Survival, remission, and quality of life in diabetic cats.

BACKGROUND: Remission is documented in a substantial proportion of cats with diabetes. The effects of diabetes mellitus (DM) on the lives of cats and their owners should be considered when evaluating treatment success. OBJECTIVES: To study outcome in cats with DM and the impact DM has on the life situation of cat and owner. ANIMALS: Domestic and pedigree cats with a diagnosis of DM (n = 477) insured by a Swedish insurance company during 2009 to 2013. METHODS: Retrospective cross-sectional study. A questionnaire was sent to 1369 owners of cats diagnosed with DM. The questions concerned the cat, treatment, owner perceptions of the disease and treatment and disease outcome. Data were analyzed using multiple linear and logistic regression, with outcomes set as survival for more than 4 weeks after diagnosis, survival time, achieving remission, remission without relapse and quality of life (QoL) for the cat. RESULTS: The response rate was 35%, leaving 477 questionnaires for analysis. The remission rate among treated cats was 29% (118/405). Feeding a commercially available wet diet was associated with both remission (OR 3.16, 95% confidence interval 1.27-8.12) and remission without relapse (OR 14.8, 95% confidence interval 2.25-153.8). Remission was associated with a better QoL for the cat. CONCLUSIONS AND CLINICAL IMPORTANCE: The association between feeding a commercially available wet diet and remission is important and strengthens the role of diet in treatment of DM in cats. Linking remission and a better QoL for the cat emphasizes remission as a goal in disease management.

Investigation of interference from canine anti-mouse antibodies in hormone immunoassays.

BACKGROUND: Canine anti-mouse antibodies are a potential source of immunoassay interference, but erroneous immunoassay results are not always easily identifiable. Anti-Müllerian hormone (AMH) is a marker for the presence of gonads in dogs, but elevated AMH concentrations in neutered dogs could also be caused by antibody interference. For other assays, a discrepant result obtained after antibody precipitation might indicate antibody interference. OBJECTIVES: We aimed to evaluate if canine anti-mouse antibodies are a source of erroneous results in the AMH assay and if antibody precipitation with polyethylene glycol (PEG) is a useful tool for detecting antibody interference in a variety of immunoassays used in the veterinary clinical laboratory. METHODS: Twenty-nine positive and 25 negative samples for anti-mouse antibodies were analyzed for AMH, canine total thyroxine (TT4 ), canine thyroid-stimulating hormone (TSH) and progesterone before and after treatment with PEG. Results that differed by more than four SDs from the intra-assay coefficients of variation were considered discrepant. Elevated AMH concentrations in neutered dogs with anti-mouse antibodies and no visible gonads present were considered evidence of interference. RESULTS: Evidence of antibody interference was found in two samples analyzed for AMH. The presence of anti-mouse antibodies did not lead to a higher proportion of discrepant results after PEG treatment for any of the immunoassays. The overall incidence of discrepant results for healthy controls was very high (73%). CONCLUSIONS: Canine anti-mouse antibodies are a source of erroneous AMH results. Antibody precipitation with PEG is not a useful tool for detecting interference caused by such antibodies.

Evaluation of an ELISA for metanephrines in feline urine.

Catecholamines can be used to evaluate neuroendocrine tumors, stress, and potentially pain, but catecholamines degrade rapidly. Their metabolites normetanephrine (NME) and metanephrine (ME) have better stability in urine. In cats, urine sampling in a home environment would be beneficial to reduce effects of clinical stress and simplify sampling. We evaluated a human urine ELISA for analysis of NME and ME in feline urine, and investigated the effects of acidification, cat tray pellets, and storage time at room temperature up to 8.5 h. In 26 feline urine samples, mean NME concentration was 192 ± 80 ng/mL, mean intra- and inter-assay CV was 6.5% and 4.2%, respectively, and spike recovery was 98-101%, but dilutional recovery was unsatisfactory. For ME, mean intra- and inter-assay CV was 10.2% and 4.1%, respectively. Mean urine ME concentration was 32.1 ± 18.3 ng/mL, close to the kit's lowest standard, and spike recovery was 65-90%; the ELISA could not be validated for ME. The stability study, performed for NME on 12 urine samples, did not identify differences between acidified and non-acidified samples, cat tray pellets, or storage time, and no interaction effects. The ME ELISA was not suitable for feline urine; performance of the NME ELISA was acceptable, except for dilution recovery. For analysis of NME, feline urine can be sampled at home using cat tray pellets and stored at room temperature up to 8.5 h without acidification.

Insulin-like growth factor I in cats: validation of an enzyme-linked immunosorbent assay and determination of biologic variation.

BACKGROUND: Insulin-like growth factor I (IGF-I) measurements are used in veterinary medicine for diagnosing growth hormone disorders. IGF-I assays are subject to interference by IGF-binding proteins (IGFBP) which may not be efficiently removed by standard extraction methods. Adding excess IGF-II during analysis may improve accuracy. OBJECTIVES: The purpose of the study was to validate a commercial human IGF-I ELISA which uses excess IGF-II for feline samples and to evaluate biologic variation. METHODS: Precision was determined by calculating the coefficient of variation (CV). Accuracy was determined by recovery after removal of IGFBP, addition of IGF-I, and linear dilution after the addition of IGFBP. Biologic variation was determined by repeated sampling in 7 cats. RESULTS: There was interference by IGFBP in the high measuring range, resulting in falsely low IGF-I concentrations. This was overcome by the addition of high concentrations of IGF-II. Untreated serum had a measured/expected ratio of 98-115% compared to serum where IGFBP had been removed. Recovery after the addition of IGF-I was 83-112%. Inter- and intra-assay CVs ranged from 2.4% to 5.0% which is within the minimum acceptance criteria based on biologic variation. The reference interval of IGF-I was wide (90-1207 ng/mL) and there was a significant association between body weight and ln IGF-I (P < .000001). CONCLUSIONS: This human ELISA is suitable for feline samples, but interfering IGFBP can cause falsely low concentrations. It is recommended to dilute samples such that IGF-I is < 28 ng/mL on the standard curve to grant for sufficient IGF-II for binding of interferent IGFBP.

Validation of an enzyme-linked immunosorbent assay developed for measuring cortisol concentration in human saliva and serum for its applicability to analyze cortisol in pig saliva.

BACKGROUND: The purpose of this study was to validate a commercially available enzyme-linked immunosorbent assay (ELISA) developed for measuring free cortisol in human saliva and total cortisol concentration in diluted human serum, for its applicability in measuring cortisol concentration in pig saliva. Collection of saliva is less stressful than e.g. blood sampling, and is a non-invasive method. FINDINGS: Saliva was collected by allowing sows to chew on cotton swabs held by forceps. Thereafter, the swabs were centrifuged to retrieve the saliva. The ELISA was performed according to instructions provided by the manufacturer. To validate the ELISA, determination of the intra-assay coefficient of variation (CV), inter-assay CV, recovery, linearity and parallelism was performed. The intra-assay CV was below 10% and inter-assay CV below 15% for samples of high, medium and low cortisol concentrations. The mean recovery was 117% and the linearity and parallelism showed an r2-value of 0.994 and 0.993, respectively. For biological assessment of induced social stress, two saliva samples were collected in the morning from 6 primiparous and 21 multiparous sows. One sample was collected when the sows were individually housed in a farrowing pen and a second sample was collected when the sows were group housed. The primiparous sows had a significant higher cortisol concentration compared to the multiparous sows when group housed. CONCLUSION: The results obtained in this validation study indicate that the ELISA is suitable for measuring cortisol concentration in porcine saliva.

Incidence of pyometra in Swedish insured cats.

Pyometra is a clinically relevant problem in intact female cats and dogs. The etiology is similar in both animal species, with the disease caused by bacterial infection of a progesterone-sensitized uterus. Here, we studied pyometra in cats with the aim to describe the incidence and probability of developing pyometra based on age and breed. The data used were reimbursed claims for veterinary care insurance or life insurance claims or both in cats insured in a Swedish insurance database from 1999 to 2006. The mean incidence rate (IR) for pyometra was about 17 cats per 10,000 cat years at risk (CYAR). Cats with pyometra were diagnosed at a median age of 4 years and a significant breed effect was observed. The breed with the highest IR (433 cats per 10,000 CYAR) was the Sphynx, and other breeds with IR over 60 cats per 10,000 CYAR were Siberian cat, Ocicat, Korat, Siamese, Ragdoll, Maine coon, and Bengal. Pyometra was more commonly diagnosed with increasing age, with a marked increase in cats older than 7 years. The mean case fatality rate in all cats was 5.7%, which is slightly higher than corresponding reports in dogs of 3% to 4%. Geographical location (urban or rural) did not affect the risk of developing the disease. The present study provides information of incidence and probability of developing pyometra based on age, breed, and urban or rural geographical location. These data may be useful for designing cat breeding programs in high-risk breeds and for future studies of the genetic background of the disease.

Outcome of pyometra in female dogs and predictors of peritonitis and prolonged postoperative hospitalization in surgically treated cases.

BACKGROUND: One of the most common diseases in intact bitches is pyometra--a potentially life-threatening disease associated with a variety of clinical and laboratory findings. The aims of the present study were to describe complications of the disease and to investigate clinically useful indicators associated with peritonitis and/or prolonged postoperative hospitalization. RESULTS: A retrospective study was performed using records from 356 bitches diagnosed with pyometra during the years 2006-2007 at the University Animal Hospital, Swedish University of Agricultural Sciences, Uppsala, Sweden. Of the 356 bitches, 315 were surgically treated by ovariohysterectomy, 9 were medically treated and 32 were euthanized without treatment. In the surgically treated bitches, univariable associations between clinical and laboratory data, risk for prolonged hospitalization (≥ 3 days) and/or signs of peritonitis, were analyzed by Chi-square and Fisher's exact test. Logistic regression models were used to assess multivariable associations.The most common complication observed in surgically treated bitches was peritonitis (40 bitches), followed by urinary tract infection (19 bitches), wound infection (8 bitches), uveitis (6 bitches), and cardiac arrhythmia (5 bitches). Leucopenia and fever/hypothermia were associated with increased risk for peritonitis (18-fold and three-fold, respectively). Moderate to severe depression of the general condition, pale mucous membranes and leucopenia were associated with increased risk (seven-fold, three-fold, and over three-point-five-fold, respectively) for prolonged postoperative hospitalization. CONCLUSIONS: Several clinically useful indicators were identified. Leucopenia was the most important marker, associated with 18-fold increased risk for peritonitis and an over three-point-five increased risk for prolonged hospitalization. Fever/hypothermia, depression and pale mucous membranes were associated with increased risk for peritonitis and/or prolonged hospitalization. The results of the present study may be valuable for identifying peritonitis and predicting increased morbidity in surgically treated bitches with pyometra.

Prostasomes from four different species are able to produce extracellular adenosine triphosphate (ATP).

BACKGROUND: Prostasomes are extracellular vesicles. Intracellularly they are enclosed by another larger vesicle, a so called "storage vesicle" equivalent to a multivesicular body of late endosomal origin. Prostasomes in their extracellular context are thought to play a crucial role in fertilization. METHODS: Prostasomes were purified according to a well worked-out schedule from seminal plasmas obtained from human, canine, equine and bovine species. The various prostasomes were subjected to SDS-PAGE separation and protein banding patterns were compared. To gain knowledge of the prostasomal protein systems pertaining to prostasomes of four different species proteins were analyzed using a proteomic approach. An in vitro assay was employed to demonstrate ATP formation by prostasomes of different species. RESULTS: The SDS-PAGE banding pattern of prostasomes from the four species revealed a richly faceted picture with most protein bands within the molecular weight range of 10-150kDa. Some protein bands seemed to be concordant among species although differently expressed and the number of protein bands of dog prostasomes seemed to be distinctly fewer. Special emphasis was put on proteins involved in energy metabolic turnover. Prostasomes from all four species were able to form extracellular adenosine triphosphate (ATP). ATP formation was balanced by ATPase activity linked to the four types of prostasomes. CONCLUSION: These potencies of a possession of functional ATP-forming enzymes by different prostasome types should be regarded against the knowledge of ATP having a profound effect on cell responses and now explicitly on the success of the sperm cell to fertilize the ovum. GENERAL SIGNIFICANCE: This study unravels energy metabolic relationships of prostasomes from four different species.

Shedding of chlamydiae in relation to titers of serum chlamydiae-specific antibodies and serum concentrations of two acute-phase proteins in cats without conjunctivitis.

OBJECTIVE: To investigate shedding of chlamydiae from conjunctiva and genital tracts of cats without clinical signs of conjunctivitis or other infectious disease in relation to their titers of serum antibodies against chlamydiae and to serum amyloid A (SAA) and serum α(1)-acid glycoprotein (AGP) concentrations. ANIMALS: 62 healthy cats. PROCEDURES: Serum from each cat was analyzed for antibodies against chlamydiae and for SAA and AGP concentrations. Swab samples from the conjunctival sac and genital tract were analyzed with a real-time PCR assay for Chlamydiaceae. RESULTS: 4 of 8 of cats with high antibody titers (ie, 1,600) shed chlamydiae, but only from the conjunctiva. Chlamydiae could not be detected in samples from cats with lower antibody titers nor from any genital tract samples. In cats with antibody titers of 1,600, mean ± SD SAA concentration was significantly higher when chlamydiae were detected in conjunctival swab samples (3.9 ± 1.0 mg/L) than when no chlamydiae were detected (1.4 ± 1.0 mg/L). However, SAA concentration was greater than the limit for an acute-phase response in only one of those cats. There was no significant difference in serum AGP concentrations between cats with high titers that were or were not shedding chlamydiae. Nine of 30 (30%) cats (5 with and 4 without detectable serum antibodies against chlamydiae) that had been mated developed reproductive disorders. CONCLUSIONS AND CLINICAL RELEVANCE: Clinically normal cats with high chlamydiae-specific antibody titers can shed and thus transmit chlamydiae. Venereal spread from cats without clinical signs of infection is likely not common.

Expression of interferon gamma in the brain of cats with natural Borna disease virus infection.

Borna disease virus (BDV) is a neurotropic, negative-stranded RNA virus, which causes a non-suppurative meningoencephalomyelitis in a wide range of animals. In cats, BDV infection leads to staggering disease. In spite of a vigorous immune response the virus persists in the central nervous system (CNS) in both experimentally and naturally infected animals. Since the CNS is vulnerable to cytotoxic effects mediated via NK-cells and cytotoxic T-cells, other non-cytolytic mechanisms such as the interferon (IFN) system is favourable for viral clearance. In this study, IFN-γ expression in the brain of cats with clinical signs of staggering disease (N=12) was compared to the expression in cats with no signs of this disease (N=7) by quantitative RT-PCR. The IFN-γ expression was normalised against the expression of three reference genes (HPRT, RPS7, YWHAZ). Cats with staggering disease had significantly higher expression of IFN-γ compared to the control cats (p-value ≤ 0.001). There was no significant difference of the IFN-γ expression in BDV-positive (N=7) and -negative (N=5) cats having clinical signs of staggering disease. However, as BDV-RNA still could be detected, despite an intense IFN-γ expression, BDV needs to have mechanisms to evade this antiviral immune response of the host, to be able to persist.

Morbidity of insured Swedish cats during 1999-2006 by age, breed, sex, and diagnosis.

The aim of the study was to describe patterns of morbidity, as reflected by reimbursed claims for veterinary care among cats covered by a Swedish insurance company during 1999-2006. The annual incidence rates of having at least one veterinary care event were calculated overall and stratified by sex, age, breed, diagnosis and urban/other location. The total rate was 875 (95% CI 858-892) cats with claims per 10,000 cat-years at risk. The Siamese, Burmese, and the Abyssinians had high rates of claims, compared to the Norwegian Forest cat, the Maine Coon, the European Shorthair and domestic cats. The most common causes for insurance claims were trauma, gastrointestinal and lower urinary tract problems and there was substantial breed, sex and age variation for certain problems. For example, among cats under 9 years of age, the rate for lower urinary tract problems was 2-3 times higher in males than females.

The Swedish breeding cat: population description, infectious diseases and reproductive performance evaluated by a questionnaire.

The questionnaire based study gives a combined description of management, infectious diseases and reproductive performance in breeding catteries during 1 year. The mean number of cats per cattery was 6.1, and 25% of the breeders let some of their cats have free access to outdoors. Breeders reported that infection with feline panleukopenia virus, feline immunodeficiency virus or feline leukaemia virus was uncommon, but 8% of the breeders had sold or had themselves owned a cat that died of feline infectious peritonitis. Presence of conjunctivitis was reported by 33.3% of the breeders. Mean litter size was 3.7+/-1.5, with 9.7% stillbirths and 8.3% kitten mortality week 1-12. The percentage of stillborn kittens increased with the age of the queen and litter size, and also differed among breeds. Kitten mortality differed among breeds, but did not increase with age of the queen. Seven percent of the litters were delivered by caesarean section, significantly more during winter and positively associated with presence of stillborn kittens.

Characterization of the bacterial population of the genital tract of adult cats.

OBJECTIVE: To characterize the bacteria of the genital tract in adult cats; assess the effect of estrus, mating, and administration of progestins on those microorganisms in females; and evaluate whether results of bacteriologic culture of vaginal swabs are affected by cleansing of the vulva prior to sampling or by repeated sampling. ANIMALS: 66 female and 29 male cats undergoing routine ovariohysterectomy or castration. PROCEDURE: Specimens were obtained from vaginal and uterine or preputial mucosae with swabs moistened with sterile saline (0.9% NaCl) solution. In 9 cats, vaginal specimens were obtained before and after cleansing of the vulva with ethanol; in 7 female cats, 2 vaginal speci mens were obtained in immediate succession. RESULTS: Aerobic bacteria were most commonly isolated from cats' vaginas and prepuces; anaerobic bacteria were isolated frequently from males (41%) but rarely from females (5%). Generally, culture results were not affected by cleansing of the vulva or repeated vaginal sampling. The bacterial population of the vaginas of cats was influenced by stage of the estrous cycle but not by mating or administration of progestins. Bacteria were not isolated from the uterus of any cat. CONCLUSIONS AND CLINICAL RELEVANCE: In cats, bacteria of the genital tract in females are predominantly aerobic; in males, aerobic and anaerobic bacteria are found. The bacterial population of the vagina is affected by stage of the estrous cycle. Pure growth of bacteria in culture of genital tract specimens is a normal finding; antimicrobials should only be administered if clinical signs of genital infection are present.