Molecular detection of hemoplasmas in domiciled and stray cats (Felis catus domesticus) from the states of Minas Gerais and São Paulo

• Grant number: 23/06168-2
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2023
• End date: June 30, 2024
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Preventive Veterinary Medicine
• Principal Investigator: Marcos Rogério André
• Grantee: Gabriela Veiga Oliveira
• Host Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil

Abstract

There are three species of hemoplasmas belonging to the genus Mycoplasma that are usually detected in domestic cats: Mycoplasma haemofelis, 'Candidatus Mycoplasma haemominutum' and 'Candidatus Mycoplasma turicensis'. They are cosmopolitan microorganisms, having already been identified infecting cats in all continents and in all regions of Brazil. Felids that get sick with hemoplasmosis may present acute or chronic disease, and the development of clinical signs - and their intensity - is related to several factors, such as the stage of the disease, the species of hemoplasma involved in the infection, host immunity, and the presence of co-infections. The clinical signs are quite nonspecific so the diagnosis should be made by molecular techniques such as conventional polymerase chain reaction (cPCR) or quantitative real-time quantitative (qPCR), which are more sensitive and specific than stained blood smears. The amplification of these gene fragments followed by sequencing allows phylogenetic analysis to be performed and subsequently to accurately determine the species involved in the feline infection. The present study aims to molecularly detect the occurrence of M. haemophelis, 'C. M. haemominutum' and 'C. M. turicensis' in cats from the states of Minas Gerais and São Paulo. For this 156 blood samples were obtained from cats in the cities of Araguari (MG), Uberlândia (MG) and Jaboticabal (SP) and the DNA will be extracted from them. In order to ensure that there are no inhibitors in the samples a cPCR assay targeting gapdh - the endogenous mammalian gene - will be conducted. Only positive samples will undergo a qPCR to screen for Mycoplasma spp based on the 16S rRNA gene. Samples that amplify in this screening assay will be subjected to characterization in a semi-nested PCR assay and a cPCR assay based on the 16S rRNA and 23S rRNA genes, respectively. Positive amplicons will be purified using a specific commercial kit and sequenced following the method described by Sanger. The sequences obtained will then be compared with those already deposited in Genbank with the aid of BLAST software. This will be the first study of molecular detection of hemoplasmas in cats from the state of Minas Gerais, which besides its clinical importance as possible cause of hemolytic anemia in domestic cats may also represent a public health problem due to its zoonotic potential and a concern for blood donor animals because the transmission via blood transfusion has already been reported.