SAMRC Microbial Water Quality Monitoring Centre, University of Fort Hare, Alice, South Africa.
Applied and Environmental Microbiology Research Group, Department of Biochemistry and Microbiology, University of Fort Hare, Alice, South Africa.
PLoS One. 2020 Dec 7;15(12):e0238084. doi: 10.1371/journal.pone.0238084. eCollection 2020.
The increasing occurrence of multidrug-resistant Enterobacteriaceae in clinical and environmental settings has been seen globally as a complex public health challenge, mostly in the developing nations where they significantly impact freshwater used for a variety of domestic purposes and irrigation. This paper detail the occurrence and antibiogram signatures of the Enterobacteriaceae community in Tsomo and Tyhume rivers within the Eastern Cape Province, the Republic of South Africa, using standard methods. The average distribution of the presumptive Enterobacteriaceae in the rivers ranged from 1 × 102 CFU/100ml to 1.95 × 104 CFU/100ml. We confirmed 56 (70.8%) out of 79 presumptive Enterobacteriaceae isolated being species within the family Enterobacteriaceae through the Matrix-Assisted Laser Desorption Ionization Time of Flight technique. Citrobacter-, Enterobacter-, Klebsiella species, and Escherichia coli were selected (n = 40) due to their pathogenic potentials for antibiogram profiling. The results of the antibiotic susceptibility testing gave a revelation that all the isolates were phenotypically multidrug-resistant. The resistance against ampicillin (95%), tetracycline and doxycycline (88%), and trimethoprim-sulfamethoxazole (85%) antibiotics were most prevalent. The Multiple Antibiotic Resistance indices stretched from 0.22 to 0.94, with the highest index observed in a C. freundii isolate. Molecular characterisation using the PCR technique revealed the dominance of blaTEM (30%; 12/40) among the eight groups of β-lactams resistance determinants assayed. The prevalence of others was blaCTX-M genes including group 1, 2 and 9 (27.5%), blaSHV (20%), blaOXA-1-like (10%), blaPER (2.5%), and blaVEB (0%). The frequencies of the resistance determinants for the carbapenems were blaKPC (17.6%), blaGES (11.8%), blaIMP (11.8%), blaVIM (11.8%), and blaOXA-48-like (5.9%). Out of the six plasmid-mediated AmpC (pAmpC) genes investigated blaACC, blaEBC, blaFOX, blaCIT, blaDHA, and blaMOX, only the first four were detected. In this category, the most dominant was blaEBC, with 18.4% (7/38). The prevalence of the non-β-lactamases include tetA (33.3%), tetB (30.5%), tetC (2.8%), tetD (11.1%), tetK (0%), tetM (13.9%), catI (12%), catII (68%), sulI (14.3%), sulII (22.9%) and aadA (8.3%). Notably, a C. koseri harboured 42.8% (12/28) of the genes assayed for which includes five of the ESBL genes (including the only blaPER detected in this study), two of the pAmpC resistance genes (blaACC and blaCIT), and five of the non-β-lactamase genes. This study gives the first report on C. koseri exhibiting the co-occurrence of ESBL/AmpC β-lactamase genes from the environment to the best of our knowledge. The detection of a blaPER producing Citrobacter spp. in this study is remarkable. These findings provide evidence that freshwater serves as reservoirs of antimicrobial resistance determinants, which can then be easily transferred to human beings via the food chain and water.
本文详细介绍了南非东开普省 Tsomo 和 Tyhume 河流中肠杆菌科社区的发生和抗生素图谱特征,使用了标准方法。河流中推定肠杆菌科的平均分布范围为 1×102 CFU/100ml 至 1.95×104 CFU/100ml。通过基质辅助激光解吸电离飞行时间技术,我们证实了从 79 株推定肠杆菌科中分离出的 56 株(70.8%)为肠杆菌科家族中的物种。由于其具有致病性,因此选择了柠檬酸杆菌、肠杆菌、克雷伯菌和大肠杆菌(n=40)进行抗生素图谱分析。抗生素敏感性测试的结果表明,所有分离株均表现出表型上的多药耐药性。对氨苄西林(95%)、四环素和强力霉素(88%)以及磺胺甲恶唑/甲氧苄啶(85%)的耐药性最为普遍。多重抗生素耐药指数范围为 0.22 至 0.94,在 C. freundii 分离株中观察到最高指数。使用 PCR 技术进行的分子特征分析显示,在所检测的 8 组β-内酰胺类耐药决定因子中,blaTEM(30%;12/40)最为普遍。其他的耐药决定因子的检出率分别为 blaCTX-M 基因群 1、2 和 9(27.5%)、blaSHV(20%)、blaOXA-1 样(10%)、blaPER(2.5%)和 blaVEB(0%)。碳青霉烯类耐药决定因子的检出率分别为 blaKPC(17.6%)、blaGES(11.8%)、blaIMP(11.8%)、blaVIM(11.8%)和 blaOXA-48 样(5.9%)。在所研究的 6 种质粒介导的 AmpC(pAmpC)基因 blaACC、blaEBC、blaFOX、blaCIT、blaDHA 和 blaMOX 中,仅检测到前 4 种。在这一类中,最普遍的是 blaEBC,检出率为 18.4%(7/38)。非β-内酰胺类耐药基因的检出率包括 tetA(33.3%)、tetB(30.5%)、tetC(2.8%)、tetD(11.1%)、tetK(0%)、tetM(13.9%)、catI(12%)、catII(68%)、sulI(14.3%)、sulII(22.9%)和 aadA(8.3%)。值得注意的是,一株 C. koseri 携带了 42.8%(12/28)的基因,其中包括 5 种 ESBL 基因(包括本研究中唯一检出的 blaPER)、2 种 pAmpC 耐药基因(blaACC 和 blaCIT)和 5 种非β-内酰胺类耐药基因。据我们所知,这是首次报道从环境中到肠杆菌科中存在同时携带 ESBL/AmpCβ-内酰胺酶基因的 C. koseri。本研究中检测到的产 PER 的柠檬酸杆菌属是值得关注的。这些发现表明,淡水是抗生素耐药决定因子的储库,这些因子可以通过食物链和水很容易地转移到人类身上。