State University of Maringá Agricultural Science Center Animal Science Graduate Program Gavazzoni, L.; Galhardo, D.; Pavanelli, F.M.; and Toledo, V.A.A. SEARCH OF Bacillus spp. in Nannotrigona testaceicornis (Lepeletier, 1836) Native Írai (Nannotrigona testaceicornis) Stingless bees Meliponines Family Apidae ~ 3mm © Copyright Hadel Go 2014 - Social; Subfamily - They live in colonies; Apinae - Neotropical region; Tribe Meliponini 50 cm 15 cm Colonies have from 2.000 to 3.000 bees Wide distribution in Brazil Pollination in greenhouses Maintenance of biodiversity and floristic composition. HEALTH AND PRODUCTIVITY: Ecological and Economic Role Healthy Bees Microbiota in Balance Symbiosis with Microorganisms + Benefits: - Nutritional - Immunity - Bacillus spp. - Defense OBJECTIVE Identify these microorganisms in the stingless bee Nannotrigona testaceicornis (Iraí or Black Jataí). MATERIAL AND METHODS Samples collection 1 1 1 - Maringá city 2 2 - Araruna city 3 3 - Campo Mourão Figure 1. Location of the study area. Samples Collection: Colonies of Nannotrigona testaceicornis FEI - UEM Araruna - Pr Campo Mourão - Pr ADULT BEES HONEY STERILE COLLECTION POLLEN MATERIAL BROOD and LARVAE PROPOLIS Sample Preparation: ADULT BEES, Stingless bees (n=20 units per BROOD AND colony) LARVAE Hypochlorite 2.5% Sterile water BHI broth Solution 10% Sample Preparation: Incubated at 37 °C for 6 hours 10 grams BHI Honey Beebread Propolis Pre-enrichment Microbiological evaluation: 1mL 1mL Serial Dilution 10 -1 10 -2 10 -3 10% Solution BHI Sample TSA + 100µL AN BHI broth 24/48 horas 24/48 Spread Plate Spread Plating MC Incubation 37 °C MAN MRS Pour plate Microbiological Isolation evaluation: Subcultive Streaking Individual colonies RESULTS Sporulated bacilli Gram positive staining ✓ Rods Positive microorganisms for Bacillus spp. Catalase + RESULTS Figure 2. Mean total microorganisms CONCLUSIONS Bacillus spp. SYMBIOSIS ✓Antimicrobial potential; Colony health ; ✓Aid in the pre-digestion of pollen; New means of research! Probiotics Acknowledgments State University of Maringá MY MOM Higher Education Personnel Improvement Coordination National Council for Scientific and Technological Development.