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，并评估其用于药用及动物生产的拮抗潜力。蜂蜜样本采集自Serra Talhada-PE地区，从4个不同蜂箱各采集3份重复样本，总计12份样本。将样本稀释后置于80℃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（Colony Forming Units，CFU）进行定量检测。同时将样本涂布于麦芽琼脂及沙氏葡萄糖琼脂（Sabouraud agar）平板，于30℃培养14天以培养酵母与霉菌。采用最大似然数法（Most Probable Number，MPN）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，但仅4株具备益生潜力。其中分离株I-5的益生潜力最为突出，可抑制大肠杆菌（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.）及金黄色葡萄球菌（Staphylococcus aureus）的生长。所有分离株均未对八叠球菌属（Sarcina sp.）的生长产生抑制作用。未检出酵母、霉菌及大肠菌群。Melipona属无刺蜂蜂蜜是产孢细菌的优质来源，其对污染蜂蜜的微生物具有拮抗活性，且具备优良的微生物学品质。