Isolation and identification of exosomes from feline plasma, urine and adipose-derived mesenchymal stem cells.

BACKGROUND: Exosomes, internal proteins, lipids, and nucleic acids coated by phospholipid bilayer membranes, are one type of small extracellular vesicles, which can mediate cell-cell communication. In recent years, exosomes have gained considerable scientific interest due to their widely applied prospect in the diagnosis and therapeutics of human and animal diseases. In this study, we describe for the first time a feasible method designed to isolate and characterize exosomes from feline plasma, urine and adipose-derived mesenchymal stem cells. RESULTS: Exosomes from feline plasma, urine and adipose-derived mesenchymal stem cells were successfully isolated by differential centrifugation. Quantification and sizing of exosomes were assessed by transmission electron microscopy, flow nano analysis and western blotting. Detected particles showed the normal size (30-100 nm) and morphology described for exosomes, as well as presence of the transmembrane protein (TSG101, CD9, CD63, and CD81) known as exosomal marker. CONCLUSIONS: The results suggest that differential centrifugation is a feasible method for isolation of exosomes from different types of feline samples. Moreover, these exosomes can be used to further diagnosis and therapeutics in veterinary pre-clinical and clinical studies.

Comparative pharmacokinetic profile of cimicoxib in dogs and cats after IV administration.

Cimicoxib is a selective COX-2 inhibitor (coxib) registered for the treatment of pain and inflammation in dogs. Pharmacokinetics of some coxibs have been described in dogs and cats. In cats, total body clearance values are lower and terminal half-lives of the coxibs are longer than those in dogs. The aim of this work was to evaluate if this is also the case for cimicoxib. For this purpose, blood pharmacokinetics and urinary excretion after IV administration were compared between these species. The in vitro metabolism of cimicoxib was also evaluated using canine and feline microsomes. In canine and feline microsomes, the formation rate of demethyl-cimicoxib, a phase 1 metabolite, was decreased in presence of quinidine, a specific human cytochrome P450 (CYP)2D6 inhibitor. IC50 values were 1.6 µM and 0.056 µM with canine and feline microsomes, respectively. As quinidine was about 30 times more potent in feline microsomes, the affinity of cimicoxib to the enzyme was considered to be about 30 times lower than that in canine microsomes. Total body clearance (ClB) of cimicoxib, was 0.50 L/h kg in dogs and 0.14 L/h kg in cats (P < 0.01) and terminal half-life, T½λz, was 1.92 and 5.25 h, respectively (P < 0.01). Dose eliminated in urine was 12.2% in dogs and 3.12% in cats (P < 0.01). Conjugated demethyl-cimicoxib represented 93.7% of this amount in dogs and 67.5% in cats. Thus cimicoxib, like other veterinary coxibs, was eliminated more slowly in cats. Both CYP2D15 (the canine ortholog of CYP2D6) and UDP-glucuronyltransferase enzyme systems have reduced ability to produce metabolites of cimicoxib in cats.

Avoidance behaviour in laboratory house mice (Musmusculus) and Norway rats (Rattus norvegicus) towards predator odours.

Mus musculus and Rattus sp. are considered pest species because they reach high densities in urban areas, crop fields and food storage and productive systems such as breeding farms and orchards. Their control relies mainly on rodenticide application, but the effectiveness of this application is reduced due to behavioural responses and resistance. Novel methods are based on the use of chemical signals as odours that may be attractants, repellents or may reduce the reproductive success of pest species. The aim of this paper is to study the aversive effect of TMT, cat urine and cat body odour on predator-inexperienced Mus musculus and Rattus norvegicus under laboratory conditions. The experimental apparatus comprised three boxes connected by PVC pipes in a linear arrangement. In lateral boxes, odour sources or distilled water were introduced, while animals were placed in the central box at the beginning of the experiment. Rats showed freezing behaviour, reduced visits in the presence of TMT and cat fur. Mice reduced their visits with cat body and cat urine. This study provides evidence of the usefulness of using fear responses as a way to control rodent pests, which must be adapted to the environment and species to be applied.

Desempenho de diferentes marcas de tiras reagentes relativo à densidade e à análise química da urina de cães e gatos / Performance of different brands of reagent strips for density and chemical analysis of dog and cat urine

Objetivou-se verificar a compatibilidade entre diferentes marcas de tiras reagentes para urinálise, tanto de uso veterinário, como de uso humano, e confrontar os parâmetros semiquantitativos desse instrumento com métodos quantitativos. Para isso, foram analisadas 77 amostras frescas de urina de cães e gatos e testados 04 modelos de tiras reagentes. Quanto à densidade urinária, houve correlação razoável entre os métodos quantitativo e semiquantitativo naquelas amostras com pH ácido, mas não naquelas com pH neutro ou alcalino. Quanto à concentração proteica, houve similaridade de 53,3% a 83,3% entre as marcas testadas e quando comparadas com a análise fotométrica houve uma correlação razoável (rs = 0,69752 a 0,75074). Em ponto de corte de 15mg/dL de proteína, a sensibilidade da tira reagente foi 82,5% e 100% para urina canina e felina, respectivamente. No tocante à hematúria, houve divergência razoável entre a sedimentoscopia e as diferentes marcas de tiras reativas. Quanto à piúria, há uma baixa sensibilidade das tiras em relação às amostras caninas com muitos resultados falso-negativos (33% a 75%), enquanto em amostras felinas a sensibilidade foi de 100%. Assim, independente da marca, as tiras reagentes devem servir apenas como teste rápido de triagem, sendo mais apropriado o uso de métodos quantitativos na avaliação clínica do paciente a partir da urinálise.

The aim was to verify the compatibility between different brands of urinary dipsticks, for both human and veterinary use, and to compare the semiquantitative parameters of this instrument with quantitative methods. For this, 77 fresh samples of urine from dogs and cats were analyzed e and 04 models of reagent strips were tested. Regarding urinary density, a reasonable correlation was observed between the quantitative and semiquantitative methods in those samples with acidic pH, which did not occur in those with neutral or alkaline pH. Regarding the protein concentration, there was similarity from 53.3% to 83.3% between the brands and in the comparative analysis between the control strip and the photometric analysis, there was a reasonable correlation (rs = 0.69752 to 0.75074). In cut-off point of 15mg/dL protein, the sensitivity of the reagent strip was 82.5% and 100% for canine and feline urine, respectively. Regarding hematuria, there was a reasonable divergence of results between sedimentation and tested dipsticks. As for pyuria, there is a low sensitivity of the strips in relation to canine samples with many false negative results (33% to 75%), while in feline samples the sensitivity was 100%. Thus, regardless of the brands, the reagent strips should serve only as a rapid screening test, while the use of quantitative methods in the clinical evaluation of the patient from urinalysis is more appropriate.

A randomized cross-over trial assessing salivary and urinary cortisol concentrations after alfaxalone and propofol administration in healthy cats.

The aim of this study was to assess the effects of commonly used anaesthetics alfaxalone and propofol on salivary and urinary cortisol in healthy cats. Fifteen male castrated research-purposed cats received randomly intravenous continuous rate infusions of 8 mg/kg/h of alfaxalone, 12 mg/kg/h of propofol and 2 ml/kg/h of Lactated Ringer's solution for 30 min, with intervals of 6 days between treatments. Saliva samples were collected for 24 h before each infusion and for 24 h from the start of each infusion. Urine was collected as single pooled samples over each 24 h period. Mean integrated saliva cortisol responses in cats treated with alfaxalone were greater than responses of cats treated with propofol (P = 0.034) and controls (P = 0.017). Integrated responses in cats treated with propofol did not differ from controls. The mean urinary cortisol/creatinine ratio (UCCR) was higher on the day of treatment than the day before treatment in cats treated with alfaxalone (P < 0.0001) and in cats treated with propofol (P = 0.0168) and did not differ between days in cats treated with lactated Ringer's solution. The mean UCCR was higher in cats treated with alfaxalone than in cats treated with lactated Ringer's solution (P = 0.0020) on the day of treatment. Mean total urinary cortisol over 24 h was greater in cats treated with alfaxalone than controls (P = 0.0267). In conclusion, alfaxalone increased short-term salivary and urinary cortisol concentrations in healthy cats as compared to propofol and a control group of non-anesthetised cats.

The profile of urinary lipid metabolites in cats.

Polyunsaturated fatty acids including arachidonic acid (AA), docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), are converted to lipid mediators by oxidation. Unlike other mammals, cats cannot synthesize AA. Since their lipid metabolic features remain unknown, we qualitatively analyzed 118 types of urinary lipid metabolites in healthy neutered cats. Using LC-MS, we found 26 lipid metabolites in urines of all individuals. In detail, 20 AA-, 5 EPA- and 1 DHA-derived lipid mediators were detected. Focusing on oxidative pathway, 17 cyclooxygenase-metabolites and 5 metabolites produced by non-enzymatic pathway were detected. Of interest, few lipoxygenase- or cytochrome P450-metabolites were excreted. Thus, AA-derived cyclooxygenase-metabolites mainly composed the urinary lipid metabolites in cats.

Increasing dietary sodium chloride promotes urine dilution and decreases struvite and calcium oxalate relative supersaturation in healthy dogs and cats.

Urolithiasis is highly prevalent in dogs and cats, with struvite and calcium oxalate being most commonly diagnosed. Some commercial diets aimed at reducing the risk of urolithiasis are based on inclusion of sodium chloride (NaCl) in an attempt to dilute the urine and the risk of crystallization, but more information on the effect of differing levels of sodium inclusion is needed. The objective of this study was to compare the short-term effect of four diets differing only in NaCl content (base diet with 0.3% sodium and diets with added NaCl to achieve 0.7, 1.0 and 1.3% sodium as fed) on urinary ion concentrations and relative supersaturation (RSS) of struvite and calcium oxalate in dogs and cats. In both species, there was a significant increase in water intake and urine volume as dietary NaCl increased. Urine sodium concentration increased with increasing dietary NaCl. The highest sodium diet increased urinary calcium excretion in dogs only, while decreasing urinary calcium concentration. Calcium oxalate RSS and struvite RSS both significantly decreased, with the lowest RSS values reported on the highest sodium diet in both dogs and cats (p < .001). These results suggest that an increase in dietary NaCl decreases RSS values in both dogs and cats. Despite an increase in urinary calcium excretion in dogs, urinary calcium concentration and calcium oxalate RSS were lower on high sodium diets due to urine dilution. Long-term studies are needed to confirm the relationship between RSS and stone occurrence and recurrence.

Influence of preanalytic and analytic variables in canine and feline urine specific gravity measurement by refractometer.

Urine specific gravity (USG), which is usually measured by refractometry, is an important indicator of renal concentrating ability. Few studies have evaluated refractometers with separate scales for canine and feline urine. Variables such as protein content or storage time may influence the USG. We compared the effects of measuring USG with a refractometer with single or separate scales for canine and feline urine, investigated inter- and intra-observer variability, and measured agreement between whole urine and supernatant. We evaluated the correlation between USG and osmolality, the influence of urinary protein on USG and osmolality, and the impact of storage time up to 6 mo. We examined 252 canine and 126 feline samples. Bland-Altman analysis revealed higher USG values of the single-scale refractometer than the dual-scale refractometer, with a mean difference (bias) of < 0.001 for canine and 0.003 for feline specimens. Inter- and intra-observer variability were acceptable. Good agreement was shown between USG of whole urine and supernatant. Correlations between USG and osmolality were excellent (0.98-0.99, p < 0.001). Proteinuria up to 1 g/L had no major impact on USG or osmolality. Storage time had no significant effect on USG. The difference between the refractometers is clinically irrelevant, and the use of a refractometer with separate feline and canine scales is unnecessary. Whole urine and supernatant stored up to 6 mo can both be used for USG measurement. The influence of proteinuria <1 g/L on USG and osmolality is negligible.

Evaluation of the accuracy of urine analyzers in dogs and cats.

The accuracy of urine analyzers used for dogs and cats has remained uncertain. This study examines the agreement between results of urine analysis obtained using two devices marketed for animals and for humans and the results of quantitative biochemical analysis. The degrees of concordance for bilirubin and ketones in the same category were ~80%, but for pH these were only ~60% in dogs and cats. Degrees of concordance for protein and the UP/C ratio clearly differed between the devices for animals and humans. We found that values for bilirubin and ketones obtained using urine analyzers may be reliable, but pH is unlikely to be accurate enough to be clinically useful for dogs and cats.

Effects of different odours on the reproductive success of Mus musculus as an alternative method of control.

BACKGROUND: The house mouse (Mus musculus) is a cosmopolitan rodent that has become adapted to living in close association with humans and is considered a serious pest because it poses a risk to human health, and causes economic losses due to food and crop consumption and damage to buildings. Its control in livestock farms is achieved mainly through the application of anticoagulant rodenticides, but the effect of these compounds is limited due to the presence of resistant individuals and aversive behaviours. A potential alternative method is the use of chemical signals to reduce rodent reproductive success. In this study, we assessed the effects of odours from an unfamiliar male, 17ß-oestradiol, overcrowding, cat urine and 2,5-dihydro-2,4,5-trimethylthiazoline (TMT) on the reproductive success of laboratory Mus musculus females. RESULTS: According to the generalized linear mixed models, cat urine odour increased the proportion of abortions per female, unfamiliar male odour decreased the mean number of offspring born per female, and TMT had an overall negative effect on mean offspring production at birth and at weaning. The other odours had no significant effects on reproductive success. CONCLUSIONS: TMT seems to be the best candidate for population control because it caused a decrease in the mean number of offspring born and the mean number of live offspring at weaning. TMT also has the advantage of being available in commercial forms. To be useful for rodent management in field conditions, these results should be confirmed using wild house mice females. © 2019 Society of Chemical Industry.

Basal glucosuria in cats.

Objective of this study was to demonstrate the ubiquitous presence of glucose in urine of euglycemic cats by a highly sensitive glucose assay. The local electronic database was searched for results of quantitative urine glucose measurements in cats. A total of 325 feline urine glucose measurements were identified, of which 303 (93%) had been submitted by one of the co-authors working in a near-by small animal practice. After the exclusion of patients with kidney disease (n = 60), hyperthyroidism (n = 15), diabetes mellitus (n = 11), multiple diseases (n = 9) or steroid treatment (n = 3), as well as serial measurements (n = 87) and outliers (n = 8), the final study population consisted of 132 cats. Urine creatinine concentration was unavailable in five patients. Whereas all but one cat had glucose concentrations above the detection limit of the assay (0.11 mmol/L, Gluco-quant Enzyme Kit/Roche Diagnostics), no positive glucose dipstick test result (Combur 9-Test, Roche Diagnostics) was observed. The median (range) of urinary glucose concentration and the glucose-to-creatinine ratio (UGCR) was 0.389 (<0.11-1.665) mmol/L and 0.0258 (0.007-0.517) respectively. The UGCR was not affected by age, gender, breed or leukocyturia, whereas cats with hematuria had slightly higher values. Data show that so-called "basal glucosuria" is present in the majority of cats and by no means diagnostic for diabetes mellitus or renal glucosuria. This has to be considered when using bio-analytical methods with a low limit of quantification.

Comparison of the performance of the IDEXX SediVue Dx® with manual microscopy for the detection of cells and 2 crystal types in canine and feline urine.

BACKGROUND: Microscopic evaluation of urine is inconsistently performed in veterinary clinics. The IDEXX SediVue Dx® Urine Sediment Analyzer (SediVue) recently was introduced for automated analysis of canine and feline urine and may facilitate performance of urinalyses in practice. OBJECTIVE: Compare the performance of the SediVue with manual microscopy for detecting clinically relevant numbers of cells and 2 crystal types. SAMPLES: Five-hundred thirty urine samples (82% canine, 18% feline). METHODS: For SediVue analysis (software versions [SW] 1.0.0.0 and 1.0.1.3), uncentrifuged urine was pipetted into a cartridge. Images were captured and processed using a convolutional neural network algorithm. For manual microscopy, urine was centrifuged to obtain sediment. To determine sensitivity and specificity of the SediVue compared with manual microscopy, thresholds were set at ≥5/high power field (hpf) for red blood cells (RBC) and white blood cells (WBC) and ≥1/hpf for squamous epithelial cells (sqEPI), non-squamous epithelial cells (nsEPI), struvite crystals (STR), and calcium oxalate dihydrate crystals (CaOx Di). RESULTS: The sensitivity of the SediVue (SW1.0.1.3) was 85%-90% for the detection of RBC, WBC, and STR; 75% for CaOx Di; 71% for nsEPI; and 33% for sqEPI. Specificity was 99% for sqEPI and CaOx Di; 87%-90% for RBC, WBC, and nsEPI; and 84% for STR. Compared to SW1.0.0.0, SW1.0.1.3 had increased sensitivity but decreased specificity. Performance was similar for canine versus feline and fresh versus stored urine samples. CONCLUSIONS AND CLINICAL IMPORTANCE: The SediVue exhibits good agreement with manual microscopy for the detection of most formed elements evaluated, but improvement is needed for epithelial cells.

Effect of calcium sources in the diets of adult cats on urinary parameters and acid-base balance / Efeitos das fontes de cálcio na dieta de gatos adultos sobre parâmetros urinários e equilíbrio ácido-básico

Calcium is a macroelement that is part of the mineral composition of the diet of companion animals, and is considered a cation of strong alkalizing power, increasing urinary pH. Calcium salts have different solubilities and depending on the anion to which calcium is associated with, it can be more or less absorbed, modifying the pH of the urine. The aim of this study was to evaluate the efficiency of calcium sources on alkalinization of urinary pH, as well as excretion of urinary electrolytes and acid-base balance of adult cats. An extruded diet for cats was selected, and had 160mEq/kg of calcium from the sources of either calcium carbonate (CaCO3) or calcium gluconate (C12H22CaO14) added. In the control treatment there was no addition of calcium sources, resulting in three treatments. Nine adult cats were used, mixed breed, in two experimental periods, with six replicates per treatment. Animal average age was 4±1.3 years old and average weight was 3.96±0.71kg. The cats remained in metabolic cages for an adaptation period of seven days, followed by six days of urine total collection, with volume, density, pH and calcium concentration (g/d) measurements. The acid-base balance was studied by blood gas analysis of venous blood. The two sources of calcium alkalinized the urine (P<0.001). However, calcium gluconate had less alkalinization power compared to the calcium carbonate (P<0.05). Urinary calcium was not affected by treatments, and represented less than 0.5% of calcium intake. The experiment showed that calcium, although an alkaline cation and considered strong influencer of the EB of the diet, cannot be evaluated individually, because depending on its associated anion it may have greater or lesser influence on cats urine pH.(AU)

O cálcio (Ca) é um macroelemento que faz parte da composição mineral da dieta de animais de companhia. Este macroelemento é considerado um cátion de forte capacidade alcalinizante e, de acordo com a fonte e quantidade inclusa, pode aumentar o pH urinário. Os sais de cálcio têm diferentes solubilidades e dependendo do ânion ao qual o cálcio está associado, pode ser mais ou menos absorvido e assim, alterar o pH da urina. O objetivo deste estudo foi avaliar os efeitos de duas fontes de cálcio na alcalinização do pH urinário, bem como a excreção de eletrólitos urinários e o equilíbrio ácido-básico de felinos. Foi selecionada uma dieta extrusada para gatos e adicionados 160mEq/kg de cálcio das fontes carbonato de cálcio (CaCO3) ou gluconato de cálcio (C12H22CaO14). No tratamento controle, não houve adição de fontes de cálcio. Foram utilizados nove gatos adultos, de raças mistas, em dois períodos experimentais, com seis repetições por tratamento. Os animais apresentavam idade média de 4,0±1,3 anos e peso corporal médio de 3,96±0,71kg. Estes permaneceram em gaiolas metabólicas em período de adaptação durante sete dias, seguido de coleta total de urina durante seis dias. Nestas amostras foram aferidos o volume, densidade, pH e concentração de cálcio (g/d). O equilíbrio ácido-básico foi avaliado por hemogasometria em amostras de sangue venoso. As duas fontes de cálcio alcalinizaram a urina (P<0,001). No entanto, o gluconato de cálcio apresentou menor potencial de alcalinização em comparação ao carbonato de cálcio (P<0,05). O cálcio urinário não foi afetado pelos tratamentos e representou menos de 0,5% da ingestão de Ca. O experimento demonstrou que o cálcio, apesar de ser um cátion alcalinizante e influenciador do EB da dieta, não pode ser avaliado individualmente, porque dependendo do ânion associado, pode apresentar maior ou menor influência no pH da urina de gatos.(AU)

Effects of a nutrient-enriched water with and without poultry flavoring on water intake, urine specific gravity, and urine output in healthy domestic cats fed a dry kibble diet.

OBJECTIVE To investigate water intake and urine measures in healthy cats provided free-choice access to a nutrient-enriched water with (NWP) or without (NW) added poultry flavoring offered at 3 different volumes in addition to tap water (TW). ANIMALS 36 domestic shorthair cats. PROCEDURES Control group cats (n = 4) received dry food with TW ad libitum throughout the study. Cats of the NW and NWP groups (n = 16/group) received the same food with TW only (period 1; 7 days) followed by TW and the assigned treatment ad libitum at 1X, 1.5X, and 2X the volume of TW consumed in period 1 during periods 2 (17 days), 3 (10 days), and 4 (10 days), respectively. Liquid consumption, food intake, and total water intake (from all sources) were measured; urine collected over 48 hours in each period was measured, and urine specific gravity (USG) was determined. Data were analyzed with mixed-effects models. RESULTS TW and food calorie intake were similar among groups in period 1; TW consumption by control cats did not differ during the study. Liquid consumed by drinking increased 18%, 57%, and 96% for the NWP group in periods 2, 3, and 4, respectively, with increases of 25% and 44% for the NW group in periods 3 and 4, respectively, compared with period 1 values for the same groups. Increased urine output and decreased USG were significantly associated with period and treatment. CONCLUSIONS AND CLINICAL RELEVANCE Increasing the volumes of NW or NWP offered to healthy cats led to increased free liquid consumption and was associated with greater urine output and dilution as measured by USG. Studies are warranted to determine whether these treatments provide health benefits for cats in need of greater water consumption.

Evaluation of the analytical variability of urine protein-to-creatinine ratio in cats.

BACKGROUND: Proteinuria quantification with the urinary protein-to-creatinine (UPC) ratio is part of the diagnostic process in feline patients suspected of chronic kidney disease (CKD). In affected cats, monitoring and substaging of the UPC according to the International Renal Interest Society (IRIS) guidelines is also necessary for appropriate patient management. No information is available about the possible effects of analytical variability on urinary proteins (UPs) and the UPC ratio in cats. OBJECTIVES: This study aimed to determine whether imprecision and method-dependent differences due to the two dye-binding methods, pyrogallol red-molybdate (PRM) and Coomassie brilliant blue (CBB), could affect IRIS substaging. METHODS: Urine samples were collected from proteinuric and nonproteinuric cats. Intra-assay and inter-assay repeatability were assessed with both the PRM and CBB methods. Urinary supernatants (n = 120) were tested using both methods. Agreement between the methods and concordance with sample classification according to IRIS guidelines were determined. RESULTS: On average, the PRM method yielded a higher CV (UP 8.4 ± 5.2%; UPC 9.5 ± 4.8%) than the CBB method (UP 5.6 ± 2.6%; UPC 7.2 ± 2.6%), but similar rates of misclassification were found in samples with UPC ratios close to the IRIS cut-off. Although the two methods were correlated, the CBB method tended to yield UPs and UPC ratios that were significantly higher (P < 0.0001) than that of the PRM method. The Passing-Bablok test also found constant and proportional errors between the PRM and CBB methods. Concordance in substaging samples according to IRIS was good (k coefficient = 0.62). CONCLUSIONS: The two methods were precise, but the higher UPC ratios obtained with the CBB methods might affect the interpretation using the IRIS guidelines and clinical decisions.

Influence of protein concentration and quality in a canned diet on urine composition, apparent nutrient digestibility and energy supply in adult cats.

BACKGROUND: Protein concentration and quality in cat food can vary considerably, and the impact on feline urine composition and nutrient supply is of high practical relevance. In the present study, 6 canned diets with varying protein concentrations and qualities were fed to 10 healthy adult cats. Protein quality in the diet differed depending on the amount of collagen-rich ingredients. Hydroxyproline concentrations were 2.56-4.45 g/kg dry matter in the high quality and 3.76-9.44 g/kg dry matter in the low quality diets. Protein levels were 36.2, 43.3 and 54.9% in the high quality and 36.7, 45.0 and 56.1% in the low quality groups. Each diet was fed for 6 weeks, using a randomized cross-over design. In the last 2 weeks of each feeding period, urine and faeces of the cats were collected. RESULTS: Renal calcium (Ca), oxalate (Ox) and citrate excretion were unaffected by the dietary protein concentration, possibly mediated by a high urine volume (24.2-34.2 ml/kg bodyweight (BW)/day) in all groups. However, renal Ox excretion was lower when the high quality diets were fed (P = 0.013). Urinary relative supersaturation (RSS) with calcium oxalate (CaOx) was low in general, but reduced in the high quality groups (P = 0.031). Urinary RSS values for magnesium ammonium phosphate (MAP) were high (2.64-5.00) among all groups. Apparent digestibility of crude protein and most minerals was unaffected by the different diets. Feed intake was higher in the low quality groups (P = 0.026), but BW of the cats did not differ depending on dietary protein quality. BW of the cats increased with increasing dietary protein concentrations (P = 0.003). CONCLUSION: In conclusion, a high protein canned diet might not be a specific risk factor for CaOx urolith formation in cats. In contrast, all diets resulted in high RSS MAP values, which might be critical concerning MAP crystallization. Protein quality had a minor, but significant impact on urine composition, necessitating further research on this subject. A lower energy supply when feeding a low protein quality can be assumed. Changes in BW were only small and require a careful interpretation.

Lack of glucuronidation products of trans-resveratrol in plasma and urine of cats.

Resveratrol has generated interest in cats due to reported health benefits. Cats have low activity of ß-glucuronidase, and we hypothesized they could not form two common resveratrol metabolites, resveratrol-3-O-glucuronide and resveratrol-4'-O-glucuronide. Resveratrol, 3 mg/cat/day, was given orally to intact male (n = 5) and female cats (n = 5) for 4 weeks. A control group (8 intact males) was used for comparison. Plasma and urine were collected weekly and analysed using high-pressure liquid chromatography coupled with tandem mass spectrometry. Resveratrol and resveratrol-3-O-sulphate, but no glucuronide metabolites, were detected in plasma and urine. Median (range 10-90th percentile) plasma resveratrol for control and treatment groups was 0.46 ng/ml (0.02-1.74 ng/ml) and 0.96 ng/ml (0.65-3.21 ng/ml). Median (range) plasma resveratrol-3-O-sulphate for control and treatment groups was 6.32 ng/ml (2.55-10.29 ng/ml) and 11.45 ng/ml (1.47-53.29 ng/ml). Plasma resveratrol differed from control in week 4, while plasma resveratrol-3-O-sulphate was different in all weeks (p < 0.05). Median (range) urine resveratrol for control and treatment groups was 0.28 ng/ml (0.05-1.59 ng/ml) and 19.98 ng/ml (8.44-87.54 ng/ml). Median (range) urine resveratrol-3-O-sulphate for control and treatment groups was 26.71 ng/ml (10.50-75.58 ng/ml) and 108.69 ng/ml (11.83-231.05 ng/ml). All time points for urine resveratrol and resveratrol-3-O-sulphate were significantly different from control (p < 0.05), except for weeks 1, 3 and 4 for resveratrol. The results support our hypothesis that cats are unlikely able to glucuronidate resveratrol, most likely due to a reduction in the activity of ß-glucuronidase.

Concentrations of thromboxane metabolites in feline urine.

OBJECTIVE To determine the predominant thromboxane (TX) metabolite in urine of healthy cats, evaluate whether the method of sample collection would impact concentration of that metabolite, and propose a reference interval for that metabolite in urine of healthy cats. ANIMALS 17 cats (11 purpose-bred domestic shorthair cats, 5 client-owned domestic shorthair cats, and 1 client-owned Persian cat). PROCEDURES All cats were deemed healthy on the basis of results for physical examination, a CBC, serum biochemical analysis, urinalysis, and measurement of prothrombin time and activated partial thromboplastin time. Voided and cystocentesis urine samples (or both) were collected. Aliquots of urine were stored at -80°C until analysis. Concentrations of TXB2, 11-dehydroTXB2, and 2,3 dinorTXB2 were measured with commercially available ELISA kits. Urinary creatinine concentration was also measured. RESULTS 11-dehydroTXB2 was the most abundant compound, representing (mean ± SD) 59 ± 18% of the total amount of TX detected. In all samples, the concentration of 11-dehydroTXB2 was greater than that of 2,3 dinorTXB2 (mean, 4.2 ± 2.7-fold as high). Mean concentration of 11-dehydroTXB2 for the 17 cats was 0.57 ± 0.47 ng/mg of creatinine. A reference interval (based on the 5% to 95% confidence interval) of 0.10 to 2.1 ng of 11-dehydroTXB2/mg of creatinine was proposed for healthy cats. CONCLUSIONS AND CLINICAL RELEVANCE In this study, 11-dehydroTXB2 was the major TX metabolite in feline urine. Measurement of this metabolite may represent a noninvasive, convenient method for monitoring in vivo platelet activation in cats at risk for thromboembolism.

Predator odours attract other predators, creating an olfactory web of information.

Many studies have reported the aversive reactions of prey towards a predator's odour signals (e.g. urine marks), a behaviour widely thought to reduce the risk of predation by the predator. However, because odour signals persist in the environment, they are vulnerable to exploitation and eavesdropping by predators, prey and conspecifics. As such, scent patches created by one species might attract other species interested in information about their enemies. We studied this phenomenon by examining red fox investigation of odours from conspecifics and competing species in order to understand what prey are responding to when avoiding the odours of a predator. Surprisingly, foxes showed limited interest in conspecific odours but were highly interested in the odours of their competitors (wild dogs and feral cats), suggesting that odours are likely to play an important role in mediating competitive interactions. Importantly, our results identify that simple, dyadic interpretations of prey responses to a predator odour (i.e. cat odour = risk of cat encounter = fear of cats) can no longer be assumed in ecological or psychology research. Instead, interactions mediated by olfactory cues are more complex than previously thought and are likely to form a complicated olfactory web of interactions.

Impact of Increasing Dietary Calcium Levels on Calcium Excretion and Vitamin D Metabolites in the Blood of Healthy Adult Cats.

BACKGROUND: Dietary calcium (Ca) concentrations might affect regulatory pathways within the Ca and vitamin D metabolism and consequently excretory mechanisms. Considering large variations in Ca concentrations of feline diets, the physiological impact on Ca homeostasis has not been evaluated to date. In the present study, diets with increasing concentrations of dicalcium phosphate were offered to ten healthy adult cats (Ca/phosphorus (P): 6.23/6.02, 7.77/7.56, 15.0/12.7, 19.0/17.3, 22.2/19.9, 24.3/21.6 g/kg dry matter). Each feeding period was divided into a 10-day adaptation and an 8-day sampling period in order to collect urine and faeces. On the last day of each feeding period, blood samples were taken. RESULTS: Urinary Ca concentrations remained unaffected, but faecal Ca concentrations increased (P < 0.001) with increasing dietary Ca levels. No effect on whole and intact parathyroid hormone levels, fibroblast growth factor 23 and calcitriol concentrations in the blood of the cats were observed. However, the calcitriol precursors 25(OH)D2 and 25(OH)D3, which are considered the most useful indicators for the vitamin D status, decreased with higher dietary Ca levels (P = 0.013 and P = 0.033). Increasing dietary levels of dicalcium phosphate revealed an acidifying effect on urinary fasting pH (6.02) and postprandial pH (6.01) (P < 0.001), possibly mediated by an increase of urinary phosphorus (P) concentrations (P < 0.001). CONCLUSIONS: In conclusion, calcitriol precursors were linearly affected by increasing dietary Ca concentrations. The increase in faecal Ca excretion indicates that Ca homeostasis of cats is mainly regulated in the intestine and not by the kidneys. Long-term studies should investigate the physiological relevance of the acidifying effect observed when feeding diets high in Ca and P.