Composition characterization of various viperidae snake venoms using MS-based proteomics N-glycoproteomics and N-glycomics.

Viperidae snake species is widely abundant and responsible for most envenomation cases in Turkey. The structural and compositional profiles of snake venom have been investigated to study the venom component variation across different species and to profile the venom biological activity variation against prey. In this context, we used proteomics, glycoproteomics and glycomics strategies to characterize the protein, glycoproteins and glycan structural and compositional profiles of various snake venoms in the Viperidae family. Moreover, we compared these profiles using the downstream bioinformatics and machine learning classification modules. The overall mass spectrometry profiles identified 144 different proteins, 36 glycoproteins and 78 distinct N-glycan structures varying in composition across the five venoms. A high amount of the characterized proteins belongs to the glycosylated protein family Trypsin-like serine protease (Tryp_SPc), Disintegrin (DISIN), and ADAM Cysteine-Rich (ACR). Most identified N-glycans have a complex chain carrying galactosylated N-glycans abundantly. The glycan composition data obtained from glycoproteomics aligns consistently with the findings from glycomics. The clustering and principal component analyses (PCA) illustrated the composition-based similarities and differences between each snake venom species' proteome, glycoproteome and glycan profiles. Specifically, the N-glycan profiles of M. xanthina (Mx) and V. a. ammodytes (Vaa) venoms were identical and difficult to differentiate; in contrast, their proteome profiles were distinct. Interestingly, the variety of the proteins across the species highlighted the impact of glycosylation on the diversity of the glycosylated protein families. This proposed high throughput approach provides accurate and comprehensive profiles of the composition and function of various Viperidae snake venoms.

Carbapenem resistance and biofilm formation status of Enterobacterales isolated from raw milk via molecular versus phenotypic methods.

Antibiotic resistance genes can easily be transferred between bacteria in the biofilm. In the dairy industry, many bacterial species forming biofilms on the surfaces of equipment are widely reported. The experiments reported in this research paper aimed to investigate the carbapenem resistance and biofilm formation properties of Enterobacterales isolates which are spoilage microorganisms obtained from raw milk. In addition, the study determined that whether there was a relationship between the biofilm formation ability or the protein spectra of these isolates. In this study, ninety-two Enterobacterales isolates collected from 173 raw milk samples were investigated. Initially, the isolates were identified as Citrobacter braakii (n = 18), Citrobacter freundii (n = 12), Enterobacter asburiae (n = 1), Enterobacter cloacae (n = 3), Escherichia coli (n = 10), Hafnia alvei (n = 18), Klebsiella oxytoca (n = 1), Serratia fonticola (n = 24), Serratia liquefaciens (n = 4), and Serratia marcescens (n = 1) using MALDI-TOF MS. As a result, carbapenem resistance was determined in 6.5% of the isolates by CIM test, MHT, and the disk diffusion methods, but none of them had blaOXA-48, blaKPC, blaNDM-1, blaOXA23, blaOXA-58, blaOXA-51, blaVIM, and blaIMP genes. This may be due to the effect of other resistance mechanisms such as porin loss or increased flow pump activity. Furthermore, biofilm formation (weak and moderate) was detected in 97.8% of the Enterobacterales isolates. The mass spectra of the moderate biofilm producer isolate of Serratia spp. and the mass spectra of the weak biofilm producers of E.coli presented similarities.

Maldi-TOF MS identification and antibiotic resistance of Escherichia coli isolated from playground.

In this study, it was aimed to determine the antibiotic resistance of Escherichia coli strains isolated from samples taken from various children's parks of Ankara and to confirm the resistance by molecular methods. Five hundred fifty-four samples, including soil samples from 140 different parks and 414 swab samples from slides, swings, ferris wheels, seesaws, and other toys from 176 different parks, were taken. Fourty E. coli strains isolated from these samples were included in the study. Antibiotic susceptibility tests of 40 E. coli isolates were performed by EUCAST recommendations. The resistance rates of E. coli isolates were found as ciprofloxacin 5%, ampicillin 17%, trimethoprim/sulfamethoxazole 15%, streptomycin 12.5%, tobramycin 5%, gentamicin 5%, cefotaxime 2.5%, and ceftazidime 2.5%. Intermediate rates were found as 95%, 90%, and 70% for tobramycin, gentamicin, and streptomycin respectively. blaCTX-M ß-lactamase gene was investigated for an isolate determined to be resistant to both cefotaxime and ceftazidime but blaCTXM gene could not be detected. Aminoglycoside resistance of strains has been investigated because of high intermediate sensitivity rates. For this purpose, aac(6')-Ib, aac(3')-IIa, aph(3')-VI, ant(3')-I, aac(3')-IV, ant(2')-Ia genes scanned, and were detected 97.5% of our isolates ant (3')-I, %25 aac(6')-Ib', 5% aac(3')-IIa, 2.5% ant(2')-Ia. Also, aph(3')-VI, and aac(3')-IV genes could not be detected in any of the isolates. Consequently, it has been revealed that resistant E. coli strains isolated from children's parks can pose a potential risk in public health for transmission of resistant genes.

[The Epidemiology of Carbapenemases in Escherichia coli and Klebsiella pneumoniae Isolated in 2019 in Turkey]. / Türkiye'de 2019 Yili Içinde Izole Edilen Escherichia coli ve Klebsiella pneumoniae Izolatlarinda Karbapenemaz Epidemiyolojisi.

Antibiotic resistance is one of the most important public health problem and one of the most critical steps in preventing resistance is the monitorization of the resistance. Local, regional and global monitoring enables the spread of antibiotic resistance to be understood more clearly. In this study, it was aimed to evaluate the results of the pilot study for the establishment of molecular-based carbapenem surveillance system in Escherichia coli and Klebsiella pneumoniae isolates and to investigate the carbapenemase epidemiology in Turkey. Hospitals (n= 28) from 26 different statistical level II regions from Turkey were included in the study. The hospitals participated in the study submitted ten carbapenem susceptible and ten carbapenem resistant E.coli and K.pneumoniae isolates to our laboratory that were isolated in two different periiods of six-month either between 1 March-31 August or 1 April-30 September 2019. A total of 509 isolates were collected from 26 of the 28 participating hospitals in the study. Isolates were identified by matrix assisted laser desorptionization-time of flight mass spectrophotometry (MALDI TOF MS) (Bruker Daltonics, Germany) method and antibiotic susceptibility tests for imipenem, meropenem and colistin were studied by broth microdilution. Moreover, susceptibilities to amikacin, amoxicillin-clavulanic acid, ampicillin, aztreonam, cefepime, cefotaxime, ceftazidime, ciprofloxacin, ertapenem, gentamicin, piperacillin-tazobactam, tobramycin and trimethoprim-sulfamethoxazole were determined by disc diffusion method. The resistance genes were investigated in isolates which were found to be phenotypically resistant to carbapenem and colistin, in house method was used to investigate carbapenemase genes and a commercial colistin resistant real-time PCR kit (Biospeedy, Turkey) was used for colistin resistance genes. In total, 493 of the 509 isolates collected from hospitals were identified as E.coli (25.7%, n= 127) and K.pneumoniae (74.3%, n= 366) and included in the study. It was determined that 31% of the isolates evaluated were from community-acquired infections and 69% were either from healthcare-associated infections or from colonization sites. Among the tested isolates, 248 (50.3%) were susceptible to carbapenems and 245 (49.7%) were resistant. The types of carbapenemases in carbapenemase-producing were OXA-48 (52.2%), KPC (16.1%), NDM-1 (15%), OXA-48 + NDM-1 (12.6%), KPC + NDM-1 (2.8%) and VIM (0.5%) and OXA-48+VIM (0.5%). Resistance to colistin was detected in 23.3% of the isolates but mcr1-8 genes were not detected. It was found that all colistin resistant isolates are resistant to at least one of the carbapenems. The importance of a molecular-based antimicrobial resistance surveillance system in our country was demonstrated with this pilot study. It is thought that continuous monitoring of these epidemiological features will contribute to the management of infections due to carbapenemase-producing organisms.