Fecal Concentrations of Long-Chain Fatty Acids, Sterols, and Unconjugated Bile Acids in Cats with Chronic Enteropathy.

Chronic enteropathy (CE) in cats encompasses food-responsive enteropathy, chronic inflammatory enteropathy (or inflammatory bowel disease), and low-grade intestinal T-cell lymphoma. While alterations in the gut metabolome have been extensively studied in humans and dogs with gastrointestinal disorders, little is known about the specific metabolic profile of cats with CE. As lipids take part in energy storage, inflammation, and cellular structure, investigating the lipid profile in cats with CE is crucial. This study aimed to measure fecal concentrations of various fatty acids, sterols, and bile acids. Fecal samples from 56 cats with CE and 77 healthy control cats were analyzed using gas chromatography-mass spectrometry, targeting 12 fatty acids, 10 sterols, and 5 unconjugated bile acids. Fecal concentrations of nine targeted fatty acids and animal-derived sterols were significantly increased in cats with CE. However, fecal concentrations of plant-derived sterols were significantly decreased in cats with CE. Additionally, an increased percentage of primary bile acids was observed in a subset of cats with CE. These findings suggest the presence of lipid maldigestion, malabsorption, and inflammation in the gastrointestinal tract of cats with CE. Understanding the lipid alterations in cats with CE can provide insights into the disease mechanisms and potential future therapeutic strategies.

Fecal S100A12 concentrations in cats with chronic enteropathies.

OBJECTIVES: The aim of this study was to compare fecal S100A12 concentrations in cats diagnosed with chronic enteropathy (CE) with healthy control cats. METHODS: This was a prospective, cross-sectional study. Forty-nine cats that had gastrointestinal signs for >3 weeks and a complete diagnostic work-up, including bloodwork, abdominal ultrasound and upper and/or lower gastrointestinal endoscopic biopsies, were enrolled into the CE group. Nineteen cats from the CE group were diagnosed with inflammatory bowel disease (IBD) or chronic inflammatory enteropathy (CIE) and 30 with alimentary lymphoma (LSA), based on histopathology results and additional testing with immunohistochemistry or molecular clonality testing with PCR if indicated. Nineteen apparently healthy control cats were included in the study. One fecal sample was collected from each cat and S100A12 concentrations were quantified by an analytically validated in-house ELISA. RESULTS: Fecal S100A12 concentrations differed between cats with LSA (median 110 ng/g; interquartile range [IQR] 18-548) and control cats (median 4 ng/g; IQR 2-25 [P <0.001]) and between cats with IBD (median 34 ng/g; IQR 15-973) and control cats (P <0.003). S100A12 concentrations in CE cats (median 94 ng/g; IQR 16-548) were statistically significantly higher compared with control cats (P <0.001). The area under the receiver operating characteristic curve (AUROC) to separate healthy cats from CE cats was 0.81 (95% confidence interval [CI] 0.70-0.92) and was statistically significant (P <0.001). The AUROC to separate cats with IBD from cats with LSA was 0.51 (95% CI 0.34-0.68) and was not statistically significant (P = 0.9). CONCLUSIONS AND RELEVANCE: Fecal S100A12 concentrations at the time of diagnostic investigation were higher in cats with CIE and LSA than in healthy controls but did not differ between cats with LSA and those with CIE/IBD. This study is an initial step toward evaluating a novel non-invasive marker of feline CIE. Further studies are needed to determine the diagnostic utility of fecal S100A12 concentrations in cats with CE, including comparing cats with IBD/CIE and LSA, and to compare them with cats with extra-gastrointestinal disease.

Dysbiosis index to evaluate the fecal microbiota in healthy cats and cats with chronic enteropathies.

OBJECTIVES: Previous studies have identified various bacterial taxa that are altered in cats with chronic enteropathies (CE) vs healthy cats. Therefore, the aim of this study was to develop a targeted quantitative molecular method to evaluate the fecal microbiota of cats. METHODS: Fecal samples from 80 client-owned healthy cats and 68 cats with CE were retrospectively evaluated. A panel of quantitative PCR (qPCR) assays was used to measure the fecal abundance of total bacteria and seven bacterial taxa: Bacteroides, Bifidobacterium, Clostridium hiranonis, Escherichia coli, Faecalibacterium, Streptococcus and Turicibacter. The nearest centroid classifier algorithm was used to calculate a dysbiosis index (DI) based on these qPCR abundances. RESULTS: The abundances of total bacteria, Bacteroides, Bifidobacterium, C hiranonis, Faecalibacterium and Turicibacter were significantly decreased, while those of E coli and Streptococcus were significantly increased in cats with CE (P <0.027 for all). The DI in cats with CE was significantly higher compared with healthy cats (P <0.001). When the cut-off value of the DI was set at 0, it provided 77% (95% confidence interval [CI] 66-85) sensitivity and 96% (95% CI 89-99) specificity to differentiate the microbiota of cats with CE from those of healthy cats. Fifty-two of 68 cats with CE had a DI >0. CONCLUSIONS AND RELEVANCE: A qPCR-based DI for assessing the fecal microbiota of cats was established. The results showed that a large proportion of cats with CE had an altered fecal microbiota as evidenced by an increased DI. Prospective studies are warranted to evaluate the utility of this assay for clinical assessment of feline CE.

A pilot study of transdermal gabapentin in cats.

BACKGROUND: Clinical use of gabapentin has increased; transdermal delivery in cats is incompletely studied. OBJECTIVE: To evaluate if gabapentin permeates feline skin in vitro and in vivo and to determine if pain scores improve after administration. ANIMALS: In vitro: cadaver skin from 6 cats; phase 1: 8 young, healthy client-owned cats; phase 2: 15 client-owned geriatric cats. METHODS: In vitro, gabapentin applied every q12h to ear or cervical skin in diffusion cells. Samples collected at 0, 2, 4, 12, and 24 hours after application. Phase 1: Cats assigned to 1 of 4 groups: 5 mg/kg or 10 mg/kg applied q8h for 5 days to either ear or cervical skin. Serum samples collected predose, and after 1 and 5 days. Phase 2: 10 mg/kg applied q8h for 5 days. Two validated pain scores recorded predose, and after days 1, 5, and 8. Serum samples collected predose, and after days 1 and 5. Samples were frozen at -80°C for concentration analysis utilizing a validated high-performance liquid chromatography mass-spectrometry method. RESULTS: Gabapentin was identified in all samples. Significant differences in gabapentin concentrations were observed from day 1 to day 5 (P < .02) and in pain scores from predose to day 5 (P < .05) and day 1 to day 5 (P < .05). No differences in pain scores were observed from predose to day 8 (P = .3). CONCLUSIONS AND CLINICAL RELEVANCE: Gabapentin in a transdermal base penetrates feline skin in vitro, is absorbed systemically in cats, and may help decrease pain scores.

Evaluation of whisker stress in cats.

OBJECTIVES: The aim of this study was to determine if cats fed from a commercially advertised whisker-friendly dish vs their normal food dish would spend more time at the food dish, eat more and drop less food. METHODS: Forty indoor cats were enrolled in the study. Owners fasted their cats for 12 h and fed them their normal measured amount of dry food in their normal dish. Owners filmed their cats eating for up to 5 mins, and measured how much food was eaten and dropped from the dish. Owners then switched to feeding their cats from a whisker-friendly dish for a 7-day transition period. Following this transition, owners were instructed to fast their cats for 12 h and then feed them their normal food from the new dish and film them eating, as previously described. The following day the owners offered food in both dishes to determine their cat's preference. RESULTS: No evidence was found that eating from the whisker-friendly dish increased the amount of time spent eating (P = 0.8), decreased the amount of food dropped (P = 0.9) or increased the amount of food eaten (P = 0.7). The estimated probability for the cats to prefer the whisker-friendly dish was 0.74 with a 95% confidence interval. CONCLUSIONS AND RELEVANCE: Cats fed from a whisker-friendly dish did not spend more time eating, drop less food or eat more food in a 5-min period. Some cats appeared to prefer the new whisker-friendly dish over their normal food dish. Overall, food dish-associated whisker stress did not affect the eating habits of the study cats.

Assessment of Veterinary Students' Responses to Questions Regarding Small Animal Pain Recognition and Analgesic Treatment Options.

Pain recognition and treatment in companion animals are important aspects of veterinary medicine, yet the teaching of these concepts may not be adequate at all academic institutions. This study was designed to evaluate veterinary students' ability to recall signs of pain and specific analgesic drugs in dogs and cats. We hypothesized that students in the fourth, or final, year of their veterinary curriculum would have a better understanding of pain recognition and be able to recall more analgesic options. A brief, voluntary, and anonymous open question survey was made available to all veterinary students, years 1 to 4, at our institution. The questions included, "How does a cat/dog show signs of pain?" and "What pain medications are used in cats/dogs?" Survey responses were collated according to the students' year in the curriculum, and the most common responses for signs of pain and analgesic medications recalled by the students in both the cat and dog were compared for significant differences. Results showed that students in the class of 2017 (seniors) had no superior recall of analgesic medications or recognition of pain in cats or dogs compared to the other classes. Vocalization was the most common sign of pain recalled with at least 50% responses from all classes. Carprofen was the most commonly recalled analgesic for dogs (the difference between classes, p = .04). Meloxicam was the most commonly recalled analgesic for cats (the difference among classes, p < .001). Based on these results, areas of improvement were identified for our analgesic curriculum.

Pharmacometabolomics with a combination of PLS-DA and random forest algorithm analyses reveal meloxicam alters feline plasma metabolite profiles.

Repeated administration of meloxicam to cats is often limited by the potential damage to multiple organ systems. Identifying molecules that predict the adverse effects of meloxicam would help to monitor and individualize its administration, maximizing meloxicam's beneficial effects. The objectives of this study were to (a) determine if the repeated administration of meloxicam to cats alters the plasma metabolome and (b) identify plasma metabolites that may serve to monitor during the administration of meloxicam in cats. Purpose bred young adult cats (n = 12) were treated with meloxicam at 0.3 mg/kg or saline subcutaneously once daily for up to 17 days. An untargeted metabolomics approach was applied to plasma samples collected prior to and at designated time points after meloxicam or saline administration. To refine the discovery of biomarkers, the machine-learning algorithms, partial least squares discriminant analysis (PLS-DA) and random forest (RF), were trained and validated using a separate unrelated group of meloxicam- and saline-treated cats (n = 8). A total of 74 metabolites were included in the statistical analysis. Metabolomic analysis shows that the repeated administration of meloxicam alters multiple substances in plasma, including nonvolatile organic acids, aromatic amino acids, monosaccharides, and inorganic compounds as early as four days following administration of meloxicam. Seventeen plasma molecules were able to distinguish meloxicam-treated from saline-treated cats. The metabolomic changes discovered in this study may help to unveil unknown mechanisms of NSAID-induced side effects. In addition, some metabolites could be valuable for individualizing the administration of meloxicam to cats to mitigate adverse effects.

Urinary chemical fingerprint left behind by repeated NSAID administration: Discovery of putative biomarkers using artificial intelligence.

Prediction and early detection of kidney damage induced by nonsteroidal anti-inflammatories (NSAIDs) would provide the best chances of maximizing the anti-inflammatory effects while minimizing the risk of kidney damage. Unfortunately, biomarkers for detecting NSAID-induced kidney damage in cats remain to be discovered. To identify potential urinary biomarkers for monitoring NSAID-based treatments, we applied an untargeted metabolomics approach to urine collected from cats treated repeatedly with meloxicam or saline for up to 17 days. Applying multivariate analysis, this study identified a panel of seven metabolites that discriminate meloxicam treated from saline treated cats. Combining artificial intelligence machine learning algorithms and an independent testing urinary metabolome data set from cats with meloxicam-induced kidney damage, a panel of metabolites was identified and validated. The panel of metabolites including tryptophan, tyrosine, taurine, threonic acid, pseudouridine, xylitol and lyxitol, successfully distinguish meloxicam-treated and saline-treated cats with up to 75-100% sensitivity and specificity. This panel of urinary metabolites may prove a useful and non-invasive diagnostic tool for monitoring potential NSAID induced kidney injury in feline patients and may act as the framework for identifying urine biomarkers of NSAID induced injury in other species.

Pilot study of the effect of gastrointestinal diets on fecal occult blood testing in cats.

OBJECTIVES: The aim of this study was to assess the methodology of guaiac-based fecal occult blood (FOB) testing in healthy cats, and to examine the effect of feeding selected gastrointestinal diets (canned and dry) on FOB results. METHODS: Stool obtained from three cats was mixed with incremental quantities of EDTA whole blood. FOB tests were performed in triplicate and interpreted at 5 mins, and at 24, 48 and 72 h post-test set-up. Ten cats were fed their normal diets, a dry hydrolyzed protein diet, a dry gastrointestinal diet and then transitioned back to their normal diet over a 7-week period. A subsequent study was repeated with similar hydrolyzed protein and gastrointestinal canned diets. Two fecal samples were tested per cat each week using human point-of-care guaiac tests. RESULTS: Ten microliters of whole feline blood could be reliably detected in feline stool at all time points evaluated. There was no evidence of an association between the dry hydrolyzed and dry gastrointestinal diets and a positive FOB test result (P = 0.33). Cats fed a canned gastrointestinal diet had a significantly higher probability of producing a positive FOB result than when on their normal diet (P <0.01). Feeding the canned hydrolyzed diet was not associated with an increased probability of a positive FOB test vs the normal diet (P = 0.94). CONCLUSIONS AND RELEVANCE: Small amounts of blood in feline stool can be reliably and repeatedly detected as early as 5 mins post-FOB test set-up, making this a user-friendly bench-top screening test in a veterinary setting. Cats fed a single dry hydrolyzed, dry gastroenteric or canned hydrolyzed diet in this study had a low chance of false-positive FOB test results, while those fed the canned gastrointestinal diet had a significantly greater likelihood of producing false-positive results.

Pharmacokinetics of mycophenolic acid and its effect on CD4+ and CD8+ T cells after oral administration of mycophenolate mofetil to healthy cats.

BACKGROUND: Mycophenolate mofetil (MMF) is an immunosuppressant used in human and veterinary medicine. Little pharmacokinetic and pharmacodynamic information on MMF is available in cats. OBJECTIVE: To evaluate the plasma disposition of mycophenolic acid (MPA) and assess its effect on total peripheral blood mononuclear cells and CD4+ /CD8+ ratios after PO administration of MMF. ANIMALS: Healthy cats (n = 10). METHODS: Mycophenolate mofetil was administered at a dosage of 10 mg/kg q12h (n = 3), 15 mg/kg q12h (n = 3), and 15 mg/kg q8h (n = 4) for 7 days. Concentrations of MPA and derivatives were determined using ultra-high-performance liquid chromatography. Flow cytometry was used to assess CD4+ /CD8+ T-cell ratios. RESULTS: All cats biotransformed MMF into MPA. Half of the cats (5/10) had adverse effects within 1 week of MMF administration. Area under the curve limit of quantification (AUC0-LOQh ) of MPA ranged from 1.27 to 2.03 hours·µg/mL and from 1.77 to 8.54 hours·µg/mL after the first and last PO dose of 10 mg/kg. The AUC0-loqh of MPA ranged from 2.18 to 31 hours·µg/mL after the first dose of 15 mg/kg of MMF. Before the first dose of MMF, the average total number of PBMC ranged from 1.2 to 9.3 million/mL. At the last dose of MMF, the average total number of PBMC ranged from 3 to 5 million/mL. CONCLUSION: Mycophenolic acid was detected in all cats. The dose 10 mg/kg given q12h for 1 week was tolerated (n = 3). The efficacy of MMF as an immunosuppressant and long-term safety in cats of this dosage regimen is unknown.

Understanding the effect of repeated administration of meloxicam on feline renal cortex and medulla: A lipidomics and metabolomics approach.

Repeated administration of meloxicam can cause kidney damage in cats by mechanisms that remain unclear. Metabolomics and lipidomics are powerful, noninvasive approaches used to investigate tissue response to drug exposure. Thus, the objective of this study was to assess the effects of meloxicam on the feline kidney using untargeted metabolomics and lipidomics approaches. Female young-adult purpose-breed cats were allocated into the control (n = 4) and meloxicam (n = 4) groups. Cats in the control and meloxicam groups were treated daily with saline and meloxicam at 0.3 mg/kg subcutaneously for 17 days, respectively. Renal cortices and medullas were collected at the end of the treatment period. Random forest and metabolic pathway analyses were used to identify metabolites that discriminate meloxicam-treated from saline-treated cats and to identify disturbed metabolic pathways in renal tissue. Our results revealed that the repeated administration of meloxicam to cats altered the kidney metabolome and lipidome and suggest that at least 40 metabolic pathways were altered in the renal cortex and medulla. These metabolic pathways included lipid, amino acid, carbohydrate, nucleotide and energy metabolisms, and metabolism of cofactors and vitamins. This is the first study using a pharmacometabonomics approach for studying the molecular effects of meloxicam on feline kidneys.

Pharmacokinetics and pharmacodynamics of mycophenolic acid in healthy cats after twice-daily intravenous infusion of mycophenolate mofetil for three days.

OBJECTIVE To evaluate the plasma disposition of mycophenolic acid (MPA) and its derivatives MPA glucuronide and MPA glucoside after twice-daily infusions of mycophenolate mofetil (MMF) in healthy cats for 3 days and to assess the effect of MMF administration on peripheral blood mononuclear cell (PBMC) counts and CD4+-to-CD8+ ratios. ANIMALS 5 healthy adult cats. PROCEDURES MMF was administered to each cat (10 mg/kg, IV, q 12 h for 3 days). Each dose of MMF was diluted with 5% dextrose in water and then administered over a 2-hour period with a syringe pump. Blood samples were collected for analysis. A chromatographic method was used to quantitate concentrations of MPA and its metabolites. Effects of MMF on PBMC counts and CD4+-to-CD8+ ratios were assessed by use of flow cytometry. RESULTS All cats biotransformed MMF into MPA. The MPA area under the plasma concentration-time curve from 0 to 14 hours ranged from 14.6 to 37.6 mg·h/L and from 14.4 to 22.3 mg·h/L after the first and last infusion, respectively. Total number of PBMCs was reduced in 4 of 5 cats (mean ± SD reduction, 25.9 ± 15.8% and 26.7 ± 19.3%) at 24 and 48 hours after the end of the first infusion of MMF, respectively. CONCLUSIONS AND CLINICAL RELEVANCE Plasma disposition of MPA after twice-daily IV infusions for 3 days was variable in all cats. There were no remarkable changes in PBMC counts and CD4+-to-CD8+ ratios.

Safety of oral and intravenous mycophenolate mofetil in healthy cats.

Objectives The aim of this study was to evaluate the safety and clinical effects of intravenous (IV) and oral mycophenolate mofetil (MMF) in healthy cats. Methods A total of 24 healthy adult cats weighing >3.5 kg were either given IV MMF (over a 2 h infusion) or oral MMF. The dosages used were as follows: 5 mg/kg IV once (n = 2), 10 mg/kg q12h IV for 1 day (n = 1), 20 mg/kg q12h IV for 1 day (n = 6) and 10 mg/kg q12h IV for 3 days (n = 5). Blood was collected from each cat at intervals of up to 12 h from the last dose for analysis purposes. Oral MMF was given at 10 mg/kg q12h for 7 days (n = 3), 15 mg/kg q12h for 7 days (n = 3) and 15 mg/kg q8h for 7 days (n = 4). Results Side effects to MMF were minimal. There was no anorexia or vomiting noted in any of the cats during or after IV medication administration. Only 4/14 cats had diarrhea from 12-48 h after IV administration. There was hyporexia in 1/10 cats given oral MMF and no vomiting noted. In 5/10 cats given oral MMF, there was diarrhea between days 2 and 7 of the study. Conclusions and relevance Cats tolerate MMF at an IV dose of 10 mg/kg q12h for 3 days and an oral dose ⩽15 mg/kg q12h for up to 7 days. There seems to be a dose-dependent incidence of gastrointestinal side effects. MMF may be a useful alternative immunosuppressant to be considered for use in some cats.

Development and validation of an ultrafast chromatographic method for quantification of the immunosuppressant mycophenolic acid in canine, feline and human plasma.

Mycophenolic acid (MPA) is the active metabolite of the prodrug mycophenolate mofetil. In this study, we developed and validated a novel ultra-high performance liquid chromatography (UHPLC) method for the rapid quantification of MPA in plasma from dogs, cats and humans. Following the protein precipitation, calibration standards and quality controls were separated by UHPLC reversed-phase on a 1.5µm 2.1×100mmC18 column and quantified using UV detection at 215nm. The procedure produced a linear curve (r2>0.997) over the concentration range 0.4-50µg/mL and exhibited a high degree of repeatability (CV% <11%). The limit of detection (LOD) and lower limit of quantitation (LLOQ) were 0.1 and ≤0.4µg/mL, respectively and the overall recovery was ≥87%. By combining isocratic conditions with a UHPLC column containing solid core particles, we were able to elute MPA and the internal standard (mycophenolic acid carboxybutoxy ether) within 3.0min. The short total run time makes this method ideal to study the disposition of MPA in large batches of plasma samples and/or monitor plasma drug concentrations, as recommended for patients that require optimized immunosuppression.

Prevalence of Cytauxzoon felis infection in healthy free-roaming cats in north-central Oklahoma and central Iowa.

Case summary Cytauxzoonosis is a tick-borne disease of cats, and Oklahoma (OK), USA, is considered an enzootic state. To determine the prevalence of Cytauxzoon felis, blood was collected from free-roaming cats, as they are frequently exposed to tick vectors. Our objective was to determine the prevalence of C felis infection in free-roaming cats in north-central Oklahoma and central Iowa (IA). Infection with C felis was determined using DNA extracted from blood and PCR amplification. Blood was collected from 380 free-roaming cats between January and April in 2014 in OK. DNA from C felis was detected in 3/380 (0.8%; 95% confidential interval [CI] 0.22-2.3%). In IA, 292 blood samples were collected between 2012 and 2014. No C felis-infected cats were detected (0; 95% CI 0-0%). Relevance and novel information The prevalence of C felis (0.8%) in north-central OK reported herein was lower than the previously reported 3.4% in domestic cats in OK. Our study supports that the prevalence in a given enzootic area can vary by location and from the pool of cats sampled. None of 291 (0%) cats were infected with C felis in central IA. To date, only one case of cytauxzoonosis in a domestic cat has been reported in IA. It is important to monitor cats for C felis infections in northern US states, as geographic distribution of Amblyomma americanum expands northward. As free-roaming cats have more contact with the tick vectors of C felis, this population allows us to monitor the expansion of C felis distribution.

Extraskeletal Osteosarcoma Induced by a Foreign Body Granuloma.

An 8 yr old spayed female Italian greyhound was presented with a mass in the cranial abdomen. Preliminary evaluation of the dog revealed a large, cavitary, irregularly shaped mass with no definitive association with any abdominal organs. During an exploratory celiotomy, a 16 cm × 12 cm × 6 cm mass was removed. On subsequent histopathology, extraskeletal osteosarcoma induced by a foreign body granuloma was diagnosed. The foreign body granuloma, based on histopathological findings, was suspected to be secondary to a retained surgical sponge from her routine ovariohysterectomy performed 7 yr prior to presentation. Animals with granulomas induced by foreign bodies can remain asymptomatic for years; however, those granulomas can progress to extraskeletal osteosarcomas, which carry a poor prognosis.

Toxic shock syndrome in two dogs.

Two young, unrelated, spayed female Labrador retrievers were evaluated for severe, diffuse, generalized erythema and edema of the skin. Both dogs exhibited signs of disseminated intravascular coagulopathy and were euthanized. On postmortem examination, toxic shock syndrome (TSS) was diagnosed based on histopathology and supported by skin cultures. TSS is a rarely reported disease in veterinary medicine and can cause acute and profound clinical signs. Rapid recognition of this disease process and immediate treatment may improve the clinical outcome.