Departamento de Microbiología e Inmunología, Facultad de Ciencias Exactas, Físico-Químicas y Naturales, Universidad Nacional de Río Cuarto, Río Cuarto, Córdoba, Argentina.
Departamento de Microbiología e Inmunología, Instituto de Ciencias Veterinarias (INCIVET-CONICET-UNRC), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Río Cuarto, Córdoba, Argentina.
J Basic Microbiol. 2023 Aug;63(8):877-887. doi: 10.1002/jobm.202300008. Epub 2023 May 8.
Beneficial Bacillus strains can be administered to livestock as probiotics to improve animal health. Cyclic lipopeptides produced by Bacillus such as surfactins may be responsible for some of the beneficial effects due to their anti-inflammatory and immunomodulatory activity. The aim of the present study was to isolate and evaluate the biocompatibility of native Bacillus spp. strains and their surfactin-like lipopeptides in vitro and in vivo to determine their potential to be used on animals. Biocompatibility of endospore suspensions (10 UFC/mL), and different dilutions (1:10; 1:50; 1:100; 1:500, and 1:1000) of Bacillus lipopeptide extracts containing surfactin was tested on Caco-2 cells by microculture tetrazolium-based colorimetric assay. Genotoxicity was tested on BALB/c mice (n = 6) administered 0.2 mL of endospore suspensions by the bone marrow erythrocyte micronuclei assay. All the isolates tested produced between 26.96 and 239.97 µg mL of surfactin. The lipopeptide extract (LPE) from isolate MFF1.11 demonstrated significant cytotoxicity in vitro. In contrast, LPE from MFF 2.2; MFF 2.7, TL1.11, TL 2.5, and TC12 had no cytotoxic effect (V% > 70%) on Caco-2 cells, not affecting cell viability signifficantly in most treatments. Similarly, none of the endospore suspensions affected cell viability (V% > 80%). Likewise, endospores did not cause genotoxicity on BALB/c mice. This study was elementary as a first step for a new line of research, since it allowed us to choose the safest isolates to keep working on the search of new potentially probiotic strains destined to production animals to improve their performance and health.
有益的芽孢杆菌菌株可以作为益生菌施用于牲畜，以改善动物健康。芽孢杆菌产生的环状脂肽，如表面活性剂，可能因其抗炎和免疫调节活性而对一些有益作用负责。本研究的目的是分离和评估天然芽孢杆菌菌株及其表面活性剂样脂肽的体外和体内生物相容性，以确定它们在动物身上使用的潜力。通过微量培养四唑比色法，测试了芽孢杆菌脂肽提取物中含有的表面活性剂的芽孢悬浮液（10 UFC/mL）和不同稀释度（1:10；1:50；1:100；1:500 和 1:1000）对 Caco-2 细胞的生物相容性。通过骨髓红细胞微核试验在 BALB/c 小鼠（n = 6）中测试了遗传毒性，给予 0.2 mL 芽孢悬浮液。所有测试的分离株均产生 26.96 至 239.97 µg/mL 的表面活性剂。分离株 MFF1.11 的脂肽提取物（LPE）在体外显示出显著的细胞毒性。相比之下，MFF 2.2；MFF 2.7、TL1.11、TL 2.5 和 TC12 的 LPE 对 Caco-2 细胞没有细胞毒性作用（V%>70%），在大多数处理中对细胞活力没有显著影响。同样，没有一种芽孢悬浮液影响细胞活力（V%>80%）。同样，芽孢也没有在 BALB/c 小鼠中引起遗传毒性。这项研究是作为新研究的第一步进行的，因为它使我们能够选择最安全的分离株，继续寻找新的潜在益生菌菌株，用于生产动物，以提高它们的性能和健康。
