Pseudomonas aeruginosa is a major cause of nosocomial infection in children and adults, resulting in significant morbidity and mortality due to its ability to acquire drug resistance. The ability of P. aeruginosa in the environment to cause infection in individuals has been reported previously; henceforth, surveillance of the emergence and transmission of P. aeruginosa strains among patients is important for infection control in a clinical setup. Various gene-typing methods have been used for epidemiological typing of P. aeruginosa isolates for the purpose of surveillance. In this work, the suitability and comparability of two typing methods, enterobacterial repetitive intergenic consensus (ERIC)-PCR and random amplification of polymorphic DNA (RAPD)-PCR fingerprinting, were studied to characterize P. aeruginosa strains isolated from clinical and environmental sources. Forty-four clinical and environmental bacterial isolates of P. aeruginosa were collected between October 2015 and January 2016. DNA extraction, ERIC-PCR and RAPD-PCR, agarose gel electrophoresis, and phylogenetic analyses were carried using the unweighted pair-group method with mean. RAPD typing revealed less clonality among clinical isolates, whereas the ERIC method showed greater similarity in comparison with RAPD. Environmental isolates, however, showed greater similarity using RAPD compared with ERIC typing. With only a few exceptions, most clinical isolates were distinct from environmental isolates, irrespective of the typing method. In conclusion, both the RAPD and ERIC typing methods proved to be good tools in understanding clonal diversity. The results also suggest that there is no relationship between clinical and environmental isolates. The absence of clonality among the clinical isolates may indicate that most P. aeruginosa infection cases could be endemic and not epidemic and that endemic infections may be due to nonclonal strains of P. aeruginosa.
Lactic acid bacteria and their own bacteriocin are great promising in health and industry; hence, too many lactic acid bacteria species and metabolites are involve in health maintenance and treatment of infectious disease as well as food preservation and dairy production. Current study sought to produce, isolate, purify and characterize novel bacteriocin from oral Bifidobacterium adolescentis and studying its effect on antibiotic-resistant pathogenic bacteria. Saliva of 18 volunteers postmenopausal women with their age ranging from 49 to 58 years was included. Bifidobacterium spp. cultivates in anaerobic conditions and identified by API50. Bifidobacterium bacteriocin prepared in De Man, Rogosa and Sharpe broth (MRS) broth as a crude preparation then concentrated by ammonium sulphate precipitation and purified by ion exchange and gel filtration chromatography. The molecular weight of B. adolescentis bacteriocin was estimated by gel filtration chromatography. The effect of temperature and pH value on purified bacteriocin activity was estimated using Escherichia coli American Type Culture Collection (ATCC) and Staphylococcus aureus ATCC. To determine the nature of produced bacteriocin and whether or not the bacteriocin contain lipid or carbohydrate moiety, it was treated with some enzymes (pepsin, lipase, α-amylase, papain and chemotrypsin). Challenged pathogenic bacteria were surveyed for occurrence of antibiotic resistance genes by PCR and to verify the inhibitory effect of B. adolescentis bacteriocin on challenged pathogenic bacteria, disc diffusion method was used. The result of anaerobic culture showed that, 35 Bifidobacterium isolates were obtained, the predominant species was B. adolescentis other species was lesser, namely Bifidobacterium dentium, Bifidobacterium longum and Bifidobacterium urinalis. The purified B. adolescentis bacteriocin was 14 600-Da protein and was active at wide range of pH value (3–8), thermostable and has no lipid or carbohydrate moiety. Almost all of pathogenic bacteria, whether or not they carry antibiotic resistance genes, appeared to be sensitive to crude and purified bacteriocin but purified bacteriocin was more effective as antimicrobial agent. The results suggest that Bifidobacterium spp. was dominant in oral cavities of postmenopausal women who have no caries it may prevent dental caries via antimicrobial activities of their own bacteriocin. Bifidobacterium bacteriocin is protein in nature, without lipid or carbohydrate moiety, heat-stable and active at wide range of pH values and can be classified as type II bacteriocin and so-called bifidoadocin. The antimicrobial activity of bifidoadocin makes it probable food preservative and antagonistic agent against pathogens of oral cavity and causative agents of dental caries as well as other pathogens.
A. baumannii has an ability to produce lectin protein; Lec gene was detected in A. baumannii, and the sequence was recorded under accession number KX766405.1 and KX766406.1.; Lectin was extracted by glass beads and purified by chromatographyic technique; Lectin had strong effect against biofilm formation.
Lactic acid bacteria (LAB) have reported as antifungal, antibacterial, and immunostimulatory agents. Exopolysaccharide (EPS) of lactic acid bacteria effectively could stimulate the production of cytokines by macrophages. This study was aimed to extract, purified, and characterize the EPS from Leuconostoc mesenteroides subsp. Cremoris and to evaluate the immunostimulatory and antibacterial activities of EPS against extended-spectrum beta-lactamase (ESBL) producing Burkholderia cepacia strains. Nine EPSs producing L. mesenteroides subsp. cremoris strains were isolated from local dairy products and the isolated bacteria were identified by using API 50. Eight B. cepacia strains were isolated from different specimens in the hospitals of Medical city, Baghdad. Furthermore, genotypic and phenotypic detection of antibiotic resistance were determined including ESBL genes. EPS of L. mesenteroides subsp. Cremoris was extracted and purified by gel filtration chromatography. EPS physical and chemical analysis were performed to characterize it. Antibacterial and immunomodulatory effect of EPS were studied in vivo using mice and ELISA was used to determine the levels of IL-10 in the mice sera. The extracted EPS was found to have a maximum relative viscosity in water (3.51 dl/g) and maximum specific viscosity (2.93 dl/g), while the intrinsic viscosity recorded 1.41 dl/g. The chemical analysis of the extracted polysaccharide was found to contain the following components, carbohydrates, protein, uronic acids, hexosamines, acetyl groups, ketal linked pyruvate groups, phosphate groups, and sulfate groups, also show the following functional groups under infrared (IR) spectra (hydroxyl, alkanes, carbonyl, carbonyl of carboxylic acid, phosphates, and aliphatic amines). HPLC analysis revealed the presence of mannose as a major component with a calculated molecular weight of 1.71 × 103 g/mol. Genotypic detection of the blaPER-1 gene among ESBL producing B. cepacia strains showed the presence of blaPER-1 gene in three (42.86%) strains. Furthermore, to confirm the biological potential, the EPS was evaluated for its antibacterial activity against multidrug resistance B. cepacia strains in vitro and the result showed that the purified EPS was more effective than crude EPS in all concentrations. The protective activities of L. mesenteroides and EPS were observed when administered 7 days before and after B. cepacia infection, whereas therapeutic activities were monitored by administering EPS for 7 days after B. cepacia induction. This results revealed that the administration of L. mesenteroides and EPS significantly decreased the number of B. cepacia in liver, spleen, and lung (P < 0.05). Furthermore, they enhanced production of IL-10. In conclusion, the L. mesenteroides and its EPSs possess antibacterial and immunostimulator properties and are nontoxic with medicinal importance. Therefore, further studies in human participants should determin the role of L. mesenteroides and its EPS as an immunomodulatory and its relationship in the host protection to pathogens.
Background: Insertion sequence is a short DNA sequence encode for proteins implicated in the transposition activity. Transposase catalyzes the enzymatic reaction allowing the insertion sequence to +9*lo2 move. ;qqa;. Objective: To study the sequencing of transposase gene, tnp, IS1216V of S. aureus isolated from food and then compared with that documented in National Center for Biotechnology Information (NCBI). Methods: Food samples of animal and plant origin were collected, and screened for presence of S. aureus, IS1216V was identified in the Tn1546-like elements in the genomes of all Staphylococcus aureus isolates. Results: About 75% of total food samples were positive to S. aureus especially in the food of animal origin. tnp amplification showed that, 85% of isolates gave positive result. Sequencing of amplified part of IS1216V tnp of S. aureus isolates showed that, tnp gene had high identity (78-79%) with the reference strains of NCBI. Conclusion: High percentage of local food samples were contaminated with S. aureus especially of animal origin. Most of the S. aureus isolates showed the presence of transposase gene (tnp) of IS1216V. Sequencing showed some dissimilarity between the sequence of transposase gene (tnp) of IS1216V S. aureus isolated from local foods and strains recorded in database of NCBI.
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