Juan D. García-Martínez

Serum ferritin and paraoxonase-1 in canine leishmaniosis

Ferritin and paraoxonase-1 (PON-1) were measured in dogs experimentally infected by Leishmania infantum (during experimental infection and following treatment) and also in naturally-infected dogs which presented different degrees of proteinuria. Experimentally-infected dogs were monitored for 7 months post-infection, then treated for 3 months with allopurinol, and their response to therapy was followed for 11 additional months. Naturally-infected dogs were staged based on the urine protein/creatinine (UPC) ratio into three groups as follows: group 1 (non-proteinuric; UPC ratio: <0.2), group 2 (borderline proteinuric; UPC ratio: 0.2-0.5) and group 3 (proteinuric; UPC ratio>0.5). An increase in serum ferritin values and a decrease in PON-1 activity were observed 2 months after infection. Both analytes returned to preinfection values following treatment. Significantly higher concentrations of ferritin were observed in dogs classified as either borderline or proteinuric when compared with non-proteinuric dogs whereas serum PON-1 activity was decreased only in proteinuric dogs.

Urinary C reactive protein levels in dogs with leishmaniasis at different stages of renal damage

The objectives of the study were to validate a time-resolved immunofluorometric assay for C reactive protein (CRP) quantification in urine of dogs and to investigate the influence that the presence of proteinuria and azotemia could have on serum and urinary CRP (uCRP) values in dogs with leishmaniasis. Samples obtained from dogs naturally infected with Leishmania infantum were classified into four groups on the basis of the results of urinary protein/creatinine ratio and serum creatinine (sCr). In addition, 7 dogs were monitored at initial diagnosis and after a follow up visit. The assay showed good analytical performance based on precision, accuracy and limit of detection results. Results of the study suggested that CRP is present in urine of dogs with leishmaniasis and renal damage since uCRP/creatinine ratio was significantly increased in dogs with proteinuria, being the highest values observed in dogs with proteinuria and elevated sCr, and that the measurement of uCRP could be a tool to detect and evaluate the possible kidney damage associated with this disease.

Urinary clusterin as a renal marker in dogs

A validation of a species-specific enzyme immunoassay for urinary clusterin measurement in dogs was performed, and the use of urinary clusterin as a marker of renal damage was evaluated in a population of dogs with leishmaniasis. Urine was obtained from 75 dogs; 64 dogs had leishmaniasis and 11 were healthy. The dogs with leishmanias were divided into 5 groups: I (n = 9; serum creatinine [SCr] < 1.4 mg/dl, urinary protein-to-creatinine [UPC] ratio ≤ 0.5); II (n = 29; SCr < 1.4 mg/dl, UPC > 0.5); III (n = 6; SCr ≥ 1.4 mg/dl to <2 mg/dl, UPC > 0.5); IV (n = 13; SCr ≥ 2 mg/dl to <5 mg/dl, UPC > 0.5); and V (n = 7; SCr ≥ 5 mg/dl, UPC > 0.5). The urinary clusterin concentration was measured, and the urinary clusterin-to-creatinine ratio was calculated. Canine urinary clusterin assay showed good analytical performance based on precision accuracy and limit-of-detection results. There was a statistically significant increase in urinary clusterin and clusterin-to-creatinine ratio in groups II–V compared with group I and healthy group. The results of the current study showed that urinary clusterin concentration and urinary clusterin-to-creatinine ratios are increased in dogs with analytical evidences of renal damage and that the urinary clusterin-to-creatinine ratio might be used as a potential early biomarker of chronic kidney disease.

Urinary ferritin and cystatin C concentrations at different stages of kidney disease in leishmaniotic dogs

Traditional analytes do not detect early renal disease; therefore there is a need to find new early markers of chronic kidney disease (CKD) in dogs to avoid the progression to irreversible renal damage. Our objective was to evaluate the presence of ferritin and cystatin C in urine of dogs with CKD and to relate their concentrations with the severity of the disease. Samples obtained from dogs naturally infected with Leishmania infantum were classified into four groups on the basis of the results of urinary protein/creatinine ratio and serum creatinine. This study shows that ferritin and cystatin C concentrations were increased in the urine of dogs with renal damage. Cystatin C value in urine only increased in severe stages of CKD with serum creatinine values >1.4 mg/dL, while the urinary ferritin concentration increased in dogs with proteinuria and serum creatinine <1.4 mg/dL, being, therefore, a renal biomarker earlier than creatinemia.

Canine C-reactive protein measurements in cerebrospinal fluid by a time-resolved immunofluorimetric assay

In the current study, the quantification of C-reactive protein (CRP) in cerebrospinal fluid (CSF) of dogs using an adapted time-resolved immunofluorimetric assay (TR-IFMA) was investigated, as well as whether the assay could be used to detect the range of CRP concentrations found in different clinical situations. Intra- and interassay coefficients of variation were below 15% in all cases. The TR-IFMA measured the CRP values in a proportional and linear manner (r = 0.99); also CRP concentrations measured in CSF and in serum were significantly correlated (r = 0.80, P = 0.003). The limit of detection of the method was 7.1 × 10−6 mg/l. The assay was able to detect differences in CRP concentrations in CSF of dogs with inflammatory disorders compared with dogs with spinal cord compression or idiopathic epilepsy. In conclusion, TR-IFMA constitutes a very sensitive, precise, and accurate method for the measurement of CRP concentrations in CSF.

Serum paraoxonase 1 (PON1) activity in acute pancreatitis of dogs.

OBJECTIVES Serum paraoxonase 1 is considered a marker of inflammation and oxidative damage. The aims of this study were to evaluate changes in serum paraoxonase 1 activity in dogs with acute pancreatitis, to correlate serum paraoxonase 1 activity and other analytes known to be altered in dogs with pancreatitis and to assess the relationship between serum paraoxonase 1 activity and disease severity in dogs with acute pancreatitis. MATERIALS AND METHODS Retrospective analysis of dogs with acute pancreatitis and healthy dogs in which serum paraoxonase 1 activity was measured were compared. RESULTS Median serum paraoxonase 1 activity was significantly lower in dogs with pancreatitis (n = 19) compared to healthy ones (n = 19). Serum paraoxonase 1 activity was negatively correlated with serum lipase and amylase activities, and C-reactive protein and haptoglobin concentrations and was positively correlated with total cholesterol and glucose concentration. Disease severity was negatively correlated with serum paraoxonase 1 activity and positively correlated with triglyceride and C-reactive protein concentration. CLINICAL SIGNIFICANCE Serum paraoxonase 1 activity is lower in dogs with acute pancreatitis and together with triglyceride and C-reactive protein concentrations is a potential marker of disease severity.

ACUTE PHASE PROTEIN RESPONSE IN THE CAPYBARA (HYDROCHOERUS HYDROCHAERIS)

We evaluated the acute phase protein response in capybaras (Hydrochoerus hydrochaeris). Three animal groups were used: 1) healthy animals (n=30), 2) a group in which experimental inflammation with turpentine was induced (n=6), and 3) a group affected with sarcoptic scabies (n=14) in which 10 animals were treated with ivermectin. Haptoglobin (Hp), acid-soluble glycoprotein (ASG) and albumin were analyzed in all animals. In those treated with turpentine, Hp reached its maximum value at 2 wk with a 2.7-fold increase, whereas ASG increased 1.75-fold and albumin decreased 0.87-fold 1 wk after the induction of inflammation. Capybaras affected with sarcoptic scabies presented increases in Hp and ASG of 4.98- and 3.18-fold, respectively, and a 0.87-fold decrease in albumin, compared with healthy animals. Haptoglobin and ASG can be considered as moderate, positive acute phase proteins in capybaras because they showed less than 10-fold increases after an inflammatory process and reached their peak concentrations 1 wk after the induction of inflammation. Conversely, albumin can be considered a negative acute phase protein in capybaras because it showed a reduction in concentration after inflammatory stimulus.

Serum and urinary adiponectin in dogs with renal disease from leishmaniasis

The objective of this study was to perform an analytical validation of a commercially available ELISA kit (human adiponectin) for urinary adiponectin determination in dogs, and to evaluate urinary adiponectin in dogs with glomerular injury. For this purpose, urine samples from three healthy dogs and three dogs with diagnosed kidney disease were used for analytical validation of the method. In order to evaluate possible influence of kidney damage on urinary adiponectin, serum and urine samples from six healthy and 58 dogs with leishmaniasis were included. The diseased dogs were allocated to three groups according to their urine protein/creatinine (UPC) ratio as non-proteinuric (NP), borderline proteinuric (BP), and proteinuric (P). Intra- and inter-assay coefficients of variation (CV) were lower than 10 per cent and 12 per cent, respectively. Dilutions of canine urine samples resulted in linear regression equations close to 1. Mean recovery was of 112 per cent. The detection limit was 0.75 ng/ml. Urinary adiponectin and urinary adiponectin/creatinine (UAC) ratio showed significantly higher values in urine of P group dogs compared with healthy, NP and BP dogs. In conclusion, an ELISA kit can be used for precise and accurate urinary adiponectin measurement in dogs. Urinary adiponectin is increased in dogs with proteinuria suggesting its possible use as a marker of kidney damage.

Evaluation of various biomarkers for kidney monitoring during canine leishmaniosis treatment

BackgroundThe objective of this study was to evaluate and compare the evolution of the profile currently recommended by the International Renal Interest Society (IRIS) (sCr, UPC and sSDMA) with a panel of other different kidney biomarkers during treatment for canine leishmaniosis. This panel included three urinary glomerular biomarkers (uIgG, uCRP and uferritin) and three urinary tubular biomarkers (uGGT, uNAG and uRBP).These biomarkers were measured in two groups of dogs with canine leishmaniosis at IRIS stage I. Group 1: dogs showing proteinuria (UPC > 0.5) before treatment which did not decrease after treatment; Group 2: dogs showing proteinuria before treatment which decreased after treatment.ResultsGroup 1 showed no significant changes in any biomarker after treatment. In group 2, among the biomarkers recommended by the IRIS, only UPC showed a significant decrease after treatment. However all biomarkers of glomerular damage showed a significant decrease after treatment, with uIgG/Cr and uCRP/Cr showing the greater decreases. In addition uRBP/Cr and uNAG/Cr showed significant decreases after treatment.ConclusionsIn dogs with leishmaniosis at IRIS stage I that reduced UPC after treatment, there were no significant changes in serum creatinine and sSDMA. However, all the urine biomarkers evaluated with exception of uGGT showed a significant decrease. These decreases were more evident in those markers related with glomerular function, being uIgG/Cr the biomarker more associated with UPC. Further studies involving a larger number of animals and histological analysis of the kidney would be recommended to confirm these findings and evaluate the routine practical use of these urine biomarkers in canine leishmaniosis.

An Atypical Case of Leishmaniasis Associated with Chronic Duodenitis in a Dog

We describe an atypical case of duodenal leishmaniasis in a boxer dog presenting with chronic diarrhea and poor general condition. Antidiarrheic therapy was previously administered without success and inflammatory bowel disease localized to the small intestine was suspected, given the chronic clinical signs and by ruling out other known causes of gastrointestinal inflammation. Endoscopic biopsy of duodenum showed a moderate increase in lamina propria lymphocytes, plasma cells, and macrophages. Basophilic bodies were seen in the cytoplasm of numerous macrophages, suggestive of Leishmania spp, confirmed by immunostaining, and a diagnosis of granulomatous duodenitis associated to Leishmania infection was made. After 7 mo of therapy, a significant clinical improvement and weight gain were observed, and endoscopic histology showed no evidence of Leishmania. A progressive decline of anti-leishmanial antibody titer was also observed during follow-up. This report emphasizes the importance of atypical symptoms and the unusual location of visceral leishmaniasis, suggesting the need to consider leishmaniasis in the differential diagnosis of canine chronic enteritis, especially in endemic areas.