cats following gonadectomy. However, since genistein affects immunological responses in other species it is important to determine if there is any immunological effect of genistein in cats, before long-term feeding of it could be considered. It was hypothesized that the minimum estrogenic dose of genistein (peak serum concentration 0.276+/-0.055 microg/mL) would be associated with altered innate and acquired immunity in cats. Cats (n=8 per group) were surgically neutered then treated daily with either 0.5 microg estradiol subcutaneously, 100mg/kg genistein orally, or vehicle only for 38 days. Circulating CD4+ and CD8+ T cells, and CD21+ B cells were quantified using flow cytometry. Concanavalin-A induced lymphocyte proliferation was assayed using incorporation of tritiated thymidine. Indirect ELISA was used to assay serum IgG in response to a commercial vaccine. Neutrophil respiratory burst in response to opsonized zymosan was evaluated using flow cytometric detection of oxidized dihydrorhodamine. Delayed-type hypersensitivity was evaluated with formalin-killed Yersinia pseudotuberculosis. Genistein treatment decreased circulating CD8+ cells (P=0.006), increased neutrophil respiratory burst (P=0.034), and increased wheal formation at 24h (P=0.038) but decreased wheal formation at 72 h (P=0.012) following intradermal challenge with killed Y. pseudotuberculosis. There were no significant differences between genistein treated cats and the other treatment groups in any other parameters. It is concluded that the minimum estrogenic dose of genistein alters selected leukocyte responses in cats. Unexpected effects of genistein suggest that extrapolation from one species to other species may not be appropriate in regards to the effects of genistein on immunity.

Diet-induced (taurine deficiency) dilated cardiomyopathy is reported more in large than small dogs possibly because taurine biosynthesis rate (TBR) is lower in large than small dogs. The TBR in 6 mongrels (37.9 +/- 2.1 kg) and 6 beagles (12.8 +/- 0.4 kg) was determined from the fractional dilution rate of urinary [1,2-(2)H(2)]-taurine, (d4-tau). All dogs were given a 15.6% protein, 0.60% sulfur amino acid (SAA) diet in amounts to maintain an ideal body condition score. After 3 mo, 14.6 mg/kg body weight of d4-tau was given orally and TBR determined from d4-tau to taurine ratio in urine collected each d for 6 d. Enrichments of d4-tau were determined by GC-MS. Thereafter, mongrels and beagles were paired by ranking of SAA intake per metabolic body weight per kg(0.75). Each pair received the same amount of diet/kg(0.75) for 2 wk, then TBR was again determined. Concentrations of taurine in plasma, blood, and urine and concentrations of plasma thiols were measured during each TBR determination. In Expt. 1, TBR and taurine concentrations in plasma and urine of mongrels were lower (P < 0.05) than those of beagles. In Expt. 2, TBR and taurine concentrations in blood and plasma of mongrels were lower (P < 0.05) than beagles. Together, the results support the hypothesis that large compared with small dogs have lower TBR when fed diets near-limiting in dietary SAA, but adequate to maintain ideal body condition.

Normal adiposity occurs in humans and mice deficient of adipose lipoprotein lipase (LPL) activity. Subnormal adiposity found in LPL-deficient cats is indicative of limited de novo synthesis of fatty acids (FAs). In 14 LPL-deficient (3.0 +/- 0.1 kg) and 8 normal (3.7 +/- 0.1 kg) queens, FAs in triacylglycerol (TAG), phospholipid (PL), and nonesterified FAs (NEFAs) of plasma and inguinal subcutaneous adipose were determined before and after (d 38, 61, 110, 117, and 251) dietary linseed oil supplementation (30 g/kg). By d 60, LPL-deficient queens gained body weight (+0.4 +/- 0.1 kg), developed normal body fat mass (25 +/- 2%), and were enriched in 18:3(n-3) in their plasma and adipose lipids. Adipose TAG 18:3(n-3) enrichment in LPL-deficient queens was subnormal at all sampling times and, as observed in normal queens, apparently not equilibrated by d 251. Adipose FA profiles in TAG but not PL were substantially different (P < 0.05) between LPL-deficient and normal queens; the 16:0 to 18:2(n-6) ratio was high in LPL-deficient (2.4-4.4) relative to normal queens (1.0-1.4). In LPL-deficient queens, fed-state plasma NEFA (n-6) and (n-3) enrichments were similar to those in adipose TAG, and plasma NEFA concentration was high (0.62 +/- 0.05 mmol/L) and similar to that in normal queens after withholding diet for 16 h. These data indicate that LPL deficiency in cats reduces dietary FA storage efficiency, favors storage of saturated over unsaturated FAs, and stimulates de novo FA synthesis substantive enough to support normal adiposity.

Although taurine is not dietarily essential for dogs, taurine deficiency and dilated cardiomyopathy (DCM) are sporadically reported in large-breed dogs. Taurine status and husbandry were examined in 216 privately owned Newfoundlands, a giant dog breed with high incidence of idiopathic DCM (1.3-2.5%). Plasma taurine concentration was positively correlated (P < 0.01) with plasma cyst(e)ine (r = 0.37) and methionine (r = 0.35) concentrations and was similar across age, sex, neutering status, body weight, and body-condition scores. Plasma taurine concentration was low (< or =40 micromol/L) in 8% of dogs. Dogs with low plasma taurine were older, less active, had more medical problems and treatments, and had lower plasma albumin, cyst(e)ine, tryptophan, and alpha-amino-n-butyric acid concentrations than the other dogs (P < 0.05). Of 9 taurine-deficient, clinically evaluated dogs, 3 had DCM that was reversed by taurine supplementation and 1 had retinal degeneration. When given a diet apparently adequate in sulfur amino acids (5.4 g/kg) for 3 wk, 6 Newfoundlands (52.5 +/- 2.3 kg, 3.5-7 y), compared with 6 Beagles (13.2 +/- 2.3 kg, 5.5 y), had lower (P < 0.01) concentrations of plasma taurine (49 +/- 16 vs. 97 +/- 25 micromol/L) and cyst(e)ine and blood glutathione, lower (P < 0.01) de novo taurine synthesis (59 +/- 15 vs. 124 +/- 27 mg x kg(-0.75) x d(-1)), and greater (P < 0.05) fecal bile acid excretion (1.7 +/- 0.2 vs. 1.4 +/- 0.2 micromol/g). Newfoundlands would appear to have a higher dietary sulfur amino acid requirement than Beagles, a model breed used in nutrient requirement determinations.

Evidence suggests that amino terminal extensions of CCK-8 affect the carboxyl terminal bioactive region of CCK. Cat CCK-58 was purified by low pressure reverse phase and ion-exchange chromatography steps and several reverse phase HPLC steps. The purified peptide and its tryptic fragments were characterized by mass spectral analysis and microsequence analysis. The structure of cat CCK-58 is: AVQKVDGEPRAHLGALLARYIQQARKAPSGRMSVIKNLQSLDPSHRISDRDY(SO3) MGWMDF-amide. Cat and dog CCK-58 are identical except for position 40 which is serine in cat and asparagine in dog. Radioimmunoassay detected cat CCK-58 about 1/10th as well as dog CCK-58, indicating a marked effect on C-terminal immunoreactivity. Cat CCK-58 with a serine at position 40, the same residue found in pig, mouse, cow and rabbit CCK-58, can be used as a unique bioprobe for defining how amino terminal amino acids influence the structure and bioactivity of the carboxyl terminal region of CCK.

An overweight or obese body condition commonly develops after gonadectomy (GX) in domestic cats. The cause appears to be a rapid, quantal (approximately 12%), increase in food intake that is sustained and probably mediated by withdrawal of gonadal hormone. Recently, an interaction of gonadal hormone and cholecystokinin (CCK) effectiveness has been suggested. A reduction in the satiating potency of intestinal CCK was presently hypothesized to contribute to the disturbance of food intake control caused by GX in domestic cats. Pre- and post-prandial intestinal CCK secretion as indicated by plasma CCK concentrations were determined in 16 adult male cats (5.1 +/- 0.1 kg) 8 weeks before and 57 weeks after eight of the cats were gonadectomized. During ad libitum intake of a commercial dry, expanded diet, body weight increased from 22% to 28% in gonadectomized cats and was unchanged in intact cats. Baseline CCK concentrations were not different between gonadectomized and intact cats. Amounts of diet ingested during CCK determinations were 15-19% of daily metabolizable energy requirement and were not different between gonadectomized and intact cats. The post-prandial area under the curve (AUC; 0-400 min) CCK concentration increased linearly with meal size (p < 0.01) and was not correlated with body weight. Area under the curve CCK concentration, when normalized for meal size, was 34% greater (p < 0.01) in gonadectomized cats than that in intact cats. The findings indicate GX increases meal-induced intestinal CCK secretion and therefore, do not support the study hypothesis. The findings indicate GX may slow digestion and absorption and attenuate inhibition of food intake by CCK.

Objective: To determine taurine status in a large group of Newfoundlands related by environment, diet, or breeding to a dog with dilated cardiomyopathy and taurine deficiency., Design: Prospective study., Animals: 19 privately owned Newfoundlands between 5 months and 11.5 years old that had been fed commercial dry diets meeting established nutrient recommendations., Procedure: Diet histories were obtained, and blood, plasma, and urine taurine concentrations and plasma methionine and cysteine concentrations were measured. In 8 dogs, taurine concentrations were measured before and after supplementation with methionine for 30 days. Ophthalmic examinations were performed in 16 dogs; echocardiography was performed in 6 dogs that were taurine deficient., Results: Plasma taurine concentrations ranged from 3 to 228 nmol/mL. Twelve dogs had concentrations < 40 nmol/mL and were considered taurine deficient. For dogs with plasma concentrations < 40 nmol/mL, there was a significant linear correlation between plasma and blood taurine concentrations. For dogs with plasma concentrations > 40 nmol/mL, blood taurine concentrations did not vary substantially. Taurine-deficient dogs had been fed lamb meal and rice diets. Retinal degeneration, dilated cardiomyopathy, and cystinuria were not found in any dog examined for these conditions. The taurine deficiency was reversed by a change in diet or methionine supplementation., Conclusions and Clinical Relevance: Results indicate a high prevalence of taurine deficiency among an environmentally and genetically related cohort of Newfoundlands fed apparently complete and balanced diets. Blood taurine concentrations indicative of taurine deficiency in Newfoundlands may be substantially less than concentrations indicative of a deficiency in cats.

Objective: To determine signalment, history, clinical signs, blood and plasma taurine concentrations, electrocardiographic and echocardiographic findings, treatment, and outcome of dogs with low blood or plasma taurine concentrations and dilated cardiomyopathy (DCM)., Design: Retrospective study., Animals: 12 client-owned dogs with low blood or plasma taurine concentrations and DCM., Procedure: Medical records were reviewed, and clinical data were obtained., Results: All 12 dogs were being fed a commercial dry diet containing lamb meal, rice, or both as primary ingredients. Cardiac function and plasma taurine concentration improved with treatment and taurine supplementation. Seven of the 12 dogs that were still alive at the time of the study were receiving no cardiac medications except taurine., Conclusions and Clinical Relevance: Results suggest that consumption of certain commercial diets may be associated with low blood or plasma taurine concentrations and DCM in dogs. Taurine supplementation may result in prolonged survival times in these dogs, which is not typical for dogs with DCM. Samples should be submitted for measurement of blood and plasma taurine concentrations in dogs with DCM, and taurine supplementation is recommended while results of these analyses are pending.

Taurine (Tau) deficiencies have been associated with the feeding of commercial lamb-meal and rice diets to dogs. We hypothesized that the poor digestibility of some lamb-meals may limit sulphur amino acids availability for Tau synthesis and/or increase of Tau degradation in the gut. Growing dogs were fed either a lamb-meal-based (Diet A) or poultry by-product-based (Diet B) commercial diet. Plasma, whole blood and urinary Tau were measured for 22 weeks. Plasma and whole blood Tau concentrations were similar between the groups throughout the study. Urinary excretion of Tau in dogs fed diet A was 3.2 times greater than that from dogs fed Diet B, suggesting greater renal reabsorption and the need for conservation of Tau in the Diet A group. Food restriction affected Tau status as indicted by a positive correlation of food intake and urinary Tau. Dogs fed Diet A were given antibiotics to inhibit bacterial activity in the gut. Increases in breath hydrogen, indicative of increased bacterial activity, correlated negatively with urinary Tau. Urinary Tau increased by 54% when methionine (Met) was supplemented to Diet A, supporting the suggestion of a low bioavailability of sulphur amino acids and/or an increased fecal loss of Tau in dogs consuming Diet A.

The purpose of this study was to determine the selenium (Se) requirement in kittens. Thirty-six specific-pathogen-free kittens (9.8 weeks old) were utilized in a randomized complete block design to determine the Se requirement in cats with gender and weight used as blocking criteria. Kittens were fed a low Se (0.02 mg/kg Se) torula yeast-based diet for 5 weeks (pre-test) after which an amino acid-based diet (0.027 mg Se/kg diet) was fed for 8 weeks (experimental period). Six levels of Se (0, 0.05, 0.075, 0.10, 0.20 and 0.30 mg Se/kg diet) as Na2SeO3 were added to the diet and were used to construct a response curve. Response variables included Se concentrations and Se-dependent glutathione peroxidase activities (GSHpx) in plasma and red blood cells (RBC) as well as plasma total T3 (TT3) and total T4 (TT4). No significant changes in food intake, weight gain or clinical signs of Se deficiency were noted. Estimates of the kitten's Se requirement (i.e. breakpoints) were determined for RBC and plasma GSHpx (0.12 and 0.15 mg Se/kg diet, respectively), but no definitive breakpoint was determined for plasma Se. Plasma TT3 increased linearly, whereas plasma TT4 and the ratio of TT4 : TT3 decreased in a quadratic fashion to dietary Se concentration. The requirement estimate determined in this study (0.15 mg Se/kg) for kittens is in close agreement with other species. As pet foods for cats contain a high proportion of animal protein with a Se bioavailability of 30%, it is recommended that commercial diets for cats contain 0.5 mg Se/kg DM.

Gonadectomy predisposes domestic cats to undesired body weight gain and obesity. The disturbance responsible for this disregulation of energy balance has not been clearly identified. Energy intake and expenditure, body composition and plasma concentrations of leptin, insulin, glucose and triacylglycerol were determined during a 36-wk period in adult male (2-5 y) gonadectomized (n = 8) and intact (n = 8) normal cats and gonadectomized (n = 8) and intact (n = 8) lipoprotein lipase (LPL)-deficient cats. Cats were housed individually in temperature- and light-controlled rooms and continuously provided a commercial dry-type diet. In normal and LPL-deficient cats, body weight increased (P < 0.05) after gonadectomy by 27 to 29%, mostly as a result of fat accretion. There was a rapid increase (P < 0.05) in food intake of approximately 12% after gonadectomy of normal and LPL-deficient cats. The metabolic rate (kJ.kg(-1).d(-1)), determined in normal intact (319 +/- 20, n = 5) and gonadectomized (332 +/- 36, n = 5) cats, did not differ after gonadectomy. After gonadectomy, plasma concentrations of glucose and triacylglycerol did not change, whereas plasma insulin and leptin concentrations increased (P < 0.05), but not coincidentally with body weight gain. A stair-step increase in energy intake, and not decreased energy expenditure, appears to drive the weight gain associated with gonadectomy. Body fat mass appears to increase until the energy intake supports no further expansion. Adiposity signaling through insulin or leptin does not appear to mediate the energy intake effect. LPL deficiency did not preclude development of the overweight body condition. Therefore, gonadectomy-induced weight gain in cats is not a result of changed adipose LPL activity, as previously suggested.

Breath hydrogen (H(2)) concentration, an indicator of intestinal microbial abundance, was determined in cats given purified and commercial canned and dry-type diets. Before measurements, the cats were fed diets for more than 2 wk and habituated to a daily feeding interval of 4 hr. Breath H(2) concentrations were determined before a meal (approximately 25% daily MER) and then every 20 min for 8 hr or hourly for 10 hr. A clear rise above baseline breath H(2) concentrations, 1-2 ppm, was not observed in 6 males given a casein-based purified diet. A mean (+/- SEM) peak breath H(2) concentration of 22 +/- 4 ppm was observed in 6 other males, 6.3 hr after ingestion of a canned diet with protein, fat, and carbohydrate proportions similar to those of the purified diet. Area-under-the-curve (AUC) breath H(2) responses to the canned diet were substantially greater (p < 0.05) than responses observed in 5 males given a dry-type diet, but similar to responses observed in 12 males given an uncooked form of the canned diet. Gamma irradiation to inactivate microbes in the uncooked diet did not affect the breath H(2) response. Breath H(2) responses to 2 other canned and 2 other dry-type diets were evaluated in 8 adult females using a 4 x 4 Latin-square design. Peak and AUC responses to the canned diets were similar but approximately 2 times greater (p < 0.05) than responses to the dry diets. Relative to dry-type diets, canned diets induce a substantially greater breath H(2) production, and therefore appear to support a greater intestinal microbial population.

Adult domestic cats homozygous with a naturally occurring Gly412Arg LPL gene mutation are good models for the study of LPL deficiency. Previous studies report that homozygous LPL deficient kittens have reduced growth rates and develop subnormal body fat mass. It was hypothesized in the present study that homozygote kittens would have normal growth if provided a standard low fat, highly digestible diet at weaning and that their body fat would be increased by provision of a diet high in protein. When fed a nutritionally complete, 10% fat, purified or commercial extruded diet, the body weights of homozygous (n = 24), heterozygous (n = 25) and normal (n = 16) kittens were determined at birth, 2, 3, 4, 6, 12 and 18 weeks of age. Male homozygote kittens from homozygote dams had reduced weight gains (p < 0.05) compared to normal males at 2, 3 and 4 weeks. Female heterozygotes and homozygotes from homozygote and heterozygote dams had reduced weight gains (p < 0.05) compared to normal females at 2, 3, 4 and 6 weeks. By 6 weeks for males and 18 weeks for females, genotype related differences in weight gain were not observed. At 30 weeks, homozygotes and heterozygotes were given either a 60 or 30% (dry matter) protein diet for two months. As indicated by deuterium dilution estimation of body composition, cats eating the 30% protein diet (n = 12) tended to have a lower increase in lean body mass (p = 0.057) and a greater increase in fat mass (p = 0.092) compared to cats eating the 60% protein diet (n = 12). Increase in lean body mass among homozygotes tended to be not as great as that observed in heterozygotes (p = 0.057). Poor postweaning gains previously reported in homozygotes probably reflected inappropriate selection of diet for this genotype. The high protein diet increased the rate of lean body mass development but not body fat mass.

The objective of the present study was to examine the interaction between graded levels of leucine and dietary crude protein. Dose-response curves were generated using four 3 x 3 Latin squares (two dogs/square). Each square represented one of two concentrations of crude protein (140 or 280 g/kg diet) and one of two combinations of three concentrations of leucine (5.0, 7.0 and 9.0 g/kg diet or 9.0, 11 and 13 g/kg diet). An additional experiment was performed by feeding crude protein at 210 g/kg diet with either 7.0 or 11 g leucine/kg diet. Weight gain, food intake, nitrogen retention, plasma albumin and plasma amino acids were measured. The requirement was determined to be the minimum leucine concentration required to maximize weight gain and nitrogen retention. For 8-14-week-old male Beagle dogs, 140 g crude protein/kg diet in a diet containing 18 kJ metabolizable energy/g does not appear to support maximal growth. The leucine requirement was not affected by doubling the dietary crude protein level from 140 to 280 g/kg diet. From these results, the leucine requirement of 8-14-week-old Beagle dogs appears to be 11 g leucine/kg diet independent of the level of dietary crude protein, whereas dogs over 14 weeks require only 7 g leucine/kg diet for maximal nitrogen retention.