National Culture Collection of Pakistan (NCCP), Bio-resources Conservation Institute (BCI), National Agricultural Research Centre (NARC), Park Road, Islamabad, Pakistan.
Department of Zoology, University of Science and Technology, Bannu, Pakistan.
J Glob Antimicrob Resist. 2021 Dec;27:91-94. doi: 10.1016/j.jgar.2021.08.011. Epub 2021 Sep 5.
Here we describe the first draft genome analysis of a CRISPR-carrying, multidrug-resistant, candidate novel Pseudomonas sp. NCCP-436 isolated from faeces of a neonatal diarrhoeic calf.
The genome of strain NCCP-436 was sequenced using an Illumina NovaSeq PE150 platform and analysed using various bioinformatic tools. The virulence factors and resistome were identified using PATRIC and CARD servers, while CGView Server was used to construct a circular genome map. Antimicrobial susceptibility was determined by the disk diffusion technique.
The draft genome of strain NCCP-436 contains 43 contigs with a total genome size of 3,683,517 bp (61.4% GC content). There are 3,452 predicted genes, including 60 tRNAs, 7 rRNAs and 12 sRNAs. CRISPR analysis revealed two CRISPR arrays with lengths of 1103 bp and 867 bp. Strain NCCP-436 was highly resistant to fluoroquinolone, β-lactam, cephalosporin, aminoglycoside, penicillin, rifamycin, macrolide, glycopeptide, trimethoprim/sulfonamide and tetracycline antibiotic classes. Additionally, 22 antibiotic resistance genes, 313 virulence genes and 253 pathogen-host interactor genes were predicted. Comparison of the average nucleotide identity and digital DNA-DNA hybridisation values with the closely-related strain Pseudomonas khazarica (TBZ2) was found to be 82.08% and 34.90%, respectively, illustrating strain NCCP-436 as a potentially new species of Pseudomonas.
Substantial number of antibiotic resistance and virulence genes and homology with human pathogens were predicted, exposing the pathogenic and zoonotic potential of strain NCCP-436 to public health. These findings may be used to better understand the genomic epidemiological features and drug resistance mechanisms of pathogenic Pseudomonas spp. in Pakistan.
本研究描述了从新生腹泻犊牛粪便中分离的携带 CRISPR 的多药耐药候选新型假单胞菌 NCCP-436 的首个基因组草案分析。
使用 Illumina NovaSeq PE150 平台对 NCCP-436 菌株进行基因组测序,并使用各种生物信息学工具进行分析。使用 PATRIC 和 CARD 服务器鉴定毒力因子和耐药组,同时使用 CGView Server 构建环形基因组图谱。通过纸片扩散技术测定抗菌药物敏感性。
NCCP-436 菌株的基因组草案包含 43 个重叠群,基因组大小为 3683517 bp(GC 含量为 61.4%)。共有 3452 个预测基因,包括 60 个 tRNA、7 个 rRNA 和 12 个 sRNA。CRISPR 分析显示有两个长度分别为 1103 bp 和 867 bp 的 CRISPR 数组。NCCP-436 菌株对氟喹诺酮类、β-内酰胺类、头孢菌素类、氨基糖苷类、青霉素类、利福霉素类、大环内酯类、糖肽类、复方磺胺甲噁唑类和四环素类抗生素高度耐药。此外,还预测了 22 个抗生素耐药基因、313 个毒力基因和 253 个病原体-宿主相互作用基因。与密切相关的假单胞菌 khazarica(TBZ2)相比,菌株 NCCP-436 的平均核苷酸同一性和数字 DNA-DNA 杂交值分别为 82.08%和 34.90%,表明 NCCP-436 菌株可能是一种新的假单胞菌。
大量的抗生素耐药基因和毒力基因与人类病原体同源,这揭示了 NCCP-436 菌株对公共卫生的潜在致病性和人畜共患病性。这些发现可用于更好地了解巴基斯坦致病性假单胞菌的基因组流行病学特征和耐药机制。