INHIBITION OF PATHOGENS BY SPOROGENIC BACTERIA ISOLATED FROM HONEY OF Melipona sp. (APIDAE: APINAE: MELIPONINI)

ABSTRACT The aim of this study was to isolate sporogenic bacteria from the honey of stingless bees Melipona sp., in dry forest, and to evaluate their antagonistic potential for medicinal employment purposes and animal production. The honey samples were collected in Serra Talhada-PE, where honey was taken from four different hives (in triplicate), totaling 12 samples. The samples were diluted and subjected to 80 ºC for 20 minutes to eliminate vegetative cells. The dilutions were plated onto nutrient agar and incubated at 30 ºC for 72 hours. Then the colony forming units (CFU) were quantified. The samples were also plated onto malt agar and Sabouraud agar, and incubated at 30 ºC for 14 days for the growth of yeast and molds. Total and fecal coliforms were quantified by the most probable number method (MPN). Seven isolates (I) of sporogenic bacteria (Bacillus) were obtained, however only four showed probiotic potential. Isolate I-5 showed the greatest probiotic potential and inhibited the growth of Escherichia coli, Klebsiella sp., Pseudomonas aeruginosa, Salmonella sp., and Staphylococcus aureus. The growth of the Sarcina sp. was not inhibited by any isolate. No yeast, molds or coliforms were found. The Melipona sp. honey is a source of spore-forming bacteria and is antagonistic to microorganisms that contaminate honey. It has good microbiological quality.

摘要 本研究旨在从干旱森林中的无刺蜂Melipona属（Melipona sp.）蜂蜜中分离产孢细菌，并评估其在医药领域与动物生产中的拮抗应用潜力。蜂蜜样本采集自巴西塞拉塔尔哈达-佩（Serra Talhada-PE），从4个不同蜂箱各采集3份重复样本，总计12份样本。将样本经稀释后置于80℃环境下处理20分钟以杀灭营养体细胞。将稀释液涂布于营养琼脂平板，于30℃培养72小时后对菌落形成单位（CFU, Colony Forming Units）进行定量计数。此外，将样本分别涂布于麦芽琼脂（malt agar）与沙氏葡萄糖琼脂（Sabouraud agar）平板，于30℃培养14天以分离培养酵母与霉菌。采用最大概率数法（MPN, Most Probable Number）对总大肠菌群与粪大肠菌群进行定量检测。本研究共获得7株产孢细菌（芽孢杆菌属，Bacillus）分离株，其中仅4株表现出益生潜能。其中I-5分离株的益生潜能最为显著，可抑制大肠埃希氏菌（Escherichia coli）、克雷伯氏菌属（Klebsiella sp.）、铜绿假单胞菌（Pseudomonas aeruginosa）、沙门氏菌属（Salmonella sp.）以及金黄色葡萄球菌（Staphylococcus aureus）的生长。所有分离株均未抑制八叠球菌属（Sarcina sp.）的生长。本研究未检出酵母、霉菌及大肠菌群。Melipona属蜂蜜是产孢细菌的潜在来源，且对污染蜂蜜的微生物具有拮抗活性，其微生物学质量优良。