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Hypervirulent and hypermucoviscous extended-spectrum β-lactamase-producing Klebsiella pneumoniae and Klebsiella variicola in Chile.

ABSTRACT: Convergence of virulence and antibiotic-resistance has been reported in Klebsiella pneumoniae, but not in Klebsiella variicola. We, hereby, report the detection and genomic characterization of hypervirulent and hypermucoviscous K. pneumoniae and K. variicola recovered in Chile from health-care associated infections, which displayed resistance to broad-spectrum cephalosporins. One hundred forty-six K. pneumoniae complex isolates were screened by hypermucoviscosity by the "string test." Two hypermucoid isolates, one hypermucoviscous K. pneumoniae (hmKp) and one K. variicola (hmKv), were further investigated by whole-genome sequencing. In vivo virulence was analyzed by the Galleria mellonella killing assay. In silico analysis of hmKp UCO-494 and hmKv UCO-495 revealed the presence of multiple antibiotic-resistance genes, such as bla _CTX-M-1, bla _DHA-1 and bla _LEN-25 among others clinically relevant resistance determinants, including mutations in a two-component regulatory system related to colistin resistance. These genetic features confer a multidrug-resistant (MDR) phenotype in both strains. Moreover, virulome in silico analysis confirmed the presence of the aerobactin gene iutA, in addition to yersiniabactin and/or colicin V encoding genes, which are normally associated to high virulence in humans. Furthermore, both isolates were able to kill G. mellonella and displayed higher virulence in comparison with the control strain. In summary, the convergence of virulence and the MDR-phenotype in K. pneumoniae complex members is reported for the first time in Chile, denoting a clinical problem that deserves special attention and continuous surveillance in South America.

SUBMITTER: Morales-Leon F

PROVIDER: S-EPMC7781644 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundKlebsiella pneumoniae is a conditional pathogen related to several infectious diseases. Few studies reported Klebsiella pneumoniae meningitis in the Chinese population, guidelines on diagnosis and treatment of Klebsiella pneumoniae meningitis should be considered due to its high lethality. Here, we report a case of adult intracranial infection caused by extended-spectrum-beta-lactamase (ESBL)-producing hypervirulent Klebsiella pneumoniae (hvKP) in a 65-year-old female, providing new insight for clinical awareness and epidemiological surveillance for ESBL-producing hvKP infection.Case descriptionA 65-year-old female who had a recurrent fever for more than 1 month, and vomiting for 1 week was admitted to our hospital. The computed tomography (CT) results and laboratory results indicated systematic infection, and the blood culture confirmed the infection of Klebsiella pneumoniae. A combination of antibiotics including vancomycin, caspofungin, dexamethasone, and posaconazole oral suspension was given to the patient. Further, she exhibited a convulsion with unconsciousness, the CT revealed lacunar infarction and encephalomalacia. The following physical examination showed slight neck resistance, a weak light response of the eye, low muscle tension, suspicious left Babinski sign (+), and right Babinski sign (-). The CT and cerebrospinal fluid (CSF) analyses confirmed the diagnosis of intracranial infection caused by Klebsiella pneumoniae. We employed CSF microbial metagenomic next-generation sequencing (mNGS) was employed and the results suggested the high sequence of Klebsiella pneumoniae with drug-resistant gene SHV-type beta-lactamases (blaSHV). Subsequently, 2 g meropenem every 8 hours (q8h) prolonged for 3 hours was applied to treat intracranial infection, and her body temperature and infectious manifestations were gradually relieved. The CT results represented that pulmonary edema and pleural effusion were gradually dissipated and absorbed. Based on the improvement of clinical manifestations, the patient was discharged from the hospital and a close follow-up was conducted.ConclusionsAn ESBL-producing strain of hvKP could lead to invasive infection such as severe intracranial infection, with a relatively favorable prognosis. The outcome of the disease caused by Klebsiella pneumoniae infection is firmly related to the phenotypic features, for instance, virulence factors and antibiotic susceptibility. Due to its high lethality, timely empiric anti-infection therapy and close surveillance are necessary for patients with Klebsiella pneumoniae infection in the clinic.

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.

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Hypervirulent and hypermucoviscous extended-spectrum β-lactamase-producing Klebsiella pneumoniae and Klebsiella variicola in Chile.

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