The emergence of blaCTX-M-15-carrying Escherichia coli of ST131 and new sequence types in Western China

Abstract Background bla CTX-M-15, the most widely distributed gene encoding extended-spectrum β-lactamases globally, was not common in China. This study was performed to characterize bla CTX-M-15-carrying Escherichia coli in western China. Findings Out of 144 Escherichia coli isolates from 20 hospitals in western China, 8 were found carrying bla CTX-M-15. bla CTX-M-15 was carried by isolates of ST131and 5 new STs (ST3342, ST3513, ST3516, ST3517 and ST3518). The 5 new STs shared 5 identical alleles out of 7 but only had up to 2 alleles identical to ST131. bla CTX-M-15 was located on plasmids of IncI1 (ST16) or IncFII-related group (four replicon types). The co-transfer of a few antimicrobial resistance genes including qnrA, qnrB, qnrS, qepA, aac (6′)-Ib-cr, aac (3)-II, tetA, bla TEM and bla OXA-1 with bla CTX-M-15 were examined but only bla TEM-1 was found co-transferring with bla CTX-M-15. Conclusions Five new STs of E. coli and three new types of IncFII-related plasmids carrying bla CTX-M-15 were identified. This study together with several reports suggested that bla CTX-M-15 has emerged in China and the interruption of both vertical and horizontal transmission of bla CTX-M-15 is required to hurdle its further spread. </jats:sec

Ackermannviridae bacteriophage against carbapenem-resistant Klebsiella pneumoniae of capsular type 64

Lytic bacteriophages (phages) are promising clinically viable therapeutic options against carbapenem-resistant Klebsiella pneumoniae (CRKP). In China, the predominant strains are those assigned to sequence type 11 and capsular type 64 (ST11-KL64). The emergence of phage resistance is a major bottleneck hindering effective phage therapy, requiring more new phages to provide the flexibility for creating different phage cocktails. However, the majority of phages against ST11-KL64 CRKP belong to the genus Przondovirus of the family Autographiviridae, which limits the options for constructing cocktails. We recovered a novel lytic phage of the genus Taipeivirus within the family Ackermannviridae against ST11-KL64 CRKP from a river in China. We phenotypically characterized this phage and obtained its genome sequence for analysis. This phage can inhibit the growth of ST11-KL64 CRKP for 6.5 h at a 0.1 multiplicity of infection and exhibits a narrow host range, being unable to attack CRKP strains of the other 30 capsular types. This phage carries no genes encoding antimicrobial resistance, virulence, or lysogeny. It is stable across a wide range of temperatures and pH values, making it suitable for phage therapy. Unlike other Taipeivirus phages, P01 has two tail spike proteins and a unique tail fiber protein. The distinct tail composition of this phage contributes to its activity against ST11-KL64 CRKP and its narrow host range. Taken together, we recovered a phage of a novel viral species with the potential for therapy, which expands the phage biobank against CRKP

The Occurence of Colistin-Resistant Hypervirulent Klebsiellapneumoniae in China

Hypervirulent Klebsiella pneumoniae strains are usually susceptible to many antimicrobial agents including colistin. Here we report the isolation and characterization of several colistin-resistant hypervirulent K. pneumoniae clinical strains. K. pneumoniae strains recovered from blood samples were collected at a university hospital in China. MICs of colistin were determined using microdilution. Colistin-resistant strains were subjected to whole genome sequencing to reveal their clonal background, antimicrobial resistance determinants and virulence factors. Virulence assays were performed with strains carrying the mucoid phenotype regulator gene rmpA using wax moth larvae. The pmrB gene encoding a P344L substitution was cloned into a colistin-susceptible K. pneumoniae strain to examine whether the substitution confers colistin resistance. Five colistin-resistant hypervirulent K. pneumoniae were recovered from blood samples of patients in China, belonging to four sequence/capsular types (ST23:K1, ST412:K57, ST660:K16, and ST700:K1) and carried the virulence factor rmpA. Three strains had the known colistin-resistant D150G substitution in PhoQ including one ST700:K1 strain also carrying mcr-1. The remaining two isolates had a P344L substitution of PmrB but cloning of pmrB encoding the substitution into a colistin-susceptible isolate did not alter MICs of colistin, suggesting that such a substitution did not confer resistance to colistin. In conclusion, the convergence of colistin resistance and hypervirulence in K. pneumoniae of multiple clonal backgrounds has emerged and may warrant further surveillance

Anti-hepatotoxic and anti-oxidant effects of extracts from Piper nigrum L. root

The aim of this study was to investigate the effect of Piper nigrum L. root extracts on carbon tetrachloride (CCl4)-induced rat liver injury. Among the three different extracts (water, ethanol and chloroform extract), ethanol extract exhibits the highest hepatoprotective activity (p &lt; 0.05). When using the ethanol extract at a dose of 120 mg/ kg to treat the CCl4-intoxicated rat, the activities of alanine transaminase (ALT) and aspartate transanimase (AST) in rat serum decreased to 65.7 and 84.5%, respectively. At the same time, the lipid peroxidation (MDA) decreased to 52.3% and glutathione (GSH) increased to 55.8% in the rats liver homogenate, as compared with those of the CCl4 positive control rats. The hepatoprotective effect of ethanol extract was also supported by the histopathological observations. Moreover, the ethanol extract was studied for its in vitro antioxidant activity using the methods of ferric thiocyanate (FTC) and thiobarbituric acid (TBA). The findings indicate that the ethanol extract of P. nigrum L. root is an efficient hepatoprotective and antioxidant agent against CCl4-induced liver injury.Keywords: Piper nigrum L. root, ethanol extract, carbon tetrachloride (CCl4), hepatoprotective, antioxidan

The co-transfer of plasmid-borne colistin-resistant genes mcr-1 and mcr-3.5, the carbapenemase gene blaNDM-5 and the 16S methylase gene rmtB from Escherichia coli

We found an unusual Escherichia coli strain with resistance to colistin, carbapenem and amikacin from sewage. We therefore characterized the strain and determined the co-transfer of the resistance determinants. Whole genome sequencing was performed using both Illumina HiSeq X10 and MinION sequencers. Short and long reads were subjected to de novo hybrid assembly. Sequence type, antimicrobial resistance genes and plasmid replicons were identified from the genome sequences. Phylogenetic analysis of all IncHI2 plasmids carrying mcr-1 available in GenBank was performed based on core genes. Conjugation experiments were performed. mcr-3.5 was cloned into E. coli DH5α. The strain belonged to ST410, a type with a global distribution. Two colistin-resistant genes, mcr-1.1 and mcr-3.5, a carbapenemase gene blaNDM-5, and a 16S methylase gene rmtB were identified on different plasmids of IncHI2(ST3)/IncN, IncP, IncX3 and IncFII, respectively. All of the four plasmids were self-transmissible and mcr-1.1, mcr-3.5, blaNDM-5 and rmtB were transferred together. mcr-1-carrying IncHI2 plasmids belonged to several sequence types with ST3 and ST4 being predominant. MIC of colistin (4 μg/ml) for DH5α containing mcr-3.5 was identical to that containing the original mcr-3 variant. In conclusion, carbapenem resistance, colistin resistance and high-level aminoglycoside resistance can be transferred together even when their encoding genes are not located on the same plasmid. The co-transfer of multiple clinically-important antimicrobial resistance represents a particular challenge for clinical treatment and infection control in healthcare settings. Isolates with resistance to both carbapenem and colistin are not restricted to a given sequence type but rather are diverse in clonal background, which warrants further surveillance. The amino acid substitutions of MCR-3.5 have not altered its activity against colistin.</p

The coexistence of two ^blaNDM-5 genes on an IncF plasmid as revealed by nanopore sequencing

ABSTRACT In a carbapenem-resistant Escherichia coli clinical isolate of sequence type 167, two copies of bla NDM-5 were found on a 144,225-bp IncF self-transmissible plasmid of the F36:A4:B − type. Both bla NDM-5 genes were located in 11,065-bp regions flanked by two copies of IS 26 . The two regions were identical in sequence but were present at different locations on the plasmid, suggesting a duplication of the same region. This study highlights the complex genetic contexts of bla NDM-5 . </jats:p

Safety and efficacy of phage application in bacterial decolonisation:a systematic review

Colonisation by bacterial pathogens typically precedes invasive infection and seeds transmission. Thus, effective decolonisation strategies are urgently needed. The literature reports attempts to use phages for decolonisation. To assess the in-vivo efficacy and safety of phages for bacterial decolonisation, we performed a systematic review by identifying relevant studies to assess the in-vivo efficacy and safety of phages for bacterial decolonisation. We searched PubMed, Embase (Ovid), MEDLINE (Ovid), Web of Science, and the Cochrane Library to identify relevant articles published between Jan 1, 1990, and May 12, 2023, without language restrictions. We included studies that assessed the efficacy of phage for bacterial decolonisation in humans or vertebrate animal models. This systematic review is registered with PROSPERO, CRD42023457637. We identified 6694 articles, of which 56 (51 animal studies and five clinical reports) met the predetermined selection criteria and were included in the final analysis. The gastrointestinal tract (n=49, 88%) was the most studied bacterial colonisation site, and other sites were central venous catheters, lung, nose, skin, and urinary tract. Of the 56 included studies, the bacterial load at the colonisation site was reported to decrease significantly in 45 (80%) studies, but only five described eradication of the target bacteria. 15 studies reported the safety of phages for decolonisation. No obvious adverse events were reported in both the short-term and long-term observation period. Given the increasing life-threatening risks posed by bacteria that are difficult to treat, phages could be an alternative option for bacterial decolonisation, although further optimisation is required before their application to meet clinical needs.</p