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Characterization of extended-spectrum-β-lactamase producing Klebsiella pneumoniae phage KP1801 and evaluation of therapeutic efficacy in vitro and in vivo.

ABSTRACT: Extended spectrum β lactamase-producing Klebsiella pneumoniae (ESBL-KP) is being reported with high morbidity and mortality rates and is considered as the highest priority for new antimicrobial strategies. To develop an alternative antimicrobial agent, phage KP1801 with broad lytic activity was isolated. The genome of phage KP1801 was double stranded DNA of 49,835 base pairs, with a GC content of 50.26%. There were 75 putative open reading frames. Phage KP1801 was classified as being in the order Caudovirales, belonging to the Siphoviridae family. About 323 proteins were detected by shotgun proteome analysis. The phage inhibited biofilm formation and reduced pre-formed biofilm in a dose dependent manner. Scanning electron microscopic studies demonstrated a membrane damage of bacterial cells treated with phage, resulting in cell death. Prophylactic and therapeutic efficacies of the phage were evaluated in Galleria mellonella. Administration of ESBL-KP infection with phage significantly improved the survival of G. mellonella. The number of intracellular bacteria in larvae showed a significant decrease compared with untreated control while the number of phage increased. These studies suggested that phage KP1801 has the potential for development as an alternative for antibiotics and biocontrol agents against ESBL-KP infection.

SUBMITTER: Wintachai P

PROVIDER: S-EPMC7367294 | biostudies-literature | 2020 Jul

REPOSITORIES: biostudies-literature

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Characterization of extended-spectrum-β-lactamase producing Klebsiella pneumoniae phage KP1801 and evaluation of therapeutic efficacy in vitro and in vivo.

Wintachai Phitchayapak P Naknaen Ampapan A Thammaphet Jirapath J Pomwised Rattanaruji R Phaonakrop Narumon N Roytrakul Sittiruk S Smith Duncan R DR

Scientific reports 20200716 1

Extended spectrum β lactamase-producing Klebsiella pneumoniae (ESBL-KP) is being reported with high morbidity and mortality rates and is considered as the highest priority for new antimicrobial strategies. To develop an alternative antimicrobial agent, phage KP1801 with broad lytic activity was isolated. The genome of phage KP1801 was double stranded DNA of 49,835 base pairs, with a GC content of 50.26%. There were 75 putative open reading frames. Phage KP1801 was classified as being in the orde ...[more]

PMID: 32678251

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Project description:Background. Klebsiella pneumoniae is part of the ESKAPE (Enterococcus faecium, Staphylococcus aureus, K. pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter spp.) group of multidrug-resistant (MDR) pathogens. K. pneumoniae is the leading cause of antimicrobial resistance-associated mortality and the second leading cause of nosocomial bloodstream infections (BSIs), globally and in sub-Saharan Africa. Therefore, it was aimed to determine the antibiotic resistance patterns of K. pneumoniae isolated from blood cultures of patients with features of sepsis at Mulago National Referral Hospital, Uganda. Methods. The cross-sectional study on patients with features of sepsis utilized K. pneumoniae (n=30) isolated from positive blood culture specimens. The antibiotic resistance profile was determined by the Clinical and Laboratory Standards Institute's Kirby-Bauer disc diffusion method, which was used to classify the isolates as susceptible, intermediate and resistant. K. pneumoniae isolates that were resistant to third-generation cephalosporins were subjected to extended-spectrum β-lactamase (ESBL) screening and confirmation using the double-disc synergy test using cefotaxime, ceftazidime, ceftriaxone, cefotaxime-clavulanic acid and ceftazidime-clavulanic acid. The results were analysed for frequencies. Results. K. pneumoniae isolates showed emerging resistance to imipenem at 13% (4 out of 30) followed by amikacin at 17% (5 out of 30). There was intermediate resistance to gentamycin at 60% (18 out of 30). However, K. pneumoniae showed the highest resistance to piperacillin at 100% (30 out of 30) followed by sulphamethoxazole-trimethoprim and cefepime, both showing a percentage of 97% (29 out of 30). Up to 16 out of 30 (53.3%) of K. pneumoniae were positive for ESBL production, whilst 14 out of 30 (46.7%) were negative. Conclusion. There was a high prevalence of antibiotic-resistant ESBL-producing K. pneumoniae isolates from BSI of patients with features of sepsis in Uganda's Mulago National Referral Hospital.

Project description:Background and aimThe World Health Organization considers multidrug-resistant (MDR) Klebsiella pneumoniae a major global threat. Horses harbor commensal isolates of this bacterial species and potentially serve as reservoirs for human MDR bacteria. This study investigated antimicrobial resistance in horses caused by extended-spectrum β-lactamase (ESBL)-producing K. pneumoniae.Materials and methodsOne hundred fifty-nine nasal swab samples were collected from horses with respiratory distress not treated with cefotaxime and erythromycin. Biochemical and serological identification was performed on all samples. Polymerase chain reaction (PCR) was used to detect 16S-23S ITS, mucoviscosity-associated gene (magA), uridine diphosphate galacturonate 4-epimerase gene (uge), and iron uptake system gene (kfu), bla TEM, bla SHV, and bla CTX genes. Sequence analysis and phylogenetic relatedness of randomly selected K. pneumoniae isolates carrying the bla TEM gene were performed.ResultsTen isolates of Klebsiella spp. were obtained from 159 samples, with an incidence of 6.28% (10 of 159). Based on biochemical and serological identification, K. pneumoniae was detected in 4.4% (7 of 159) of the samples. Using PCR, all tested K. pneumoniae isolates (n=7) carried the 16S-23S ITS gene. By contrast, no isolates carried magA, uge, and kfu genes. The bla TEM gene was detected in all test isolates. Moreover, all isolates did not harbor the bla SHV or bla CTX gene. Sequence analysis and phylogenetic relatedness reported that the maximum likelihood unrooted tree generated indicated the clustering of the test isolate with the other Gram-negative isolate bla TEM. Finally, the sequence distance of the bla TEM gene of the test isolate (generated by Lasergene) showed an identity range of 98.4-100% with the bla TEM gene of the different test isolates.ConclusionThe misuse of antimicrobials and insufficient veterinary services might help generate a population of ESBL-producing K. pneumoniae in equines and humans, representing a public health risk.

Dataset Information

Characterization of extended-spectrum-β-lactamase producing Klebsiella pneumoniae phage KP1801 and evaluation of therapeutic efficacy in vitro and in vivo.

Publications

Characterization of extended-spectrum-β-lactamase producing Klebsiella pneumoniae phage KP1801 and evaluation of therapeutic efficacy in vitro and in vivo.

Similar Datasets

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