Particulate matter from wildfire smoke in northern Colorado appears to be associated with conjunctivitis in dogs.

OBJECTIVE: To evaluate ocular surface parameters in dogs with normal eyes when exposed to 3 different air quality index (AQI) categories corresponding to levels of normal air pollutants ("good," 0 to 50; "moderate," 51 to 100) and wildfire smoke ("smoke," 101 to 150). ANIMALS: 15 privately owned dogs. METHODS: A prospective cohort study with dogs living in northern Colorado. Ocular surface parameters (conjunctival chemosis and hyperemia, Schirmer tear test-1, tear film break-up time, fluorescein stain, conjunctival microbiology, etc) were evaluated when the AQI was reported in 1 of the 3 categories (good, moderate, and smoke) for 3 consecutive days. The AQI and air pollutant levels (particulate matter < 2.5 µm in diameter [PM2.5], ozone, etc) were retrieved from the AirNow database. RESULTS: Due to scheduling conflicts, only 7 dogs were examined during the smoke category. Average AQI in the 3 categories were good, 44.1; moderate, 73.7; and smoke, 103.7. The odds for more severe hyperemia and more severe chemosis for smoke were 5.39 and 7,853.02 times the odds, respectively, when compared to good AQI. Additionally, the odds for more severe chemosis were 34,656.62 times the odds for smoke when compared to moderate AQI. A significant relationship was found between chemosis and PM2.5. CONCLUSION: Exposure to increased AQI related to wildfire smoke caused a significant increase in conjunctivitis. The significant relationship between chemosis and PM2.5 could indicate that PM2.5 in wildfire smoke is associated with an inflammatory factor. CLINICAL RELEVANCE: Preventive measures (eg, use of eyewash, artificial tears, or eye protection) for dogs that are exposed to wildfire smoke should be instituted to decrease the risk of ocular irritation.

Fipronil and (S)-methoprene can lessen the risk of transmission of Bartonella clarridgeiae among cats with exposure to Ctenocephalides felis.

OBJECTIVE: To cohouse cats experimentally infected with Bartonella clarridgeiae (Bc) with naive cats in a flea-free environment or with Ctenocephalides felis, Bartonella henselae (Bh), Mycoplasma haemofelis, and Candidatus Mycoplasma haemominutum to determine which flea could be a vector and to assess whether transmission of the infectious agents could be blocked by fipronil and (S)-methoprene. ANIMALS: Specific pathogen-free cats (n = 34). METHODS: In experiment 1, Bc was inoculated in 1 cat that was housed with 9 naive cats without C felis. In experiment 2, the 2 cats inoculated with Bc were housed with 6 other cats (2 inoculated with Bh, 2 inoculated with M haemofelis, and 2 inoculated with Candidatus M haemominutum) in the center (enclosure 2) of 3 housing enclosures separated by mesh walls that allow passage of fleas but precludes fighting. C felis were placed only on cats in enclosure 2 (5 times). Cats in enclosures 1 (n = 8) and 2 (8) were untreated, and cats in enclosure 3 (8) were administered fipronil and (S)-methoprene. Blood was collected from all cats for PCR assays for the pathogens. RESULTS: None of the cats housed with the cat inoculated with Bc became PCR positive in the absence of C felis. All cats in enclosure 2 became Bc DNA positive. While 2 of 8 cats in enclosure 1 became Bc PCR positive, none of the treated cats in enclosure 3 became infected. CLINICAL RELEVANCE: The study demonstrated that C felis can be a vector for Bc. The results support the recommendation that flea control products can reduce the risk of transmission of flea-borne pathogens.

Comparison of Antiviral Immune Responses in Healthy Cats Induced by Two Immune Therapeutics.

BACKGROUND: Effective immunotherapeutic agents for use in cats are needed to aid in the management of intractable viral diseases, including feline infectious peritonitis (FIP) infection. The objectives of this study were to compare two different immune stimulants for antiviral activity in cats: (1) TLR 2/6-activating compound polyprenyl immunostimulant; (PI) and (2) liposome Toll-like receptor 3/9 agonist complexes (LTCs) to determine relative abilities to stimulate the induction of type I (IFN-α, IFN-ß) and type II (IFN-γ) interferon immune responses in vitro and to study the effects of treatment on immune responses in healthy cats. METHODS: Cytokine and cellular immune responses to PI and LTC were evaluated using peripheral blood mononuclear cells (PBMCs) from healthy cats incubated with LTC and PI at indicated concentrations using reverse transcriptase polymerase chain reaction assays and ELISA assays. The effects of the immune stimulants on inhibiting FIPV replication were assessed using a feline macrophage cell line (fcwf-4). Cytokine and cellular immune responses to PI and LTC were evaluated in blood samples from healthy cats treated with PI and LTC, using reverse transcriptase polymerase chain reaction (RT-PCR) and ELISA assays. RESULTS: In the in vitro studies, both compounds triggered the upregulated expression of IFN-α, IFN-γ, and IL-1ß genes in cat PBMC, whereas treatment with LTC induced significantly greater expression of IFN-α and IFN-γ on Day 1 and IL-1b on Day 3. There was significant protection from FIPV-induced cytopathic effects when fcwf-4 cells were treated with conditioned medium from LTC-activated leukocytes. In the healthy cat study (in vivo), both PI and LTC increased the mRNA signal for IFN-α, IFN-γ, and IL-1ß above baseline at multiple time points with statistically greater increases in the LTC group on either Day 1 (IFN-α, IFN-γ) or Day 3 (IL-1ß). In addition, RANTES increased over time in cats treated with the LTC. CONCLUSIONS: Both LTC and PI protocols induced immune-enhancing effects, suggesting a possible clinical use for the management of chronic infectious diseases like FIP. Activating the TLR 3 and 9 pathways (LTC) induced superior broad interferon production in vitro than the activation of the TLR 2 and 6 pathways (PI).

Single cell transcriptomic analysis of the canine duodenum in chronic inflammatory enteropathy and health.

Chronic inflammatory enteropathy (CIE) is a common condition in dogs causing recurrent or persistent gastrointestinal clinical signs. Pathogenesis is thought to involve intestinal mucosal inflammatory infiltrates, but histopathological evaluation of intestinal biopsies from dogs with CIE fails to guide treatment, inform prognosis, or correlate with clinical remission. We employed single-cell RNA sequencing to catalog and compare the diversity of cells present in duodenal mucosal endoscopic biopsies from 3 healthy dogs and 4 dogs with CIE. Through characterization of 35,668 cells, we identified 31 transcriptomically distinct cell populations, including T cells, epithelial cells, and myeloid cells. Both healthy and CIE samples contributed to each cell population. T cells were broadly subdivided into GZMAhigh (putatively annotated as tissue resident) and IL7Rhigh (putatively annotated as non-resident) T cell categories, with evidence of a skewed proportion favoring an increase in the relative proportion of IL7Rhigh T cells in CIE dogs. Among the myeloid cells, neutrophils from CIE samples exhibited inflammatory (SOD2 and IL1A) gene expression signatures. Numerous differentially expressed genes were identified in epithelial cells, with gene set enrichment analysis suggesting enterocytes from CIE dogs may be undergoing stress responses and have altered metabolic properties. Overall, this work reveals the previously unappreciated cellular heterogeneity in canine duodenal mucosa and provides new insights into molecular mechanisms which may contribute to intestinal dysfunction in CIE. The cell type gene signatures developed through this study may also be used to better understand the subtleties of canine intestinal physiology in health and disease.

Pradofloxacin for Treatment of Bartonella henselae in Experimentally Inoculated Cats.

Bartonella henselae is associated with numerous clinical syndromes in people. Cats are the definitive hosts for B. henselae, develop high levels of bacteremia, and are associated with human infections, particularly in the presence of Ctenocephalides felis. Several antibiotic protocols used for the treatment of B. henselae infection in cats have failed to clear bacteremia. The purpose of this study was to assess the safety and efficacy of a high-dose pradofloxacin protocol to eliminate B. henselae bacteremia. Bartonella henselae infection was initiated in 8 cats by intravenous inoculation of infected feline blood and then pradofloxacin was administered at 7.5 mg/kg, PO, twice daily for 28 days, starting 12 weeks after inoculation. Complete blood cell counts were performed prior to pradofloxacin administration and then every 2 weeks for 10 weeks. Bartonella PCR assay was performed prior to pradofloxacin administration and approximately every 2 weeks for 10 weeks and then weekly for 4 weeks. Methylprednisolone acetate (5 mg/kg) was administered by intramuscular injection to all cats on week 10. The cats remained normal and none developed a hematocrit, platelet count, lymphocyte count, or neutrophil count outside of the normal reference ranges. In the one month prior to pradofloxacin administration, all cats were PCR-positive for Bartonella DNA on at least two of four sample dates; after pradofloxacin administration, all cats were negative for B. henselae DNA in blood on all nine sample dates. The protocol appears to be safe and failure to amplify B. henselae DNA from the blood after the administration of pradofloxacin and one dose of methylprednisolone acetate suggests either an antibiotic effect or the organism was cleared spontaneously.

Total thyroxine and thyroid-stimulating hormone responses of healthy cats to different doses of thyrotropin-releasing hormone.

Thyrotropin-releasing hormone (TRH) stimulation can be used as a test of thyroid function and pituitary thyrotropin (thyroid-stimulating hormone, TSH) reserve, but optimal stimulation testing protocols in cats are unreported. We randomly divided 6 healthy young adult cats into 3 groups of 2 and administered 3 different intravenous doses of TRH (0.01, 0.05, 0.1 mg/kg) at weekly intervals in our crossover study. Serum TSH and thyroxine (T4) concentrations were measured using chemiluminescent immunoassay before, and at 30 and 60 min after, TRH administration. All cats were monitored for 4 h post-TRH administration for side effects. All 3 TRH doses induced significant TSH (0.01 mg/kg, p = 0.001; 0.05 mg/kg, p = 0.002; 0.1 mg/kg, p = 0.006) and total T4 (0.01 mg/kg, p = 0.008; 0.05 mg/kg, p = 0.006; 0.1 mg/kg, p = 0.001) responses. Lower TRH doses (0.01 and 0.05 mg/kg) caused fewer side effects (1 of 6 cats) than did the highest dose (3 of 6 cats), and may be safer in cats than the previously reported higher dose (0.1 mg/kg) of TRH. Our results do not support the use of maropitant to prevent side effects of a TRH stimulation test in cats.

Efficacy of an elemental diet in achieving clinical remission in dogs with chronic enteropathy.

BACKGROUND: Diet may induce clinical remission in dogs with chronic enteropathy (CE). Elemental diets (EDs), providing protein as amino acids, modulate intestinal immunity and microbiome in rodents and humans. HYPOTHESIS: Evaluate the impact of an amino acid-based kibble (EL) on CE clinical activity and gastrointestinal (GI)-relevant variables. ANIMALS: Client-owned dogs (n = 23) with inadequately controlled CE. METHODS: Prospective, uncontrolled clinical trial. Diagnostic evaluation including upper and lower GI endoscopy was performed before study entry. Canine chronic enteropathy clinical activity index (CCECAI), serum biomarkers, and fecal microbiome were evaluated before and after 2 weeks of EL. Dogs with stable or improved CE remained in the study for another 6 weeks. Pre- and post-EL clinical and microbiological variables were compared statistically using a mixed model. RESULTS: After 2 weeks of EL, 15 of 22 dogs (68%; 95% confidence interval [CI], 47%-84%) consuming the diet were classified as responders with a median (range) decrease in CCECAI from 6 (3-12) to 2 (0-9; P < .001). Fourteen of 15 responders and 2/7 nonresponders at 2 weeks completed the trial; all 16 were experiencing adequate control at week 8 with a median CCECAI of 2 (0-3). In total, 16/23 dogs (70%; 95% CI, 49%-84%) were responders. Feeding EL caused shifts in fecal bacterial communities, which differed between responders and nonresponders. Serum biomarker concentrations were unchanged throughout the study apart from serum alkaline phosphatase activity. CONCLUSIONS: Exclusive feeding of EL improved clinical signs in 16 of 23 dogs with uncontrolled CE. Fecal microbiome shifts were associated with response to diet and may represent a mechanism for clinical improvement.

Effect of repeated administration of a parenteral feline herpesvirus-1, calicivirus, and panleukopenia virus vaccine on select clinicopathologic, immunological, renal histologic, and immunohistochemical parameters in healthy adult cats.

OBJECTIVE: To assess whether hyperinoculation of cats with a feline herpesvirus-1, calicivirus, and panleukopenia virus (FVRCP) vaccine could be used as a model to study interstitial nephritis and to assess humoral and cell-mediated immune responses toward vaccinal α-enolase. ANIMALS: 6 healthy young adult purpose-bred research cats. PROCEDURES: Baseline renal cortical biopsies, whole blood, serum, and urine were collected prior to administration of a commercial FVRCP parenteral vaccine. Vaccine hyperinoculation was defined as a total of 8 vaccinations given at 2-week intervals over a 14-week period. Blood samples were collected immediately prior to each vaccination, and a second renal biopsy was performed 2 weeks after hyperinoculation (week 16). Renal histopathology, renal α-enolase immunohistochemistry, and assays to detect humoral and cell-mediated immune reactions against Crandell-Rees feline kidney (CRFK) cell lysates and α-enolase were performed. An α-enolase immunoreactivity score for renal tubules and glomeruli based on signal intensity was determined by a blinded pathologist. RESULTS: Hyperinoculation with the vaccine was not associated with clinicopathologic evidence of renal dysfunction, and interstitial nephritis was not recognized by light microscopy in the time studied. The mean serum absorbance values for antibodies against CRFK antigen and α-enolase were significantly (P < 0.001) higher at weeks 4, 8, and 16 versus week 0. Renal tubular and glomerular α-enolase immunoreactivity scores were higher at week 16 compared to baseline. CLINICAL RELEVANCE: Findings suggested that systemic immunological reactions occurred and renal tissues were affected by vaccine hyperinoculation; however, short-term FVRCP vaccine hyperinoculation cannot be used to study interstitial nephritis in cats.

Pro-inflammatory cytokines in aqueous humor from dogs with anterior uveitis and post-operative ocular hypertension following phacoemulsification, primary glaucoma, and normal healthy eyes.

BACKGROUND: The aim of this study was to evaluate the levels of pro-inflammatory cytokines in aqueous humor (AH) from dogs with anterior uveitis and post-operative ocular hypertension (POH) following phacoemulsification, in AH from dogs with primary glaucoma, and in normal healthy eyes with no signs of anterior uveitis or other ocular diseases. METHODS: An exploratory study including 21 samples of AH collected from 15 dogs; post-phacoemulsification with anterior uveitis and POH ('POH group', n = 10 samples), primary glaucoma ('glaucoma group', n = 6 samples), and normal ('normal group', n = 5 samples). Target mass spectrometry via multiple reaction monitoring (MRM-MS) with the Canine Cytokine SpikeMix™ as internal standard was used to measure the pro-inflammatory cytokine levels. RESULTS: The MRM-MS method measured 15 pro-inflammatory cytokines. Tumor-necrosis-factor-alpha (TNFα) and interleukin-18 (IL-18) levels in AH were different between all three groups (glaucoma>POH>normal) (p = .05, p = .02, respectively). Additionally, IL-6 was higher in the 'POH group' compared to the 'glaucoma group' (p = .04) and IL-4 was higher in the 'POH group' compared to the 'normal group' (p = .04). Intraocular pressure (IOP) was positively associated with increased AH levels of IL-18 (Spearman correlation = .64, p = .03). CONCLUSIONS: MRM-MS using the Canine Cytokine SpikeMix™ as an internal standard was established as a method to detect pro-inflammatory cytokine levels in canine AH. The study demonstrated increased levels of IL-4, IL-6, IL-18, and TNFα in AH from canines with POH following phacoemulsification. Primary glaucomatous eyes had the highest levels of IL-18 and TNFα which may indicate that inflammation plays a role in the pathogenesis of primary glaucoma in dogs.

Efficacy of feeding a diet containing a high concentration of mixed fiber sources for management of acute large bowel diarrhea in dogs in shelters.

BACKGROUND: Use of diets with increased concentrations of dietary fiber is thought to be beneficial in the management of dogs with large bowel diarrhea. OBJECTIVE: To determine whether feeding a diet with high concentrations of soluble and insoluble fiber to dogs with acute colitis would be superior to feeding a diet with typical fiber levels. ANIMALS: A total of 52 dogs with acute signs of large bowel diarrhea housed in an animal shelter were entered into the study; 11 dogs per diet completed the protocol. METHODS: In this randomized, prospective study, dogs with a fecal score of 4, 5, 6, or 7 and signs of acute colitis were fed a high fiber diet (4.54% soluble; 15.16% insoluble fiber) or a standard diet (0.6% soluble; 5.33% insoluble fiber) and fecal scores compared over the course of the study with significance defined as P < .05. RESULTS: All dogs fed the high fiber diet (11/11; 100%) had a fecal score <5 on the day of adoption or day 9, which was statistically different (P < .04) than dogs fed the standard diet (6/11 dogs; 55%; 95% CI: 23-83). The proportions of stools with a fecal score >4 were greater (P = .0001) in the dogs fed the standard diet (29/48 samples; 60%; 95% CI: 45-74) compared to the high fiber diet (8/50 samples; 16%; 95% CI: 7-29). CONCLUSIONS AND CLINICAL IMPORTANCE: The results support feeding the high fiber diet described herein to dogs with acute large bowel diarrhea.

Associations between presence of Bartonella species deoxyribonucleic acid and complete blood cell count and serum biochemical changes in client-owned cats.

BACKGROUND: Infection with Bartonella species is common in cats but reported effects of bacteremia on laboratory variables differ. OBJECTIVES: Evaluate for associations between Bartonella bacteremia and CBC and serum biochemical changes in sick and healthy cats throughout the United States. ANIMALS: A total of 3964 client-owned cats. METHODS: Retrospective cohort study using submissions to a commercial laboratory between 2011 and 2017. Serum biochemistry and CBC abnormalities (categorized as above or below reference intervals), age, and location (high- or low-risk state for Ctenocephalides felis) in presumed healthy and sick cats were evaluated for associations with presence of Bartonella spp. DNA, detected by PCR. Univariate and multivariable logistic regression analyses were performed. RESULTS: Bartonella spp. DNA was amplified from 127 (3.2%) of 3964 cats; 126 (99.2%) of 127 were from high flea risk states and 121 (95.3%) of 127 were presumed sick. Fever of unknown origin was the most common PCR panel requested. In the multivariable analysis, neutrophilia, decreased ALP activity, clinical status (presumed sick), and young age (≤2 years) each were positively associated whereas neutropenia and hyperproteinemia both were negatively associated with Bartonella spp. bacteremia. Presence of Bartonella spp. DNA had no association with test results for other infectious disease agents. CONCLUSIONS AND CLINICAL IMPORTANCE: In both healthy and sick cats, active Bartonella infections had minimal association with clinically relevant laboratory abnormalities. However, based on these results, in areas considered high risk for C. felis, active infection with Bartonella spp. is a reasonable differential diagnosis for cats presented with unexplained fever and neutrophilia, particularly if the cat is young.

Increase in infected corneal ulcerations in dogs during the northern Colorado's 2020 wildfire season.

OBJECTIVE: In the fall of 2020, Colorado experienced the two largest wildfires in state history. The smoke blanketed the college town of Fort Collins, Colorado, the location of the Veterinary Teaching Hospital at Colorado State University (CSU-VTH). The objective for this cross-sectional observational study was to evaluate how these wildfires and the corresponding elevated air quality index (AQI)) was associated with infected corneal ulcerations in dogs when compared to the two previous years. ANIMALS: Seventeen dogs were included in this study. PROCEDURES: Medical records from dogs presented to the CSU-VTH ophthalmology service with infected corneal ulcerations in August, September, and October of 2020, 2019, and 2018 were evaluated. Only corneal ulcerations with growth on their microbial cultures were included in this study. RESULTS: The study revealed a significant increase in prevalence of infected corneal ulcerations in dogs presented to the CSU-VTH during the three wildfire months of 2020 that is, 3.5% (9/255) when compared with the two previous years, 2019: 1.0% (4/383, p = 0.04), and 2018: 0.9% (4/457) (p = .01). The AQI (mean ± standard error) was also significantly elevated for dogs that presented with infected corneal ulcerations in 2020 (70.2 ± 5.8) compared with 2019 (19.7 ± 8.7) and 2018 (45.6 ± 8.7) (p < .01). CONCLUSIONS AND CLINICAL RELEVANCE: Elevation of AQI from wildfires seems to be correlated with an increased prevalence of infected corneal ulceration in dogs. As the duration and frequency of wildfires continues to rise globally, the effects of these wildfires on animal health should be investigated further.

Experimental infection of cats with Cryptosporidium felis.

OBJECTIVES: The aims of this study were to experimentally inoculate cats with Cryptosporidium felis oocysts and compare fecal detection by fluorescent antibody assay (FA) and quantitative PCR (qPCR), and document clinical signs associated with infection. METHODS: Cryptosporidium felis oocysts were concentrated from the feces of a naturally infected cat and orally inoculated into six cats that tested negative for C felis by an FA and fecal flotation (FF). Cats were observed daily for the presence of clinical signs consistent with infection. Fecal samples from all cats on days 0 and 9, and one sample per cat (days 18-21), were evaluated by all assays. On day 31, two cats negative for C felis by FF and FA were administered methylprednisolone acetate and all assays were repeated on days 34, 36 and 38. Samples from all cats were tested by FF and FA on days 41, 43, 45 and 48. RESULTS: A total of 41 samples were tested, 25 of which were compared by FA and qPCR. Cryptosporidium felis was detected in 2/25 (8%) and in 19/25 (76%) samples by FA and by qPCR, respectively; the other 16 samples were tested by FF and FA. None of the cats was positive for C felis by FF or FA in samples collected on days 0, 9 or 18-21. One, five and six samples tested positive by qPCR on days 0, 9 and 18-21, respectively. The cats administered methylprednisolone acetate tested positive for C felis by FA on day 36 and by qPCR on days 31, 34, 36 and 38. None of the cats showed clinical signs of disease. CONCLUSIONS AND RELEVANCE: Clinical signs were not recognized in any of the cats for the duration of the study. FA was insensitive compared with qPCR for detecting cats with subclinical C felis infection.

Evaluation for associations amongst Giardia duodenalis assemblages and diarrhea in dogs.

Giardia duodenalis is a species complex comprising at least eight assemblages. Most dogs harbor the host-adapted assemblages C and D and approximately 30 % harbor the zoonotic assemblages. Humans and dogs with giardiosis can exhibit a variety of clinical manifestations ranging from the absence of clinical signs to acute or chronic diarrhea. Human studies report conflicting results concerning associations between clinical signs and assemblage type. The objective of this study was to use results of molecular and phylogenetic analyses to evaluate associations between G. duodenalis assemblages and diarrhea in client-owned dogs from the United States. Fecal samples that were positive for Giardia cysts were classified as normal or diarrheal. Samples were analyzed by PCR assays of the beta-giardin (bg), glutamate dehydrogenase (gdh), and triose phosphate isomerase (tpi) genes. Sequences of the three genes were analyzed by BLAST analysis and phylogenetic analysis was performed by Neighbor-Joining analysis. Two hundred and eighty-eight Giardia-positive fecal samples were evaluated by the three PCRs. One or more genes were amplified from 95 normal samples and 93 diarrheal samples, 27 samples were positive for one or more genes but could not be sequenced due to low quality DNA, and 73 samples tested negative. Ninety seven percent of the samples (182/188) in both the diarrheal and normal groups typed as dog-specific assemblages (D or C) by at least one gene. Phylogenetic analysis of the three genes placed the isolates from assemblages A, B, C and D separated from each other with strong bootstrap support. Diarrhea was not associated with the Giardia assemblage or other parasitic co-infection in this sample set. Other factors, such as the role of gut microbiota in giardiosis should be considered in future studies.

Evaluation of hair and nail cortisol concentrations and associations with behavioral, physical, and environmental indicators of chronic stress in cats.

BACKGROUND: Chronic stress is implicated in behavioral and health issues in cats, but methods for recognition, evaluation, and measurement of stress are lacking. Cortisol concentration is typically used as an indicator of stress. OBJECTIVES: To evaluate use of an enzyme immunoassay to quantitate hair and nail cortisol concentrations (HCC and NCC) in cats and evaluate associations between HCC and NCC and behavioral, physical, and environmental correlates of chronic stress in cats. ANIMALS: Forty-eight adult, owned or community cats. METHODS: Cross-sectional study. Nail clippings and hair were collected from cats. Medical history and cat daily lifestyle questionnaires were completed by owners or caretakers. A commercial laboratory performed cortisol extraction and quantification using a validated enzyme immunoassay kit. Correlational and regression analyses were used to evaluate associations between HCC and NCC and behavioral, environmental, and medical factors. RESULTS: Hair and nail cortisol concentrations were significantly associated (rs  = 0.70; P < .001), but HCCs varied widely within and among cats. Cats with litterbox issues had significantly increased HCC (P = .02) and NCC (P = .001) as compared to cats without litterbox issues. Cats with groomed coats had lower HCCs (P = .02) as compared to cats without groomed coats, whereas cats with dander and mats had higher NCCs (P = .01) as compared to cats without dander and mats. CONCLUSIONS AND CLINICAL IMPORTANCE: The quantification of NCCs might improve identification and evaluation of chronic stress in cats. The variability of HCCs in individual cats warrants caution using this measurement in chronic stress studies.

Evaluation of the role of Babesia species and Cytauxzoon felis in feline anemia cases in Colorado, USA.

OBJECTIVES: The aim of this study was to evaluate the blood of cats in Colorado, USA, with suspected infectious causes of anemia for the presence of Babesia species and Cytauxzoon felis DNA. Results of PCR testing for other common vector-borne diseases potentially associated with anemia are also reported. METHODS: Samples from 101 cats were tested using a PCR assay that coamplified the DNA of C felis and Babesia species mitochondrial DNA. PCR testing for DNA of hemoplasmas, Bartonella species, Ehrlichia species, Anaplasma species, Neorickettsia risticii and Wolbachia genera was also performed if not carried out previously. RESULTS: Twenty-two cats (21.8%) were positive for DNA of an infectious agent. DNA from hemoplasma species were amplified from 14 cats (13.9%). Bartonella species DNA was amplified from four cats (4%) and Ehrlichia canis, Anaplasma platys, Anaplasma phagocytophilum and Wolbachia genera DNA were amplified from one cat each. Babesia species and C felis mitochondrial DNA were not amplified from any sample. CONCLUSIONS AND RELEVANCE: Based on the results of this study, it does not appear that Babesia species or C felis are clinically relevant in anemic cats in Colorado, USA. For C felis, this suggests that the vector Amblyomma americanum is still uncommon in this geographic area.

Serum 25-hydroxyvitamin D concentration and infectious respiratory disease complex in shelter dogs.

BACKGROUND: Hypovitaminosis D is a risk factor for the development of respiratory infections in humans and repletion can be protective. OBJECTIVES: Determine if serum 25-hydroxyvitamin (OH)D concentrations are lower in shelter dogs and if 25(OH)D concentrations are associated with clinical signs of canine infectious respiratory disease complex (CIRDC) or with time in the shelter. ANIMALS: One hundred forty-six shelter dogs (clinically ill n = 36, apparently healthy n = 110) and 23 nonshelter control dogs. METHODS: Prospective cohort study. Shelter dogs were grouped as clinically ill or apparently healthy based on the presence or absence, respectively, of clinical signs associated with CIRDC. Serum 25(OH)D concentrations were measured with a competitive chemiluminesence immunoassay. Nucleic acids of agents associated with the CIRDC were amplified by polymerase chain reaction assays. RESULTS: The concentration of 25(OH)D was 7.3 ng/mL (4.5-9.9, 95% confidence interval [CI]) lower in dogs with signs of CIRDC than apparently healthy shelter dogs (t(142) = 2.0, P = .04). Dogs positive for DNA of canine herpesvirus (CHV)-1 had serum 25(OH)D concentrations 14.9 ng/mL (-3.7 to 29.6, 95% CI) lower than dogs that were negative (t(137) = 2.0, P = .04). Serum 25(OH)D concentrations in shelter dogs were not different from control dogs (t(45) = -1.4, P = .17). Serum 25(OH)D concentration was not associated with duration of time in the shelter (F(1, 140) = 1.7, P = .2, R2 = 0.01). CONCLUSION AND CLINICAL IMPORTANCE: Vitamin D could have a role in acute respiratory tract infections in shelter dogs.

Experimental infection of cats with Cystoisospora felis.

BACKGROUND: Cystoisospora felis is a common parasite of cats and is diagnosed by fecal flotation, but false-negative results can be common. HYPOTHESIS/OBJECTIVES: To experimentally inoculate cats with C. felis oocysts, to compare fecal flotation and polymerase chain reaction (PCR) results, and to describe any clinical signs consistent with infection. ANIMALS: Six cats. METHODS: Cystoisospora felis oocysts were identified morphologically from feces of a naturally infected kitten with diarrhea, sporulated oocysts (5000) were inoculated to 6 cats that were negative for fecal parasites by fecal flotation and by a fluorescent antibody assay (FA) for Giardia spp. and Cryptosporidium spp. Cats were observed daily for the presence of clinical signs consistent with infection. Fecal samples were evaluated by fecal flotation and FA up to 3 times per week post inoculation (PI) to Day 27. Thirty-six samples collected before inoculation and from Days 8, 10, 13, 15, and 20 PI were assayed using an internal transcribed spacer 1 (ITS1) PCR that amplifies DNA of C. felis. RESULTS: All cats were negative for C. felis by both assays before inoculation. All cats shed C. felis oocysts by Day 10 PI, oocysts were not detected by fecal flotation after Day 15 PI. Cystoisospora felis DNA was amplified from 24/36 (66.6%) fecal samples from 6/6 (100%) of the cats. Oocysts were not detected by fecal flotation in 4 of the samples that were positive for C. felis DNA by PCR. Clinical signs were not recognized in any of the study cats. CONCLUSIONS AND CLINICAL IMPORTANCE: Fecal flotation is a convenient assay for detection of C. felis but could occasionally give false-negative results when compared to this ITS1 PCR.

Ehrlichia canis in dogs experimentally infected, treated, and then immune suppressed during the acute or subclinical phases.

BACKGROUND: Concerns for recrudescence of Ehrlichia canis infection arise when immunosuppressive drugs are used to treat immune-mediated diseases in dogs previously infected with E. canis. OBJECTIVES: Determine whether administration of prednisolone and cyclosporine would reactivate E. canis infection in dogs previously treated with doxycycline during the acute or subclinical phases. ANIMALS: Seven beagles previously experimentally infected with E. canis and administered doxycycline for 4 weeks were included. Three of the 7 dogs were incidentally concurrently infected with Anaplasma platys and Babesia vogeli and were administered 2 doses of imidocarb 2 weeks apart before enrollment in the current study. METHODS: Experimental study. Each dog was administered prednisolone and cyclosporine for 6 weeks. Clinical signs, complete blood cell count (CBC), polymerase chain reaction (PCR) assays for E. canis, A. platys, and B. vogeli DNA in blood, E. canis indirect fluorescent antibodies (IFA) titers, and flow cytometry for antiplatelet antibodies were monitored. RESULTS: All dogs completed the immunosuppressive protocol. No evidence for recrudescence of E. canis, A. platys, or B. vogeli were detected based on clinical signs or results of CBC, PCR, IFA, and flow cytometry for antiplatelet antibodies. E. canis IFA titers were negative in 5/7 dogs at the end of immunosuppressive protocol and were negative 6 months after the protocol in 5/5 dogs available for testing. CONCLUSIONS AND CLINICAL IMPORTANCE: Dogs administered with a 4-week course of doxycycline with or without imidocarb failed to show evidence of activation of E. canis infection after administration of a commonly used immune suppressive protocol.