Antibiotic-resistant Bacterial Pathogens Research Articles

Targeting the Essential Transcription Factor HP1043 of Helicobacter pylori: A Drug Repositioning Study.

Antibiotic-resistant bacterial pathogens are a very challenging problem nowadays. Helicobacter pylori is one of the most widespread and successful human pathogens since it colonizes half of the world population causing chronic and atrophic gastritis, peptic ulcer, mucosa-associated lymphoid tissue-lymphoma, and even gastric adenocarcinoma. Moreover, it displays resistance to numerous antibiotics. One of the H. pylori pivotal transcription factors, HP1043, plays a fundamental role in regulating essential cellular processes. Like other bacterial transcription factors, HP1043 does not display a eukaryote homolog. These characteristics make HP1043 a promising candidate to develop novel antibacterial strategies. Drug repositioning is a relatively recent strategy employed in drug development; testing approved drugs on new targets considerably reduces the time and cost of this process. The combined computational and in vitro approach further reduces the number of compounds to be tested in vivo. Our aim was to identify a subset of known drugs able to prevent HP1043 binding to DNA promoters. This result was reached through evaluation by molecular docking the binding capacity of about 14,350 molecules on the HP1043 dimer in both conformations, bound and unbound to the DNA. Employing an ad hoc pipeline including MMGBSA molecular dynamics, a selection of seven drugs was obtained. These were tested in vitro by electrophoretic mobility shift assay to evaluate the HP1043–DNA interaction. Among these, three returned promising results showing an appreciable reduction of the DNA-binding activity of HP1043. Overall, we applied a computational methodology coupled with experimental validation of the results to screen a large number of known drugs on one of the H. pylori essential transcription factors. This methodology allowed a rapid reduction of the number of drugs to be tested, and the drug repositioning approach considerably reduced the drug design costs. Identified drugs do not belong to the same pharmaceutical category and, by computational analysis, bound different cavities, but all display a reduction of HP1043 binding activity on the DNA.

Characterization and Genomic Analysis of a Novel Jumbo Bacteriophage vB_StaM_SA1 Infecting Staphylococcus aureus With Two Lysins.

The development of new antimicrobial agents is critically needed due to the alarming increase in antibiotic resistance in bacterial pathogens. Phages have been widely considered as effective alternatives to antibiotics. A novel phage vB_StaM_SA1 (hereinafter as SA1) that can infect multiple Staphylococcus strains was isolated from untreated sewage of a pig farm, which belonged to Myoviridae family. At MOI of 0.1, the latent period of phage SA1 was 55 min, and the final titer reached about 109 PFU/mL. The genome of phage SA1 was 260,727 bp, indicating that it can be classified as a jumbo phage. The genome of SA1 had 258 ORFs and a serine tRNA, while only 53 ORFs were annotated with functions. Phage SA1 contained a group of core genes that was characterized by multiple RNA polymerase subunits and also found in phiKZ-related jumbo phages. The phylogenetic tree showed that phage SA1 was a phiKZ-related phage and was closer to jumbo phages compared with Staphylococcus phages with small genome. Three proteins (lys4, lys210, and lys211) were predicted to be associated with lysins, and two proteins with lytic function were verified by recombinant expression and bacterial survival test. Both lys210 and lys211 possessed efficient bactericidal ability, and lys210 could lyse all test strains. The results show that phage SA1 and lys210/lys211 could be potentially used as antibiotic agents to treat Staphylococcus infection.

Seaweed-associated heterotrophic bacteria: are they future novel sources of antimicrobial agents against drug-resistant pathogens?

Emergence of multidrug-resistant microorganisms and requirements for novel antimicrobial compounds necessitate exploring newer habitats to develop potential bioactive leads. Culture-contingent analysis of heterotrophic bacterial flora from the seaweeds led to the isolation of bioactive strains possessing potential antibacterial properties against wide-ranging clinical pathogens viz., methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecalis (VREfs). Seven of the most active strains belonging to the phylum Firmicutes isolated from a brown seaweed (Phaeophyceae) Sargassum wightii exhibited spot-over-lawn assay guided inhibition zone of larger than 30mm. Integrated phenotypic and genotypic studies have led to the characterization of the seaweed-associated bacteria particularly belonging to the phylum Firmicutes. The organic extracts of the studied bacteria exhibited promising antibacterial properties againstMRSA and VREfs with minimum inhibitory concentration ranging between 6.25 and 12.50μg/mL. Time-kill kinetic profiles of those bacteria displayed rapid bactericidal activity against both E. coli and MRSA, showing a ≥ 3log10 reduction in viable cell count than the initial. Among the studied bioactive Bacillus spp, B. tequilensis MTCC13043 and B. altiitudinis MTCC13046 were found to possess functional polyketide synthase (pks) gene (MW027664 and MW027660) that could be amplified. The outcome of amplified genes encrypting for polyketide synthase in conjunction with antibacterial activities unveiled the broad-spectrum antimicrobial activities of the marine heterotrophic Firmicutes, which could be further used against the emergent problem of antibiotic-resistant bacterial pathogens.

Changes in resistance pattern of ESKAPE pathogens between 2010 and 2020 in the clinical center of University of Szeged, Hungary.

The acronym ESKAPE stands for six antibiotic-resistant bacterial pathogens namely, Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp. Monitoring their resistance is an important task for clinical microbiology laboratories. Our aim was to analyze the resistance patterns of these bacteria over ten years in clinical samples of our department. We examined the sample types from which these pathogens were most frequently isolated. The incidence of tests with resistant results for each pathogen in aggregate and the most important subgroups of each was also analyzed. We have also intended to predict the local priorities amongst these pathogens. The results of 1,268,126 antibiotic susceptibility tests performed on a total of 70,099 isolates over this period were examined. Most strains were derived from urine, blood culture, trachea, vagina, wounds, and abscesses. Prevalence of ESKAPE bacteria increased between 2011 and 2020 however, the steepest intensifications were seen in the cases of K. pneumoniae and P. aeruginosa. The number of antibiotic susceptibility tests with resistant results has also increased over the decade but the most notable increase was detected in E. faecium and A. baumannii. Based on the calculation of antimicrobial resistance index for each pathogen, the most serious challenges for us at present are A. baumannii, P. aeruginosa, and E. faecium and their multi-resistant forms. The theoretical prediction of proportion of resistant tests between 2020 and 2030 in our care area draws attention to a worrying trend in the cases of vancomycin-resistant E. faecium and carbapenem-resistant A. baumannii strains.

Decay and damage of therapeutic phage OMKO1 by environmental stressors.

Antibiotic resistant bacterial pathogens are increasingly prevalent, driving the need for alternative approaches to chemical antibiotics when treating infections. One such approach is bacteriophage therapy: the use of bacteria-specific viruses that lyse (kill) their host cells. Just as the effect of environmental conditions (e.g. elevated temperature) on antibiotic efficacy is well-studied, the effect of environmental stressors on the potency of phage therapy candidates demands examination. Therapeutic phage OMKO1 infects and kills the opportunistic human pathogen Pseudomonas aeruginosa. Here, we used phage OMKO1 as a model to test how environmental stressors can lead to damage and decay of virus particles. We assessed the effects of elevated temperatures, saline concentrations, and urea concentrations. We observed that OMKO1 particles were highly tolerant to different saline concentrations, but decayed more rapidly at elevated temperatures and under high concentrations of urea. Additionally, we found that exposure to elevated temperature reduced the ability of surviving phage particles to suppress the growth of P. aeruginosa, suggesting a temperature-induced damage. Our findings demonstrate that OMKO1 is highly tolerant to a range of conditions that could be experienced inside and outside the human body, while also showing the need for careful characterization of therapeutic phages to ensure that environmental exposure does not compromise their expected potency, dosing, and pharmacokinetics.

Metabolic reprogramming of Pseudomonas aeruginosa by phage-based quorum sensing modulation.

The Pseudomonas quinolone signal (PQS) is a multifunctional quorum sensing molecule of key importance to P.aeruginosa. Here, we report that the lytic Pseudomonas bacterial virus LUZ19 targets this population density-dependent signaling system by expressing quorum sensing targeting protein (Qst) early during infection. We demonstrate that Qst interacts with PqsD, a key host quinolone signal biosynthesis pathway enzyme, resulting in decreased levels of PQS and its precursor 2-heptyl-4(1H)-quinolone. The lack of a functional PqsD enzyme impairs LUZ19 infection but is restored by external supplementation of 2-heptyl-4(1H)-quinolone, suggesting that LUZ19 exploits the PQS system for successful infection. We establish a broad functional interaction network of Qst, which includes enzymes of cofactor biosynthesis pathways (CoaC/ThiD) and a non-ribosomal peptide synthetase pathway (PA1217). Qst therefore represents an exquisite example of intricate reprogramming of the bacterium by a phage, which may be further exploited as tool to combat antibiotic resistant bacterial pathogens.

USE OF BACTERIOPHAGES AS A TARGET SPECIFIC THERAPY AGAINST FOOD-BORNE PATHOGENS IN FOOD INDUSTRY- A REVIEW

Within the context of food safety, the use of modern antimicrobial technologies to inactivate the food-borne pathogens is not infallible, as proven by the ceaseless increase in the food-borne illnesses. The extensive use of antibiotics to minimize the microbial contamination further results in the evolution of antibiotic-resistant bacterial pathogens. Moreover, some of the decontamination methods frequently used in the food industry cannot be directly applied to fresh fruits and vegetables, and RTE meals. Therefore, efforts are being made by researchers in developing a new approach so as to control the bacterial contamination. Consequently, bacteriophages have evolved as the safe, green, effective and natural alternative for treatment and complete destruction of pathogens in food industry. The review provides a general description about bacteriophages and has mainly focused on their potential use as antimicrobials during the postharvest processing of foods. Numerous research papers, review papers, book chapters and other relevant literature was used for writing this review paper.

Integrative Reverse Genetic Analysis Identifies Polymorphisms Contributing to Decreased Antimicrobial Agent Susceptibility in Streptococcus pyogenes.

ABSTRACTIdentification of genetic polymorphisms causing increased antibiotic resistance in bacterial pathogens traditionally has proceeded from observed phenotype to defined mutant genotype. The availability of large collections of microbial genome sequences that lack antibiotic susceptibility metadata provides an important resource and opportunity to obtain new information about increased antimicrobial resistance by a reverse genotype-to-phenotype bioinformatic and experimental workflow. We analyzed 26,465 genome sequences of Streptococcus pyogenes, a human pathogen causing 700 million infections annually. The population genomic data identified amino acid changes in penicillin-binding proteins 1A, 1B, 2A, and 2X with signatures of evolution under positive selection as potential candidates for causing decreased susceptibility to β-lactam antibiotics. Construction and analysis of isogenic mutant strains containing individual amino acid replacements in penicillin-binding protein 2X (PBP2X) confirmed that the identified residues produced decreased susceptibility to penicillin. We also discovered the first chimeric PBP2X in S. pyogenes and show that strains containing it have significantly decreased β-lactam susceptibility. The novel integrative reverse genotype-to-phenotype strategy presented is broadly applicable to other pathogens and likely will lead to new knowledge about antimicrobial agent resistance, a massive public health problem worldwide.

Fine-Scale Reconstruction of the Evolution of FII-33 Multidrug Resistance Plasmids Enables High-Resolution Genomic Surveillance.

ABSTRACTWe examined 185 complete, publicly available FII-33 plasmid sequences, characterizing their backbone and various insertions. The variable characteristic insertions facilitated evolutionary reconstruction for this plasmid group, beginning with the acquisition of a primary resistance region (PRR) over 10 years ago. FII-33 plasmids have evolved by acquiring additional resistance genes in the PRR via translocatable elements and by forming cointegrates with plasmids of other types. In all cases, IS26 is suspected to have mediated cointegration. Plasmid cointegration has contributed to the accumulation of resistance genes and may have increased the transmissibility, stability, and host range of the original FII-33 lineage. A particularly important sublineage was formed by a replicative IS26 cointegration event that fused an FII-33 plasmid with a blaKPC-2-containing R-type plasmid, interrupting the FII-33 traI gene encoding the conjugative relaxase. The FII-33:R cointegrate arose in the Klebsiella pneumoniae ST11 clone and remains largely confined there due to the abolition of transfer ability by the FII-33:R cointegration event. However, in some cases FII-33:R cointegrates have fused with additional plasmids and acquired complete transfer regions or oriT sequences that might restore their ability to transfer horizontally. Cointegration events across FII-33 plasmid sublineages have involved plasmids of at least 15 different types. This suggests that plasmid cointegration occurs readily and is more common than previously appreciated, raising questions about the effects of cointegrate formation on plasmid host range, stability, and capacity for horizontal transfer. Resources are provided for detecting and characterizing FII-33 plasmid sublineages from complete or draft genome sequences.IMPORTANCE Effective genomic surveillance of antibiotic-resistant bacterial pathogens must consider plasmids, which are frequently implicated in the accumulation and transfer of resistance genes between bacterial strains or species. However, the evolution of plasmids is complex, and simple typing or comparison tools cannot accurately determine whether plasmids belong to the same sublineages. This precludes precise tracking of plasmid movement in bacterial populations. We have examined the FII-33 group, which has been associated with multidrug resistance and particularly carbapenem resistance in clinically significant members of the Enterobacterales in China. Our analysis has provided insight into the evolution of this important plasmid group, allowing us to develop resources for rapidly typing them to the sublineage level in complete or draft genome sequences. Our approach will improve detection and characterization of FII-33 plasmids and facilitate surveillance within and outside China. The approach can serve as a model for similar studies of other plasmid types.

Antibacterial activity of Myrtus communis L. and Melaleuca leucadendron var. cajaputi essential oils against antibiotic-resistant bacterial strains

Background: Antimicrobial resistance (AMR) is a global threat to public health. There is a dire need for new antibiotics as AMR threatens our last-resort antibiotics efficacy.1,2 Myrtle plants have been used in folk medicine for centuries. Essential oils from these plants demonstrated therapeutics effects. This study aims to examine the antibacterial activity of essential oils extracted from Myrtus communis L and Melaleuca leucadendron against clinically relevant bacterial pathogens.3Methods: Bacterial growth curves and bactericidal assays were performed on antibiotic-sensitive Staphylococcus aureus (SA) and antibiotic low and high resistant Methicillin-resistant Staphylococcus aureus (MRSA) strains; sensitive and resistant Klebsiella pneumoniae strains (KPS and KPR). Minimum bactericidal concentration (MBC) was performed on 4 strains of colistin sensitive E. coli and 4 colistin resistant E. coli strains. Results: Myrtle essential oils demonstrated a dose-dependent antibacterial activity against all tested strains and inhibited growth even after 24 hours. The tested oils dilutions ranged from 6.15 μl/ml up to 50 μl/ml and inhibited bacterial growth of both antibiotic sensitive and resistant strains of SA, MRSA (Figure 1), and Klebsiella. Further, the incubation of colistin sensitive and colistin-resistant E. coli strains with 50 μl/ml of Myrtle oil for one hour inhibited the growth of all tested strains. The viability of bacteria was tested by spotting on agar plates and further incubation overnight. The data suggest that Myrtle essential oils can effectively kill bacterial pathogens. Conclusion: Essential oils from Myrtus communis L. and Melaleuca leucadendron var. cajaputi possess potent antibacterial activity against antibiotic sensitive and resistant bacterial pathogens. There is a potential for utilizing these antibacterial oils as topical treatment of wound infections.

Antibiotic resistance and potential bacterial pathogens identified in red deer's faecal DNA.

In the last decades, the wildlife-human interface has been increasing due to several anthropogenic factors. Therefore, it is crucial to be aware of the impact of these new dynamics on the health of wild animals and their associated zoonotic disease risks. This study aimed to characterize the faecal microbiota of two populations of red deer (Cervus elaphus) by metabarcoding, with a particular focus on potential human and veterinary pathogens, and to perform an assessment of antibiotic resistance genes (ARGs) occurrence. The faecal microbiota of red deer was assessed by metabarcoding using the 16S rRNA marker, and OTUs of the genera Treponema, Yersinia, Clostridium, Mycobacterium, and Rickettsia were identified. Two of them affiliated with species more commonly regarded as pathogens (Clostridiumpiliforme and Yersinia enterocolitica). The quantification of ARGs was performed by quantitative real-time PCR, using a metagenomic approach, and the most abundant genes were found to be blaTEM , sul1, tetracycline resistance genes (tetW, tetO, and tetQ) and ermF. From these, tetO and tetW are rank II ARGs, which were recently considered future threats for human health. Our results suggest the need for screening programs for the occurrence of pathogens and ARGs in wildlife and particularly in-game species.

Dynamics of the Catalytic Active Site of Isoleucyl tRNA Synthetase from Staphylococcus aureus bound with Adenylate and Mupirocin.

The development of new antimicrobial drugs is critically needed due to the alarming increase in antibiotic resistance in bacterial pathogens. The active sites of different bacterial aminoacyl tRNA synthetases (aaRS) are validated targets of antibiotics. At present, the only aaRS inhibitor approved is mupirocin (MRC) which targets bacterial isoleucyl tRNA synthetase (IleRS). The present work is aimed at understanding the lacunae of knowledge concerning the active site conformational dynamics in IleRS in the presence of inhibitor mupirocin. With this end in view, we have carried out classical molecular dynamics simulation and metadynamics simulations of the open state of IleRS from Staphylococcus aureus (SaIleRS), the closed state tripartite complex bound with cognate adenylate (Ile-AMP) and tRNA, the closed state tripartite complex bound with noncognate MRC, and the closed state tripartite complex bound with tRNA and MRC with mutated SaIleRS (V588F). The present simulation established a dynamic picture of SaIleRS complexed with cognate and the noncognate substrates which are completely consistent with crystallographic and biochemical studies and explain the existing lacunae of knowledge. The active site is significantly more compact in the Ile-AMP bound complex compared to the open state due to the closure of the KMSKS and HMGH loops and clamping down of the tRNA acceptor end near the active site gate. The present result shows that the unusual open conformational state of the KMSKS loop observed in the cognate substrate-bound complex in the crystal is due to crystallographic constraints. Although the mupirocin tightly fits the catalytic active site in the MRC-bound complex, the nonanoic acid moiety is partly exposed to the water. The KMSKS loop is pushed open in the MRC-bound complex to accommodate the noncognate MRC. New tunnels open up, extending to the editing site in the complex. Out of its three broad segments, the C12 to C17 segment, the conjugated segment, and the nonanoic moiety, the conjugated part of MRC binds most effectively with the active site of the MRC-bound complex. The aromatic residues packing around the C12 to C17 segment of MRC stabilize the tRNA hairpin conformation in a similar way as observed in the TrpRS. The V588F mutation is weakening the interaction between this region of the active site and weakens the binding of MRC in the active site. This result explains why the V588F mutation is responsible for low-level mupirocin resistance. The free energy of unbinding the conjugated segment (and C12 to C17 segment, as well) largely contributes to the total free energy of unbinding the MRC. The active site organization of IleRS from eukaryotic Candida albicans is compared with the bacterial IleRS active site to understand the low binding affinity in eukaryotic IleRS. The present study could be a starting point of future studies related to the development of effective drug binding in the SaIleRS.

Engineered Liposomes Protect Immortalized Immune Cells from Cytolysins Secreted by Group A and Group G Streptococci.

The increasing antibiotic resistance of bacterial pathogens fosters the development of alternative, non-antibiotic treatments. Antivirulence therapy, which is neither bacteriostatic nor bactericidal, acts by depriving bacterial pathogens of their virulence factors. To establish a successful infection, many bacterial pathogens secrete exotoxins/cytolysins that perforate the host cell plasma membrane. Recently developed liposomal nanotraps, mimicking the outer layer of the targeted cell membranes, serve as decoys for exotoxins, thus diverting them from attacking host cells. In this study, we develop a liposomal nanotrap formulation that is capable of protecting immortalized immune cells from the whole palette of cytolysins secreted by Streptococcus pyogenes and Streptococcus dysgalactiae subsp. equisimilis—important human pathogens that can cause life-threatening bacteremia. We show that the mixture of cholesterol-containing liposomes with liposomes composed exclusively of phospholipids is protective against the combined action of all streptococcal exotoxins. Our findings pave the way for further development of liposomal antivirulence therapy in order to provide more efficient treatment of bacterial infections, including those caused by antibiotic resistant pathogens.

Антибиотикорезистентность бактерий, выделенных из маточного содержимого свиноматок при послеродовом эндометрите в условиях свиноводческих хозяйств Республики Казахстан

The paper presents the results of determining the sensitivity of the bacterial flora of the uterine contents of sows with postpartum endometritis to the most common groups of antibacterial drugs used in veterinary medicine. The following pathogenic bacterial strains have been identified: Escherichia coli, Trueperella pyogenes, Aeromonas salmonicida, Proteus vulgaris. The sensitivity of the bacterial flora isolated from the uterine contents of pigs with postpartum endometritis to antibacterial drugs of the following groups was determined: carbapenems, cephalosporins, lincosamides, polymyxins, tetracyclines, beta-lactams, penicillins, sulfonamides, aminoglycosides, tetracyclines. To a large number of studied drugs, all types of isolated bacteria showed persistent resistance. Such high rates of antibiotic resistance in bacterial pathogens suggest that alternative therapies need to be sought.

Genome-assisted Identification, Purification, and Characterization of Bacteriocins.

Bacteriocins are antimicrobial peptides with activity against antibiotic resistant bacterial pathogens. Here, we describe a set of methods aimed at purifying, identifying, and characterizing new bacteriocins. The purification consists of ammonium sulphate precipitation, cation-exchange chromatography, and reversed-phase chromatography. The yield of the bacteriocin is quantified by bacteriocin antimicrobial activity in a microtiter plate assay after each purification step. The mass of the purified bacteriocin is assessed by MALDI TOF MS analysis of the active fractions after reversed-phase chromatography. The mass is compared with the theoretical mass based on genetic information from the whole genome sequencing of the bacteriocin producer strain. Physicochemical characterization is performed by assessing antimicrobial activity following heat and protease treatments. Fluorescent techniques are used to examine the capacity of the bacteriocin to disrupt membrane integrity. Herein a set of protocols for purification and characterization of the bacteriocin nisin Z is used as a typical example in this paper.

Gut microbiome in the emergence of antibiotic-resistant bacterial pathogens.

The human gastrointestinal tract is home to a complex and dynamic community of microorganisms known as gut microbiota, which provide the host with important metabolic, signaling, and immunomodulatory functions. Both the commensal and pathogenic members of the gut microbiome serve as reservoirs of antimicrobial-resistance genes (ARG), which can cause potential health threats to the host and can transfer the ARGs to the susceptible microbes and into the environment. Antimicrobial resistance is becoming a major burden on human health and is widely recognized as a global challenge. The diversity and abundance of ARGs in the gut microbiome are variable and depend on the exposure to healthcare-associated antibiotics, usage of antibiotics in veterinary and agriculture, and the migration of the population. The transfer frequency of the ARGs through horizontal gene transfer (HGT) with the help of mobile genetic elements (MGEs) like plasmids, transposons, or phages is much higher among bacteria living in the GI tract compared to other microbial ecosystems. HGT in gut bacteria is facilitated through multiple gene transfer mechanisms, including transformation, conjugation, transduction, and vesicle fusion. It is the need of the hour to implement strict policies to limit indiscriminate antibiotic usage when needed. Developing rapid diagnostic tests for resistance determination and alternatives to antibiotics like vaccination, probiotics, and bacteriophage therapy should have the highest priority in the research and development sectors. Collective actions for sustainable development against resistant pathogens by promoting endogenous gut microbial growth and diversity through interdisciplinary research and findings are key to overcoming the current antimicrobial resistance crisis.

Bacterial Isolates and Antibiotic Susceptibility of Ear Infections in Al-Kindy Teaching Hospital, Baghdad, Iraq

Background: Ear infections can manifest in many forms depending on site of infection whether external, middle or internal ear and the culprit pathogen whether viral, bacterial or fungal. Acute middle ear infections are usually accompanied by aural discharge. Objective: 1. To get an overview on the bacterial pathogens involved in ear infections. 2. To assess the antibiotic resistance of bacterial pathogens. Methods: A cross sectional study conducted in Al-Kindy Teaching Hospital / Baghdad /Iraq. Swabs taken from 225 patients suffering from aural discharge were tested for culture and sensitivity for the duration of two years 2018-2019. Aural discharge is cultured by inoculating it into blood, MacConkey agar, chocolate agars and Sabouraud agar (for fungi). Then the antibiotic susceptibility and resistance is assessed by (Kirby-Bauer Method). Results: Then, by analyzing the percentage of pathogens involved in ear infections we have found that the highest percentage is for Pseudomonas aeruginosa (51%), followed by Staph, aureus (20%), Proteus vulgaris (11%). Discussion: Cefotaxime, which was known to be an efficient antibiotic against pseudomonas previously, has lost its effectiveness. Similarly, gentamycin is no longer effective against E.coli. Conclusion: Choosing the proper antibiotic in any bacterial infection is of tremendous importance. However, reassessment of antibiotic resistance profiles is vital and should be regarded as a routine task on regular intervals.

Acinetobacter baumannii: Virulence Strategies and Host Defense Mechanisms.

Acinetobacter baumannii is a highly antibiotic-resistant bacterial pathogen known to cause severe life-threatening infections, including pneumonia, meningitis, and sepsis. Recent emergence of this bacterium as a serious nosocomial pathogen has led to categorization of A. baumannii as a “high-priority” pathogen by the World Health Organization (WHO), for which research efforts are urgently required to develop therapeutic interventions. Some of the properties that make A. baumannii a serious pathogen include its capacity to tolerate high levels of stress and enhanced expression of efflux pumps that enable high degrees of antibiotic resistance. Virulence mechanisms employed by A. baumannii to establish successful infection and host responses elicited against A. baumannii to counter the infection are discussed in detail in this article.

An integrated multidisciplinary-based framework for characterizing environmental risks of heavy metals and their effects on antibiotic resistomes in agricultural soils

In this study, a new integrated multidisciplinary-based framework has been proposed to better understand the environmental risks of heavy metals (HMs) in agricultural soils. The source apportionment results revealed by a multilinear engine model were incorporated into the geochemical indexes and the probabilistic health risk assessment models for identifying the source-oriented risks of HMs in the environment. High-throughput sequencing-based metagenomic assembly analysis was used for characterizing the prevalence and dissemination risk of antibiotic resistomes and their associations with the geochemical enrichment of HMs in the soils. Results showed agricultural and industrial activities were the main sources of HMs in the environment. Although the soils were contaminated moderately by HMs and the health risks posed by soil metals were negligible for both adult and children, source-oriented risk evaluation suggested agricultural activities contributed relatively higher contamination and health risks than the other sources. Notably, abundant and diverse antibiotic resistant genes, mobile gene elements, virulence factors, and antibiotic-resistant bacterial pathogens were identified in the agricultural soils, as well as their co-occurrences on the same contigs, implying a non-negligible resistome risk. Further, statistical and network analyses showed the geochemical enrichment of HMs exerted significant effects on the antibiotic resistomes in the environment.

Identification of bacterial antibiotic resistance genes in next-generation sequencing data (review of literature).

The spread of antibiotic-resistant human bacterial pathogens is a serious threat to modern medicine. Antibiotic susceptibility testing is essential for treatment regimens optimization and preventing dissemination of antibiotic resistance. Therefore, development of antibiotic susceptibility testing methods is a priority challenge of laboratory medicine. The aim of this review is to analyze the capabilities of the bioinformatics tools for bacterial whole genome sequence data processing. The PubMed database, Russian scientific electronic library eLIBRARY, information networks of World health organization and European Society of Clinical Microbiology and Infectious Diseases (ESCMID) were used during the analysis. In this review, the platforms for whole genome sequencing, which are suitable for detection of bacterial genetic resistance determinants, are described. The classic step of genetic resistance determinants searching is an alignment between the query nucleotide/protein sequence and the subject (database) nucleotide/protein sequence, which is performed using the nucleotide and protein sequence databases. The most commonly used databases are Resfinder, CARD, Bacterial Antimicrobial Resistance Reference Gene Database. The results of the resistance determinants searching in genome assemblies is more correct in comparison to results of the searching in contigs. The new resistance genes searching bioinformatics tools, such as neural networks and machine learning, are discussed in the review. After critical appraisal of the current antibiotic resistance databases we designed a protocol for predicting antibiotic resistance using whole genome sequence data. The designed protocol can be used as a basis of the algorithm for qualitative and quantitative antimicrobial susceptibility testing based on whole genome sequence data.