Holý Ondrej, Parra-Flores Julio, Lepuschitz Sarah, Alarcón-Lavín María Paula, Cruz-Córdova Ariadnna, Xicohtencatl-Cortes Juan, Mancilla-Rojano Jetsi, Ruppitsch Werner, Forsythe Stephen
Department of Public Health, Palacký University Olomouc, 77515 Olomouc, Czech Republic.
Department of Nutrition and Public Health, Universidad del Bío-Bío, Chillán 3800708, Chile.
Foods. 2020 Dec 23;10(1):20. doi: 10.3390/foods10010020.
spp. are opportunistic pathogens of the family. The organism causes infections in all age groups, but the most serious cases occur in outbreaks related to neonates with meningitis and necrotizing enterocolitis. The objective was to determine the in silico and in vitro putative virulence factors of six strains isolated from powdered milk (PM) in the Czech Republic. Strains were identified by MALDI-TOF MS and whole-genome sequencing (WGS). Virulence and resistance genes were detected with the Ridom SeqSphere+ software task template and the Comprehensive Antibiotic Resistance Database (CARD) platform. Adherence and invasion ability were performed using the mouse neuroblastoma (N1E-115 ATCCCRL-2263) cell line. The CRISPR-Cas system was searched with CRISPRCasFinder. Core genome MLST identified four different sequence types (ST1, ST145, ST245, and ST297) in six isolates. Strains 13755-1B and 1847 were able to adhere in 2.2 and 3.2 × 10 CFU/mL, while 0.00073% invasion frequency was detected only in strain 1847. Both strains 13755-1B and 1847 were positive for three (50.0%) and four virulence genes, respectively. The gene was not detected. Twenty-eight genes were detected by WGS and grouped as flagellar or outer membrane proteins, chemotaxis, hemolysins, and invasion, plasminogen activator, colonization, transcriptional regulator, and survival in macrophages. The colistin-resistance-encoding and cephalothin-resis-encoding genes and IncFII(pECLA) and IncFIB(pCTU3) plasmids were detected. All strains exhibited CRISPR matrices and four of them two type I-E and I-F matrices. Combined molecular methodologies improve spp. decision-making for health authorities to protect the population.
某属细菌是该菌科的机会致病菌。该生物体可在所有年龄组引发感染，但最严重的病例发生在与新生儿脑膜炎和坏死性小肠结肠炎相关的疫情中。目的是确定从捷克共和国奶粉（PM）中分离出的6株菌株的计算机模拟和体外推定毒力因子。通过基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）和全基因组测序（WGS）对菌株进行鉴定。使用Ridom SeqSphere+软件任务模板和综合抗生素抗性数据库（CARD）平台检测毒力和抗性基因。使用小鼠神经母细胞瘤（N1E-115 ATCC CRL-2263）细胞系检测黏附和侵袭能力。使用CRISPRCasFinder搜索CRISPR-Cas系统。核心基因组多位点序列分型（MLST）在6株分离株中鉴定出4种不同的序列类型（ST1、ST145、ST245和ST297）。菌株13755-1B和1847能够分别以2.2×10和3.2×10 CFU/mL的浓度黏附，而仅在菌株1847中检测到0.00073%的侵袭频率。菌株13755-1B和1847分别有3个（50.0%）和4个毒力基因呈阳性。未检测到某基因。通过WGS检测到28个基因，并分为鞭毛或外膜蛋白、趋化性、溶血素、侵袭、纤溶酶原激活剂、定植、转录调节因子和在巨噬细胞中的存活相关基因。检测到编码黏菌素抗性的某基因和编码头孢菌素抗性的某基因以及IncFII(pECLA)和IncFIB(pCTU3)质粒。所有菌株均表现出CRISPR阵列，其中4株有两种I-E型和I-F型阵列。联合分子方法可改善某属细菌的情况，为卫生当局保护民众提供决策依据。
