Multi Platforms Strategies and Metabolomics Approaches for the Investigation of Comprehensive Metabolite Profile in Dogs with Babesia canis Infection.

Ivana Rubić,Richard Burchmore,Predrag Novak,Peter David Eckersall,Renata Barić Rafaj,Martina Crnogaj,Stefan Weidt,Clement Regnault,Josipa Kuleš,Vladimir Mrljak,Tomislav Mašek,Anita Horvatić

doi:10.3390/ijms23031575

Abstract

Canine babesiosis is an important tick-borne disease worldwide, caused by parasites of the Babesia genus. Although the disease process primarily affects erythrocytes, it may also have multisystemic consequences. The goal of this study was to explore and characterize the serum metabolome, by identifying potential metabolites and metabolic pathways in dogs naturally infected with Babesia canis using liquid and gas chromatography coupled to mass spectrometry. The study included 12 dogs naturally infected with B. canis and 12 healthy dogs. By combining three different analytical platforms using untargeted and targeted approaches, 295 metabolites were detected. The untargeted ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) metabolomics approach identified 64 metabolites, the targeted UHPLC-MS/MS metabolomics approach identified 205 metabolites, and the GC-MS metabolomics approach identified 26 metabolites. Biological functions of differentially abundant metabolites indicate the involvement of various pathways in canine babesiosis including the following: glutathione metabolism; alanine, aspartate, and glutamate metabolism; glyoxylate and dicarboxylate metabolism; cysteine and methionine metabolism; and phenylalanine, tyrosine, and tryptophan biosynthesis. This study confirmed that host–pathogen interactions could be studied by metabolomics to assess chemical changes in the host, such that the differences in serum metabolome between dogs with B. canis infection and healthy dogs can be detected with liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC-MS) methods. Our study provides novel insight into pathophysiological mechanisms of B. canis infection.

Highlights

Canine babesiosis is a tick-borne disease of worldwide importance caused by intraerythrocytic protozoa of different Babesia species [1]
A total of 22 metabolites were identified using more than one plat- than one platform: 14 metabolites were measured by liquid chromatography-mass spectrometry (LC-MS)
We found that metabolomic profiles of dogs infected with B. canis distinguish significantly from healthy dogs

Summary

Introduction

Canine babesiosis is a tick-borne disease of worldwide importance caused by intraerythrocytic protozoa of different Babesia species [1]. Babesiosis in dogs is caused by both large and small forms of Babesia (B. canis, B. vogeli, B. gibsoni, and B. microti-like isolates) [2]. The large Babesia, previously considered to be B. canis, is currently split into three distinct species, namely B. canis, B. rossi, and B. vogeli [2]. Canine babesiosis caused by B. canis is the most common infection of dogs in certain regions of Europe [3]. The occurrence of canine babesiosis is associated with clinical cases, mostly in spring and autumn, because the relatively mild and wet weather is ideal for ticks [4]. The disease can be clinically classified into uncomplicated and complicated forms

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