Detection and molecular characterization of two canine circovirus genotypes co-circulating in Vietnam

BackgroundCanine circovirus is reported in dogs in many countries, including the USA, China and Thailand. It has been detected in healthy dogs and dogs with diarrhea, hemorrhagic gastroenteritis, and vasculitis. It comprises five genotypes and is frequently found as a coinfection with canine parvovirus-2 (CPV-2).AimTo characterize canine circovirus genotypes co-circulating with CPV-2 in Vietnam.MethodPCR assessment of 81 CPV-2-positive fecal samples from Vietnamese diarrheic dogs up to seven months of age for other viral enteric pathogens, including canine bocavirus, canine adenovirus, paramyxovirus, canine coronavirus, porcine circovirus-3 and canine circovirus. In addition, eight selected full genome sequences of Vietnamese canine circovirus were analyzed and used for phylogeny.ResultsIn total 19.8% of samples were found to be positive for canine circovirus. Phylogeny revealed that the Vietnamese canine circovirus strains were clustered in two different genotypes (genotype-1 and -3). The genetic diversity among Vietnamese canine circovirus was 86.0–87.2%. The nucleotide discrepancy among both genotypes altered the deduced amino acid sequence in 14 and ten residues of the replicase and capsid proteins, respectively. Genetic recombination analysis revealed that the Vietnamese canine circovirus-6 strain has the American and Chinese canine circovirus as its major and minor parents, respectively. Only a single dog revealed triple detections of CPV-2c, Canine circovirus and canine adenovirus (1.2%).ConclusionThe co-circulation of two different genotypes of canine circovirus and CPV-2c in dogs in Vietnam has been illustrated.Clinical relevanceThe mortality rate with CPV-2 only (22%) doubled in dogs with canine circovirus and CPV-2 co-infection.     

Molecular epidemiological and phylogenetic analyses of canine parvovirus in domestic dogs and cats in Beijing, 2010–2013

Fifty-five samples (15.62%) collected from dogs and cats were identified as canine parvovirus (CPV) infection in Beijing during 2010–2013. The nucleotide identities and aa similarities were 98.2–100% and 97.7–100%, respectively, when compared with the reference isolates. Also, several synonymous and non-synonymous mutations were also recorded for the first time. New CPV-2a was dominant, accounting for 90.90% of the samples. Two of the 16 samples collected from cats were identified as new CPV-2a (12.5%), showing nucleotide identities of 100% with those from dogs. Twelve samples (15.78%) collected from completely immunized dogs were found to be new CPV-2a, which means CPV-2 vaccines may not provide sufficient protection for the epidemic strains.     

Blood neutrophil-to-lymphocyte ratio (NLR) as a diagnostic marker in dogs with chronic enteropathy

Few routinely available biomarkers are clinically useful in assessing dogs with chronic enteropathy (CE) and aid in CE subclassification. The diagnostic potential of the blood neutrophil-to-lymphocyte ratio (NLR) has not been evaluated in canine CE. We evaluated the NLR in 93 dogs with CE (no steroid treatment for ≥2 wk prior) and tested for an association with clinical, clinicopathologic, and histologic characteristics and also with CE subclassification. NLR was significantly higher in CE dogs with severe clinical disease than dogs with mild clinical disease (p = 0.047). Hypoalbuminemia (p < 0.001), but not hypocobalaminemia, was associated with higher NLRs. NLR was correlated with fecal alpha₁-proteinase inhibitor concentrations (ρ = 0.47) and the serum-to-fecal alpha₁-proteinase inhibitor ratio (ρ = –0.48; both p < 0.001) but not with serum or fecal inflammatory markers nor with the overall histologic score (all p > 0.05). Dogs with steroid- or other immunosuppressant-responsive (IRE) or nonresponsive enteropathy (NRE) had significantly higher NLRs (median: 7.3) than dogs with food-responsive enteropathy (FRE; median: 3.0; p = 0.003), and a NLR ≥5.5 best distinguished both groups of dogs. No difference in NLR was detected between dogs with IRE and dogs diagnosed with NRE. These findings suggest that leukogram changes (i.e., NLR) could be clinically useful in canine CE, and that neutrophils might play a role in the systemic inflammatory response associated with canine CE. The NLR can be easily assessed on routine hematology and can potentially aid in the subclassification of dogs with CE based on the response to treatment.     

Central nervous system lesions caused by canine distemper virus in 4 vaccinated dogs

We examined the cerebellum and cerebrum of 4 vaccinated dogs, 3–60-mo-old, that displayed clinical signs of canine distemper virus (CDV) infection, and died 7–40 d after developing neurologic signs. The main histologic lesions were demyelination, gliosis, meningitis, perivascular lymphocytic cuffing, and inclusion bodies. These lesions were similar in all 4 cases regardless of the time since vaccination, except that meningoencephalitis and gliosis were subacute in 3 dogs and chronic in 1 dog. However, these differences did not appear to be related to their vaccination status. Immunohistologically, a CDV-positive immunoreaction was seen mainly in astrocytes, neurons and their axons, lymphocytes around and in the blood vessels of the pia mater and choroid plexus, ependymal cells of each ventricle, and the cells of the choroid plexus. The histologic and immunohistologic changes were similar in the cerebellum and cerebrum. The genetic characterization of the virus strains in 2 of these naturally occurring canine distemper cases confirmed that they were South American wild-type strains (Kiki and Uy251) belonging to the EU1/SA1 lineage. These strains are not included in the commercial CDV vaccines available in Uruguay.     

Urinary corticoid to creatinine ratios using IMMULITE 2000 XPi for diagnosis of canine hypercortisolism

The urinary corticoid to creatinine ratio (UCCR) is one of the most commonly used screening tests for canine hypercortisolism (HC). In this study, a reference interval was established for UCCR using IMMULITE 2000 XPi, the latest chemiluminescence enzyme immunoassay. The diagnostic performance of this method for UCCR in canine HC was also evaluated. The median UCCR was 1.06 × 10⁻⁵ (range: 0.28–2.49) for 58 healthy dogs, and an upper reference limit of 1.98 × 10⁻⁵ (90% confidence interval: 1.76–2.15) was determined. The median UCCR in the 12 dogs with HC (7.38 × 10⁻⁵, range 1.86–29.98) was significantly higher than that in the 16 dogs with mimic-HC (1.59 × 10⁻⁵, range 0.47–3.42, P<0.001). The area under the curve for UCCR to differentiate HC dogs from mimic-HC dogs was 0.971, with a sensitivity of 91.7% and specificity of 100% when the cut-off value was set at 3.77 × 10⁻⁵. The UCCR of 16 paired urine samples collected at home and in hospital showed that the UCCR of samples collected in the hospital was significantly higher than that of samples collected at home (mean difference 3.30 × 10⁻⁵, 95% confidence interval: 0.70–5.90, P=0.001). In summary, we established the upper reference limit for UCCR using IMMULITE 2000 XPi in dogs and confirmed that UCCR is a useful diagnostic test for HC in dogs if urine samples are collected at home.     

Stray dogs as sentinels for canine parvovirus

An attempt to determine the prevalence of canine parvovirus in the stray dog population of Franklin County, Ohio (U.S.A.) was made by sampling dogs during the first 6 months of 1981. Serum and fecal samples, which were collected from 209 strays at time of euthanasia, were tested by serum hemagglutination inhibition (HI) and fecal hemagglutination (HA) techniques to determine canine parvovirus experience (seropositive) or fecal virus shedding, respectively. Sera collected from 93 strays for an unrelated study conducted in 1979 were used as the comparison group. All of the 1979 sera were HI negative (< 1:80) whereas, 139 of 201 (69.2%) sera suitable for testing from the 1981 group of strays were HI positive (? 1:80). The fecal HA results from the 1981 group revealed 26 of the 209 (12.4%) dogs were shedding parvovirus at time of euthanasia (HA titers ? 1:64). Of these 26 of the 209 (12.4%) dogs were shedding parvovirus at time of euthanasia were found to be seropositive. These results indicate that the stray population of Franklin County, Ohio, was not exposed in 1979, but by 1981 had experienced and for the most part, had recovered from canine parvovirus as indicated by a 69.2% seropositive dog population with 12.4% active virus shedders. The stray dog population, if sampled regularly, could thus serve as a sentinel for canine parvovirus activity in a community.     

Quantification of normetanephrine in canine urine using ELISA: evaluation of factors affecting results

Catecholamine release increases in dogs with pheochromocytomas and in situations of stress. Although plasma catecholamines degrade rapidly, their metabolites, normetanephrine (NME) and metanephrine (ME), are stable in acidified urine. Our aim was to verify a human urine ELISA kit for the quantification of NME and ME in canine urine and to determine the effects on metabolite stability of sampling time (morning or midday) and day (ordinary or day spent in a clinic). We analyzed 179 urine samples from 17 healthy dogs. For NME, the mean intra-assay CV was 6.0% for all samples and 4.3% for the canine control; inter-assay CVs were 3.3, 3.8, and 12% for high and low concentration human urine positive controls supplied in the ELISA kit and a positive canine control, respectively; spike-recovery was 90–101%. For ME, mean intra-assay CV was 6.5% for samples and 9.0% for the canine control; inter-assay CVs were 12.7, 7.2, and 22.5% for high and low concentration human urine positive controls supplied in the ELISA kit and a positive canine control, respectively; spike-recovery was 85–89%. Dilution recovery was unsatisfactory for both metabolites. Based on our verification results, NME was selected for remaining analyses. We found no effect on NME concentrations of acidification or room temperature storage for up to 24 h. The NME:creatinine ratio was higher after the first of 3 clinic days compared to the same morning (111.2 ± 5.5 vs. 82.9 ± 5.3; p < 0.0001), but not on the other days. NME verification results were generally superior to ME. Dilution studies were unsatisfactory for both metabolites. Given that NME was stable without acidification at room temperature, urine samples can be collected at home. The clinic environment can cause higher NME:creatinine ratios, especially in unaccustomed dogs.     

Verification of the fCAL turbo immunoturbidimetric assay for measurement of the fecal calprotectin concentration in dogs and cats

The concentration of calprotectin in feces (fCal) is a clinically useful marker of chronic gastrointestinal inflammation in humans and dogs. No commercial assay is widely available to measure fCal in small animal medicine, to date. Thus, we verified the immunoturbidimetric fCAL turbo assay (Bühlmann) of fCal for canine and feline fecal extracts by determining linearity, spiking and recovery, and intra-assay and inter-assay variability. We determined RIs, temporal variation over 3 mo, and effect of vaccination and NSAID treatment. Observed:expected (O:E) ratios ($\bar{x}$ ± SD) for serial dilutions of feces were 89–131% (106 ± 9%) in dogs and 77–122% (100 ± 12%) in cats. For spiking and recovery, the O:E ratios were 90–118% (102 ± 11%) in dogs and 83–235% (129 ± 42%) in cats. Intra- and inter-assay CVs for canine samples were ≤19% and ≤7%, and for feline samples ≤22% and ≤21%. Single-sample RIs were <41 μg/g for dogs and <64 μg/g for cats. With low reciprocal individuality indices, using population-based fCal RIs is appropriate, and moderate fCal changes between measurements (dogs 44.0%; cats: 43.2%) are considered relevant. Cats had significant (but unlikely relevant) fCal increases post-vaccination. Despite individual fCal spikes, no differences were seen during NSAID treatment. The fCAL turbidimetric assay is linear, precise, reproducible, and sufficiently accurate for measuring fCal in dogs and cats. Careful interpretation of fCal concentrations is warranted in both species during the peri-vaccination period and for some patients receiving NSAID treatment.     

Efficacy of two low-dose oral tylosin regimens in controlling the relapse of diarrhea in dogs with tylosin-responsive diarrhea: a prospective,single-blinded,two-arm parallel,clinical field trial

Background

Despite its wide acceptance as a treatment for canine chronic enteropathies, the macrolide antibiotic tylosin lacks official oral dosage recommendations. Not even textbooks share consensus about the dose; daily recommendations vary from 25 to 80 mg/kg and dosing intervals from one to three times daily.The objective of this prospective, single-blinded, two-arm parallel, clinical field trial was to determine whether doses of 5 mg/kg or 15 mg/kg tylosin administered orally once daily for seven days would have a similar effect on fecal consistency in diarrhea relapses to that of a 25 mg/kg dose of tylosin administered once daily for seven days, a dosage that has proved effective in controlling canine tylosin-responsive diarrhea (TRD). A further objective was to compare the efficacy of the 5 mg/kg and 15 mg/kg tylosin dosages. Fifteen client-owned dogs diagnosed with TRD that had responded to a dose of 25 mg/kg tylosin once daily for seven days were enrolled in the study. After a relapse of diarrhea the dogs were allocated into two groups receiving tylosin orally in doses of either 5 mg/kg or 15 mg/kg once daily for seven days. The owners were blinded to the dosage. The elimination of diarrhea was the main criterion in assessing treatment success. The mean fecal consistency score of the last three treatment days for all dosages, including 25 mg/kg, as evaluated by the owners according to a standardized fecal scoring system, served as the primary outcome measures.

Results

All eight dogs responded to the 5 mg/kg dose, and six of seven dogs responded to the 15 mg/kg dose. The mean fecal consistency scores at the 25 mg/kg tylosin dosage were no significantly different from scores at the 5 mg/kg or 15 mg/kg tylosin dosages (P = 0.672, P = 0.345).

Conclusions

Interestingly, 14/15 (93%) of the dogs responding to a dose of 25 mg/kg tylosin once daily for seven days also responded to the lower dosages at diarrhea relapse. The data indicate that a suitable dose of tylosin for treating diarrhea relapse in canine TRD could be as low as 5 mg/kg once daily for seven days.     

Urinary free metanephrines measurement in dogs with adrenal gland diseases using a new simple liquid chromatography tandem mass spectrometry method

Measurement of urinary metanephrines in spot samples is used for the diagnosis of canine pheochromocytoma (PC). We describe a simple analytical method based on liquid chromatography tandem mass spectrometry (LC-MS/MS) for measuring free metanephrine (MN) and normetanephrine (NMN) in spot urine samples. Using the developed method, we evaluated the stability of urinary free-MN and free-NMN at various storing conditions. In addition, we assessed the feasibility of urinary free-MN and -NMN measurement for diagnosing PC. Urine samples were mixed with stable isotope internal standards and thereafter purified by ultrafiltration. The purified samples were analyzed by LC-MS/MS in the multiple reaction monitoring mode after separation on a multimode octa decyl silyl column. The coefficient of variation of free-MN and -NMN measurement was 7.6% and 5.5%, respectively. The linearity range was 0.5–10 µg/l for both analytes. Degradation was less than 10% for both analytes under any of the storage conditions. The median free-NMN ratio to creatinine of 9 PC dogs (595, range 144–47,961) was significantly higher (P<0.05) than that of 13 dogs with hypercortisolism (125, range 52–224) or 15 healthy dogs (85, range 50–117). The developed method is simple and may not require acidification of spot urine. The results of this preliminary retrospective study suggest that the measurement of urinary free metanephrines is a promising tool for diagnosing canine PC.     

Serum vascular endothelial growth factor in dogs with various proliferative diseases

Angiogenesis plays an important role in the proliferation and metastasis mechanisms of malignant tumors. Vascular endothelial growth factor (VEGF), a group of cytokines that contribute to angiogenesis and vasculogenesis. This study aimed to investigate the serum VEGF-A concentrations in dogs with various proliferative diseases. A total of 202 dogs that were histopathologically diagnosed with proliferative diseases were included in the study. Serum VEGF-A concentrations were measured using enzyme-linked immunosorbent assay. Median serum VEGF-A concentrations in dogs were as follows: healthy dogs, 4 pg/ml [0–21 pg/ml]; hepatocellular carcinoma, 30 pg/ml [0–158 pg/ml, P=<0.001]; hepatocellular adenoma, 32 pg/ml [0–49 pg/ml, P=0.003]; hepatic nodular hyperplasia, 18 pg/ml [0–51 pg/ml, P=0.595]; adrenal pheochromocytoma, 32 pg/ml [0–187 pg/ml, P=<0.001]; adrenocortical carcinoma, 32 pg/ml [3–161 pg/ml, P=0.002]; adrenocortical adenoma, 27 pg/ml [0–106 pg/ml, P=0.005]; colorectal adenocarcinoma, 36 pg/ml [0–75 pg/ml, P=0.002]; colorectal adenoma, 43 pg/ml [0–48 pg/ml, P=0.144]; inflammatory colorectal polyps, 37 pg/ml [0–111 pg/ml, P=<0.001]; pulmonary adenocarcinoma, 35 pg/ml [4–107 pg/ml, P=0.002]; pulmonary histiocytic sarcoma, 35 pg/ml [0–131 pg/ml, P=0.016]; and follicular thyroid carcinoma, 35 pg/ml [0–106 pg/ml, P=0.009]. The serum VEGF-A concentrations were significantly higher in dogs with neoplastic lesions compared to healthy dogs, except for colorectal adenoma. High serum VEGF-A concentrations were observed in dogs with proliferative diseases. The present study suggests that angiogenesis-inhibiting therapy, which targets VEGF-A, may be useful for canine neoplastic diseases.     

Evaluation of an automated colorimetric assay for the measurement of lipase activity in canine sera.

An automated colorimetric method for determining lipase activity in canine sera was evaluated for precision, linearity and correlation to existing assay methods. The colorimetric method was a commercial reagent that used a series of enzymatic reactions based on the hydrolysis of 1,2 diglyceride by pancreatic lipase. Within-run and between-run coefficients of variation were < 6.8% and < 8.3%, respectively. Linearity was determined to be at least 1366 U/L. Canine serum lipase concentrations attained using the colorimetric method were compared to both titrimetric and dry-film methods for measuring serum lipase activity, resulting in significant (P < or = 0.05) correlation coefficients of 0.92 and 0.77, respectively. Canine serum lipase concentrations measured using the colorimetric assay on 2 different automated analyzers had a significant (P < or = 0.05) correlation coefficient of 0.92. A laboratory reference range using serum samples from 56 healthy dogs (0-561 U/L) was established. There were no significant (P < or = 0.05) differences in mean serum lipase concentrations comparing male and female dogs or comparing young dogs (< or = 3 y) to mature (4-7 y) and older (> 7 y) dogs using this assay. It was concluded that the automated colorimetric assay was a reliable indicator of canine serum lipase activity and offered several advantages, including small sample volume and short analysis time.     

Nutrient digestibility and fecal characteristics,microbiota, and metabolites in dogs fed human-grade foods

Human-grade (HG) pet foods are commercially available, but they have not been well studied. Our objective was to determine the apparent total tract digestibility (ATTD) of HG pet foods and evaluate their effects on fecal characteristics, microbiota, and metabolites, serum metabolites, and hematology of dogs. Twelve dogs (mean age = 5.5 ± 1.0; BW = 11.6 ± 1.6 kg) were used in a replicated 4 × 4 Latin square design (n = 12/treatment). The diets included 1) Chicken and Brown Rice Recipe (extruded; Blue Buffalo); 2) Roasted Meals Tender Chicken Recipe (fresh; Freshpet); 3) Beef and Russet Potato Recipe (HG beef; JustFoodForDogs); and 4) Chicken and White Rice Recipe (HG chicken; JustFoodForDogs). Each period consisted of 28 d, with a 6-d diet transition phase, 16 d of consuming 100% of the diet, a 5-d phase for fecal collection, and 1 d for blood collection. All data were analyzed using the Mixed Models procedure of SAS 9.4. Dogs fed the extruded diet required a higher (P < 0.05) daily food intake (dry matter basis, DMB) to maintain BW. The ATTD of dry matter (DM), organic matter (OM), energy, and acid-hydrolyzed fat (AHF) were greater (P < 0.05) in dogs fed the HG diets than those fed the fresh diet, and greater (P < 0.05) in dogs fed the fresh diet than those fed the extruded diet. Crude protein ATTD was lower (P < 0.05) for dogs fed the extruded diet than those fed all other diets. Dogs fed the extruded diet had greater (P < 0.05) fecal output (as-is; DMB) than dogs fed fresh (1.5–1.7 times greater) or HG foods (2.0–2.9 times greater). There were no differences in fecal pH, scores, and metabolites, but microbiota were affected by diet. Dogs fed HG beef had higher (P < 0.05) relative abundance of Bacteroidetes and lower (P < 0.05) relative abundance of Firmicutes than dogs fed the fresh or HG chicken diets. The Actinobacteria, Fusobacteria, Proteobacteria, and Spirochaetes phyla were unchanged (P > 0.05), but diet modified the relative abundance of nearly 20 bacterial genera. Similar to previous reports, these data demonstrate that the fecal microbiota of dogs fed HG or fresh diets is markedly different than those consuming extruded diets, likely due to ingredient, nutrient, and processing differences. Serum metabolites and hematology were not greatly affected by diet. In conclusion, the HG pet foods tested resulted in significantly reduced fecal output, were highly digestible, maintained fecal characteristics, serum chemistry, and hematology, and modified the fecal microbiota of dogs.     

Plasma and Urine Neutrophil Gelatinase–Associated Lipocalin (NGAL) in Dogs with Acute Kidney Injury or Chronic Kidney Disease

Background

Neutrophil gelatinase–associated lipocalin (NGAL) is a protein that is used in human medicine as a real‐time indicator of acute kidney injury (AKI).

Hypothesis

Dogs with AKI have significantly higher plasma NGAL concentration and urine NGAL‐to‐creatinine ratio (UNCR) compared with healthy dogs and dogs with chronic kidney disease (CKD).

Animals

18 healthy control dogs, 17 dogs with CKD, and 48 dogs with AKI.

Methods

Over a period of 1 year, all dogs with renal azotemia were prospectively included. Urine and plasma samples were collected during the first 24 hours after presentation or after development of renal azotemia. Plasma and urine NGAL concentrations were measured with a commercially available canine NGAL Elisa Kit (Bioporto® Diagnostic) and UNCR was calculated. A single‐injection plasma inulin clearance was performed in the healthy dogs.

Results

Median (range) NGAL plasma concentration in healthy dogs, dogs with CKD, and AKI were 10.7 ng/mL (2.5–21.2), 22.0 ng/mL (7.7–62.3), and 48.3 ng/mL (5.7–469.0), respectively. UNCR was 2 × 10⁻⁸ (0–46), 1,424 × 10⁻⁸ (385–18,347), and 2,366 × 10⁻⁸ (36–994,669), respectively. Dogs with renal azotemia had significantly higher NGAL concentrations and UNCR than did healthy dogs (P < .0001 for both). Plasma NGAL concentration was significantly higher in dogs with AKI compared with dogs with CKD (P = .027).

Conclusions and Clinical Importance

Plasma NGAL could be helpful to differentiate AKI from CKD in dogs with renal azotemia.     

Interobserver reliability of canine urine specific gravity assessed by analog or digital refractometers

Refractometry is utilized routinely to evaluate canine urine specific gravity (USG) in veterinary clinical settings. We aimed to determine if the magnitude of interobserver reliability when assessing canine USG via refractometry could impact clinical judgment. USG was determined in 38 dogs by 3 registered veterinary technicians (RVTs) using both an optical analog refractometer and a digital refractometer. Summary statistics were reported, interobserver reliability was assessed via intraclass correlation coefficient (ICC) analysis through a 2-way mixed-effects model, and agreement between RVT pairs was compared through Bland–Altman plots. The median analog refractometer USG measurement was 1.018 (range: 1.004–1.040) and for the digital refractometer was 1.0176 (1.0035–1.0357). The analog refractometer average measure ICC was 0.995 (95% CI: 0.992, 0.997; p < 0.001). The digital refractometer average measure ICC was 0.999 (95% CI: 0.999, 1.000; p < 0.001). Strong agreement between all pairs of RVTs was seen via Bland–Altman plots for both analog and digital refractometers, with 95% CIs spanning no more than 0.002 in either the positive or negative direction for all pairings. The interobserver variability in canine USG measurements by RVTs was trivial and did not impact clinical judgment and decision-making.     

Effect of native Lactobacillus murinus LbP2 administration on total fecal IgA in healthy dogs

The objective of the present work was to determine the effect of Lactobacillus murinus strain LbP2 on canine fecal immunoglobulin A (IgA) levels. Seven dogs were orally treated with a 3-mL suspension of L. murinus LbP2 containing 5 × 10⁹ colony-forming units on alternate days for 2 wk. Six dogs were treated with 3 mL of phosphate-buffered saline as placebo. Fecal samples were taken from the rectal ampulla on days 0 and 16, and the total canine fecal IgA concentration was determined with an immunoperoxidase assay kit. The IgA levels of individual dogs were compared with the nonparametric Wilcoxon test. Differences were considered significant when the P-value was less than 0.05. An increase in the total fecal IgA concentration was observed in the 7 dogs after treatment with L. murinus LbP2 (P = 0.01796). No differences were detected between the initial total fecal IgA values and those obtained at the end of placebo treatment. Thus, after oral administration L. murinus LbP2 showed potential immunomodulatory effects, an important property to assess in a microorganism being considered for use as a probiotic.     

Increased canine pancreatic lipase immunoreactivity (cPLI) and 1,2-o-dilauryl-rac-glycero-3-glutaric acid-(6′-methylresorufin) ester (DGGR) lipase in dogs with evidence of portal hypertension and normal pancreatic histology: a pilot study

The clinical presentations of both liver disease and pancreatitis are nonspecific and overlapping, which may cause difficulty in diagnosis. In our retrospective pilot study, we assessed whether dogs with evidence of portal hypertension and absence of pancreatitis on pancreatic histology have increases in canine pancreatic lipase immunoreactivity (cPLI) and 1,2-o-dilauryl-rac-glycero-3-glutaric acid-(6′-methylresorufin) ester (DGGR) lipase. We included dogs that had been presented between 2008 and 2019 if they had normal pancreatic histology, histologically confirmed hepatopathy, and if canine pancreas-specific lipase (Spec cPL; Idexx) or DGGR lipase had been measured. Only dogs with portal hypertension were included. Six dogs fulfilled the inclusion criteria. Four of 6 and 2 of 6 dogs had Spec cPL and DGGR lipase exceeding the upper reference limit, respectively. From the 4 dogs with increased Spec cPL, 2 had concentrations of 200–400 µg/L and 2 had concentrations ≥ 400 µg/L. Our results suggest that canine portal hypertension might lead to increased Spec cPL and DGGR lipase values in the absence of pancreatitis on histology. Until more evidence in a larger number of dogs with portal hypertension is available, both tests should be interpreted cautiously in the presence of portal hypertension.     

Association between breed,gender and age in relation to cardiovascular disorders in insured dogs in Japan

The association between breed, gender and age and cardiovascular disorders in the insured dog population in Japan was investigated, using multiple logistic regression analysis and data from 299,555 dogs insured between April 2010 and March 2011. The overall annual prevalence of cardiovascular disorder diagnosis was 2.1%. Using the Miniature Dachshund as the reference breed, Cavalier King Charles Spaniel had the highest odds of cardiovascular disorder with a ratio of 16.2 (95% confidence interval: 14.4–18.2), followed by Maltese, Pomeranian, Chihuahua and Shih Tzu. Male dogs had increased odds of 1.2 (1.1–1.3). The dogs had increased odds of having cardiovascular disorder by 1.5 times as their age increased by one year.     

Evaluation of five enzyme immunoassays compared with the cytotoxicity assay for diagnosis of Clostridium difficile-associated diarrhea in dogs.

Clostridium difficile-associated-diarrhea (CDAD) is a nosocomial infection in dogs. Diagnosis of this infection is dependent on clinical signs of disease supported by laboratory detection of C. difficile toxins A or B, or both, in fecal specimens via enzyme-linked immunosorbent assay (ELISA). Unfortunately, to the authors' knowledge, commercially available ELISAs have not been validated in dogs to date. We evaluated 5 ELISAs done on 143 canine fecal specimens (100 diarrheic and 43 nondiarrheic dogs) and on 29 C. difficile isolates. The results of each ELISA were compared with the cytotoxin B tissue culture assay (CTA). Clostridium difficile was isolated from 23% of the fecal specimens. Eighteen of the 143 fecal specimens were toxin positive (15 diarrheic and 3 nondiarrheic dogs). On the basis of multiplex polymerase chain reaction (PCR) analysis for toxin-A and -B genes, 72% of the isolates were toxigenic. The carriage rate of toxigenic isolates in diarrheic dogs was higher than that in the nondiarrheic dogs; however, these differences were not statistically significant. A good correlation was found between CTA, PCR, and culture results. The ELISAs done on fecal specimens collected from diarrheic dogs had low sensitivity (7-33%). In contrast, ELISA for toxin A or B, or both, performed on toxigenic isolates had high sensitivity (93%). These results suggest that commercially available human ELISAs are inadequate for the diagnosis of canine C. difficile-associated diarrhea when tested on fecal specimens. In contrast, the Premier ToxinA/B and Techlab ToxinA/B ELISAs may be useful for the diagnosis of canine CDAD when used on toxigenic isolates.     

Development and Evaluation of an ELISA for the Quantitation of Anti‐Lagenidium giganteum forma caninum Antibodies in Dogs

Background

Lagenidium giganteum forma caninum infection causes severe cutaneous and disseminated disease in dogs. Currently, diagnosis requires culture and rRNA gene sequencing.

Objective

To develop and evaluate an ELISA for quantitation of anti‐L. giganteum f. caninum IgG in canine serum.

Animals

Sera were evaluated from 22 dogs infected with L. giganteum f. caninum, 12 dogs infected with Paralagenidium karlingii, 18 dogs infected with Pythium insidiosum, 26 dogs with nonoomycotic fungal infections or other cutaneous or systemic diseases, and 10 healthy dogs.

Methods

Antigen was prepared from a soluble mycelial extract of L. giganteum f. caninum. Optimal antigen and antibody concentrations were determined by checkerboard titration. Results were expressed as percent positivity (PP) relative to a strongly positive control serum.

Results

Medians and ranges for PP for each group were: L. giganteum f. caninum (73.9%, 27.9–108.9%), P. karlingii (55.0%, 21.0–90.6%), P. insidiosum (31.3%, 15.8–87.5%), nonoomycotic fungal infection or other cutaneous or systemic diseases (19.2%, 3.2–61.0%), and healthy dogs (9.9%, 7.6–24.6%). Using a PP cutoff value of 40%, sensitivity and specificity (with 95% CI) of the ELISA for detecting L. giganteum f. caninum infection in clinically affected dogs were 90.9% (72.2–97.5%) and 73.2% (60.4–83.0%), respectively. Specificity in dogs infected with P. karlingii was 41.7% (19.3–68.1%) and with P. insidiosum was 66.7% (43.8–83.7%).

Conclusions and Clinical Importance

Quantitation of anti‐L. giganteum f. caninum antibodies for detection of this infection in dogs has moderately high sensitivity but poor specificity, the latter because of substantial cross‐reactivity with anti‐P. karlingii and anti‐P. insidiosum antibodies.