The purpose of this study was the identification of genetic lineages and antimicrobial resistance (AMR) and virulence genes in Klebsiella pneumoniae isolates associated with severe infections in the neuro-ICU. Susceptibility to antimicrobials was determined using the Vitek-2 instrument. AMR and virulence genes, sequence types (STs), and capsular types were identified by PCR. Whole-genome sequencing was conducted on the Illumina MiSeq platform. It was shown that K. pneumoniae isolates of ST14^K2, ST23^K57, ST39^K23, ST76^K23, ST86^K2, ST218^K57, ST219^KL125/114, ST268^K20, and ST2674^K47 caused severe systemic infections, including ST14^K2, ST39^K23, and ST268^K20 that were associated with fatal incomes. Moreover, eight isolates of ST395^K2 and ST307^{KL102/149/155} were associated with manifestations of vasculitis and microcirculation disorders. Another 12 K. pneumoniae isolates of ST395^K2,KL39, ST307^{KL102/149/155}, and ST147^K14/64 were collected from patients without severe systemic infections. Major isolates (n = 38) were XDR and MDR. Beta-lactamase genes were identified: bla_SHV (n = 41), bla_CTX-M (n = 28), bla_TEM (n = 21), bla_OXA-48 (n = 21), bla_NDM (n = 1), and bla_KPC (n = 1). The prevalent virulence genes were wabG (n = 41), fimH (n = 41), allS (n = 41), and uge (n = 34), and rarer, detected only in the genomes of the isolates causing severe systemic infections—rmpA (n = 8), kfu (n = 6), iroN (n = 5), and iroD (n = 5) indicating high potential of the isolates for hypervirulence.

Klebsiella pneumoniae is an increasingly important hospital pathogen. Classical K. pneumoniae (cKp) and hypervirulent K. pneumoniae (hvKp) are two distinct evolutionary genetic lines. The recently ongoing evolution of K. pneumoniae resulted in the generation of hybrid hvKP-MDR strains. K. pneumoniae distinct isolates (n = 70) belonged to 20 sequence types with the prevalence of ST395 (27.1%), ST23 (18.6%), ST147 (15.7%), and ST86 (7.1%), and 17 capsular types with the predominance of K2 (31.4%), K57 (18.6%), K64 (10.0%), K1 (5.7%) were isolated from patients of the Moscow neurosurgery ICU in 2014–2019. The rate of multi-drug resistant (MDR) and carbapenem-resistant phenotypes were 84.3% and 45.7%, respectively. Whole-genome sequencing of five selected strains belonging to cKp (ST395^K47 and ST147^K64), hvKp (ST86^K2), and hvKp-MDR (ST23^K1 and ST23^K57) revealed bla_SHV, bla_TEM, bla_CTX, bla_OXA-48, and bla_NDM beta-lactamase genes; acr, oqx, kpn, kde, and kex efflux genes; and K. pneumoniae virulence genes. Selective pressure of 100 mg/L ampicillin or 10 mg/L ceftriaxone induced changes of expression levels for named genes in the strains belonging to cKp, hvKp, and hybrid hvKp-MDR. Obtained results seem to be important for epidemiologists and clinicians for enhancing knowledge about hospital pathogens.

Acinetobacter baumannii is one of the significant healthcare-associated meningitis agents characterized by multidrug resistance and a high mortality risk. Thirty-seven A. baumannii strains were isolated from thirty-seven patients of Moscow neuro-ICU with meningitis in 2013–2020. The death rate was 37.8%. Strain susceptibility to antimicrobials was determined on the Vitek-2 instrument. Whole-genome sequencing was conducted using Illumina technology; the sequence types (ST), capsular types (KL), lipooligosaccharide outer core locus (OCL), antimicrobial resistance genes, and virulence genes were identified. The prevalent ST was ST2, belonging to the international clone IC2, and rarer, ST1, ST19, ST45, ST78, ST106, and ST400, with prevalence of KL9 and OCL1. Twenty-nine strains belonged to multidrug-resistant (MDR) and eight extensively drug-resistant (XDR) categories. Genes conferring resistance to beta-lactams (bla_PER, bla_GES, bla_ADC, bla_CARB, bla_CTX-M, bla_TEM, and bla_OXA-types), aminoglycosides (aac, aad, ant, aph, and arm), tetracyclines (tet), macrolides (msr and mph), phenicols (cml, cat, and flo), sulfonamides (dfr and sul), rifampin (arr), and antiseptics (qac) were identified. Virulence genes of nine groups (Adherence, Biofilm formation, Enzymes, Immune evasion, Iron uptake, Regulation, Serum resistance, Stress adaptation, and Antiphagocytosis) were detected. The study highlights the heterogeneity in genetic clones, antimicrobial resistance, and virulence genes variability among the agents of A. baumannii meningitis, with the prevalence of the dominant international clone IC2.

The spread of multidrug-resistant Gram-negative bacteria, which is associated with the distribution of beta-lactamase genes and class 1 and 2 integrons, is a global problem. In this study, in the Moscow neurosurgery intensive care unit (neuro-ICU), the high prevalence of the above-stated genes was found to be associated with intestinal and tracheal carriage. Seven-point prevalence surveys, which included 60 patients in the neuro-ICU, were conducted weekly in the period from Oct. to Nov. 2019. A total of 293 clinical samples were analyzed, including 146 rectal and 147 tracheal swabs; 344 Gram-negative bacteria isolates were collected. Beta-lactamase genes (n = 837) were detected in the isolates, including beta-lactamase bla_TEM (n = 162), bla_SHV (n = 145), cephalosporinase bla_CTX–M (n = 228), carbapenemase bla_NDM (n = 44), bla_KPC (n = 25), bla_OXA–48 (n = 126), bla_{OXA–51–like} (n = 54), bla_{OXA–40-like} (n = 43), bla_{OXA–23-like} (n = 8), and bla_VIM (n = 2), as well as class 1 (n = 189) and class 2 (n = 12) integrons. One extensively drug-resistant Klebsiella pneumoniae strain (sequence type ST39 and capsular type K23), simultaneously carried beta-lactamase genes, bla_SHV–40 and bla_TEM–1B, three carbapenemase genes, bla_NDM, bla_KPC, and bla_OXA–48, the cephalosporinase gene bla_CTX–M, and two class 1 integrons. Before this study, such heavily armed strains have not been reported, suggesting the ongoing evolution of antibiotic resistance.

The freeze dried samples of bovine serum albumin (BSA) and BSA—saccharose, BSA—lactose, and BSA—trehalose complexes were studied by the proton NMR relaxation technique in the humidity range from 1 to 20%. The disaccharide molecules were found to assume translational mobility at the humidity higher than 12% per dry weight. It was established that, at the same water content in the samples, saccharose, lactose, and trehalose possess, respectively, high, intermediate, and low mobility. To display water “dissolving” properties in these systems at a humidity higher than 12%, two, three, and four water molecules are required additionally per each sugar molecule for, respectively, saccharose, lactose, and trehalose.