EFFECT OF ANTIBIOTIC CONCENTRATION ON THE TRANSPORT MEDIUM OF RED BROCKET DEER (Mazama rufa) SKIN: CONTAMINATION PREVENTION STRATEGY FOR CYTOGENETIC USE.

• Grant number: 23/14577-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: January 01, 2024
• End date: December 31, 2024
• Field of knowledge: Biological Sciences - Genetics - Animal Genetics
• Principal Investigator: José Maurício Barbanti Duarte
• Grantee: Isadora Beraldo Giacometti
• Host Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil

Abstract

Cytogenetic studies are an extremely important tool for revalidating taxa and distinguishing between cryptic species of deer, as occurred with the Mazama rufa species, previously considered to be Mazama americana. In order for these studies to be carried out, it is necessary to collect tissues from the specimens and transport the material to the laboratory for cell cultivation, which requires a suitable transport medium containing the correct concentrations of antibiotics and antifungals so that there is no contamination or damage to the cells. The aim of this study is to test different concentrations of Penicillin-Streptomycin 10,000ug/mL and Amphotericin B 1mg/L that inhibit the proliferation of microorganisms in the skin transport medium without damaging the tissue. To do this, skin fragments of the Mazama rufa species will be collected, divided into 10 smaller fragments and then deposited in a culture medium supplemented with different concentrations of the antibiotics and antifungals mentioned above. Three treatments will be tested: 9% Penicillin-Streptomycin solution 10,000ug/mL and 10mg/mL Amphotericin B 1mg/L; 5% Penicillin-Streptomycin solution 10. 000ug/mL and 5mg/mL of Amphotericin B 1mg/L; 2.5% solution of Penicillin-Streptomycin 10,000ug/mL and 2.5 mg/mL of Amphotericin B 1mg/L and DMEM medium without the addition of antimicrobials will be used as a control. The tissues will remain in the tested media for 8 hours before starting cell cultivation. In order to evaluate the effects of the different concentrations of the combination of antibiotic and antifungal tested on the cells, the speed of cell growth, cell confluence, cell viability using trypan blue dye and the mitotic index will be analyzed. In order to evaluate the potential effect of inhibiting contamination, DNA will be quantified and extracted from the tested media to determine the presence of bacterial DNAs and then electrophoresed in agarose gels.