Araújo Carlos, Muñoz-Atienza Estefanía, Poeta Patrícia, Igrejas Gilberto, Hernández Pablo E, Herranz Carmen, Cintas Luis M
Grupo de Seguridad y Calidad de los Alimentos por Bacterias Lácticas, Bacteriocinas y Probióticos (Grupo SEGABALBP), Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040 Madrid, Spain.
Dis Aquat Organ. 2016 May 3;119(2):129-43. doi: 10.3354/dao02992.
The use of lactic acid bacteria (LAB) as probiotics constitutes an alternative or complementary strategy to chemotherapy and vaccination for disease control in aquaculture. The objectives of this work were (1) the in vitro safety assessment of 8 Pediococcus acidilactici strains isolated from rainbow trout (Oncorhynchus mykiss, Walbaum) feed and larvae; (2) the evaluation of their genetic relatedness; (3) the study of their antimicrobial/bacteriocin activity against fish pathogens; and (4) the biochemical and genetic characterization of the bacteriocin produced by the strain displaying the greatest antimicrobial activity. Concerning the safety assessment, none of the pediococci showed antibiotic resistance nor produced hemolysin or gelatinase, degraded gastric mucin, or deconjugated bile salts. Four strains (50%) produced tyramine or putrescine, but the corresponding genes were not amplified by PCR. Enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) fingerprinting allowed clustering of the pediococci into 2 well-defined groups (68% similarity). From the 8 pediococci displaying direct antimicrobial activity against at least 3 out of 9 fish pathogens, 6 strains (75%) were identified as bacteriocin producers. The bacteriocin produced by P. acidilactici L-14 was purified, and mass spectrometry and DNA sequencing revealed its identity to pediocin PA-1 (PedPA-1). Altogether, our results allowed the identification of 4 (50%) putatively safe pediococci, including 2 bacteriocinogenic strains. ERIC-PCR fingerprinting was a valuable tool for genetic profiling of P. acidilactici strains. This work reports for the first time the characterization of a PedPA-1-producing P. acidilactici strain isolated from an aquatic environment (rainbow trout larvae), which shows interesting properties related to its potential use as a probiotic in aquaculture.
使用乳酸菌(LAB)作为益生菌是水产养殖中疾病控制的一种替代或补充策略,可替代化疗和疫苗接种。本研究的目的是:(1)对从虹鳟(Oncorhynchus mykiss,Walbaum)饲料和幼体中分离出的8株嗜酸乳杆菌进行体外安全性评估;(2)评估它们的遗传相关性;(3)研究它们对鱼类病原体的抗菌/细菌素活性;(4)对表现出最大抗菌活性的菌株所产生的细菌素进行生化和遗传特征分析。关于安全性评估,所有嗜酸乳杆菌均未表现出抗生素抗性,也未产生溶血素或明胶酶,未降解胃粘蛋白或使结合胆汁盐解偶联。四株菌株(50%)产生酪胺或腐胺,但相应基因未通过PCR扩增。肠杆菌重复基因间共有序列PCR(ERIC-PCR)指纹图谱分析将嗜酸乳杆菌聚类为两个明确的组(相似度为68%)。在对9种鱼类病原体中的至少3种具有直接抗菌活性的8株嗜酸乳杆菌中,有6株(75%)被鉴定为细菌素产生菌。嗜酸乳杆菌L-14产生的细菌素被纯化,质谱和DNA测序显示其与植物乳杆菌素PA-1(PedPA-1)相同。总之,我们的结果鉴定出4株(50%)推定安全的嗜酸乳杆菌,其中包括2株产细菌素的菌株。ERIC-PCR指纹图谱分析是嗜酸乳杆菌菌株遗传分析的一种有价值的工具。本研究首次报道了从水生环境(虹鳟幼体)中分离出的一株产PedPA-1的嗜酸乳杆菌的特征,该菌株具有一些有趣的特性,表明其在水产养殖中作为益生菌具有潜在用途。