OCCURRENCE OF CLOSTRIDIOIDES DIFFICILE IN FOALS FECES

• Grant number: 24/01072-0
• Support Opportunities: Scholarships in Brazil - Master
• Start date: June 01, 2024
• End date: April 30, 2026
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Clinics and Surgery
• Principal Investigator: Alexandre Secorun Borges
• Grantee: Thais Fernanda Ribeiro
• Host Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

Clostridioides difficile is one of the main etiological agents responsible for gastrointestinal diseases in humans and animals, known as Clostridioides difficile infection (CDI). It is a gram-positive, anaerobic, flagellated, and spore-forming bacterium that affects different species of animals and humans. Virulence is attributed to two main toxins, toxin A (TcdA) and toxin B (TcdB), with some strains producing binary toxin (CDT). In foals, CDI has clinical manifestations ranging from mild diarrhea to hyperacute hemorrhagic enterocolitis, and profuse colitis in adult horses. Diagnosis is based on the isolation of C. difficile, followed by the detection of genes encoding TcdA and TcdB. The importance of C. difficile in foals is well recognized, but in Brazil, there are still few studies investigating this bacterium in horses. The aim of the present study is to determine the shedding of C. difficile in the feces of neonatal foals over time. Fecal samples will be collected from foals (n=100) from five different stud farms in the Botucatu region, São Paulo, at three different time points: 7, 30, and 60 days of life. The samples will be inoculated in non-selective fructose broth supplemented with 0.1% sodium taurocholate for eight days. Subsequently, they will be plated on CDMN agar under anaerobic conditions for five days. Colonies with characteristic morphology and odor will undergo confirmation, and positive ones will be subcultured. Bacterial DNA extraction will be performed, and multiplex PCR will be carried out to determine constituent genes (16sRNA) and toxin producers (tcdA, tcdB, cdtA, and cdtB). For ribotyping, samples will undergo conventional PCR with capillary electrophoresis fragment analysis, and the results will be submitted to an already established database. Statistical analysis will be conducted to determine the influence of age and epidemiological factors on the detection of the agent and its toxins.