engleski

Targeted metabolomics approach for investigation of the metabolome in dogs infected with Babesia canis

Canine babesiosis is an important worldwide tick- borne disease caused by the intra-erythrocyte protozoa of different Babesia species [1]. Although the disease process primarily affects erythrocytes, it may also have multisystemic consequences [2]. The emergence of innovative - omics technology, such as metabolomics, has led to the development of strategies aimed at identifying specific and sensitive metabolites among thousands of small molecules present in biological fluids and tissues [3]. Metabolomics methodologies have been divided into two distinct groups: untargeted metabolomics and targeted metabolomics [4]. Targeted metabolomics identifies and quantifies the abundance of defined groups of known, chemically characterized and biochemically annotated metabolites. Blood and urine contains a multitude of unstudied and unknown biomarkers that may reflect physiological and pathological states of tissues and organs. The goal of the study was to obtain serum metabolom of dogs naturally infected with Babesia canis and healthy dogs using targeted metabolomics approach on the liquid chromatography coupled to mass spectrometry (LC- MS) platform. Metabolites were analyzed using the Absolute IDQ p400 kit (Biocrates Life Science AG, Innsbruck, Austria), for the targeted metabolomic analysis of up to 408 metabolites divided into 11 metabolite classes. According to the manufacturer instructions provided with the kit, metabolites were extracted from dog serum on the specific 96- well plate system for protein-removal, internal standard normalization and derivatization. Analyses were made according to the manufacturer’s protocol using the Biocrates MetIDQ software (Biocrates Life Science AG, Innsbruck, Austria) for data processing, quality assessment, data export and mapping of measurements with chemical and biochemical background information. Targeted metabolomics analysis of serum samples was performed for 24 dogs through combined LC-MS/MS and FIA-MS/MS analysis. Univariate analysis resulted in detection of 68 significant metabolites in healthy dogs versus dogs with babesiosis. The most significant metabolites are serotonin, methionine sulfoxide, citrulline, proline and others. Biological functions of differently abundant metabolites indicate the involvement of various pathways in canine babesiosis including glutathione metabolism, alanine, aspartate and glutamate metabolism, glyoxylate and dicarboxylate metabolism and others pathways. In conclusion, the research of serum samples of dogs infected with B. canis demonstrated potential metabolites and pathways significantly changed in dogs with canine babesiosis.The targeted LC-MS metabolomic’s approach profiled the metabolic change in serum of dogs infected with B.canis.

serum samples ; babesiosis ; targeted metabolomics ; chromatography ; mass spectrometry

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