Andritsos Nikolaos D, Mataragas Marios
Department of Food Science and Technology, School of Agricultural Sciences, University of Patras, 2 G. Seferi Str., GR-301 00 Agrinio, Greece.
Department of Dairy Research, Institute of Technology of Agricultural Products, Hellenic Agricultural Organization "DIMITRA", 3 Ethnikis Antistaseos Str., GR-452 21 Ioannina, Greece.
Foods. 2023 Mar 12;12(6):1200. doi: 10.3390/foods12061200.
Listeriosis is a serious infectious disease with one of the highest case fatality rates ( 20%) among the diseases manifested from bacterial foodborne pathogens in humans, while dairy products are often implicated as sources of human infection with . In this study, we characterized phenotypically and genetically by whole-genome sequencing (WGS) 54 strains isolated from Myzithra, a traditional Greek soft whey cheese (48 isolates), and swabs collected from surfaces of a cheese processing plant (six isolates) in the Epirus region of Greece. All but one strain of belonged to the polymerase chain reaction (PCR) serogroups IIa (16.7%) and IIb (81.5%), corresponding to serotypes 1/2a, 3a and 1/2b, 3b, 7, respectively. The latter was identified as a PCR-serogroup IVb strain (1.8%) of serotypes 4b, 4d, 4e. Bioinformatics analysis revealed the presence of five sequence types (STs) and clonal complexes (CCs); ST1, ST3, ST121, ST 155, ST398 and CC1, CC3, CC121, CC155, CC398 were thus detected in 1.9, 83.3, 11.0, 1.9, and 1.9% of the isolates, respectively. Antibiograms of the pathogen against a panel of seven selected antibiotics (erythromycin, tetracycline, benzylpenicillin, trimethoprim-sulfamethoxazole, ampicillin, ciprofloxacin, and meropenem) showed that 50 strains (92.6%), the six surface isolates also included, were intermediately resistant to ciprofloxacin and susceptible to the rest of the six antimicrobial agents tested, whereas strong resistance against the use of a single from three implicated antibiotics was recorded to four strains (7.4%) of the pathogen isolated from Myzithra cheese samples. Thence, the minimum inhibitory concentrations (MICs) were determined for erythromycin (MIC = 0.19 μg/mL), ciprofloxacin (MIC ≥ 0.19 μg/mL), and meropenem (MIC = 0.64 μg/mL), and finally, just one strain was deemed resistant to the latter antibiotic. The phylogenetic positions of the strains and their genetic variability were determined through WGS, whilst also stress response and virulence gene analysis for the isolates was conducted. Findings of this work should be useful as they could be utilized for epidemiological investigations of in the food processing environment, revealing possible contamination scenarios, and acquired antimicrobial resistance along the food production chain.
李斯特菌病是一种严重的传染病,在人类由食源性病原体引起的疾病中,其病死率是最高的(20%)之一,而乳制品常被认为是人类感染的来源。在本研究中,我们通过全基因组测序(WGS)对从希腊传统软乳清奶酪米兹特拉(Myzithra)分离出的54株菌株(48株分离株)以及从希腊伊庇鲁斯地区一家奶酪加工厂表面采集的拭子(6株分离株)进行了表型和基因特征分析。除一株菌株外,所有菌株均属于聚合酶链反应(PCR)血清群IIa(16.7%)和IIb(81.5%),分别对应血清型1/2a、3a和1/2b、3b、7。后者被鉴定为血清型4b、4d、4e的PCR血清群IVb菌株(1.8%)。生物信息学分析揭示了五种序列类型(STs)和克隆复合体(CCs);因此,在分离株中分别检测到ST1、ST3、ST121、ST155、ST398以及CC1、CC3、CC121、CC155、CC398,其比例分别为1.9%、83.3%、11.0%、1.9%和1.9%。该病原体对一组七种选定抗生素(红霉素、四环素、苄青霉素、甲氧苄啶 - 磺胺甲恶唑、氨苄青霉素、环丙沙星和美罗培南)的药敏试验表明,50株菌株(92.6%,包括6株表面分离株)对环丙沙星呈中度耐药,对其余六种测试抗菌药物敏感,而从米兹特拉奶酪样品中分离出的该病原体的四株菌株(7.4%)对三种相关抗生素中的一种呈现强耐药性。因此,测定了红霉素(MIC = 0.19μg/mL)、环丙沙星(MIC≥0.19μg/mL)和美罗培南(MIC = 0.64μg/mL)的最低抑菌浓度(MICs),最后,只有一株菌株被认为对后一种抗生素耐药。通过WGS确定了分离株的系统发育位置及其遗传变异性,同时还对分离株进行了应激反应和毒力基因分析。这项工作的结果应该是有用的,因为它们可用于食品加工环境中李斯特菌病的流行病学调查,揭示可能的污染情况以及食品生产链中获得性抗菌药物耐药性。