In vitro glucuronidation of the angiotensin II receptor antagonist telmisartan in the cat: a comparison with other species

Ebner, T., Schänzle, G., Weber, W., Sent, U., Elliott, J. In vitro glucuronidation of the angiotensin II receptor antagonist telmisartan in the cat: a comparison with other species. J. vet. Pharmacol. Therap.  36 , 154–160. Glucuronidation of telmisartan comprises nearly its entire metabolic clearance in several mammalian species including human. However, data were lacking for the cat, a species noted for its inability to glucuronidate some drugs. Therefore, the glucuronidation of telmisartan was investigated using feline liver microsomes and compared to liver microsomes of rats, dogs, and human, intestinal human microsomes and cell lines expressing human UDP‐glucuronosyltransferases (UGT). Incubation of telmisartan with cat liver microsomes readily yielded telmisartan glucuronide, and pooled (N = 3 for each gender) cat liver microsomes even showed the highest glucuronidation rate (cat > dog >> human > rat). Michaelis Menten kinetics were observed with K_m of 7.5 and 10 μm and V_max of 3.9 and 3.3 nmol/min/mg for male and female cats, respectively. Confirming the in vitro data, telmisartan glucuronide was detected as the major circulating metabolite in cat plasma. To elucidate which UGT enzymes are involved, telmisartan was incubated with cell lines expressing human UGTs. The highest glucuronidation activity was observed for UGT1A8, UGT1A7, and UGT1A9. In conclusion, telmisartan was effectively glucuronidated in cats. Defects of the UGT1A6 gene in cats do not affect the glucuronidation of telmisartan as it is not a substrate of human UGT1A6.     

Cytochrome P450-mediated hepatic metabolism of new fluorescent substrates in cats and dogs

This study aimed to investigate the biotransformation of cat liver microsomes in comparison to dogs and humans using a high throughput method with fluorescent substrates and classical inhibitors specific for certain isozymes of the human cytochrome P450 (CYP) enzyme family. The metabolic activities associated with CYP1A, CYP2B, CYP2C, CYP2D, CYP2E and CYP3A were measured. Cat liver microsomes metabolized all substrates selected for the assessment of cytochrome P450 activity. The activities associated with CYP3A and CYP2B were higher than the activities of the other measured CYPs. Substrate selectivity could be demonstrated by inhibition studies with α-naphthoflavone (CYP1A), tranylcypromine/quercetine (CYP2C), quinidine (CYP2D), diethyldithiocarbamic acid (CYP2E) and ketoconazole (CYP3A) respectively. Other prototypical inhibitors used for characterization of human CYP activities such as furafylline (CYP1A), tranylcypromine (CYP2B) and sulfaphenazole (CYP2C) did not show significant effects in cat and dog liver microsomes. Moreover, IC50-values of cat CYPs differed from dog and human CYPs underlining the interspecies differences. Gender differences were observed in the oxidation of 7-ethoxy-4-trifluoromethylcoumarin (CYP2B) and 3-[2-(N, N-diethyl-N-methylamino)ethyl]-7-methoxy-4-methylcoumarin (CYP2D), which were significantly higher in male cats than in females. Conversely, oxidation of the substrates dibenzylfluorescein (CYP2C) and 7-methoxy-4-trifluoromethylcoumarin (CYP2E) showed significant higher activities in females than in male cats. Overall CYP-activities in cat liver microsomes were lower than in those from dogs or humans, except for CYP2B. The presented difference between feline and canine CYP-activities are useful to establish dose corrections for feline patients of intensively metabolized drugs licensed for dogs or humans.     

Comparative oxidative metabolic profiles of clomipramine in cats, rats and dogs: preliminary results from an in vitro study

The objectives of this in vitro study were to describe cytochrome-dependent metabolism of clomipramine in canine and feline microsomes, compare metabolic profiles between cats, rats and dogs, and investigate a potential gender-related difference in metabolic activity between male and female cats. Pooled liver microsomes were incubated with clomipramine, where species and gender-specific reactions were initiated by the addition of a nicotinamide adenine dinucleotide phosphate regenerating system and quenched with methanol at 0, 5, 15, 30, 45 and 60 min, and 0, 30, 60, 90, 120, 180, 240 and 360 min respectively. Liquid chromatography tandem mass spectrometry was used to measure clomipramine and its metabolites. Preliminary results showed that cat microsomes biotransformed clomipramine slower and less efficiently than rat and dog microsomes. Moreover, gender differences in metabolic profiles suggested that male cat microsomes may be less efficient demethylators and hydroxylators than female cat microsomes. As gender metabolic differences may carry clinical significance for this antidepressant, further studies are warranted.     

Comparing the glucuronidation capacity of the feline liver with substrate‐specific glucuronidation in dogs

This study aimed to assess the overall glucuronidation capacity of cats, using prototypic substrates identified for human UDP‐glucuronosyltransferases (UGTs). To this end, Michaelis–Menten kinetics were established for the substrates using feline hepatic microsomal fractions, and results were compared with similar experiments carried out with dog liver microsomes. Cats are known for their low capacity of glucuronide formation, and UGT1A6 was found to be a pseudogene. However, functional studies with typical substrates were not performed and knowledge of the enzymology and genetics of other glucuronidation enzymes in felidae is lacking. The results of this study showed extremely low formation of naphthol‐1‐glucuronide (1.7 ± 0.4 nmol/mg protein/min), estradiol‐17‐glucuronide (<0.7 nmol/mg protein/min), and morphine‐3‐glucuronide (0.2 ± 0.03 nmol/mg protein/min), suggesting a lack of functional UGT1A6 and UGT2B7 homologues in the cat's liver. Dog liver microsomes were producing these glucuronides in much higher amounts. Glucuronide capacity was present for the substrates 17β‐estradiol (estradiol‐3‐glucuronide, 2.9 ± 0.2 nmol/mg protein/min) and 4‐methylumbelliferone (31.3 ± 3.3 nmol/mg protein/min), assuming that cats have functional homologue enzymes to at least the human UGT1A1 and probably other UGT1A isozymes. This implies that for new drugs, glucuronidation capacity has to be investigated on a substance‐to‐substance base. Knowledge of the glucuronidation rate of a drug provides the basis for pharmacokinetic modeling and as a result proper dosage regimens can be established to avoid undesirable drug toxicity in cats.     

Characterization of urinary metabolites of testosterone, methyltestosterone, mibolerone and boldebone in greyhound dogs

Androgenic steroids are used in female greyhound dogs to prevent the onset of estrus; moreover, these steroids also have potent anabolic activity. As anabolic steroids increase muscle mass and aggression in animals, the excessive use of these agents in racing greyhounds gives an unfair performance advantage to treated dogs. The biotransformation of most anabolic steroids has not been determined in greyhound dogs. The objective of the present study was to identify the urinary metabolites of testosterone, methyltestosterone, mibolerone, and boldenone in greyhound dogs. These steroids were administered orally (1 mg/kg) to either male or female greyhound dogs and urine samples were collected pre-administration and at 2, 4, 8, 12, 24, 72, and 96 h post-administration. Urine extracts were analyzed by high-performance liquid chromatography/mass spectrometry (HPLC/MS) to identify major metabolites and to determine their urinary excretion profiles. Major urinary metabolites, primarily glucuronide, conjugated and free, were detected for the selected steroids. Sulfate conjugation did not appear to be a major pathway for steroid metabolism and excretion in the greyhound dog. Phase I biotransformation was also evaluated using greyhound dog liver microsomes from untreated dogs. The identification of several in vivo steroid metabolites generated in this study will be useful in detecting these steroids in urine samples submitted for drug screening.     

Ex vivo binding of the immunosuppressant mycophenolic acid to dog and cat plasma proteins and the effect of co‐incubated dexamethasone and prednisolone

Mycophenolic acid (MPA) has been shown to be promising for the treatment of autoimmune diseases in dogs and cats. In humans, MPA is highly bound to plasma proteins (~97%). It has been recommended to monitor free drug plasma concentrations because the free MPA correlates with its immunosuppressive effect. However, it is unknown if MPA is highly bound to plasma proteins in dogs and cats. The objectives of this study were to determine the extent of plasma protein binding of MPA and evaluate the effect of prednisolone and dexamethasone on the extent of protein binding of MPA in dogs and cats. The extent of plasma protein binding of MPA was determined in plasma collected from clinically healthy adult cats (n = 13) and dogs (n = 14) by combining high‐throughput dialysis and ultra‐high‐liquid chromatography. This study reveals that MPA is highly bound to plasma proteins (>90%) in dogs and cats, mean extent of binding of MPA at 15 μg/ml to plasma proteins being 96% (range, 95%–97%) and 92% (range, 90%–93%) for dogs and cats, respectively. In dog plasma, MPA is primarily bound to albumin. In vitro, prednisolone increased the unbound MPA in dogs (p < .01) but not in cats (p = .07) while dexamethasone had no effect on MPA plasma binding in either species (p > .05). Results of this study provide valuable information for designing future pharmacokinetic and pharmacodynamic studies and also therapeutic monitoring programs for dogs and cats.     

Phylogenetic identification of Cystoisospora spp. from dogs, cats, and raccoon dogs in Japan

Cystoisospora spp. from feces in dogs, cats, and raccoon dogs were isolated, sequenced at the small subunit ribosomal RNA gene locus and compared to other Cystoisospora spp. Cystoisospora oocysts from dogs and raccoon dogs were morphologically similar with those of C. ohioensis, and cat isolates were similar with those of C. felis. The sequences from dogs and raccoon dogs, and cats have a homology with C. ohioensis and C. felis, respectively. Phylogenetic analysis of the DNA sequences showed that the dog and raccoon dog isolates were nested in a clade with other Cystoisospora spp. including C. ohioensis, C. belli, and C. orlovi. The cat isolate formed a sister group with C. felis that was a separate clade from the dog and raccoon dog group. We report sequence variation in these Cystoisospora sequences and have identified raccoon dogs as another carnivore host for Cystoisospora spp. infecting dogs.     

Blood vitamin and choline concentrations in healthy domestic cats, dogs, and horses

Blood concentrations of thiamin, biotin, nicotinates, pantothenates, folates, riboflavin, vitamins A, B6, B12, C, E, beta-carotene and choline were analyzed in healthy animals (23 horses, 25 dogs, and 29 cats). B-Complex vitamins and choline also were analyzed in the liver of the dogs and cats. Vitamin concentrations in the blood and livers of dogs were similar; however, blood vitamin A and beta-carotene concentrations were lower in the cat than in the dog. Horses had a higher B12 blood concentration than did the dogs and cats. These data can be useful for detecting overt and hidden vitamin deficits in these species due to various conditions.     

Review of thymic pathology in 30 cats and 36 dogs

Data are presented from 30 cats and 36 dogs in which thymic disease was recognised clinically or on postmortem examination. The diagnoses included thymic lymphoma (19 cats, l 2 dogs), thymoma (five cats, 18 dogs), thymic branchial cyst formation or cystic change (one cat, four dogs), thymic hyperplasia (two cats), congenital hypoplasia (one cat, one dog), thymic haemorrhage (one cat, one dog) and thymic amyloidosis (one cat). Thymic lymphoma occurred in younger dogs and cats, and was recorded equally among domestic shorthaired and purebred (especially Siamese) cats. Eight cats with thymic lymphoma were tested for feline leukaemia virus and four were positive. Thymoma occurred more frequently in older cats and dogs, and in Labradors and German shepherd dogs. Thymic tumours were associated with paraneoplastic hypercalcaemia (six dogs), megaoesophagus (two dogs) or interface dermatitis with basement membrane immune complex deposition (one cat). Non-neoplastic thymic diseases were associated with myasthenia gravis (one cat), pemphigus foliaceus (one cat) and superficial necrolytic dermatitis (one cat).     

Equine uridine diphospho-glucuronosyltransferase 1A1, 2A1, 2B4, 2B31: cDNA cloning,expression and initial characterization of morphine metabolism

ObjectiveUridine diphospho-glucuronosyltransferases (UGTs) are membrane-bound enzymes that catalyze the conjugation of glucuronic acid onto a diverse set of xenobiotics. Horses efficiently and extensively glucuronidate a number of xenobiotics, including opioids, making UGTs an important group of drug-metabolizing enzymes for the clearance of drugs. Recombinant enzymes have allowed researchers to characterize the metabolism of a variety of drugs. The primary objective was to clone, express and characterize equine UGTs using drugs characterized as UGT substrates in other species. A secondary objective was to characterize the in vitro metabolism of morphine in horses.Study designIn vitro drug metabolism study using liver microsomes and recombinant enzyme systems.AnimalsLiver microsomes and RNA from tissue collected from two Thoroughbred mares euthanized for other reasons.MethodsBased on homology to the human UGT2B7, four equine UGT variants were expressed: UGT1A1, UGT2A1, UGT2B31 and UGT2B4. cDNA sequences were cloned and resulting protein expressed in a baculovirus expression system. Functionality of the enzymes was assessed using 4-methylumbelliferone, testosterone, diclofenac and ketoprofen. Recombinant enzyme, control cells, equine liver microsomes and human UGT2B7 supersomes were then incubated with morphine. Concentrations of metabolites were measured using liquid chromatography–tandem mass spectrometry and enzyme kinetics determined.Results4-Methylumbelliferone was glucuronidated by all expressed equine UGTs. Testosterone glucuronide was not produced by any of the expressed enzymes, and diclofenac glucuronide and ketoprofen glucuronide were produced by UG2A1 and UGT1A1, respectively. UGT2B31 metabolized morphine to morphine-3-glucuronide and low concentrations of morphine-6-glucuronide.Conclusions and clinical relevanceThis is the first successful expression of functional recombinant equine UGTs. UGT2B31 contributes to the glucuronidation of morphine; however, it is probably not the main metabolizing enzyme. These results warrant further investigation of equine UGTs, including expression of additional enzymes and further characterization of UGT2B31 as a contributor to morphine metabolism.     

New perspectives about Hemotrophic mycoplasma (formerly, Haemobartonella and Eperythrozoon species) infections in dogs and cats.

The new perspectives about hemotrophic mycoplasma infections in cats and dogs can be summarized as follows: Haemobartonella and Eperythrozoon species infecting the dog and cat have been reclassified as mycoplasmal parasites and given the names M haemofelis (Ohio or large form of H felis), M haemominutum (California or small form of H felis), and M haemocanis (H canis). The prevalence of hemotrophic mycoplasma infections in anemic cats in the United States is about 25% and usually involves M haemofelis. However, nonanemic cats may also be infected most commonly with M haemominutum. Chronic infections with hemotrophic mycoplasmas may promote myeloproliferative disorders in FeLV-infected cats. M haemocanis infection in dogs may be a widespread latent disease in kennel-raised dogs and is being investigated. The PCR assay is exquisitely sensitive for detection of M haemofelis and M haemominutum, and testing of blood donor cats and perhaps dogs should be done regularly. Fleas are involved in the transmission of M haemofelis to the cat, whereas R sanguines may be involved with transmission of M haemocanis to the dog. Treatment with doxycycline effectively controls acute infection in the cat and dog, and enrofloxacin may also be effective in the cat, but none of the antibiotics tested to date consistently clears the parasites.     

闽清并殖对终末宿主的感染性试验及并殖吸虫对人体致病性分型的讨论

探讨闽清并殖对人体的致病性。将闽清并殖新鲜囊蚴多次分别感染猫科、犬科和啮齿科的家猫、家狗及大鼠、小鼠、豚鼠。结果 ,只在猫、狗粪便中检及虫卵并在肺脏检及成虫 ,而鼠类未获感染。结合对已知并殖吸虫宿主的选择性的分析 ,发现对猫狗感染而对鼠类不适宜的虫种通常对人体致病 ,相反 ,对鼠类适宜而猫狗不适宜的虫种对人体大多不致病或致病力较弱 ,表明闽清并殖可能类似斯氏并殖对人体引起游走性皮下结节的病变     

Capnocytophaga canimorsus

Capnocytophaga canimorsus is a commensal bacterium in the oral flora of dogs and cats. The bacterium is a zoonotic agent and has been isolated from humans, infected by dog or cat bites, scratches, licks or simply exposure to dogs or cats. Here the infectious agent, its pathogenicity and potential virulence factors, infection in animals and humans, diagnostic methods, prevalence, therapy and prevention are described. Suggestions for future research are given.     

Vitamin A concentrations in commercial foods for dogs and cats

The vitamin A concentration was determined in 89 Australian brands of commercial foods for dogs and cats. It was found that 8% of the dog foods and 14% of the cat foods had concentrations of vitamin A below the minimum recommended 1.1 mg/kg dry matter (dm) for dogs and 1.8 mg/kg dm for pregnant or lactating cats. Canned and fish-labelled cat foods were the only varieties with less than the minimum recommended concentration of Vitamin A, of which 71% were the same brand. The minimum recommended concentration of vitamin A was exceeded in all canned dog food tested. Concentrations of vitamin A in dry (ca. 6% moisture) dog and cat foods and semi-moist dog foods (ca. 23% moisture) never exceeded 10 mg vitamin A/kg dm. In contrast, canned pet foods stated to contain liver or kidney showed vitamin A concentrations from 13 to 284 mg/kg dm.     

Sporotrichosis: a retrospective evaluation of 23 cases seen in northern California (1987–2007)

Sporotrichosis is an uncommon to rare cutaneous and subcutaneous mycosis of animals and humans caused by the dimorphic fungus Sporothrix schenckii . Twenty-three mammalian cases of sporotrichosis examined between 1987 and 2007 at the University of California, Davis – Veterinary Medical Teaching Hospital, were retrospectively evaluated with regard to the historical, clinical, diagnostic and treatment findings. Cats were the most common species affected ( n  = 14). In addition, sporotrichosis was diagnosed in four dogs, four horses and a donkey. Six of 23 cases were diagnosed with the localized cutaneous form of sporotrichosis (four cats, one dog, one horse), 10 with the cutaneous-lymphatic form (four cats, two dogs, three horses and a donkey), and seven with the disseminated form (six cats, one dog). Two of 23 cases did not have skin lesions at the time of diagnosis (one cat, one dog). The most common mode of diagnosis was demonstration of S. schenckii on histopathological evaluation of tissue. In contrast with most previously described sporotrichosis infections in cats, few to no fungal organisms were seen in histopathological samples (haematoxylin and eosin and special stains) in five of the 14 cats. Treatments received included itraconazole (12 cats, one dog), ketoconazole (three dogs), fluconazole (one cat, one donkey), sodium iodide (four horses, one cat) and potassium iodide (one cat, one horse, one donkey). The prognosis for successful treatment was good in all species. Fluconazole was successful in inducing resolution of the cutaneous lesions and controlling the infection in one cat with disseminated sporotrichosis.     

Evaluation of risk factors for bite wounds inflicted on caregivers by dogs and cats in a veterinary teaching hospital

OBJECTIVE: To identify factors associated with increased risk of being bitten by a dog or cat in a veterinary teaching hospital. DESIGN: Unmatched case-control study. STUDY POPULATION: 207 animal caregivers. PROCEDURE: Case subjects (n = 75) were any caregiver that reported being bitten by a dog or cat. Control subjects (n = 132) were randomly selected from a list of all caregivers interacting with dogs or cats. Information on the characteristics of the caregivers, characteristics of the dogs and cats, and the nature of the interaction between the dog or cat and the caregiver was obtained by use of self-administered questionnaires. RESULTS: Caregivers were more likely to be bitten by dogs or cats that had warning signs on their cages indicating the potential to bite or that were considered difficult to handle. Caregivers interacting with cats or with older dogs and cats were more likely to be bitten. Only 37 to 55% of dogs and cats that had characteristics traditionally associated with biting or were considered likely to bite were muzzled. CONCLUSIONS AND CLINICAL RELEVANCE: Muzzling dogs and cats should be considered more frequently. Dogs and cats considered to have the propensity to bite frequently do bite, and precautions, such as muzzling, should be taken if the medical condition or conformation of the dog or cat is amenable to this type of restraint.     

Total selenium concentrations in canine and feline foods commercially available in New Zealand

AIMS: To determine the total selenium concentrations in petfoods commercially available in New Zealand and to establish whether these meet the current minimum recommended requirements of selenium in foods for cats and dogs. METHODS: Samples (n=89) from petfoods commercially available in New Zealand were analysed for total selenium concentration using a fluorometric method. Data, expressed on a dry matter (DM) basis, were analysed according to petfood type (dog or cat, and wet or dry), predominant flavour (chicken, seafood, chicken and seafood, beef, meat mix, other), manufacturer and country of manufacture. RESULTS: Fifty percent of petfoods purchased for this study were manufactured in Australia, and the remainder were produced in the United States of America (USA), New Zealand or Thailand. Mean total selenium concentrations were similar (0.61-0.80 mg/kg DM) in petfoods produced in Australia, New Zealand and the USA, but higher (mean 3.77 mg/kg DM; p<0.05) in petfoods produced in Thailand. Petfoods produced in Australia, New Zealand and the USA contained a variety of predominant flavours, whereas petfoods from Thailand contained only seafood flavour. Seafood-based flavours had the highest selenium concentrations in both cat and dog foods. Wet and dry dog foods had similar concentrations of selenium to dry cat foods, but wet cat foods had higher and more variable concentrations of selenium than these others (p<0.05). The mean selenium concentrations in cat and dog foods were 1.14 and 0.40 mg/kg DM, respectively, and there were no significant differences between manufacturers. CONCLUSIONS: Selenium concentrations in commercial petfoods sold in New Zealand appeared to meet recommended dietary requirements, although the range of concentrations was highly variable. Whether these recommendations are adequate for the maintenance of optimal health in cats and dogs has yet to be determined. CLINICAL RELEVANCE: Overt selenium deficiency disorders are unlikely in dogs and cats in New Zealand fed commercial petfoods unless the bioavailability of selenium in particular petfoods is low.     

Middle ear disease associated with congenital palatine defects in seven dogs and one cat

Medical records of eight dogs and one cat with congenital palatine defects were reviewed retrospectively. Five of the dogs had nasal discharge and seven had radiographic signs of middle ear disease, but no clinical signs of ear disease were identified in any of the dogs, nor were any reported by their owners during a one- to five-year follow-up period. One dog had an ipsilateral impairment of hearing detected by brainstem auditory evoked responses. The cat had clinical and radiographic signs of middle ear disease. These findings suggest that, as in humans, congenital palatine defects in dogs and cats may predispose to middle ear disease. Any associated deafness could cause problems for working dogs.     

Effect of vaccination on fecal cortisol metabolites in cats and dogs

The aim of this study was to apply a cortisol metabolite determination in the faeces of cats and dogs for monitoring disturbances. In this experiment faeces from every spontaneous defecation of 10 cats and 10 dogs (5 of each sex) were collected starting from one day before until two days after the yearly vaccination. Concentrations of 11,17-dioxoandrostanes (cat) and cortisol equivalents (dog) were determined by enzyme immunoassay (EIA). Faecal cortisol metabolites increased and reached peak concentrations (median: 412% in cats and 417% in dogs, respectively above baseline values) in one of the next two samples following the vaccination. This indicated an activation of the adrenocortex, the degree to which the different parts (physical and psychological components) of the whole vaccination procedure contributed to it was not evaluated. From this experiment we conclude that measuring cortisol metabolites in the faeces is a non-invasive method to monitor stressful conditions in cats and dogs and thus is a valuable tool for evaluating animal welfare.     

Comparative evaluation of net digestive and absorptive efficiency in dogs and cats fed a variety of contrasting diet types

Comparative net digestive and absorptive efficiency was assessed with adult Beagles and domestic cats by apparent digestibility assays. Eight foods were used comprising two canned dog foods; canned cat foods; two samples of semipurified diet; and single samples of experimental dry cat food and fresh mince. Apparent digestibility percentages of crude protein, fat, nitrogen-free extract (NFE) and gross energy were all significantly higher in dogs than cats. Mean values obtained for dogs were (per cent): crude protein 87; fat 92; NFE 70 and energy 89. Respective values for cats were 82, 76, 67 and 79. On average, dogs obtained 11 and 9 per cent more digestible energy and protein per unit food eaten than cats. Most digestibility characteristics of foods measured in cats were significantly correlated with those for dogs and regression equations are presented predicting digestibility in cats from dog data.