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Background A canine-specific immunoturbidimetric CRP assay, Gentian Canine CRP Immunoassay) with species-specific controls and calibrators was introduced and recently evaluated on the clinical chemistry analyzer Abbott Architect c4000 as well as on the Olympus AU600. Aims of our study were 1) to independently evaluate the canine-specific CRP assay on the ABX Pentra 400 clinical chemistry analyzer in comparison to the previously validated human-based immunoturbidimetric assay (Randox Canine CRP assay) and 2) to assess the impact of different sample types (serum versus heparinized plasma) on the results. Imprecision, accuracy, interference and the prozone effect were determined using samples from healthy and diseased dogs ( n  = 278). The Randox Canine CRP assay calibrated with canine specific control calibration material served as a reference method. Additionally, the impact of the sample type (serum and lithium heparin) was evaluated based on samples of healthy and diseased dogs ( n  = 49) in a second part of the study. Results Linearity was present for CRP concentrations ranging from 4 to 281 mg/l. For clinically relevant CRP concentrations of 7–281 mg/l, recovery ranged between 90 and 105% and intra- and inter-assay CVs ranged between 0.68% - 12.12% and 0.88% - 7.84%, respectively. CV was thus lower than 12.16%, i.e. the desired CV% based on biological variation. Interference was not present up to a concentration of 5 g/l hemoglobin, 800 mg/l bilirubin and 10 g/l triglycerides. No prozone effect occurred up to 676 mg/l CRP. Method comparison study revealed a Spearman’s rank correlation coefficient of r s  = 0.98 and a mean constant bias of 5.2%. The sample type had a significant ( P  = 0.008) but clinically not relevant impact on the results (median CRP of 30.9 mg/l in lithium heparin plasma versus 31.4 mg/l in serum). Conclusions The species-specific Gentian Canine CRP Immunoassay reliably detects canine CRP on the ABX Pentra 400 clinical chemistry analyzer whereby both serum and heparin plasma can be used. The quality criteria reached on the Abbott Architect c4000 and Olympus AU600 could be met.

Similar to human medicine, attempts have been made in veterinary medicine to assess the severity of respiratory disorders using methods other than respiratory function evaluation; however, such approaches remain insufficient. Medical records at a single small animal private referral center for emergency care were reviewed to identify dogs with respiratory disorders diagnosed by radiography during 2016-2019. The variables of screening test evaluated in this study included patient characteristics, physical examination, and blood test findings. The cases were also divided into Survivors, which were defined as dogs surviving over 7 days from the first consultation day, and Non-survivors, including the dogs euthanized and died naturally within 7 days from the consultation day. In univariate analysis, heart rates, body temperature, white blood count (WBC), glucose, blood urea nitrogen (BUN), phosphate and lactate were significantly different between Survivors and Non-survivors. Multiple logistic regression model with these significant variables revealed that only phosphate was associated with a poor prognosis. This study has demonstrated several parameters of physical examination and blood test, especially plasma phosphate concentration, could be related with mortality in canine respiratory disorders. Although further studies are needed, these parameters may enable more accurate assessment of the severity of respiratory disorders in dogs by combining with the conventional assessments of respiratory functions including oxygenation and ventilation.

Appendicular central osteosarcoma (OSA) is a common and highly aggressive tumour in dogs. Metastatic disease to the lungs is common and even with chemotherapy the prognosis is generally poor. However, few cases survive well beyond reported median survival times. Current methods, including histologic grading schemes, have fallen short in their ability to predict clinical outcome. MicroRNAs (miRNAs) are small molecules present in all tissues and bodily fluids and are dysregulated in cancer. Previous studies have demonstrated the diagnostic and prognostic potential of miRNAs in canine OSA. We sought to investigate multiple miRNA and multiple variable models for diagnosis and prognosis of canine OSA using plasma samples across three populations of dogs from two veterinary biobanks. Fifty-six miRNAs were analyzed by real-time quantitative polymerase chain reaction. MiR-214-3p was the only miRNA with increased expression across all OSA populations compared to controls. Using a decision tree model for diagnosis, miR-214-3p was the first step in this multi-miRNA model. High expression of miR-214-3p alone was also a predictor of shorter overall survival and disease-free interval across all populations. In both multiple miRNA and multiple variable models, miR-214-3p was always the first decision point with high expression consistently predicting a worse prognosis. Additional miRNAs in combination with low expression of miR-214-3p similarly had a worse prognosis demonstrating better outcome prediction using multiple miRNAs compared to using miR-214-3p alone. Multiple variable models only need to use miRNAs to be predictive although clinical parameters such as age, sex, and tumour location were considered. MiR-214-3p is clearly an important prognostic predictor of canine OSA in plasma as supported by previous studies and across our multiple sample populations. Multiple miRNA models provided superior categorization of patients in predicting clinical outcome parameters compared to the single miRNAs.

A surge in Food and Drug Administration (FDA) consumer complaints identified concerns that legume-rich, grain-free diets were associated with nutritionally-mediated dilated cardiomyopathy (DCM). Golden retrievers represent the most reported breed affected by this condition and previous studies documented the disease is responsive to dietary change and taurine supplementation. Although dietary findings across cases are compelling, prospective studies with control groups are lacking. The role of diet in developing taurine deficiency and echocardiographic changes consistent with DCM in healthy dogs is unknown. We hypothesized that golden retrievers eating non-traditional diets are at a higher risk of having taurine deficiency and nutritionally-mediated DCM compared with those eating traditional commercial diets. We aimed to compare taurine concentrations and echocardiographic indices of systolic function between golden retrievers in each diet group and elucidate associations between diet and these variables. Additionally, we aimed to generate breed-specific reference intervals for whole blood and plasma taurine concentrations. 86 golden retrievers. Golden retrievers eating traditional or non-traditional diets were evaluated and diet history, taurine concentrations and echocardiographic data were collected. Dietary features, taurine concentrations and echocardiographic findings were compared between diet groups. Relative risks were calculated for the likelihood of echocardiographic abnormalities and taurine deficiency in each diet group. Breed-specific reference intervals were constructed for taurine concentrations in dogs from the traditional diet group. Golden retrievers eating non-traditional diets had significantly lower taurine concentrations and more frequent systolic dysfunction. Breed specific reference intervals are higher than previously reported across breeds. Non-traditional diets, which were typically grain-free and contained legumes in this study, were significantly associated with and have increased relative risk for the identification of taurine deficiency and echocardiographic abnormalities consistent with nutritionally-mediated DCM. These findings were identifiable in the absence of clinical signs and support the findings of multiple previous studies and the ongoing FDA investigation.

This study aimed to explore the potential relationship between the severity of cardiovascular disease and evidence of gastrointestinal bacterial translocation with systemic and cardiac inflammation in dogs with myxomatous mitral valve disease (MMVD). Thirty-six client-owned dogs, including 25 dogs with American College of Veterinary Internal Medicine Stage B1 or B2 disease but without increased left atrial pressure (Group 1) and 11 dogs with stage B2 disease with increased left atrial pressure or stage C disease (Group 2), were enrolled prospectively with an echocardiographic diagnosis of untreated MMVD. Serum lipopolysaccharide (LPS), interleukins (i.e., IL-2, IL-6, IL-8), tumor necrosis factor-alpha, and cardiac troponin were measured at enrollment. Presence of gastrointestinal clinical signs, fecal scores, body condition score, and muscle condition score were recorded. Linear regression models were used to compare LPS and inflammatory biomarkers based on MMVD severity. Spearman’s correlation was used to evaluate possible associations between inflammatory markers and LPS. The Fisher Exact test was used to compare proportions of dogs with gastrointestinal signs between Groups 1 and 2. Twenty percent of dogs in Group 1 had gastrointestinal clinical signs compared to 66.7% in Group 2 (P < 0.0001). Serum IL-6 (P = 0.037) and LPS (P = 0.024) concentrations were significantly associated with MMVD stage severity. Serum LPS and IL-6 were positively correlated (r s = 0.81; P < 0.0001). This study indicates that dogs with MMVD have evidence of loss of gastrointestinal barrier function as evidenced by bacterial translocation as the disease progresses in severity, which may be associated with systemic inflammation. These findings warrant further evaluation of gastrointestinal barrier function and maybe even the gastrointestinal microbiome as therapeutic targets in dogs with MMVD.

Chronic inflammatory enteropathies (CIE) in dogs are a group of disorders that are characterized by chronic persistent or recurrent signs of gastrointestinal disease and histologic evidence of mucosal inflammation. These CIEs are classified as either food‐responsive, antibiotic‐responsive, or immunosuppressant‐responsive enteropathy. Patients not clinically responding to immunomodulatory treatment are grouped as nonresponsive enteropathy and dogs with intestinal protein loss as protein‐losing enteropathy. Disease‐independent clinical scoring systems were established in dogs for assessment of clinical disease severity and patient monitoring during treatment. Histopathologic and routine clinicopathologic findings are usually not able to distinguish the subgroups of CIE. Treatment trials are often lengthy and further diagnostic tests are usually at least minimally invasive. Biomarkers that can aid in defining the presence of disease, site of origin, severity of the disease process, response to treatment, or a combination of these would be clinically useful in dogs with CIE. This article summarizes the following biomarkers that have been evaluated in dogs with CIE during the last decade, and critically evaluates their potential clinical utility in dogs with CIE: functional biomarkers (cobalamin, methylmalonic acid, folate, α1‐proteinase inhibitor, immunoglobulin A), biochemical biomarkers (C‐reactive protein, perinuclear anti‐neutrophilic cytoplasmic antibodies, 3‐bromotyrosine, N‐methylhistamine, calprotectin, S100A12, soluble receptor of advanced glycation end products, cytokines and chemokines, alkaline phosphatase), microbiomic biomarkers (microbiome changes, dysbiosis index), metabolomic biomarkers (serum metabolome), genetic biomarkers (genomic markers, gene expression changes), and cellular biomarkers (regulatory T cells). In addition, important performance criteria of diagnostic tests are briefly reviewed.

ABSTRACT Background It is unclear if dogs with acute pancreatitis differ clinically from dogs with non‐pancreatic acute gastrointestinal disease (aGId). Objectives Compare clinical findings in dogs with acute gastrointestinal signs suspected of having acute pancreatitis (sAP) based on increased DGGR‐lipase activity versus those with presumptive aGId. Animals Twenty‐six dogs with sAP, 48 dogs with aGId based on acute signs, lipase activity > 450 U/L (RI, 17–156 U/L) and within/minimally (20 U/L) > RI, respectively. Methods Prospective study. Clinical signs were graded using a simplified modified clinical activity index (MCAI). CBC, biochemistry, C‐reactive protein (CRP), pancreatic, and gastrointestinal ultrasonographic findings were compared between groups. Results Median (range) disease duration before presentation (sAP 36 h [3–96 h], aGId 48 h [3–168 h]) did not differ. Diarrhea was significantly more frequent in aGId; MCAI did not differ between groups. Median (range) lipase activities in sAP and aGId dogs were 1280 U/L (451–6712) and 49.5 U/L (14–176), respectively. Alkaline phosphatase activity and bilirubin were significantly higher in sAP. Pancreatic ultrasonographic abnormalities were significantly more common in sAP. In aGId, a mixed‐echoic (17/44, 39%), hyperechoic (9/44, 20%), hypoechoic pancreas (3/44, 7%), and hyperechoic mesentery (4/44, 9%) were found. Only a distended stomach was significantly more common in sAP. Multivariable logistic regression analysis only identified pancreatic enlargement and ultrasonographic diagnosis of pancreatitis to increase the odds of sAP. Hospitalization (median, range) did not differ (sAP 3, 1–8 days; aGId 2.5, 1–5 days). Conclusion and Clinical Importance Both groups do not differ in clinical severity; diarrhea is less prevalent, and mild cholestasis is more common in sAP. Pancreatic ultrasonographic changes suggestive of AP are rare in aGId.

Trypanosoma cruzi is the etiologic agent of Chagas disease throughout the Americas. Few population-level studies have examined the epidemiology of canine infection and strain types of T. cruzi that infect canines in the USA. We conducted a cross-sectional study of T. cruzi infection in working hound dogs in south central Texas, including analysis of triatomine vectors collected within kennel environments. Paired IFA and Chagas Stat-Pak serological testing showed an overall seroprevalence of 57.6% (n = 85), with significant variation across kennels. Dog age had a marginally significant effect on seropositivity, with one year of age increase associated with a 19.6% increase in odds of being seropositive (odds ratio 95% CI 0.996-1.435; p = 0.055). PCR analyses of blood revealed 17.4% of dogs harbored parasite DNA in their blood, including both seronegative and seropositive dogs. Molecular screening of organs from opportunistically sampled seropositive dogs revealed parasite DNA in heart, uterus, and mammary tissues. Strain-typing showed parasite discrete typing units (DTU) TcI and TcIV present in dog samples, including a co-occurrence of both DTUs in two individual dogs. Bloodmeal analysis of Triatoma gerstaeckeri and Triatoma sanguisuga insects collected from the kennels revealed exclusively dog DNA. Vector infection with T. cruzi was 80.6% (n = 36), in which T. gerstaeckeri disproportionately harbored TcI (p = 0.045) and T. sanguisuga disproportionately harbored TcIV (p = 0.029). Tracing infection status across dog litters showed some seropositive offspring of seronegative dams, suggesting infection of pups from local triatomine vectors rather than congenital transmission. Canine kennels are high-risk environments for T. cruzi transmission, in which dogs likely serve as the predominant parasite reservoir. Disease and death of working dogs from Chagas disease is associated with unmeasured yet undoubtedly significant financial consequences because working dogs are highly trained and highly valued.

ABSTRACT Background No data after hospitalization for acute pancreatitis (AP) in dogs comparing clinical signs to lipase results exists. Objectives Evaluate disease severity, lipase activity, and pancreatic lipase immunoreactivity (PLI) after hospitalization for suspected AP. Animals One hundred and six client‐owned dogs with a minimum of one re‐check 2 weeks after hospitalization for AP. Methods Combined retrospective and prospective study. Clinical signs graded using a clinical disease activity score (CDAS = CIBDAI complemented by abdominal pain) were compared to DGGR‐lipase activity (LIPC Roche) and PLI (SpecPL) at 2 weeks (t2, n = 106) after discharge. Additional re‐checks were available 6 weeks (t3, n = 56), 12 weeks (t4, n = 24), and 24 weeks (t5, n = 13) after discharge. Results Lipase activity and PLI correlated strongly at all time points (rs 0.863–0.937, p < 0.0001). Discordant results in regard to published reference intervals (RI) were rare (2.8% at t2, 1.7% at t3, 4.2% at t4, 0% at t5) and seemed clinically irrelevant. Dogs with still elevated lipase activity and PLI at t2 (24/106.22.6%) and t3 (21/56.37.5%) were significantly older compared to dogs with lipase within RI. Weak and moderate correlation between CDAS and lipase activity/PLI was found only at t2 (rs 0.391, p = 0.0009; rs 0.279, p = 0.004) and t5 (rs 0.603, p = 0.032; rs 0.57 p = 0.045). Most dogs (79.2%) with still elevated lipase at t2 had no or minimal clinical signs (CDAS 0–3). The same applied to all later re‐checks. Conclusion and Clinical Importance Both lipase assays did not differ when compared to clinical status. Most dogs with hyperlipasemia after hospitalization for AP have no or minimal clinical signs.

Dogs diagnosed with chronic enteropathy (CE) or small-cell lymphoma (SCL) exhibit marked differences in faecal microbiota and organic acid profiles compared with healthy dogs, as well as immune abnormalities in intestinal mucosal tissue. However, few studies have analysed trace organic acids, such as succinic acid, which have been suggested to be associated with IBD in humans. Therefore, in this study, we compared the faecal microbiota and organic acid profiles as well as serum inflammatory markers between dogs with disease (n = 11; 6 with CE and 5 with SCL) and healthy controls (n = 16). We also performed machine learning and correlation analysis to obtain more detailed insights into the characteristics of affected dogs. These results revealed that dogs with CE and SCL had lower levels of Erysipelotrichaceae (e.g. Turicibacter and Allobaculum ), exhibited abnormalities in the succinic acid metabolism (i.e. succinic acid accumulation and decreased levels of Phascolarctobacterium as succinic acid-utilising bacteria) and increased levels of pathobiont bacteria such as Escherichia-Shigella . Additionally, the presence of Dubosiella was significantly negatively correlated with Canine Inflammatory Bowel Disease Activity Index scores. These findings are expected to aid the development of microbiome-based medications and/or supplements, although further verification is needed.