The emergence of carbapenem-resistant Klebsiella pneumoniae isolates producing OXA-48 and NDM in the Southern (Asir) province, Saudi Arabia

Background/Objectives: Monkeypox is a re-emerging viral disease with features of infectiously transmitted zoonoses. It is now considered a public health priority because of its rising incidence and transmission from person to person. Monkeypox virus (MPXV) VP39 protein is identified as an essential protein for replication of the virus, and therefore, it is a potential target for antiviral drugs. Methods: This work analyzes the binding affinities and the differential conformational stability of three target compounds and one control compound with the VP39 protein through multiple computational methods. Results: The re-docking analysis revealed that the compounds had high binding affinities towards the target protein; among these compounds, compounds 1 and 2 showed the highest binding energies in the virtual screening, and thus, these were considered as the most active inhibitor candidates. Intermolecular interaction analysis revealed distinct binding mechanisms. While compound 1 had very strong hydrogen bonds and hydrophobic interactions, compound 2 had numerous water-mediated interactions, and compound 3 had only ionic and hydrophobic contacts. In molecular dynamic simulations, compounds 1 and 2 showed that the protein–ligand complexes had a stable conformation, with protein RMSD values around 2 Å for both compounds. In contrast, compound 3 was slightly flexible, and the control compound was more flexible. MM/GBSA analysis again supported these results, which gave the binding free energies that were also supportive for these compounds. Conclusions: Notably, all the selected compounds, especially compounds 1 and 2, demonstrate high binding affinity. Therefore, these compounds can be further tested as antiviral agents against monkeypox treatment.

Saudi Arabia is mostly barren except the southwestern highlands that are susceptible to environmental changes, a hotspot for biodiversity, but poorly studied for microbial diversity and composition. In this study, 454-pyrosequencing of 16S rRNA gene hypervariable region V6 was used to analyze soil bacterial community along elevation gradients of the southwestern highlands. In general, lower percentage of total soil organic matter (SOM) and nitrogen were detected in the analyzed soil samples. Total 33 different phyla were identified across the samples, including dominant phyla Proteobacteria, Actinobacteria and Acidobacteria. Representative OTUs were grouped into 329 and 508 different taxa at family and genus level taxonomic classification, respectively. The identified OTUs unique to each sample were very low irrespective of the altitude. Jackknifed principal coordinates analysis (PCoA) revealed, overall differences in the bacterial community were more related to the quantity of specific OTUs than to their diversity among the studied samples. Bacterial diversity and soil physicochemical properties did not show consistent changes along the elevation gradients. The large number of OTUs shared between the studied samples suggest the presence of a core soil bacterial community in the southwestern highlands of Saudi Arabia.

This study presents a comprehensive genomic analysis of NDM and OXA-48-producing Klebsiella pneumoniae in the Western region of Saudi Arabia, traversed by tens of millions of Muslims from various countries annually. This significant influx of visitors invariably leads to the spread and diversity of MDR bacteria. Genome sequencing was performed using MiSeq system of 29 CPKP isolates that were NDM and OXA-48-positive isolated from nosocomial infections and demonstrated resistance to most antibiotics, including carbapenems. WGS analysis showed that 12 (41.3%) isolates co-harbored bla_OXA-48,bla_CTX-M-15 and bla_NDM genes. Notably, 16 (55.1%) isolates were identified as high-risk clone ST14, with 50% of these isolates co-harbored bla_OXA-48, bla_NDM and bla_CTX-M-15 genes. All ST14 isolates were identified as capsular genotype KL2 and O1/O2v1 antigen with yersiniabactin locus ypt 14 carried by ICEKp5. The two isolates were identified as ST2096/KL64 hypervirulent K. pneumoniae (hvKp) clone harboring several virulence factors, including the regulator of the mucoid phenotype rmpA2 and aerobactin (iuc-1). Interestingly, two of the hvKp ST383/KL30 isolates were resistant to all tested antimicrobials except colistin and tigecycline, and simultaneously carried numerous ESBLs and carbapenemase genes. These isolates also harbor several virulence factors such as rmpA1, rmpA2, carried on KpVP-1, and aerobactin (iuc-1). this study provides insights into the spread and prevalence of high-risk clones of CPKP in the Western region of Saudi Arabia. The ST14 high-risk clone appears to be the predominant CPKP clone in this region, posing a significant threat to public health. This study also reports the presence of two globally disseminated hypervirulent K. pneumoniae (hvKp) clones, namely ST2096 and ST383. Therefore, it is essential to improve surveillance and implement strict infection control measures in this region, which receives a substantial number of visitors to effectively monitor and reduce the spread of high-risk clones of antimicrobial-resistant bacteria, including CPKP.

Carbapenem-resistant Klebsiella pneumoniae (CRKP)-associated infections have become a major concern and life-threatening worldwide. Understanding the epidemiology of CRKP using a reliable molecular technology can help to develop an effective infection control policies. In the western region of Saudi Arabia, there are no sufficient data on the prevalence of CRKP and its carbapenem-resistant determinants. Therefore, this study aimed to determine the molecular epidemiology of CRKP and identify the most common carbapenemase genes. In the current study, a total of 191 CRKP isolates were collected and obtained from clinical specimens of patients at King Fahad Armed Forces Hospital (KFAFH), Jeddah, Saudi Arabia. All isolates that were resistant or intermediately susceptible to either of the carbapenem antimicrobials (imipenem, meropenem, or ertapenem) were included. All CRKP showed resistance to ceftazidime, cefepime, and piperacillin/tazobactam, whereas low (14%) and moderate (37.7%) levels of resistance were reported against tigecycline and colistin, respectively. The most common carbapenemase genes identified were bla_OXA-48 (n = 157 [82.2%]), followed by bla_NDM in 27 (14%) isolates. The bla_VIM and bla_KPC were reported in only one isolate each and no bla_IMP producers were detected among all tested isolates. The high prevalence of OXA-48 among K. pneumoniae isolates reported in the current study may reflect that OXA-48 has become an endemic in Saudi Arabian hospitals. The second major finding was that the identification of CRKP co-harbors both bla_NDM and bla_OXA-48, and such isolates can be threating for healthcare societies (patients and healthcare workers) due to their high level of resistance to carbapenems. These results suggest that the use of molecular diagnostic methods and proper surveillance programs are required to monitor and control the spread of all multidrug-resistant (MDR) bacteria, including CRKP. Therefore, further research is recommended to expand the study and further analyze the genotyping of the most common clones of CRKP in other hospitals in the western regions of Saudi Arabia.