Identification of serum biomarkers in dogs naturally infected with Babesia canis canis using a proteomic approach

Abstract

BackgroundCanine babesiosis is a tick-borne disease that is caused by the haemoprotozoan parasites of the genus Babesia. There are limited data on serum proteomics in dogs, and none of the effect of babesiosis on the serum proteome. The aim of this study was to identify the potential serum biomarkers of babesiosis using proteomic techniques in order to increase our understanding about disease pathogenesis.ResultsSerum samples were collected from 25 dogs of various breeds and sex with naturally occurring babesiosis caused by B. canis canis. Blood was collected on the day of admission (day 0), and subsequently on the 1st and 6th day of treatment.Two-dimensional electrophoresis (2DE) of pooled serum samples of dogs with naturally occurring babesiosis (day 0, day 1 and day 6) and healthy dogs were run in triplicate. 2DE image analysis showed 64 differentially expressed spots with p ≤ 0.05 and 49 spots with fold change ≥2. Six selected spots were excised manually and subjected to trypsin digest prior to identification by electrospray ionisation mass spectrometry on an Amazon ion trap tandem mass spectrometry (MS/MS). Mass spectrometry data was processed using Data Analysis software and the automated Matrix Science Mascot Daemon server. Protein identifications were assigned using the Mascot search engine to interrogate protein sequences in the NCBI Genbank database.A number of differentially expressed serum proteins involved in inflammation mediated acute phase response, complement and coagulation cascades, apolipoproteins and vitamin D metabolism pathway were identified in dogs with babesiosis.ConclusionsOur findings confirmed two dominant pathogenic mechanisms of babesiosis, haemolysis and acute phase response. These results may provide possible serum biomarker candidates for clinical monitoring of babesiosis and this study could serve as the basis for further proteomic investigations in canine babesiosis.

Oxidative status and lipid profile in mono- and co-infection with canine monocytic ehrlichiosis

“…Ehrlichiosis is known as a catabolic disease -like most haemoparasites, the agent of the disease cannot synthesize its own lipids -and is predominantly bound to HDL derived from the host's plasma (20,22). In a present study, HDL values were significantly lower in both groups infected with vector-borne diseases compared to healthy dogs.…”

Canine Babesiosis: Where Do We Stand?

“…Excessive proinfl ammatory activity with increased concentrations of CRP was also detected in natural infection with B. canis [91] and experimental one with B. gibsoni [108]. In our proteomic study of serum changes in canine babesiosis, a number of differentially expressed proteins involved in infl ammation mediating the acute phase response, including CRP, were identifi ed in dogs with babesiosis [84].…”

“…Other studies employing two dimensional electrophoresis and liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) showed potential for using proteomic approach not only for diagnosis, but for monitoring and outcome prediction in canine babesiosis [84,107].…”

“…The destruction of the erythrocyte is caused by multifactorial components, including direct parasite damage to the erythrocyte membrane, splenic removal of damaged and parasitized erythrocytes, as well as activation of the immune system such as complement cascade and/or presence of anti-erythrocyte antibodies [83]. Using a proteomic approach, the role of haemolysis in the course of babesiosis was demonstrated by changes in expression of haptoglobin, hemopexin and serotransferrin [84]. Erythrocyte lysis is associated with a broad spectrum of clinical manifestations, which are not always proportional to the degree of anaemia and are not correlated with the level of parasitemia, which usually remains low [85].…”

Vitamin D metabolism in dogs and cats and its relation to diseases not associated with bone metabolism

“…This protein function is to carry vitamin D and its plasma metabolites (Kuleš et al, 2014). Concerning dogs with babesiosis, a study was developed evaluating proteomics, including the proteins which bind to vitamin D in the metabolism pathway.…”