Phenotypical and molecular characterization of Bulkholderia mallei in Equidae with glanders

• Grant number: 17/14434-3
• Support Opportunities: Regular Research Grants
• Start date: November 01, 2017
• End date: October 31, 2019
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Preventive Veterinary Medicine
• Principal Investigator: Alessandra Figueiredo de Castro Nassar
• Grantee: Alessandra Figueiredo de Castro Nassar
• Host Institution: Instituto Biológico (IB). Agência Paulista de Tecnologia dos Agronegócios (APTA). Secretaria de Agricultura e Abastecimento (São Paulo - Estado). São Paulo , SP, Brazil
• Associated scholarship(s): 18/08777-8 - Phenotypical and molecular characterization of Bulkholderia mallei in Equidae with glanders, BP.TT

Abstract

Glanders is a disease caused by Burkholderia mallei, a Gram negative bacterium, which affects the equines triggering disease of chronic or acute character, being able to reach the man, the carnivores and eventually small ruminants. It is a notifiable disease in the world, and is a major cause of morbidity and mortality in horses. Currently, glanders are endemic in countries such as Brazil, the Middle East, Asia and Africa. Horses presenting with symptoms are: high fever, depression, nasal discharge, cough, nodular lesions that develop into ulcers assuming a stellate form after healing, which occur more frequently in the chronic phase of the disease. The objective of this work is to perform the phenotypic and molecular identification of B. mallei in organs (lung, lymph nodes, spleen, liver and kidney) of seropositive animals for Glanders (complement fixation), real-time PCR (qPCR) standardization, and to correlate PCR with the qPCR, besides performing the genetic sequencing of the bacteria in different samples. The profile of the macroscopic and microscopic lesions will be evaluated as a complementary bacteriological and molecular diagnosis of the circulating B. mallei strains. Fifty samples of equidae with positive serological diagnosis of Glanders will be collected in the State of São Paulo, with 35 horses housed in the quarantine station of Cananéia / SP, and the others in outbreaks of the disease under sanitary interdiction of the Agriculture and Livestock Defense Coordination of the State of São Paulo. The animals will be necropsied and the organs submitted to macroscopic and histopathological analyzes (haematoxylin and eosin staining). Microbiological tests of the potato and Mueller Hinton agar samples, plus glycerol, sheep's blood and with or without penicillin will be carried out with a view to the growth and identification of colonies of B. mallei by biochemical tests. PCR from the colonies and organs will be performed to detect the B. mallei Flic gene, PCR Real Time, and later genetic sequencing of the Flip gene. The methods of direct identification of the agent (bacterial isolation, PCR and qPCR) aim to be incorporated into the laboratory diagnosis of the Biological Institute and thus to assist in the sanitary, epidemiological and disease control measures by the animal health defense organs in Brazil. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

ABREU, DIEGO CANDIDO; GOMES, ALINE SILVA; TESSLER, DANIELLE KLEIN; CHIEBAO, DANIELA PONTES; DEL FAVA, CLAUDIA; DE CAMPOS NOGUEIRA ROMALDINI, ADRIANA HELLMEISTER; ARAUJO, MATEUS CARVALHO; POMPEI, JULIO; MARQUES, GUILHERME FIGUEIREDO; HARAKAVA, RICARDO; et al. Systematic monitoring of glanders-infected horses by complement fixation test, bacterial isolation, and PCR. VETERINARY AND ANIMAL SCIENCE, v. 10, DEC 2020. (18/08777-8, 17/14434-3)