Suppressive effect of culture supernatant of erythrocytes and serum from dogs infected with Babesia gibsoni on the morphological maturation of canine reticulocytes in vitro.

he present study evaluated the effects of erythrocytes infected culture supernatant, fractionation of culture supernatant and serum of dogs infected with Babesia gibsoni (B. gibsoni) on in vitro maturation of reticulocytes dog. SDS-PAGE shows that a significantly wider band produced by both culture supernatant of infected erythrocytes and serum from chronically infected with B. gibsoni dog. 

Culture supernatant of erythrocytes infected Hamster Serum with B. gibsoni greatly suppressed the maturation of reticulocytes. previous studies have shown that chronically infected serum has an inhibitory effect on the maturation of reticulocytes and dogs pyrimidine 5'-nucleotidase special subclass I and purine 5'-nucleotidase activity. In addition, serum-free culture supernatant of infected erythrocytes have an inhibitory effect on the morphological maturation of reticulocytes. 

These results indicate that the infected serum and erythrocyte culture supernatant may accumulate excess protein and / or metabolites as a result of inhibiting the maturation of reticulocytes and decreased erythrocyte 5'-nucleotidase activity. In addition, the fraction was observed in 150 kDa- and 150-70 >> kDa- in the infected culture supernatant and serum inhibits the in vitro maturation of reticulocytes dog. Results obtained from in vitro assays, in this study, suggesting that B. gibsoni itself and / or its metabolites may release certain proteins in the infected culture supernatant and serum from an infected dog and as a result of delays in the morphological maturation of reticulocytes dogs.

Validation of human immunoturbidimetric assays are commercially available for the determination of serum haptoglobin in the sample dog.

Haptoglobin is a positive acute phase protein with a valuable role as a marker of inflammation in both human and veterinary medicine. The purpose of this study was to validate the commercially available immunoturbidimetric method designed for the determination of human haptoglobin (Izasa SA, Barcelona, ​​Spain) for use in a sample of dogs. Antisera cross-reactivity between anti-human haptoglobin and haptoglobin dog found when agarose gel immunodiffusion and ELISA tests performed. 

The use of dogs collected serum haptoglobin concentration of 6.3 g / L as supplied various analytical standard higher than the standard commercially available. Intra-assay and inter-assay coefficients of variation were 2.49% and 4.60%, respectively. Linear regression model between the results immunoturbidimetric and spectrophotometric method Hamster Serum Albumin previously validated (TRIDELTA Development Limited, Ireland) produces the slope at 95% confidence interval 0.94 (0.86, 1.02) and the y-intercept at the 95% confidence interval 0.11 ( -0.59, 0.82).

No significant differences were produced by an anticoagulant, lipemia and bilirubinaemia, although hemolysis significantly decreased haptoglobin. A significant increase of haptoglobin concentration was detected in inflammatory conditions such as pyometra and leishmaniasis, in neoplastic conditions and after the administration of glucocorticoids.