Westerdijk Fungal Biodiversity Institute, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands.
Westerdijk Fungal Biodiversity Institute, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands.
Int J Food Microbiol. 2019 Oct 2;306:108258. doi: 10.1016/j.ijfoodmicro.2019.108258. Epub 2019 Jul 8.
Propionic acid is widely used as a preservative in (poultry) feed. In this study we have isolated and identified fungal strains from nine samples poultry feed originating from different countries. The majority of the strains were Aspergilli with a eurotium-morph, such as Aspergillus proliferans and A. chevalieri. These and three other species were selected and tested for their sensitivity towards the feed preservative propionic acid, among them Penicillium lanosocoeruleum. The determined MIC values of 6.1-31 mM of these poultry feed specific fungi were well in the range as described in literature. Propionic acid (at 31 mM) damages conidia (spores) in a species dependent fashion after a 24-hour-treatment. The majority of the conidia (over 70%) of P. lanosocoeruleum germinated within 60 h on agar medium, while 50 and 80% of the A. chevalieri and A. proliferans conidia did not, respectively. Dependent on the species, cell damage was visible after incubation with propionic acid. Germ tubes of P. lanosocoeruleum in a biofilm showed extensive (85%) cell death after a 30 min treatment with propionic acid and slightly lower sensitivity was observed with A. proliferans (62% cell death). Microscopic analysis of these fungal biofilms revealed extensive damage to the cell membrane and showed distorted intracellular structures. Fluorescent life-dead staining of the germ tubes showed a clear dose response of propionic acid indicating a fungicidal effect on these growing cells. These results show that conidia can be inactivated by propionic acid, but that germ tubes show a much higher sensitivity. These observations shed new light on the mode of action of this important preservative to prevent fungal contamination of feed.
丙酸被广泛用作(家禽)饲料的防腐剂。在这项研究中,我们从来自不同国家的九种家禽饲料样品中分离和鉴定了真菌菌株。大多数菌株为曲霉属,具有 Eurotium 形态,如增殖曲霉和 A. chevalieri。这些和另外三个物种被选中并测试了它们对饲料防腐剂丙酸的敏感性,其中包括青霉 lanosocoeruleum。这些特定于家禽饲料的真菌的 MIC 值(6.1-31mM)与文献中描述的范围非常吻合。丙酸(在 31mM 时)在 24 小时处理后以依赖于物种的方式破坏分生孢子(孢子)。青霉 lanosocoeruleum 的大多数分生孢子(超过 70%)在琼脂培养基上 60 小时内发芽,而 A. chevalieri 和 A. proliferans 的分生孢子分别有 50%和 80%没有发芽。根据物种的不同,在用丙酸孵育后可以看到细胞损伤。在丙酸处理 30 分钟后,生物膜中的青霉 lanosocoeruleum 的生芽管显示出广泛的(85%)细胞死亡,而增殖曲霉的敏感性略低(62%细胞死亡)。对这些真菌生物膜的显微镜分析显示细胞膜受到广泛损伤,并显示出扭曲的细胞内结构。生芽管的荧光死活染色显示出丙酸的明确剂量反应,表明对这些生长细胞具有杀菌作用。这些结果表明,分生孢子可以被丙酸灭活,但生芽管显示出更高的敏感性。这些观察结果为这种重要防腐剂防止饲料真菌污染的作用模式提供了新的见解。