Production and Characterization of an Antifungal Compound (3-Phenyllactic Acid) Produced by <Emphasis Type="Italic">Lactobacillus plantarum</Emphasis> Strain

The Lactobacillus plantarum strain was isolated from grass silage that produces a broad spectrum of antifungal compound, active against food and feed-borne filamentous fungi in agar plate assay. Aspergillus fumigatus and Rhizopus stolonifer were the most sensitive among molds. No inhibitory activity could be detected against mold Penicillium roqueforti. Enhanced antifungal activity was observed at 30 °C in pH 6.5. Minimum inhibitory concentration values against fungal cultures were ranged from 6.5 to 12.0 mg/ml for commercial 3-phenyllactic acid. The production of antifungal compound phenyllactic acid (PLA), lactic acid, and acetic acid by L. plantarum strain was also investigated. Structure characterization of the antifungal compound was carried out by nuclear magnetic resonance spectroscopy, infrared spectroscopy, and gas chromatography. The produced compound (PLA) acted as a fungistatic and delayed the growth of a variety of fungal contaminants.     

Antimicrobial effects of probiotic bacteria against selected species of yeasts and moulds in cheese-based dips

The antimicrobial properties of selected probiotic bacteria against Aspergillus niger , Penicillium roqueforti , Fusarium spp., Candida albicans and Saccharomyces cerevisiae were examined. Well diffusion and spot and streak methods showed strong inhibition effect of probiotic bacteria and their metabolites against moulds and minimal effect against yeasts. Among the moulds species tested, the inhibitory effect was strongest against Fusarium spp., moderate against Penicillium roqueforti and minimal against A. niger . All strains of Lactobacillus rhamnosus and L. paracasei subsp. paracasei showed maximum inhibitory effect. When probiotic bacteria and yeasts and moulds were co-cultured in broth media, strains of L. rhamnosus showed maximum inhibitory effect, whereas L. paracasei subsp. paracasei , L. acidophilus , Bifidobacterium animalis and Propionibacterium showed moderate inhibitory effect against C. albicans . Saccharomyces cerevisiae was minimally controlled by probiotic bacteria. Pre-grown probiotic bacterial culture metabolites controlled yeasts and moulds more effectively than their freeze-dried or frozen forms. Adding metabolites of probiotic bacteria (5% w/w) showed an effective control against A. niger , Fusarium spp. and C. albicans during the shelf life of 10 weeks at 4 °C and no colonies of yeasts and moulds were formed on the surface of the dip.     

白酒窖泥中乳酸菌分离鉴定及其发酵产挥发性风味物质比较

从白酒窖泥中分离得到11株乳酸菌,经16S rDNA基因序列同源性分析,鉴定为1株短乳杆菌、1株鼠李糖乳杆菌、2株干酪乳杆菌和7株铅黄肠球菌。分析了11株菌MRS培养基发酵产乳酸性能,结果表明:乳酸产量与菌群生长呈正相关,鼠李糖乳杆菌L9、干酪乳杆菌L10与L11发酵液乳酸含量较高,分别为15.74、15.58、14.74 g/L。4株代表菌相同条件下发酵高粱培养液产挥发性风味组分,共有物质13种,包含了白酒中重要香气成分:乙酸、己酸、乙酸苯乙酯和苯乙醇,且乙酸、己酸相对含量较高。各菌产可挥发性组分差异明显,相对含量较高的化合物种类为:短乳杆菌L5产烷烃类化合物(27.91%),铅黄肠球菌L8产醇类化合物(43.14%),鼠李糖乳杆菌L9产酯类(7.35%)、酮类(3.61%)和吡嗪类(3.3%)化合物,干酪乳杆菌L11产酸类(27.98%)和芳香类(5.96%)化合物。仅有短乳杆菌L5产四甲基吡嗪,短乳杆菌L5和铅黄肠球菌L8可明显产乙醇,鼠李糖乳杆菌L9和干酪乳杆菌L11产3-羟基-2-丁酮。这些物质对白酒风味和口感有重要影响。研究显示,窖泥中各种乳酸菌特征不一,对白酒酿造有不同影响。本研究旨在为进一步挖掘白酒酿造功能微生物、拓展乳酸菌的应用提供理论依据。     

Selective enumeration of Lactobacillus delbrueckii ssp. bulgaricus,Streptococcus thermophilus,Lactobacillus acidophilus,bifidobacteria, Lactobacillus casei,Lactobacillus rhamnosus,and propionibacteria

Nineteen bacteriological media were evaluated to assess their suitability to selectively enumerate Lactobacillus delbrueckii ssp. bulgaricus, Streptococcus thermophilus, Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus acidophilus, bifidobacteria, and propionibacteria. Bacteriological media evaluated included Streptococcus thermophilus agar, pH modified MRS agar, MRS-vancomycine agar, MRS-bile agar, MRS-NaCl agar, MRS-lithium chloride agar, MRS-NNLP (nalidixic acid, neomycin sulfate, lithium chloride and paramomycine sulfate) agar, reinforced clostridial agar, sugar-based (such as maltose, galactose, sorbitol, manitol, esculin) media, sodium lactate agar, arabinose agar, raffinose agar, xylose agar, and L. casei agar. Incubations were carried out under aerobic and anaerobic conditions at 27, 30, 37, 43, and 45 degrees C for 24, 72 h, and 7 to 9 d. S. thermophilus agar and aerobic incubation at 37 degrees C for 24 h were suitable for S. thermophilus. L. delbrueckii ssp. bulgaricus could be enumerated using MRS agar (pH 4.58 or pH 5.20) and under anaerobic incubation at 45 degrees C for 72 h. MRS-vancomycine agar and anaerobic incubation at 43 degrees C for 72 h were suitable to enumerate L. rhamnosus. MRS-vancomycine agar and anaerobic incubation at 37 degrees C for 72 h were selective for L. casei. To estimate the counts of L. casei by subtraction method, counts of L. rhamnosus on MRS-vancomycine agar at 43 degrees C for 72 h under anaerobic incubation could be subtracted from total counts of L. casei and L. rhamnosus enumerated on MRS-vancomycine agar at 37 degrees C for 72 h under anaerobic incubation. L. acidophilus could be enumerated using MRS-agar at 43 degrees C for 72 h or Basal agar-maltose agar at 43 degrees C for 72 h or BA-sorbitol agar at 37 degrees C for 72 h, under anaerobic incubation. Bifidobacteria could be enumerated on MRS-NNLP agar under anaerobic incubation at 37 degrees C for 72 h. Propionibacteria could be enumerated on sodium lactate agar under anaerobic incubation at 30 degrees C for 7 to 9 d. A subtraction method was most suitable for counting propionibacteria in the presence of other lactic acid bacteria from a product. For this method, counts of lactic bacteria at d 3 on sodium lactate agar under anaerobic incubation at 30 degrees C were subtracted from counts at d 7 of lactic bacteria and propionibacteria.     

Apparent antifungal activity of several lactic acid bacteria against Penicillium discolor is due to acetic acid in the medium

Fifty-six dairy bacteria belonging to the genera Lactococcus, Lactobacillus, Pediococcus, Propionibacterium, Streptococcus, Enterococcus, Leuconostoc, and Brevibacterium were screened for antifungal activity against four species of fungi relevant to the cheese industry (Penicillium discolor, Penicillium commune, Penicillium roqueforti, and Aspergillus vesicolor). Most of the active strains belonged to the genus Lactobacillus, whereas Penicillium discolor was found to be the most sensitive of the four fungi investigated. Further studies on P. discolor showed antifungal activity only below pH 5. This effect of pH suggests that organic acids present in the culture could be involved in the detected activity. Determination of acid composition revealed lactic acid production for active dairy strains and the presence of acetic acid in active as well as inactive strains. It was demonstrated that the undissociated acetic acid originates from the bacterial growth medium. The synergistic effect of the acetic acid present and the lactic acid produced was likely the main factor responsible for the antifungal properties of the selected bacteria. These results could explain some discrepancies in reports of the antifungal properties of lactic acid bacteria, since the role of acetic acid has not been considered in previous studies.     

抗食品腐败真菌的乳酸菌筛选、复配及混合发酵工艺优化

应用化学防腐剂控制食品中腐败微生物,防腐剂残留对人体具有一定的潜在性危害。本文筛选抗食品腐败真菌的乳酸菌,并对其进行复配及混合发酵以提高其发酵液抗真菌活性。通过24孔板双层琼脂法及分级抑菌浓度法从多株乳酸菌和丙酸杆菌中筛选抗真菌较强的菌株并确定混合菌最优组合。通过Plackett-Burman试验,最陡爬坡试验,对其发酵工艺进行优化,用96孔板酶标仪法测其发酵上清液的抗真菌活性。结果表明:最优混合菌组合为植物乳杆菌L9和费氏丙酸杆菌D5;发酵培养基优化配方:葡萄糖55 g/L,碳酸钙6.7 g/L,酵母浸粉14.8 g/L,磷酸氢二钾 0.25 g/L,硫酸锰 0.1 g/L,乙酸钠 5.0 g/L,柠檬酸铵 2.0 g/L,接种比例为5:1（D5:L9）,发酵温度为37 ℃。对筛选出的L9和D5在优化后的发酵培养基进行混合发酵验证,其发酵液抗真菌活性可高达47.07 AU。     

宜宾多粮型白酒窖房空气中产糖化酶的霉菌

对分离自窖房空气中的104株霉菌产糖化酶能力进行检测,发现其中有14株具备产糖化酶能力。结合形态、特征和基于ITS nu-rDNA的系统发育分析,此14株菌分属于Aspergillus（6株）、Mucor（3株）、Penicillium（2株）、Rhizopus（1株）、Gibberella（1株）、Cladosporium（1株）6个属,Aspergillus为优势属,显示浓香型白酒窖房中空气存在多样性较为明显的产糖化酶霉菌。     

Identification of fungal contamination and determination of mycotoxigenic molds by micellar electrokinetic capillary chromatography in smoked paprika

The purpose of this work was to analyze the fungal contamination in smoked and unsmoked paprika processed from different cultivars of pepper and to investigate the ability of these and other mycotoxigenic molds to grow and synthesize mycotoxins in smoked paprika. Eighteen mycotoxins were evaluated using micellar electrokinetic capillary chromatography. No relevant differences were found in fungal contamination between smoked and unsmoked paprika. The number of yeasts obtained was low, ranging from 0.4 to 3.29 log CFU g(-1); most of the yeast strains were identified as Cryptococcus spp. followed by Candida spp. All mold counts were <4 log CFU g(-1). Aspergillus, Cladosporium, Penicillium, and Fusarium were the predominant hyphomycete genera. Six mycotoxins were identified in the extracts of several strains isolated from paprika and incubated on malt extract agar. Penicillium expansum followed by Penicillium citrinum and Penicillium raistrickii were the dominant mycotoxigenic fungi isolated. Most of themycotoxin-producing fungi produced detectable amounts of mycotoxins when grown on paprika agar.     

不同储藏条件下玉米真菌多样性研究

研究不同含水率玉米在温度为20、25、303、5℃及相对湿度为70%～80%、80%～90%、≥90%条件下储藏过程中霉菌数量、种群组成、Simpson优势度指数和优势微生物种群变化动态。结果表明:从不同含水率玉米中分离到的优势霉菌主要是青霉属(Penicillium)、曲霉属(Aspergillus)、根霉属(Rhizopus)和镰刀霉属(Fusarium),玉米含水率为10.04%和14.72%时,优势霉菌为青霉属和曲霉属;玉米含水率为19.41%时,优势霉菌为曲霉属中灰绿曲霉和黄曲霉。霉菌多样性特征与玉米的储藏环境有关,温度低且玉米含水率低时,霉菌的多样性大,优势度小;而高含水率玉米的霉菌多样性小,优势度大。含水率和温度对玉米脂肪酸值变化有显著影响,玉米含水率和储藏温度越高,脂肪酸值含量越高。     

Mycoflora of two types of Portuguese dry-smoked sausages and inhibitory effect of sodium benzoate, potassium sorbate, and methyl p-hydroxybenzoate on mold growth rate

The mycoflora of chouriqo types Alentejano and Ribatejano, two varieties of Portuguese dry-smoked sausages, have been investigated after a producer-defined shelf life period (120 days at 20 +/- 5 degrees C) in modified atmosphere packaging (55% N2 and 45% CO2). On the basis of morphological and physiological characteristics, the isolates were identified as Penicillium, Aspergillus, Fusarium, Rhizopus, Monilia, Absidia, and Cephalosporium. The species identified were as follows: Penicillium terrestres (43.4%), Penicillium sp. (13.3%), Fusarium sp. (10%), Aspergillus glaucus (10%), Aspergillus versicolor (6.8%), Monilia fruticola (3.3%), Absidia sp. (3.3%), Cephalosporium sp. (3.3%), Rhizopus stolonifer (3.3%), and Fusarium tricinctum (3.3%). Additionally, the effects of three preservatives (potassium sorbate [PS], sodium benzoate [SB], and methyl p-hydroxybenzoate [MHB]) were studied on the growth rate of mold representative isolates. MHB showed a greater inhibitory effect than SB and PS in all fungi isolates, except in A. glaucus [Tm30(A)], in which the inhibitory effect of MHB was similar to PS. At 0.05% (wt/vol), all fungi were inhibited with MHB, except for R. stolonifer [Tm25(A)], which started to decrease the growth rate only at a concentration higher than 0.1%. PS was more effective at inhibiting mold growth than SB, except in Absidia sp. [Tm16(R)], in which both showed a similar inhibitory effect. MHB showed great promise as an application to the surface at 0.1% (wt/vol) to improve the stability and safety of the product through the inhibition of potential spoilage and toxigenic molds.     

Proteolytic activity of lactic acid bacteria strains and fungal biota for potential use as starter cultures in dry-cured ham

During the processing of dry-cured meat products, sarcoplasmic and myofibrillar proteins undergo proteolysis, which has a marked effect on product flavor. Microbial proteolytic activity is due to the action of mostly lactic acid bacteria (LAB) and to a lesser extent micrococci. The proteolytic capacity of molds in various meat products is of interest to meat processors in the Mediterranean area. Eleven LAB and mold strains from different commercial origins were tested for proteolytic activity against pork myosin, with a view to possible use of these strains as starter cultures for Iberian dry-cured ham. Proteolytic activity was tested by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The LAB strains with the highest proteolytic activity were Lactobacillus plantarum (L115), Pediococcus pentosaceus (Saga P TM), and Lactobacillus acidophilus (FARGO 606 TM). The best fungal candidate was Penicillium nalgiovense LEM 50I followed by Penicillium digitatum, Debaryomyces hansenii, and Penicillium chrysogenum.     

泡菜发酵乳酸菌的分离鉴定及耐药性分析

对吉林地区朝鲜族自制泡菜中的发酵乳酸菌进行分离鉴定,并对菌株耐药情况进行分析,以期为泡菜发酵菌的风险评估奠定基础。采集吉林省多个不同地区的泡菜制品,通过MRS培养基分离,16S rDNA测序,分析其乳酸菌种类及数量,同时采用平板琼脂稀释法进一步检测了分离菌对10 种临床常用抗生素的药物敏感性。结果表明,从15 份不同厂家泡菜中共分离得到34 株乳酸菌,分别为植物乳杆菌12 株,短乳杆菌5 株,鼠李糖乳杆菌3 株,干酪乳杆菌2 株,清酒杆菌2 株,肠膜明串珠菌2 株,嗜热链球菌2 株,食窦魏斯氏乳杆菌1 株,弯曲乳杆菌1 株,棒状乳杆菌1 株,嗜酸乳杆菌1 株,坚强肠球菌1 株,Lactobacillus namurensis 1 株。药敏实验结果表明,所有菌株对氨苄西林和氯霉素敏感,对其他抗生素均有不同程度的耐药性,其中对环丙沙星耐药率最高。     

干酪乳杆菌基因组改组菌株耐酸性表征

本实验对干酪乳杆菌基因组改组菌株的耐酸性进行了研究,结果表明,冷冻( － 80℃)和临界pH 值(pH3.2)对干酪乳杆菌基因组改组菌株影响不显著,HCl 和游离乳酸对该菌株生长有一定影响,游离乳酸影响更大。在pH5.0 条件下发酵60h,干酪乳杆菌改组菌株获得的菌体干重为6.34g/L,乳酸产量为87.6g/L,比原始菌株耐酸性有显著提高。     

核桃浆对鼠李糖乳杆菌培养特性及其富硒后抗缺氧活性的影响

为考察以核桃浆为原料开发廉价、高效的鼠李糖乳杆菌培养体系的可行性,本文用核桃浆代替MRS培养基中氮源,研究了核桃浆的颗粒度、料液比、用量、脱脂处理以及摇床转速对鼠李糖乳杆菌的生长、发酵液抗菌活性和抗氧化活性的影响,并用所得菌体经富硒处理后饲喂小鼠3 d,监测小鼠在暴露于低压缺氧条件下（海拔5000 m）6 h后的血细胞和血红蛋白含量,评价其抗缺氧潜力。结果发现,采用料液比为1:10 (g/mL),经120目筛网过滤的去脂核桃浆完全替代MRS培养基中氮源（牛肉膏、蛋白胨）,可使培养基成本降低70%,使菌体产量提高2.5倍（从2.6×109至7.6×109 CFU/mL）,但不影响发酵液的抗菌活性和抗氧化活性（P>0.05）。连续口服富硒菌体3 d,可显著提高小鼠在低压缺氧条件下的血红蛋白含量（P<0.05）,优于MRS培养基所得菌体。综上,用核桃浆能够完全代替MRS培养基中氮源用于培养鼠李糖乳杆菌,并能有效提高菌体产量和活性。     

河南烟叶仓储期优势霉菌的分离鉴定及防霉剂的筛选

对来自河南产区仓储烟叶中的主要霉菌进行了分离、鉴定,并测定了防霉剂对优势霉菌的抑制作用.结果表明,河南烟叶样品中霉菌数量以曲霉最多,其次是青霉.山梨酸钾、丙酸钙、双乙酸钠、脱氢醋酸钠、对羟基苯甲酸乙酯和富马酸二甲酯对黄曲霉、黑曲霉和桔青霉这3种优势霉菌均有明显的抑霉作用.     

一株抑制轮枝镰孢菌的鼠李糖乳杆菌的筛选及其在玉米防霉中应用

本研究以轮枝镰孢菌JF-5-1(Fusarium verticillium)为指示菌,对实验室保存的132株乳酸菌进行抑菌筛查,并对该菌株产生的抑霉菌物质进行了初步探究。结果表明,17株乳酸菌展现了良好的抑制轮枝镰孢菌特性,对轮枝镰孢菌具有较强抑制作用的为91号菌株,经过16S rDNA鉴定其为鼠李糖乳杆菌(Lactobacillus rhamnosus)。菌株上清液对木瓜蛋白酶和胰蛋白酶不敏感,且热稳定性较好,热处理并未改变抑菌性。当pH=5时,抑菌率急速下降为25.0%,当pH大于等于5.5时,91号菌株抑菌活性全部丧失。故初步推测该抑菌物质可能为有机酸,其最小抑菌浓度为8.0 mg/mL。本研究筛选出来的乳酸菌应用在玉米等粮食防霉中,其可抑制轮枝镰孢菌对粮食的侵染且绿色安全,具有非常广阔的应用前景。     

Antifungal effect of dairy propionibacteria--contribution of organic acids

Large amounts of food and feed are lost every year due to spoilage by moulds and yeasts. Biopreservation, i.e. the use of microorganisms as preservatives instead of chemicals, has gained increased interest. Lactic acid bacteria and propionibacteria might be particularly useful due to their important role in many food fermentations. Knowledge of the antifungal effects of the organic acids produced by these bacteria is necessary to understand their inhibitory activity. We evaluated the antifungal activity of the type strains of five dairy propionibacteria, Propionibacterium acidipropionici, P. jensenii, P. thoenii, P. freudenreichii subsp. freudenreichii and P. freudenreichii subsp. shermanii against eight food- and feedborne moulds and yeasts. A dual culture system assayed the inhibitory activity on three different agar media, sodium lactate (SL), de Man Rogosa Sharp (MRS) and MRS without acetate (MRS-ac). The amounts of organic acids produced during growth of propionibacteria in liquid SL, MRS and MRS-ac were also determined. The minimal inhibitory concentration (MIC) values of propionic, acetic and lactic acid were established for all fungi at pH 3, 5 and 7. Propionic acid, followed by acetic acid, was the most potent antifungal acid. Inhibition at pH 7 generally required concentrations above 500 mM for all three acids, at pH 5 the MIC values for propionic and acetic acids were 20-120 mM and above 500 mM for lactic acid. At pH 3, the MIC values were, with one exception, below 10 mM for both propionic and acetic acid and above 160 mM for lactic acid. The yeast Pichia anomala was the fungus most resistant to organic acids. The propionibacteria exhibited a pronounced species variation in antifungal activity on MRS (+/-acetate) agar, with P. thoenii being the most potent. Four of the five propionibacteria species produced more propionic and acetic acid in liquid SL medium than in MRS (+/-acetate) broth. However, when SL agar was used as the growth medium, none of the propionibacteria inhibited fungal growth.     

山东省部分地区2018年收获玉米的真菌污染状况研究

目的 对山东省部分地区2018年产玉米表面污染真菌进行调查。方法 在马铃薯葡萄糖培养基(potato dextrose agar, PDA)上通过稀释涂布的传统分离培养方法获得真菌菌株, 利用传统的形态学与分子生物学技术相结合的方法对真菌种类鉴定。结果 共获得真菌菌株169株, 经形态学鉴定隶属于8属: 镰刀菌属(Fusarium), 青霉属(Penicillum), 曲霉属(Aspergillus), 交链孢属(Alternaria), 枝顶孢属(Acremonium), 梗霉属(Lichtheimia), 枝孢属(Cladosporium)、篮状菌属(Talaromyces)。其中, 镰刀菌(Fusarium)、青霉(Penicillum)、曲霉(Aspergillus)分离获得的数量最多, 镰刀菌种类是引起玉米穗腐病的主要病原菌, 其污染主要来自于田间。随玉米仓储时间延长, 优势真菌改变为青霉属和曲霉属。结论 山东地区玉米真菌污染初期以田间污染为主, 储藏过程中优势菌群受环境影响发生改变; 需要根据真菌污染状况改进提升传统果穗囤藏技术。     

Antifungal activity of some essential oil components

Fifteen essential oil components were evaluated for antifungal activity towards five spoilage-causing fungi. In liquid shake cultures, unsaturated aldehydes (citral, cinnamic aldehyde and cittronellal) followed by geraniol, an unsaturated alcohol were most inhibitory to Aspergillus niger, Fusarium oxysporum and Penicillium digitatum; their minimal inhibitory concentrations (MIC) was 100 μg ml⁻¹. Menthol, a terpene alcohol was most inhibitory to Rhizopus stolonifer and Mucor sp. with a MIC of 200 μg ml⁻¹. Hydrocarbons like camphene, limonene and α-terpinene were least inhibitory. When incorporated in agar medium different patterns of activity were found. Thus citral, cinnamic aldehyde, citronellal, geraniol and menthol not only failed to completely inhibit A. niger, F. oxysporum and P. digitatum but were more active against R. stolonifer and Mucor sp. than in liquid medium. The differences were due to the vapour of the volatile test compounds which accumulated over the agar medium. The growth inhibition due to the vapours alone was measured by using structurally modified petri-plates. The vapours were more active against R. stolonifer and Mucor sp. than against A. niger, F. Oxysporum and P. digitatum. Fungal growth inhibition by volatile compounds in agar medium reflects the combined activity of the vapour and the compound incorporated in the medium and the inhibition is different from that obtained in liquid medium. Regarding the structure-activity relationship of citral, the-CHO group in conjugation with a carbon to carbon double bond (C=C) was found to be responsible for the antifungal activity of the molecule.     

Fungistatic Activity of Heat-Treated Flaxseed Determined by Response Surface Methodology

ABSTRACT:  The objective of this study was to evaluate the effect of heat treatment on the fungistatic activity of flaxseed ( Linum usitatissimum ) in potato dextrose agar (PDA) medium and a fresh noodle system. The radial growth of Penicillium chrysogenum , Aspergillus flavus , and a Penicillium sp. isolated from moldy noodles, as well as the mold count of fresh noodle enriched with heat treated flaxseed, were used to assess antifungal activity. A central composite design in the response surface methodology was used to predict the effect of heating temperature and time on antifungal activity of flaxseed flour (FF). Statistical analysis determined that the linear terms of both variables (that is, heating temperature and time) and the quadratic terms of the heating temperature had significant ( P < 0.05) effects on the radial growth of all 3 test fungi and the mold count log-cycle reduction of fresh noodle. The interactions between the temperature and time were significant for all dependent variables ( P < 0.05). Significant reductions in antifungal activities were found when FF was subjected to high temperatures, regardless of heating time. In contrast, prolonging the heating time did not substantially affect the antifungal activities of FF at low temperature. However, 60% of the antifungal activity was retained after FF was heated at 100 °C for 15 min, which suggests a potential use of FF as an antifungal additive in food products subjected to low to mild heat treatments.