Pneumonia Strains Research Articles

Identification and phylogenetic analysis of carbapenemase genes from clinical strains of Klebsiella pneumoniae.

Klebsiella pneumoniae is an encapsulated Gram-negative bacterium that is responsible for numerous infections in healthcare facilities worldwide and is frequently isolated. The World Health Organization has listed K. pneumoniaeas as a critical antibiotic resistant bacterial pathogen for which new antibiotics are urgently needed. This study aimed to use molecular tools to identify and examine antibiotic resistance in clinical strains of K. pneumoniae. A total of 15 unduplicated K. pneumoniae strains isolated from patient samples with multidrug-resistant (MDR) profiles were subjected to polymerase chain reaction (PCR) to amplify the most common carbapenem resistance genes. (GTG)5 PCR and phylogenetic analysis were performed to identify the genetic relationship between the strains. All strains yielded a (GTG)5-PCR profile, and this allowed us to group these strains into 8 groups according to the size and number of characteristic bands. Phylogenetic analysis was done using the free software UPGMA and a single bacterial clone with a correlation coefficient of over 97% was identified. New Delhi metallo-beta-lactamase NDM-like (blaNDM) carbapenem resistance genes were detected in three strains of K. pneumoniae, which represented a resistance rate of 20%. However, carbapenemases type A [Klebsiella pneumoniae carbapenemase (KPC) and imipenem-hydrolysing beta-lactamase (IMI), type D [oxacillinase-48 (OXA-48)], and other metallo-β-lactamase [Verona integron-encoded metallo-beta-lactamase (VIM), and enzyme active on imipenem (IMP)] were not detected. We identified and grouped the blaNDM resistance genes of Klebsiella pneumonia strains.

Susceptibility Analysis of Klebsiella Pneumoniae Strains to Team Essential Oil Emulsion and Surface Active Compounds

Susceptibility Analysis of Klebsiella Pneumoniae Strains to Team Essential Oil Emulsion and Surface Active Compounds

Characteristics of the Klebsiella Pneumoniae Strain Isolated from a Positive Blood Culture of a Premature Newborn Baby According to the Results of Whole Genome Sequencing

Relevance. K. pneumoniae is a common nosocomial pathogen in pediatric hospitals, often characterized by the presence of a wide range of virulence factors and genetic determinants of antibiotic resistance. Аim. To analyze the results obtained during wholegenome sequencing of a Klebsiella pneumoniae strain isolated from a positive blood culture of a premature newborn. Materials and methods. An ESBL-producing strain of K. pneumoniae isolated from a positive blood culture of a newborn premature infant. Sequencing was performed on the MiSeq platform (Illumina). Analysis of DNA nucleotide sequences of the complete genome of K. pneumoniae was carried out using the website of the Center for Genomic Epidemiology. The search for genetic determinants of antibiotic resistance and virulence was carried out using online services. Results and its discussion. The resulting nucleotide sequence was 5,414,099 bp in length, and the proportion of GC nitrogenous bases was 57.3%. The isolated strain belonged to the sequence type ST3559, had 4 genes encoding the synthesis of enzymes that hydrolyze antibacterial drugs from the beta-lactam group, 2 genes providing resistance to quinolones/fluoroquinolones, 1 resistance gene each to trimethoprim, chloramphenicol, fosfomycin and aminoglycoside antibiotics. Most of the virulence factor genes identified in the studied strain ensure the recognition and absorption of iron ions necessary for the competitive functioning of the bacterial cell. K. pneumoniae possesses the acrA efflux pump gene and its regulators, as well as 4 prophage particles and 1 CRISPCas IE system. Conclusions. Whole-genome sequencing of the K. pneumoniae strain isolated from a positive blood culture of a premature newborn allows us to characterize in detail the causative agent of a generalized infection and detect a wide range of genetic determinants of virulence factors and antibiotic resistance. The ESBL-producing strain of K. pneumoniae, as the etiological agent of neonatal sepsis, was characterized by the presence of virulence genes, multidrug resistance, both due to genes encoding enzymes that hydrolyze antibiotics, and due to the presence of efflux pumps and their regulators. The use of the results of traditional cultural research methods together with high-throughput sequencing data is a promising area of scientific research and has a reserve of practical application in the field of clinical medicine, genetics of microorganisms, molecular epidemiology at the local and global levels

Optimization of the production of Exopolysaccharide (EPS) from biofilm-forming bacterial consortium using different parameters

Bacterial cells produce a variety of exopolysaccharide (EPS) throughout their life cycle. EPS plays a crucial role in the formation of biofilms and the control of environmental processes in microbial communities. The bacterial community benefits from biofilms in several ways, including nutrient compartmentation and storage, environmental stress resistance and synergism. The generation of EPS by consortium-associated bacteria, which involves a variety of microorganisms, is versatile and efficient for use in industrial and environmental processes, such as biotechnology and pollution degradation. The qualitative and quantitative chemical characteristics of the consortium's EPS are mostly determined by the type of culture utilized, nutrient type, temperature, and pH. The main goal of the study was to find out how EPS biosynthesis works in a bacterial consortium culture. To achieve maximum EPS regeneration, pH, temperature, carbon, and nitrogen sources were optimized. A consortium of three bacterial strains i.e. Klebsiella pneumonia strain, Pseudomonas aeruginosa strain and Burkholderia cepacian strain that were distinguished by their capacity to produce biofilms were developed as part of the study. The maximum total EPS production was detected after 8 days of incubation at pH 7.5 (15.5 g/L) and a temperature of 35 °C (12.60 g/L). The optimum EPS production was identified as 17.84 g/L and 21.07 g/L when ammonium sulfate and glucose were employed as nitrogen and carbon supplements, respectively. The protein concentration of the EPS was 3067 µg/mL, which was more than the carbohydrate (263.10 µg/mL) and DNA (7.60 µg/mL) content. The total protein/carbohydrate ratio of the EPS detected in the present study was 11.65, which was much higher than in prior investigations. FTIR analysis of EPS confirmed the functional groups of carbohydrates, proteins, lipids and DNA, viz., CO, COOH, NH, OH, CH and OCH3. The study's findings revealed that enhancing the consortium's EPS molecules involves lowering biofilm-related infections and improving the efficacy of antimicrobial treatments in wastewater.

Distribution, characterization and antibiotic resistance of hypervirulent Klebsiella Pneumoniae (hvKp) strains versus classical strains(CKp) causing healthcare associated infections in Sohag University Hospitals

Distribution, characterization and antibiotic resistance of hypervirulent Klebsiella Pneumoniae (hvKp) strains versus classical strains(CKp) causing healthcare associated infections in Sohag University Hospitals

Gamma-Irradiated Honey: Its Medicinal Efficacy and Biological Activities for Burned Skin in a Rat Model

Honey, with its diverse chemical, physical, and antibacterial properties, exhibits significant variability depending on the floral origin of the nectar source. Likewise, its therapeutic potential in the context of healing cutaneous burn injuries shows remarkable variation. In this comprehensive two-week study, four distinct honey varieties (Nigella Sativa Honey - NSH, Moringa Honey - MOH, Sidr Honey - SIH, and Pumpkin Honey - PUH), along with Vaseline, were administered as therapeutic interventions in a rat model. The study covered a range of parameters, including wound contraction, neovascularization rates, malondialdehyde (MDA) content determined via colorimetry, and assessments of breaking strength and dry tissue weight. Agar diffusion tests were conducted to evaluate the efficacy of treatments against Streptococcus pyogenes and Klebsiella pneumonia strains. On the 15th day, the MOH-treated group exhibited the most rapid wound contraction at 84.58%, while the Vaseline-treated group showed the slowest at 68.40%. Notably, neovascularization was most pronounced in MOH-treated animals (45.01%), followed by SIH (42.73%), PUH (36.15%), and NSH (31.17%). In contrast, the Vaseline group had the lowest rate at 15.53%. Histological examinations revealed that animals treated with Vaseline had fewer oval-shaped cells compared to honey-treated counterparts. The highest weight loss occurred in the Vaseline group at 42.25% on the 15th day. MDA content was highest in SIH-treated animals (29.18 nmol/mg), followed by MOH (24.01 nmol/mg), PUH (22.88 nmol/mg), and NSH (22.51 nmol/mg), with the Vaseline-treated group having the lowest level at 7.37 nmol/mg. Furthermore, breaking strength was highest in rats treated with NSH ointment (755.29 g), followed by PUH (739.55 g), SIH (613.11 g), and MOH (600.32 g), while the Vaseline group had the lowest strength at 359.3 g. Similarly, the greatest dry tissue weight was observed in rats treated with PUH ointment (188.79 mg), followed by NSH (182.34 mg), SIH (179.15 mg), MOH (176.15 mg), while the Vaseline group had the lowest dry tissue weight at 101.17 mg. Notably, NSH-treated animals exhibited a reduction in Streptococcus pyogenes growth (5.76 mm) compared to the Vaseline group (19.68 mm), while MOH-treated animals displayed decreased Klebsiellapneumoniae growth (5.33 mm) compared to the Vaseline group (17.42 mm). These results collectively emphasize the comparable efficacy of honey in facilitating burn wound healing.

Exploring the In vitro Antibacterial Efficacy of Yemidir Embuay (Cucumis focifolius) and Endod (Phytolacca dodecandra) of Different Extracts Against Some Bacteria Pathogens

Medicinal plants are one of the most indigenous resources in the world for the treatment of different diseases. Now a day most of infectious diseases are resistant to the antimicrobial agents. Therefore, the aim of this study was to explore the efficacy of Cucumis focifolius and Phytolacca dodecandra against Staphylococcus aureus, Klebsiella Pneumonia and Neisseria gonorrhea. Ethanol, Methanol, Chloroform and Water are used as solvents for the extraction of phytochemical compounds from Cucumis focifolius and Phytolacca dodecandra plants. The effective extraction of bioactive compounds was achieved using methanol, chloroform and water as solvents. According to this study there were bioactive compounds that extracted from selected plants which were potentially effective against Neisseria gonorrhea and some other pathogens. Both Cucumis focifolius and Phytolacca dodecandra some inhibition zone on S. auerus and k. pneumonia strains. Highly statistical significant antibacterial effect against N. gonorrhea (16.6±3.05) for ethanol root extract of Cucumis focifolius followed by methanol root extract with an inhibition zone of (15±2.64) were exhibited. The extract also shows (13.3±1.25mm, 14±1.05mm by chloroform and distilled water root extract respectively. Ethanol C. focifolius root shows lowest Minimum inhibitory concentration against Neisseria gonorrhea as compared to other extracts. Therefore, we recommend people to search for new antimicrobial agents for better treatment of Neisseria Gonorrhea, which is one of causative agent of sexual transmitted disease.

Evolutionary and co-evolutionary phage training approaches enhance bacterial suppression and delay the emergence of phage resistance.

The development of phage resistance by bacteria is a major barrier that impedes the therapeutic use of phages. Phage training has been proposed as a novel tool that harnesses the evolutionary potential of phages to improve phage infectivity. Both evolutionary and co-evolutionary phage training models have been previously reported to train phages. However, both of these phage training models have been reported able to effectively suppress the emergence of phage-resistant bacteria mutants, thus presenting a contradictory phenomenon. Therefore, in this study, we set out to systematically compare the effectiveness of both evolutionary and co-evolutionary phage training models with regard to phage physiology, infectivity, and genotype. To this end, a natural lytic phage capable of infecting a Klebsiella pneumonia strain was isolated from wastewater and subjected to evolutionary and co-evolutionary phage training for 30days. After the phage training, the physiology and genomic characteristics of evolved and co-evolved phages were assessed. Our results demonstrated that both evolved and co-evolved phages exhibit improved bacterial suppression activity and are able to delay the emergence of phage resistance. Furthermore, both phages harbored unique genome mutational changes in different functionally associated phage proteins. Similarly, evolved and co-evolved phage-resistant bacteria mutants that arose post phage infection displayed varying phage resistance sensitivities, which may be correlated to the unique genome mutational change identified in cell membrane structure. In particular, co-evolved phage-resistant bacteria mutants exhibited less phage resistance compared to evolved phage-resistant bacteria mutants. These results highlighted the finding that the co-evolutionary phage training model serves as a better phage training model as it endows phage with improved infectivity, but also selects for phage-resistant bacteria with a lower phage resistance when compared to evolutionary phage training.

Rapid Semiconducting Supramolecular Mg(II)-Metallohydrogel: Exploring Its Potential in Nonvolatile Resistive Switching Applications and Antiseptic Wound Healing Properties.

An effective strategy was employed for the rapid development of a supramolecular metallohydrogel of Mg(II) ion (i.e., Mg@PEHA) using pentaethylenehexamine (PEHA) as a low-molecular-weight gelator in aqueous medium under ambient conditions. The mechanical stability of the synthesized Mg@PEHA metallohydrogel was characterized by using rheological analysis, which showed its robustness across different angular frequencies and oscillator stress levels. The metallohydrogel exhibited excellent thixotropic behavior, which signifies that Mg@PEHA has a self-healing nature. Field emission scanning electron microscopy and transmission electron microscopy images were utilized to explore the rectangular pebble-like hierarchical network of the Mg@PEHA metallohydrogel. Elemental mapping through energy-dispersive X-ray spectroscopy analysis confirmed the presence of primary chemical constituents in the metallohydrogel. Fourier transform infrared spectroscopy spectroscopy provided insights into the possible formation strategy of the metallohydrogel. In this work, Schottky diode structures in a metal-semiconductor-metal geometry based on a magnesium(II) metallohydrogel (Mg@PEHA) were constructed, and the charge transport behavior was observed. Additionally, a resistive random access memory (RRAM) device was developed using Mg@PEHA, which displayed bipolar resistive switching behavior at room temperature. The researchers investigated the switching mechanism, which involved the formation or rupture of conduction filaments, to gain insights into the resistive switching process. The RRAM device demonstrated excellent performance with a high ON/OFF ratio of approximately 100 and remarkable endurance of over 5000 switching cycles. RRAM devices exhibit good endurance, meaning they can endure a large number of read and write cycles without significant degradation in performance. RRAM devices have shown promising reliability in terms of long-term performance and stability, making them suitable for critical applications that require reliable memory solutions. Significant inhibitory activity against the drug-resistant Klebsiella pneumonia strain and its biofilm formation ability was demonstrated by Mg@PEHA. The minimum inhibitory concentration value of the metallohydrogel was determined to be 3 mg/mL when it was dissolved in 1% DMSO. To study the antibiofilm activity, an MTT assay was performed, revealing that biofilm inhibition (60%) commenced at 1 mg/mL of Mg@PEHA when dissolved in 1% DMSO. Moreover, in the mouse excisional wound model, Mg@PEHA played a crucial role in preventing postoperative wound infections and promoting wound healing.

Antibiotic Resistance in Metal-Tolerant Microorganisms from Treatment Facilities.

The study examines the antibiotic resistance of metal-tolerant bacteria isolated from the wastewater treatment plant of a large city to six antibiotics belonging to the β-lactam antibiotics, aminoglycosides and amphenicols. Resistance of bacteria from sewage sludge multitolerant to heavy metals to 18 antibiotics of the β-lactam antibiotics, tetracyclines, aminoglycosides, diaminopyrimidines, amphenicols and ansamycins was studied also. Out of 10, the metal-tolerant microorganisms isolated from wastewater treatment facilities only the Klebsiella pneumonia strain (tolerant to 3 mM Cu) from the sludge of a secondary settling tank did not show resistance to the studied antibiotics at the concentrations considered. Resistance to the maximum amount of antibiotics was typical for strains Serratia fonticola SS0-1, isolated from fresh sewage sludge and resistant to 5 mmol Cu and 3 mmol Pb, or Stenotrophomonas maltophilia SS0-5, also isolated from fresh sludge and resistant to 3 mmol Zn and Cu. It is possible that bacterial resistance to antibiotics develops not only as a result of the use of antibiotics themselves, but also as a result of environmental pollution with heavy metals, and vice versa.

Evaluation of Phytochemicals, and Pharmacological Efficiency Studies of Nyctanthes Arbor-Tristis Linn

Nyctanthes arbor-tristis is a member of the Oleaceae family with various therapeutic characteristics. Different plant components are used intraditional medicine to treat illnesses such as sciatica, chronic fever, and skin diseases. It is today regarded as a rich source of numerous uniquecompounds for creating medicines against various diseases and certain industrial products. In general, the leaves, roots, flowers, and seeds of Parijatare used to cure several ailments in various dose forms, including juice and powder. The current paper discusses the Plant Profile of N. arbor-tristisLinn., as well as the Phytochemical Screening of Extract of N. arbor-tristis Leaves, physicochemical parameters, and the Extraction Procedure andPharmacological Activities conducted on N. arbor-tristis Leaves. The study aims to analyze the antioxidant, antibacterial, and anti-inflammatory potentialof petroleum ether extract of N. arbor-tristis leaves. DPPH scavenging assays and in vitro anti‐inflammatory activity was evaluated using Albumindenaturation, Proteinase inhibitory assay and was subjected to GC-MS analysis to identify its active components. The antibacterial potential wasanalyzed using the agar well diffusion method against Gram-negative and Gram-positive bacteria. Preliminary phytochemical screening confirmed thepresence of alkaloids, flavonoids, phenols, tannins, triterpenoids, Amino acids, Carbohydrate, and the absence of alkaloids, Anthraquinone, Phenols,Saponin, and tannin. Remarkably, free radicals scavenging ability was observed in DPPH with the IC50 of 610.71±13.6μg/mL, and the anti-inflammatoryassay also observed Albumin denaturation with the IC50 of 781.93±13.6μg/mL and the proteinase inhibitory activity with the IC50 of 652.98±32.5μg/mL.In the antibacterial assay, the zone of inhibition was recorded as 20.6±1.6mm, 21.3±1.8mm, 21.3±1.8mm, and 17.9±1.5mm for the strains of Escherichiacoli, Klebsiella pneumonia, Pseudomonas spp., Staphylococcus aureus, and Bacillus spp. respectively. The highest inhibition zone was observed against E. coli(24.8±1.7 and 26.6±1.4mg/mL, respectively), and the least inhibitory was observed against L. acidophilus (19.5±1.1 and 21.7±1.7mg/mL) respectively.GC-MS profile exhibited the presence of 29 components highly accountable for its pharmaceutical activities. The results suggested the presence ofphytochemicals in Nyctanthes arbor-tristis Leaves possess antioxidant and anti-inflammatory properties in the petroleum ether extract. This informationcan be used as a starting point in the search for natural-based drugs that are effective at alleviating inflammatory symptoms and antioxidant aspects.Keywords: , , , , , ,

Enhancing the Production of Polyhydroxybutyrate, a Biodegradable Polymer by an Optimized Process Using a Novel Klebsiella Pneumonia Strain

Enhancing the Production of Polyhydroxybutyrate, a Biodegradable Polymer by an Optimized Process Using a Novel Klebsiella Pneumonia Strain

UhpTE350Q mutation along with the presence of fosA6/5 genes in the genome probably contributes to inherent fosfomycin resistance of Klebsiella pneumoniae

To investigate the molecular mechanism underlying inherent fosfomycin resistance of Klebsiella pneumoniae (K. pneumoniae). The draft genomic sequences of 14 clinical hypervirulent/hypermucoviscous K. pneumoniae (HvKP/ HmKP) isolates were obtained using the next-generation sequencing technology. The genomic sequences were analyzed using the Resistance Gene Identifier (RGI) software for predicting the resistome based on homology and SNP models in the Comprehensive Antibiotic Resistance Database (CARD) and for identification of the presence of phosphomycin resistancerelated genes uhpt and fosA and their mutations in the bacterial genomes. The results were verified by analyzing a total of 521 full-length genomic sequences of K. pneumonia strains obtained from GenBank. All the 14 clinical isolates of HvKP/ HmKP carried hexose phosphate transporter (UhpT) gene mutation, in which the glutamic acid was mutated to glutamine at 350aa (UhpTE350Q mutation); the presence of fosA6 gene was detected in 12 (85.71%) of the isolates and fosA5 gene was detected in the other 2 (14.29%) isolates. Analysis of the genomic sequences of 521 K. pneumonia strains from GenBank showed that 508 (97.50%) strains carried UhpTE350Q mutation, 439 (84.26%) strains harbored fosA6, and 80 (15.36%) strains harbored fosA5; 507 (97.31%) strains were found to have both UhpTE350Q mutation and fosA6/5 genes in the genome. Only 12 (2.30%) strains carried fosA6/5 genes without UhpTE350Q mutation; 1 (0.19%) strain had only UhpTE350Q mutation without fosA6/5 genes, and another strain contained neither UhpTE350Q mutation nor fosA6/5 genes. UhpTE350Q mutation with the presence of fosA6/5 genes are ubiquitous in K. pneumonia genomes, indicating a possible intrinsic mechanism of fosfomycin resistance in the bacterium to limit the use of fosfomycin against infections caused by K. pneumoniae, especially the multi-resistant HvKP/HmKP strains.

COMPARATIVE ANTIMICROBIAL ACTIVITY OF SOME METABIOTICS SYNTHESIZED BY LACTIC ACID BACTERIA

Abstract This paper presents the comparative characteristics of the antimicrobial activity of selected lactic acid bacteria strains and antibiotics. The metabiotics of probiotic lactic acid bacteria inhibited the growth of pathogenic, conditionally pathogenic bacteria, different etiology antibiotic resistant bacteria such as Salmonella sp., E. coli, Proteus mirabilis Pasteurella spp., Clostridium sp., Streptococcus sp., Staphylococcus aureus, Shigella sp., Yersinia enterocolitica, Bacillus cereus with different efficiency depending on pathogens isolation sources. It was shown that bacteriocins of lactic acid bacteria in the same concentration did not affect growth of the commensal microbiota strains, belonging to different genera and species. Lactobacillus and Enterococcus genera showed high sensitivity to investigated antibiotics (about 70%). Among all studied LAB strains of Enterococcus genus, some strains were shown to synthesize polysaccharides. The antimicrobial activity of isolated polysaccharides from Enterococcus faecium К Э-14, Enterococcus faecium К Э-5, Enterococcus lactic acid bacteria. Sp. К Э-9, Enterococcus lactic acid bacteria sp. К Э-6 strains was investigated. It was found that only polysaccharides isolated from Enterococcus faecium К Э-14 and Enterococcus faecium К Э-5 strains show an antimicrobial effect. The Enterococcus faecium КЭ-5 (MDC 9662) lactic acid bacteria strain was selected which produce protein-like substances and disaccharide polymers with antimicrobial activity, consist of glucose and galactose. The growth suppression of different Kl. pneumonia and St. pneumonia strains causing pneumonia by antimicrobial preparations of lactic acid bacteria was shown. The highest antimicrobial activity (100%) was observed when the antimicrobial preparations obtained after cultivation of lactic acid bacteria strains of the Enterococcus genus. The activity depends on the source of isolation of pathogens from a patient. The selected strains can be recommended for the creation of probiotic preparations with targeted purposes.

3-acetyl-coumarin-substituted thiosemicarbazones and their ruthenium, rhodium and iridium metal complexes: An investigation of the antibacterial, antioxidant and cytotoxicity activities

A series of ruthenium, iridium and rhodium half-sandwich complexes containing 3-acetyl chromen-2-one-4(N)-substituted thiosemicarbazone derivatives have been prepared for use as antibacterial and anticancer agents. These complexes have been isolated as cationic bidentate (N, S) and neutral bidentate (N, S) complexes. The complexes have been characterized by spectral techniques (FT-IR, UV-Vis, NMR and HRMS) and single-crystal X-ray crystallography methods. The structure of complex 6 has been determined by a single-crystal X-ray diffraction study, which shows that coordination of the 3-acetyl chromen-2-one-4(N)-substituted thiosemicarbazone ligand to the metal occurs via azomethine nitrogen and thiolate sulfur atoms. Vitro antibacterial activity against gram-positive Staphylococcus aureus and gram-negative Escherichia coli, Klebsiella pneumonia strains and antioxidant activity have been evaluated. Also, the said complexes were tested for anticancer efficacy (in vitro for 48h) in Dalton's lymphoma (DL) cancer cell lines using cell cytotoxicity (MTT) and apoptosis tests. The results were compared with cisplatin (standard drug) under identical experimental conditions. The cell cytotoxicity results showed that complex 5 induced higher cytotoxicity in DL cells, followed by complex 4, 1, and 2.

16SrRNA sequencing analysis for identification of Klebsiella pneumoniae isolated from the extreme kitchen environment

Swabs from dishwasher samples were collected and cultured on different media, and then a gram stain was conducted on pure colonies to find whether they were Gram-positive or negative. 32-gram negative Isolates were obtained from the dishwasher; then, we chose isolates under study depending on morphological features on previous media for further investigation. 32 Gram-negative isolates were obtained from a dishwasher, and three isolates of Klebsiella pneumoniae were diagnosed by some phenotypic characteristics and approved by using 16 SrRNA partial sequencing analyses. The 3 isolates deposited in the NCBI database under accession number OK 254156.1 for K.pneumoniae strain NPK 323, OK 247423.1 for K. pneumonia as strain CUMB SAM-61, and OK245427.1 for K. pneumonia strain PD17. The phylogenetic tree for 3 isolates was done by using MEGA II software. Many experiments have been conducted on two isolates (OK 247423.1 and OK245427.1 ) because the OK 254156.1 strain was lost during laboratory work and repeated cultures. A hemolysis test on blood agar and a lipase test on egg–yolk agar were done; both isolates showed negative results for hemolysis blood and producing lipase enzyme, while both isolates showed their ability to produce lecithinase enzyme. The two isolates gave an excellent result in the tube method for the biofilm formation test. Also, a good candidate production test was obtained for these two isolates using L.B. acetate agar medium. Conclusion: Bacterial species differ according to the environments in which they live, as the species that are isolated from clinical sources and possess many virulence factors that make them more dangerous and pathogenic to humans differ about the same species if isolated from a variety of external environments, which makes them virulent or have new characteristics that make them adapted to live in the environments from which they are isolated. Keywords: Klebsiella pneumonia, virulence factor, extreme conditions, phylogenetic tree.

Microparticulation of Levofloxacin HCl by Crystallo-co-agglomeration Technique

Particle engineering techniques have gained a unique place in the present pharma industry to improve physicochemical properties of the drugs. The aim of this research work was to formulate and evaluate directly compressible agglomerates of Levofloxacin HCl with a view to improve their micromeritic properties and thereby to reduce the cost of production. Fluoroquinolone anti-infective, is used to treat bacterial conjunctivitis, sinusitis, chronic bronchitis, community- acquired pneumonia and pneumonia caused by penicillin resistant strains of Streptococcus pneumonia. Some of the fluoroquinolone high dose’s exhibit poor compressibility and flow properties, hence may not be suitable candidate for direct compression process, but by applying the crystallo-co-agglomeration (CCA) technique, the attempt may be made to change the properties of these molecules to make them suitable candidates for direct compression. This investigation was aimed to utilize CCA process to develop spherical agglomerates of levofloxacin HCl in selected polymers in different ratio. The developed spherical agglomerates of levofloxacin HCl may exhibit improved micrometric and dissolution properties hence may be suitable for direct compression process. Results indicated that micromeritic, mechanical and compressional properties of the agglomerates were greatly influenced by nature and type of polymer incorporated. The mean release time, mean dissolution time, dissolution efficiency, Q30 and Q90 of the tablets prepared from agglomerates showed remarkable increase in CCA compared to tablets prepared by wet granulation. Observations also revealed that by varying the type and concentration of polymer, desired release rate can be obtained.

Differential Study of Antimicrobial Activity of Vancomycin and Teicoplanin (Targocid) against Strains of Staphylococcus aureus and Streptococci sp.

The glycopeptide antibiotics vancomycin and teicoplanin (targocid) are widely used in the treatment of infections caused by gram positive bacteria. Vancomyc inhibits both transglycosylation and transpeptidation reaction during peptidoglycan assembly, targocid inhibits peptidoglycan polymerization, resulting in inhibition of bacterial cell wall synthesis and cell death. In this study twenty strains of streptococcus pyogenes and streptococcus pneumoniae and twenty five strains of staphylococcus aureus were collected from different infections. MI Cs and MBCs of vancomycin and targocid were determined against the bacterial strains, the MICs of vancomycin for the streptococcus strains were 4-8 µg /ml compared with 8-16 µg/ml for targocid and MI Cs of targocid for staphylococcus aureus were 4-16 µg/ml MBC for the same strains were 8-32 µg/ml. the conclusion of this study was that targocid was less active than vancomycin against staphylococci but equal or more active against streptococci specifically it presents excellent in vitro activity against streptococcus pneumonia strains.

Phenotypic and Genetic Analysis of KPC-49, a KPC-2 Variant Conferring Resistance to Ceftazidime-Avibactam and Maintaining Resistance to Imipenem and Meropenem.

Klebsiella pneumoniae, a gram-negative bacterium, poses a severe hazard to public health, with many bacterial hosts having developed resistance to most antibiotics in clinical use. The goal of this study was to look into the development of resistance to both ceftazidime-avibactam and carbapenems, including imipenem and meropenem, in a K. pneumonia strain expressing a novel K. pneumoniae carbapenemase-2 (KPC-2) variant, referred to as KPC-49. After 1 day of incubation of K1 on agar containing ceftazidime-avibactam (MIC = 16/4 mg/L), a second KPC-producing K. pneumoniae strain (K2) was recovered. Antimicrobial susceptibility assays, cloning assays, and whole genome sequencing were performed to analyse and evaluate antibiotic resistance phenotypes and genotypes. K. pneumoniae strain (K1), that produced KPC-2, was susceptible to ceftazidime-avibactam but resistant to carbapenems. The K2 isolate harboured a novel bla KPC-49 variant, which differs from bla KPC-2 by a single nucleotide (C487A), and results in an arginine-serine substitution at amino acid position 163 (R163S). The mutant K2 strain was resistant to both ceftazidime-avibactam and carbapenems. We demonstrated the ability of KPC-49 to hydrolyse carbapenems, which may be attributed to high KPC-49 expression or presence of an efflux pump and/or absence of membrane pore proteins in K2. Furthermore, blaKPC-like was carried on an IncFII (pHN7A8)/IncR-type plasmid within a TnAs1-orf-orf-orf-orf-orf-orf-ISKpn6-bla KPC-ISKpn27 structure. The bla KPC-like gene was flanked by various insertion sequences and transposon elements, including the Tn3 family transposon, such as TnAs1, TnAs3, IS26, and IS481-ISKpn27. New KPC variants are emerging owing to sustained exposure to antimicrobials and modifications in their amino acid sequences. We demonstrated the drug resistance mechanisms of the new mutant strains through experimental whole genome sequencing combined with bioinformatics analysis. Enhanced understanding of laboratory and clinical features of infections due to K. pneumoniae of the new KPC subtype is key to early and accurate anti-infective therapy.

Detection of Klebsiella pneumonia DNA and ESBL positive strains by PCR-based CRISPR-LbCas12a system.

Klebsiella pneumonia (K. pneumonia) is a Gram-negative bacterium that opportunistically causes nosocomial infections in the lung, bloodstream, and urinary tract. Extended-spectrum β-Lactamases (ESBLs)-expressed K. pneumonia strains are widely reported to cause antibiotic resistance and therapy failure. Therefore, early identification of K. pneumonia, especially ESBL-positive strains, is essential in preventing severe infections. However, clinical detection of K. pneumonia requires a time-consuming process in agar disk diffusion. Nucleic acid detection, like qPCR, is precise but requires expensive equipment. Recent research reveals that collateral cleavage activity of CRISPR-LbCas12a has been applied in nucleic acid detection, and the unique testing model can accommodate various testing models. This study established a system that combined PCR with CRISPR-LbCas12a targeting the K. pneumoniae system. Additionally, this study summarized the antibiotic-resistant information of the past five years' K. pneumoniae clinic cases in Luohu Hospital and found that the ESBL-positive strains were growing. This study then designs a crRNA that targets SHV to detect ESBL-resistant K. pneumoniae. This work is to detect K. pneumoniae and ESBL-positive strains' nucleic acid using CRISPR-Cas12 technology. We compared PCR-LbCas12 workflow with PCR and qPCR techniques. This system showed excellent detection specificity and sensitivity in both bench work and clinical samples. Due to its advantages, its application can meet different detection requirements in health centers where qPCR is not accessible. The antibiotic-resistant information is valuable for further research.