Metabolomics study of cat small intestine during the early stage of Toxoplasma gondii oocyst formation identifies potential biomarkers -raw datas pos mode
Description
Toxoplasma gondii is a zoonotic intracellular protozoan parasite that can invade, replicate and survive in almost all cells of warm-blooded animals. Toxoplasmosis can fatally threaten immunocompromised individuals and pregnant women. As the only definitive host of T. gondii, felids spread the pathogen mainly by forming oocysts in the small intestines and discharging the oocysts into the ambient environment, consequently polluting water, vegetables, and meat products. In this study, we used untargeted metabolomics technology to study the changes in metabolites that occurred during the early stage of oocyst formation in the cat small intestine following T. gondii infection and attempted to identify metabolic biomarkers that could potentially be used as diagnostic molecular markers in the future. Domestic cats (Felis catus) were infected with T. gondii Pru tissue cysts, and samples of their small intestinal epithelium were collected at 2 and 4 days post-infection (DPI) for metabolic analysis. LC-MS/MS and multivariate statistical analysis were employed to detect metabolomic signatures that discriminated between the infected and control groups. A total of 1,673 ions and 1,201 ions were obtained in the positive and negative modes, respectively. Of these ions, 175 were up-regulated and 127 were down-regulated in the positive ion mode; whereas, 123 were up-regulated and 81 were down-regulated in the negative ion mode. Three commonly altered ions (0.74_313.0414m/z, 8.82_615.2621m/z and 8.16_325.2362m/z) were determined to have potential research value. Seventy common metabolic pathways were enriched at two time points, with arginine biosynthesis, pyrimidine metabolism, pantothenate and CoA biosynthesis being the three most significant pathways related to T. gondii. The area under the curve (AUC) of differential metabolites combined with relevant literature analysis showed that N-Methylpelletierine and 3,3-Difluoro-17-methyl-5alpha-androstan-17beta-ol have higher predictability and better potential application value than other metabolites. Our analysis of metabolic markers during the early stage of T. gondii oocyst formation in the small intestine of the definitive host (cat) provided novel insight for understanding oocyst development and a theoretical basis for the application of potential biomarkers.
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Eighteen female domestic cats (Felis catus, Chinese Li Hua breed, aged 7-9 months) were purchased from a local breeder and housed individually in a controlled environment. They were kept under laboratory conditions for a week after they had adapted to the environment and were then randomly assigned to two experimental groups (2DPI and 4DPI, corresponding to T. gondii-infected hosts at 2 and 4 days post infection, respectively) and one control group (control), with six cats in each group. All the tested animals were negative for T. gondii serology and common feline diseases. Animals from the experimental groups were challenged intragastrically with 600 Pru tissue cysts in 100 µL phosphate buffered saline (PBS) each, whereas those from the control group were fed with the corresponding dose of PBS. At 2 and 4 days post-infection (DPI) with T. gondii, tissue collection was performed as a terminal procedure from cats under deep isoflurane anesthesia, a sample of intestinal epithelium from each cat was collected, quickly frozen in liquid nitrogen, and transferred to the company BGI (Shenzhen, China) for sequencing.