奶山羊消化道源乳酸菌对黄曲霉抑制及AFB1解毒的特性分析

doi:10.12160/j.issn.1672-5190.2026.03.005

摘要/Abstract

摘要： [目的] 旨在从奶山羊消化道分离的乳酸菌中,筛选出兼具抑制产毒黄曲霉生长和降解黄曲霉毒素 B₁（aflatoxin B₁,缩写AFB₁）能力的优良菌株。[方法] 以30株奶山羊消化道源乳酸菌为材料,采用96孔板法测定无细胞发酵上清液（CFS）对黄曲霉孢子萌发的抑制率,采用ELISA法检测CFS对AFB₁的降解率,综合抑制率和降解率结果筛选优良菌株。通过琼脂扩散法、双层平板法分别评估所筛菌株的CFS和培养液的抗真菌活性,利用LC-MS分析筛选菌株CFS有机酸组成及变化,利用扫描电子显微镜（scanning electron microscope,SEM）观察菌株培养液对黄曲霉形态的影响,并比较具有活性的与灭活处理的CFS及菌体培养液对AFB₁的去除效果。[结果] 筛选获得两株优良菌株：植物乳杆菌R3和戊糖片球菌I3。R3和I3菌株的CFS对黄曲霉孢子抑制率均高于84%,对AFB₁的降解率分别为81.42%和81.96%。琼脂扩散法结果表明,I3的CFS抗真菌活性略强于R3;但双层平板法结果表明,R3菌株的培养液抗真菌活性强于I3菌株。有机酸分析表明,乳酸、丙酮酸等是CFS中的主要成分,且在与AFB₁作用后R3菌株CFS处理的毒素组的丙酮酸含量与R3的对照组相比显著（P<0.05）降低;在与AFB₁作用后,I3菌株CFS处理的毒素组的DL-3-苯基乳酸的含量与I3的对照组相比显著（P<0.05）降低。SEM观察结果表明,两株乳酸菌培养液导致黄曲霉菌丝形态畸变,孢子产生减少。AFB₁去除实验显示,经过沸水灭活处理后,R3和I3菌株的总去除率分别提升到54.36%和51.65%;经过沸水灭活后,两株菌的CFS对AFB₁的降解率均有所降低。R3菌株对AFB₁的总去除效果略优于I3。[结论] 该研究从奶山羊消化道分离得到乳酸菌菌株 R3、I3,两者均能通过产生有机酸抑制黄曲霉生长并去除AFB₁。其中,植物乳杆菌R3抗真菌作用和去除AFB₁能力更强。

关键词: 奶山羊消化道源乳酸菌, 黄曲霉, 黄曲霉毒素B₁, 有机酸

Abstract: [Objective] To screen superior strains with the ability to inhibit the growth of toxigenic Aspergillus flavus and degrade aflatoxin B1 （AFB₁） from lactic acid bacteria isolated from the digestive tract of dairy goats. [Methods] With 30 strains of lactic acid bacteria from digestive tract of dairy goats as materials, the inhibition rate of cell-free fermentation supernatant （CFS） on spore germination of A. flavus was determined using 96-well plate method, and the degradation rate of AFB₁ by CFS was detected using ELISA method. Superior strains were screened based on the combined results of inhibition rate and degradation rate. The CFS and the antifungal activities of culture broth of the screened strains were evaluated using the agar diffusion method and double-layer plate method, respectively. The composition and changes of organic acids in CFS were analyzed using LC-MS. The effect of culture broth on the morphology of A. flavus was observed by scanning electron microscope （SEM）, and the removal effect of active and inactivated CFS and bacterial culture broth on AFB₁ was compared. [Results] Two superior strains were obtained: Lactobacillus plantarum R3 and Pediococcus pentosaceus I3. The inhibition rates of CFS against A. flavus spores of R3 and I3 strains were higher than 84 %, and the degradation rates of AFB₁ were 81.42% and 81.96%, respectively. The results of agar diffusion method showed that the antifungal activity of CFS of I3 was slightly stronger than that of R3. However, the results of double-layer plate method showed that the antifungal activity of the culture broth of R3 strain was stronger than that of I3 strain. Organic acid analysis showed that lactic acid and pyruvic acid were the main components of CFS, and the pyruvic acid content in the toxin group treated with R3 CFS after interaction with AFB₁ significantly decreased （P<0.05） compared with the R3 control group; after interaction with AFB₁, the DL-3-phenyllactic acid content in the toxin group treated with I3 CFS significantly decreased （P<0.05） compared with the I3 control group. The results of SEM observation showed that the culture broth of both strains resulted in the morphological deformation of A. flavus mycelium and the decrease of spore production. The AFB₁ removal experiment showed that after boiling water inactivation, the total removal rates of R3 and I3 strains increased to 54.36% and 51.65%, respectively. After being inactivated by boiling water, the degradation rate of AFB₁ by CFS of the two strains decreased. The total removal effect of R3 strain on AFB₁ was slightly better than that of I3. [Conclusion] In this study, lactic acid bacteria strains R3 and I3 were isolated from the digestive tract of dairy goats. Both of them can inhibit the growth of A. flavus and remove AFB₁ by producing organic acids. Among them, L. plantarum R3 exhibits stronger antifungal effect and ability to remove AFB₁.

Key words: lactic acid bacteria from digestive tract of dairy goats, Aspergillus flavus, aflatoxin B₁, organic acids

中图分类号:

S827
Q939.9

马睿, 王平. 奶山羊消化道源乳酸菌对黄曲霉抑制及AFB₁解毒的特性分析[J]. 畜牧与饲料科学, 2026, 47(3): 37-49.

MA Rui, WANG Ping. Characteristic Analysis of Lactic Acid Bacteria from the Digestive Tract of Dairy Goats on Inhibition of Aspergillus flavus and Detoxification of AFB₁[J]. Animal Husbandry and Feed Science, 2026, 47(3): 37-49.

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奶山羊消化道源乳酸菌对黄曲霉抑制及AFB₁解毒的特性分析

Characteristic Analysis of Lactic Acid Bacteria from the Digestive Tract of Dairy Goats on Inhibition of Aspergillus flavus and Detoxification of AFB₁

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