Standardization of a Polymerase Chain Reaction (PCR) for Leptospira detection in tissue samples of aquatic mammals of t. inunguis and t. manatus species

• Grant number: 16/09215-8
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: June 01, 2016
• End date: February 28, 2017
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Preventive Veterinary Medicine
• Principal Investigator: Lara Borges Keid
• Grantee: Kerstin Muner
• Host Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil

Abstract

Leptospirosis is an infectious disease that affects several animal species, which includes aquatic mammals. In these animals, infection caused by L. interrogans and L. kirschneri serovars, for example, have been described. Furthermore, it was discovered recently a genetically similar strain of L. interrogans in whale (Eubalaena australis), which is sea water resistant. The microbiological culture is considered the gold standard technique for leptospirosis diagnosis, however, the polymerase chain reaction (PCR) can be used as an alternative for its diagnosis, by the detection of Leptospira DNA in tissue or body fluid samples from suspected animals, which might be more practical and feasible when dealing with aquatic mammals, due to the fact that it is not always possible to process the samples immediately after their collection, as the culture technique requires. In order to turn easier, practical, and to standardize the molecular diagnosis of leptospirosis in these animals, a review of the described primers in literature will be conducted, by checking the performance, analytical specificity, and sensitivity and specificity of these primers to amplify Leptospira DNA in marine manatee (Trichechus manatus) and Amazon manatee (Trichechus inunguis) samples. The establishment of a standard protocol to identify, practically and satisfactorily, leptospires causing disease in aquatic mammals is essential for the diagnosis and treatment of leptospirosis, identification of the infection sources, differentiation between outbreaks and sporadic cases, and for a better understanding of the disease epidemiology in the aquatic mammal population.