Functional analysis of the lysis genes of Staphylococcus aureus phage P68 in Escherichia coli

Microbiology ◽  
2005 ◽  
Vol 151 (7) ◽  
pp. 2331-2342 ◽  
Author(s):  
Marian Takáč ◽  
Angela Witte ◽  
Udo Bläsi

Double-stranded DNA phages of both Gram-positive and Gram-negative bacteria typically use a holin–endolysin system to achieve lysis of their host. In this study, the lysis genes of Staphylococcus aureus phage P68 were characterized. P68 gene lys16 was shown to encode a cell-wall-degrading enzyme, which causes cell lysis when externally added to clinical isolates of S. aureus. Another gene, hol15, was identified embedded in the −1 reading frame at the 3′ end of lys16. The deduced Hol15 protein has three putative transmembrane domains, and thus resembles class I holins. An additional candidate holin gene, hol12, was found downstream of the endolysin gene lys16 based on two predicted transmembrane domains of the encoded protein, which is a typical trait of class II holins. The synthesis of either Hol12 or Hol15 resulted in growth retardation of Escherichia coli, and both hol15 and hol12 were able to complement a phage λ Sam mutation. The hol15 gene has a dual start motif beginning with the codons Met1-Lys2-Met3…. Evidence is presented that the hol15 gene encodes a lysis inhibitor (anti-holin) and a lysis effector (actual holin). As depolarization of the membrane converted the anti-holin to a functional holin, these studies suggested that hol15 functions as a typical dual start motif class I holin. The unusual arrangement of the P68 lysis genes is discussed.

Author(s):  
Rubal C Das ◽  
Rajib Banik ◽  
Robiul Hasan Bhuiyan ◽  
Md Golam Kabir

Macrophomina phaseolina is one of the pathogenic organisms of gummosis disease of orange tree (Citrus reticulata). The pathogen was identified from the observation of their colony size, shape, colour, mycelium, conidiophore, conidia, hyaline, spore, and appressoria in the PDA culture. The crude chloroform extracts from the organism showed antibacterial activity against a number of Gram positive and Gram-negative bacteria. The crude chloroform extract also showed promising antifungal activity against three species of the genus Aspergillus. The minimum inhibitory concentration (MIC) of the crude chloroform extract from M. phaseolina against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, and Shigella sonnie were 128 ?gm, 256 ?gm, 128 ?gm and 64 ?gm/ml respectively. The LD50 (lethal dose) values of the cytotoxicity assay over brine shrimp of the crude chloroform extract from M. phaseolina was found to be 51.79 ?gm/ml. DOI: http://dx.doi.org/10.3329/cujbs.v5i1.13378 The Chittagong Univ. J. B. Sci.,Vol. 5(1 &2):125-133, 2010


1988 ◽  
Vol 55 (4) ◽  
pp. 597-602 ◽  
Author(s):  
Lydia Bautista ◽  
Rohan G. Kroll

SummaryEffects of the addition of a proteinase (Neutrase 1–5S) and a peptidase (aminopeptidase DP-102) as agents for accelerating the ripening of Cheddar cheese on the survival of some non-starter bacteria (Staphylococcus aureus, Enterococcus faecalis, Escherichia coliand aSalmonellasp.) were studied throughout a 4-month ripening period. The enzymes were found to have no significant effect on the survival of the Gram-positive bacteria but some significant effects were observed, at some stages of the ripening period, with the Gram-negative bacteria in that lower levels were recovered from cheeses treated with the enzyme system.


2020 ◽  
Vol 73 (4) ◽  
Author(s):  
Courtney K Lawrence ◽  
Chris Sathianathan ◽  
Mauro Verrelli ◽  
Philippe Lagacé-Wiens ◽  
Robert Ariano ◽  
...  

Background: Given the morbidity and mortality associated with bloodstream infections in hemodialysis patients, understanding the microbiology is essential to optimizing treatment in this high-risk population.Objectives: To conduct a retrospective surveillance study of clinical blood isolates from adult hemodialysis patients, and to predict the microbiological coverage of empiric therapies for bloodstream infections in this population.Methods: Clinical blood isolate data were collected from the 4 main outpatient hemodialysis units in Winnipeg, Manitoba, from 2007 to 2014. The distribution of organisms and antimicrobial susceptibilities were characterized. When appropriate, changes over time were tested using time series analysis. Study data were used to predict and compare the microbiological coverage of various empiric therapies for bloodstream infections in hemodialysis patients.Results: The estimated annual number of patients receiving chronic hemodialysis increased steadily over the study period (p < 0.001), whereas the number of blood isolates increased initially, then decreased significantly, from 180 in 2011 to 93 in 2014 (p = 0.04). Gram-positive bacteria represented 72.6% (743/1024) of isolates, including Staphylococcus aureus (36.9%, 378/1024) and coagulase-negative staphylococci (23.1%, 237/1024). Only 26.1% (267/1024) of the isolates were gram-negative bacteria, the majority Enterobacteriaceae. The overall rate of methicillin resistance in S. aureus was 17.5%, and although annual rates were variable, there was a significant increase over time (p = 0.04). Antibiotic resistance in gram-negative bacteria was relatively low, except in Escherichia coli, where 13.5% and 16.2% of isolates were resistant to ceftriaxone and ciprofloxacin, respectively. Empiric therapy with vancomycin plus an agent for gram-negative coverage was predicted to cover 98.8% to 99.7% of blood isolates from hemodialysis patients, whereas cefazolin plus an agent for gram-negative coverage would cover only 67.5% to 68.4%.Conclusions: In an era of increasing antimicrobial resistance, data such as these and ongoing surveillance are essential components of antimicrobial stewardship in the hemodialysis population.Keywords: hemodialysis, microbiology, surveillance, resistance, antimicrobial stewardshipRÉSUMÉ Contexte : Étant donné la morbidité et la mortalité associées aux infections du sang parmi les patients en hémodialyse, la compréhension de la microbiologie est essentielle à l’optimisation du traitement de cette population exposée à un risque élevé.Objectifs : Mener une étude de surveillance rétrospective des isolats de sang cliniques des patients adultes en hémodialyse et prédire la couverture microbiologique des thérapies empiriques contre les infections du sang dans cette population.Méthodes : Les données relatives aux isolats de sang cliniques ont été recueillies dans les quatre unités ambulatoires principales d’hémodialyse à Winnipeg (Manitoba), entre 2007 et 2014. La caractérisation a porté sur la distribution des organismes et les susceptibilités aux antimicrobiens. L’évolution dans le temps a été testée au besoin à l’aide d’une analyse chronologique. Les données de l’étude ont permis de prédire et de comparer la couverture microbiologique de diverses thérapies empiriques contre les infections du sang pour les patients en hémodialyse.Résultats : On estime que le nombre annuel de patients recevant une hémodialyse chronique a augmenté régulièrement au cours de la période de l’étude (p < 0,001); le nombre d’isolats de sang a tout d’abord augmenté, puis il a grandement diminué : de 180 en 2011, il est passé à 93 en 2014 (p = 0,04). Les bactéries à Gram positif représentaient 72,6 % (743/1024) des isolats, y compris les Staphylococcus aureus (36,9 %, 378/1024) et les staphylocoques à coagulase négative (23,1 %, 237/1024). Seulement 26,1 % (267/1024) des isolats étaient des bactéries à Gram négatif, la majorité desquelles étant des Enterobacteriaceae. Le taux général de résistance à la méticilline de S. aureus était de 17,5 %, et bien que les taux annuels étaient variables, une augmentation importante a été observée avec le temps (p = 0,04). La résistance aux antibiotiques des bactéries à Gram négatif était relativement faible, sauf Escherichia coli, où respectivement 13,5 % et 16,2 % des isolats étaient résistants à la ceftriaxone et à la ciprofloxacine. On prévoyait que la thérapie empirique à la vancomycine associée à un agent pour la couverture à Gram positif couvrirait de 98,8 % à 99,7 % des isolats de sang des patients en hémodialyse, tandis que la céfazoline associée à un agent de la couverture à Gram négatif ne couvrirait que 67,5 % à 68,4 %.Conclusions : À une époque qui se caractérise par une augmentation de la résistance aux antimicrobiens, des données comme celles-ci et celles portant sur la surveillance continue sont des composantes essentielles de la bonne gestion de l’utilisation des antimicrobiens pour les patients adultes en hémodialyse.Mots-clés : hémodialyse, microbiologie, surveillance, résistance, gestion de l’utilisation des antimicrobiens


2021 ◽  
Author(s):  
Sanjay Ratan Kumavat ◽  
SATYENDRA MISHRA

Abstract Plants are emerging as a cost-effective and ecofriendly method for green synthesis of nanoparticles. The plant extract Launaea procumbens was used as a reduction agent in the green synthesis of silver nanoparticles. UV-Visible spectroscopy, HR-TEM, SAED, FE-SEM, EDAX, DLS, and FT-IR were used to study the green synthesized silver nanoparticles. UV-Vis spectroscopy of a prepared silver solution revealed maximum absorption at 435 nm. The synthesized silver nanoparticles were found to be spherical in shape with a size in the range of 24.28 to 31.54 nm. DLS analysis was used to determine the size of the green synthesized silver nanoparticles, which showed outstanding antibacterial action against Gram-positive bacteria Bacillus subtilis and Staphylococcus aureus, as well as Gram-negative bacteria Escherichia coli and Pseudomonas aeruginosa. Gram-positive Bacillus subtilis had a maximum zone of inhibition of 20 mm, Staphylococcus aureus had a zone of inhibition of 19 mm, and Gram-negative bacteria Escherichia coli and Pseudomonas aeruginosa had zones of inhibition of 13 mm.


2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Paulina L. Páez ◽  
Claudia M. Bazán ◽  
María E. Bongiovanni ◽  
Judith Toneatto ◽  
Inés Albesa ◽  
...  

The prevalence of antibiotic resistance has resulted in the need for new approaches to be developed to combat previously easily treatable infections. The main aim of this work was to establish the potential of the syntheticα-diimine chromium(III) and ruthenium(II) complexes (where theα-diimine ligands are bpy = 2,2-bipyridine, phen = 1,10-phenanthroline, and dppz = dipyrido[3,2-a:2′,3′-c]-phenazine) like [Cr(phen)3]3+, [Cr(phen)2(dppz)]3+, [Ru(phen)3]2+, and [Ru(bpy)3]2+as antibacterial agents by generating oxidative stress. The [Cr(phen)3]3+and [Cr(phen)2(dppz)]3+complexes showed activity against Gram positive and Gram negative bacteria with minimum inhibitory concentrations (MICs) ranging from 0.125 μg/mL to 1 μg/mL, while [Ru(phen)3]2+and [Ru(bpy)3]2+do not exhibit antimicrobial activity against the two bacterial genera studied at the concentration range used. When ciprofloxacin was combined with [Cr(phen)3]3+for the inhibition ofStaphylococcus aureusandEscherichia coli, an important synergistic effect was observed, FIC 0.066 forS. aureusand FIC 0.064 forE. coli. The work described here shows that chromium(III) complexes are bactericidal forS. aureusandE. coli. Our results indicate thatα-diimine chromium(III) complexes may be interesting to open new paths for metallodrug chemotherapy against different bacterial genera since some of these complexes have been found to exhibit remarkable antibacterial activities.


2012 ◽  
Vol 3 ◽  
pp. 684-691 ◽  
Author(s):  
Mayuree Jaisai ◽  
Sunandan Baruah ◽  
Joydeep Dutta

Paper with antimicrobial properties was developed through in situ growth of ZnO nanorods. The targeted application for this type of paper is in health centers as wallpaper, writing paper, facemasks, tissue paper, etc. The paper was tested on three model microbes, Gram-positive bacteriaStaphylococcus aureus,Gram-negative bacteriaEscherichia coliand common airborne fungusAspergillus niger. No viable bacterial colonies or fungal spores could be detected in the areas surrounding test samples of the antimicrobial paper. Gram-negative bacteriaEscherichia coliwere found to be inhibited in an area that is 239% and 163% the area of the paper sample under different room lighting conditions, i.e., halogen and fluorescent lamp illumination, respectively. For Gram-positive bacteriaStaphylococcus aureusthe zones of inhibition surrounding the paper samples are 102% and 70%, and forAspergillus niger, 224% and 183% of the sample area, under similar lighting conditions.


2011 ◽  
Vol 322 ◽  
pp. 160-163
Author(s):  
Yin Lu ◽  
Hong Chen

A medicinal wild kiwi in China, Actinidia valvata Dunn, has been well known for its activities against leprosy and cancers. The compositions and the antimicrobial activity of its leaf oil were reported for the first time. The oil obtained by hydrodistillation and analyzed by GC and GC-MS, was characterized by the high content of monoterpenes. Linalool (48.14%) is the major component identified, followed by 1,2-dimethyl-lindoline (7.94%), linolenic acid methylester (6.57%) and (E)-phytol (5.29%). The antimicrobial activity of the oil was evaluated against four bacterial and three fungal species. The results showed that it exhibited a mild antibacterial activity against two Gram-positive bacteria (Staphylococcus aureus and Bacillus subtilis), a significant activity against Gram-negative bacteria (Escherichia coli), and no activity on Pseudomonas aeruginosa. The test fungi were more sensitive to the oil, with a MIC range of 0.78~1.56 μL/mL than bacteria in the range which were significantly higher from 0.78 to 25.50 μL/mL.


Author(s):  
I. M. Uneze ◽  
J. O. Otonko ◽  
A. K. Adigun ◽  
S. J. Adebayo

The synthesis and application of nanoparticles is an important area of research that is gaining attention recently. In this recent project, we report the synthesis of silver nanoparticles, AgNP using aqueous solution of silver nitrate and Gnetum africanum leaf extract (reducing agent). The synthesis of AgNP was achieved by mixing aqueous solution of silver nitrate (70ml, 15.75mM) with a solution of Gnetum africanum leaf extract 100 ml) in a reaction flask and allowed to stand for 24 hours in a dark cupboard. A color change from light brown to yellowish brown was observed which indicated that synthesis of silver nanoparticles took place. The presence of AgNP was ascertained using UV-vis spectra analysis and absorption at 442 nm showed the presence of AgNP. The antioxidant assay of both the synthesized AgNP and the leaf extract was determined using DPPH. Antimicrobial activity was conducted using three different organisms which were Staphylococcus aureus, Escherichia coli and Pseudomonas respectively. The antioxidant results using DPPH scavenging ability of AgNp showed that at concentrations of 2mg/ml,1mg/ml and 0.1mg/ml, the percentage inhibition  of  DPPH  by AgNp was 61.69, 53.06 and 38.31 respectively and that of Gnetum africanum leaf extract was 81.32, 78.49, and 58.29 respectively at the same concentrations using Ascorbic acid as a standard. The antimicrobial activity of both the synthesized AgNps and Gnetum Africanum Leaf extract using one gram positive bacteria (Staphylococcus aureus) and two gram negative bacteria (Escherichia coli and Pseudomonas) revealed that the synthesized AgNps showed lesser activity than Gnetumafricanum leaf extract for both the gram positive bacteria (Staphylococcus aureus) and gram negative bacteria (Pseudomonas) and (Escherichia coli). From the above findings, it can be observed that Gnetum Africanum Leaf extract reduced Ag+ to Ag0 and also both the synthesized AgNps and the Gnetum Africanum Leaf extract showed reasonable antioxidant activity against DPPH and antimicrobial activity against the tested microorganisms. This implied that both samples have medicinal values.


2021 ◽  
Author(s):  
Samina Nazir Chaki ◽  
Levina Msuya ◽  
Deborah Mchaile ◽  
Michael Johnson Mahande ◽  
Ronald Mwitalemi Mbwasi ◽  
...  

Abstract Background: Bloodstream infections are major causes of morbidity and mortality among children in Sub-Saharan countries. This study aimed to determine the prevalence of bacteremia, etiological agents’ antibiotic susceptibility pattern and predictors of bacteremia among children with bloodstream Infection.Methodology: This hospital-based cross-section study involved children aged two months to thirteen years. All children meeting the inclusion criteria during the study period were enrolled. All consenting parents were interviewed via a questionnaire to collect data, followed by a thorough physical assessment and venipuncture was done to collect blood samples. Data were analysed using SPSS version 23.Results: Among 242 study participants, 154(63.6%) were male and blood cultures were positive in 37(15.3%). Gram-positive and gram-negative bacteria constituted 32(80%) and 8(20%), respectively. The frequent pathogen found was Staphylococcus aureus 25(62.5%), followed by Enterococcus spp. 4(10%), Escherichia coli 4(10%), Pseudomonas aeruginosa 3(7.5%), Streptococcus pyogenes 3(7.5%) and 1(2.5%) Klebsiella pneumonia. The majority of bacterial isolates showed high resistance to commonly used antibiotics in the study area. Predictors of bacteremia were severe malnutrition, hydrocephalus, hyperglycemia, lethargy and BSI with no foci of infection. Conclusion: Prevalence of bacteremia was 15.3%. Gram-positive bacteria were more prevalent than gram-negative bacteria. Staphylococcus aureus and Escherichia coli were the prevalent isolates causing BSI. Effective antibiotics for both gram-negative & gram-positive organisms are imipenem, meropenem and piperacillin-tazobactam followed by amikacin; vancomycin & clindamycin for gram-positive organisms. To curb the growing antimicrobial resistance that we see in this and other studies, continuous antimicrobial stewardship is necessary, else we risk failing to treat BSI.


Author(s):  
Adnal K. P. Husein Putra ◽  
Sri Sundari

Background: This study provides an overview of diabetic ulcer infection, antibiotic susceptibility patterns, the most common types of antibiotics, factors influence in antibiotics administrations, and verifying the guideline used of antibiotics.Methods: The research used descriptive analysis combined with interviews. The study was starting from February-March 2021. Eighty-six diabetic ulcer patients were selected considering the inclusion and exclusion criteria. We interviewed seven doctors.Results: 71 bacterias were found with a gram-negative bacteria count of 80.3%- higher than gram-positive bacteria (19.7%). The most common gram-negative bacteria found are Escherichia coli (12.7%) and show the most heightened sensitivity with imipenem (80.7%), while cefuroxime and trimethoprim-sulfamethoxazole show the highest resistance (64.9%). The most common gram-positive bacteria found are Staphylococcus aureus (14.1%), and vancomycin shows the most heightened sensitivity (100%), while penicillin shows the highest resistance (71.4%). The most common single antibiotic administration was ceftriaxone, as well as a combination of two antibiotics, namely ceftriaxone and metronidazole.Conclusions: Escherichia coli was the most gram-negative bacteria, which has the highest sensitivity with imipenem, while Staphylococcus aureus was the most gram-positive bacteria, which has the most heightened sensitivity with vancomycin. The most common single antibiotic administration was ceftriaxone, as well as a combination of two antibiotics, ceftriaxone and metronidazole. Factors that influence antibiotics administration are patient condition, susceptibility testing, training, and advice from colleagues. Every doctor uses different guidelines for diabetic foot ulcer.


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