A steam autoclave was used to sterilize bacteria in clinical solid waste in order to determine an alternative to incineration technology in clinical solid waste management. The influence of contact time (0, 5, 15, 30 and 60 min) and temperature (111 °C, 121 °C and 131 °C) at automated saturated steam pressure was investigated. Results showed that with increasing contact time and temperature, the number of surviving bacteria decreased. The optimum experimental conditions as measured by degree of inactivation of bacteria were 121 °C for 15 minutes (min) for Gram negative bacteria, 121 °C and 131 °C for 60 and 30 min for Gram positive bacteria, respectively. The re-growth of bacteria in sterilized waste was also evaluated in the present study. It was found that bacterial re-growth started two days after the inactivation. The present study recommends that the steam autoclave cannot be considered as an alternative technology to incineration in clinical solid waste management.
Matched MeSH terms: Gram-Negative Bacteria/growth & development
In the present work, the microstructure, corrosion, and bioactivity of graphene oxide (GO) coating on the laser-modified and -unmodified surfaces of TiNb shape memory alloys (SMAs) were investigated. The surface morphology and chemical composition was examined using field emission scanning electron microscopy (FE-SEM) and X-ray diffraction (XRD). The surface modification was carried out via a femtosecond laser with the aim to increase the surface roughness, and thus increase the adhesion property. FE-SEM analysis of the laser-treated Ti-30at.% Nb revealed the increase in surface roughness and oxygen/nitrogen containing groups on the Ti-30at.% Nb surface after being surface modified via a femtosecond laser. Furthermore, the thickness of GO was increased from 35μm to 45μm after the surface was modified. Potentiodynamic polarisation and electrochemical impedance spectroscopy studies revealed that both the GO and laser/GO-coated samples exhibited higher corrosion resistance than that of the uncoated TiNb SMA sample. However, the laser/GO-coated sample presented the highest corrosion resistance in SBF at 37°C. In addition, during soaking in the simulated body fluid (SBF), both the GO and laser/GO coating improved the formation of apatite layer. Based on the bioactivity results, the GO coating exhibited a remarkable antibacterial activity against gram-negative bacteria compared with the uncoated. In conclusion, the present results indicate that Ti-30at.% Nb SMAs may be promising alternatives to NiTi for certain biomedical applications.
Matched MeSH terms: Gram-Negative Bacteria/growth & development
Objective: To evaluate methanolic, ethanolic, acetone and aqueous extracts from different parts of Eurycoma longifolia (E. longifolia) (leave, stem, and root) for antibacterial activity against Gram-positive and Gram-negative bacteria and to utilize the leaves and stem parts rather than the root, which is already used for male sexual enhancement in Malaysia.
Methods: The study took place in the Laboratory of Molecular Biology of Biotechnology Engineering Department, Malaysia between January 2005 and June 2006. Methanolic, ethanolic, acetone and aqueous extracts of leaves, stems and roots of E. longifolia were investigated for their antibacterial properties using Agar-well diffusion method.
Results: The alcoholic and acetone extracts of the leaves and stem extracts were active on both Gram-positive and Gram-negative bacteria except against 2 strains of Gram-negative bacteria (Escherichia coli and Salmonella typhi). The root extracts had no antibacterial activity against Gram-positive and Gram-negative bacteria tested. Aqueous leaves extract showed antibacterial activity against Staphylococcus aureus and Serratia marscesens.
Conclusion: The alcoholic and acetone extracts from leaves and stems of E. longifolia contain potent antibacterial agent(s). This plant can serve as a potential source of antibacterial compounds.
Matched MeSH terms: Gram-Negative Bacteria/growth & development
Cinnamomum species have been widely used in many traditional systems of medicine around the world. In the Malaysian traditional system of medicine, the leaves, stem bark and stem wood of Cinnamomum iners, Cinnamomum porrectum, Cinnamomum altissimum and Cinnamomum impressicostatum have been used to treat wound infections. To study the antibacterial effects of Cinnamomum iners, Cinnamomum porrectum, Cinnamomum altissimum and Cinnamomum impressicostatum against common bacteria found in wound infections with primary focus on methicillin-resistant Staphylococcus aureus (MRSA).
Matched MeSH terms: Gram-Negative Bacteria/growth & development
Sea urchin gonad is considered as a highly prized delicacy in several countries. It is also rich in valuable bioactive compounds including polyunsaturated fatty acids (PUFAs) and β-carotene. This study was undertaken to examine the antimicrobial properties of the ovary extract from sea urchin Diadema setosum against selected Gram-negative and Gram-positive bacteria.
Matched MeSH terms: Gram-Negative Bacteria/growth & development
Natural glycopeptide antibiotics like vancomycin and teicoplanin have played a significant role in countering the threat posed by Gram-positive bacterial infections. The emergence of resistance to glycopeptides among enterococci and staphylococci has prompted the search for second-generation drugs of this class and semi-synthetic derivatives are currently under clinical trials. Antimicrobial resistance among Gram-positive organisms has been increasing steadily during the past several decades and the current development of antibiotics falls short of meeting the needs. Oritavancin (LY-333328 diphosphate), a promising novel second-generation semisynthetic lipoglycopeptide, has a mechanism of action similar to that of other glycopeptides. It has concentration-dependent activity against a variety of Gram-positive organisms specially methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-intermediate resistant Staphylococcus aureus (VISA), Streptococcus pneumoniae and vancomycin-resistant enterococcus. It is rapidly bactericidal against many species and in particular for enterococci where vancomycin and teicoplanin are only bacteriostatic even against susceptible strains. The pharmacokinetic profile of oritavancin has not been fully described; however, oritavancin has a long half-life of about 195.4 hours and is slowly eliminated by renal means. Oritavancin is not metabolized by the liver in animals. Oritavancin will most probably be prescribed as a once-daily dose and it demonstrates concentration-dependent bactericidal activity. Oritavancin has demonstrated preliminary safety and efficacy in Phase I and II clinical trials. In a Phase III clinical trial, oritavancin has achieved the primary efficacy end point in the treatment of complicated Gram-positive skin and skin-structure infections. To date, adverse events have been mild and limited; the most common being administration site complaints, headache, rhinitis, dry skin, pain, increases in liver transaminases and accumulation of free cholesterol and phospholipids in phagocytic (macrophages) and nonphagocytic (fibroblast) cells. Oritavancin appears to be a promising antimicrobial alternative to vancomycin (with additional activity against Staphylococcus and Enterococcus resistant to vancomycin) for the treatment of complicated Gram-positive skin and skin-structure infections. Additional clinical data are required to fully explore its use.
Matched MeSH terms: Gram-Negative Bacteria/growth & development
Both Gram-positive and Gram-negative bacteria can take up exogenous DNA when they are in a competent state either naturally or artificially. However, the thick peptidoglycan layer in Gram-positive bacteria's cell wall is considered as a possible barrier to DNA uptake. In the present work, two transformation techniques have been evaluated in assessing the protocol's ability to introduce foreign DNA, pBBRGFP-45 plasmid which harbors kanamycin resistance and green fluorescent protein (GFP) genes into a Gram-positive bacterium, Bacillus cereus EB2. B. cereus EB2 is an endophytic bacterium, isolated from oil palm roots. A Gram-negative bacterium, Pseudomonas aeruginosa EB35 was used as a control sample for both transformation protocols. The cells were made competent using respective chemical treatment to Gram-positive and Gram-negative bacteria, and kanamycin concentration in the selective medium was also optimized. Preliminary findings using qualitative analysis of colony polymerase chain reaction (PCR)-GFP indicated that the putative positive transformants for B. cereus EB2 were acquired using the second transformation protocol. The positive transformants were then verified using molecular techniques such as observation of putative colonies on specific media under UV light, plasmid extraction, and validation analyses, followed by fluorescence microscopy. Conversely, both transformation protocols were relatively effective for introduction of plasmid DNA into P. aeruginosa EB35. Therefore, this finding demonstrated the potential of chemically prepared competent cells and the crucial step of heat-shock in foreign DNA transformation process of Gram-positive bacterium namely B. cereus was required for successful transformation.
Matched MeSH terms: Gram-Negative Bacteria/growth & development
Enrofloxacin is a fluoroquinolone derivative used for treating urinary tract, respiratory and skin infections in animals. However, low solubility and low bioavailability prevented it from using on humans. Polyvinylpyrrolidone (PVP) is an inert, non toxic polymer with excellent hydrophilic properties, besides it can enhance bioavailability by forming drug polymer conjugates. With the aim of increasing solubility and bioavailability, enrofloxacin thin films were prepared using PVP as a polymer matrix. The obtained oral thin films exhibited excellent uniformity and mechanical properties. Swelling properties of the oral thin films revealed that the water uptake was enhanced by 21%. The surface pH has been found to be 6.8±0.1 indicating that these films will not cause any irritation to oral mucosa. FTIR data of the oral thin films indicated physical interaction between drug and polymer. SEM analysis revealed uniform distribution of drug in polymer matrix. In vitro drug release profiles showed enhanced release profiles (which are also pH dependant) for thin films compared to pure drug. Antibacterial activity was found to be dose dependent and maximum susceptibility was found on Klebsiella pneumonia making this preparation more suitable for respiratory infections.
Matched MeSH terms: Gram-Negative Bacteria/growth & development
The aim of the present research work was synthesis of some 2-furyl[(4-aralkyl)-1-piperazinyl]methanone derivatives and to ascertain their antibacterial potential. The cytotoxicity of these molecules was also checked to find out their utility as possible therapeutic agents. The synthesis was initiated by reacting furyl(-1-piperazinyl)methanone (1) in N,N-dimethylformamide (DMF) and lithium hydride with different aralkyl halides (2a-j) to afford 2-furyl[(4-aralkyl)-1-piperazinyl]methanone derivatives (3a-j). The structural confirmation of all the synthesized compounds was done by IR, EI-MS, 1H-NMR and 13C-NMR spectral techniques and through elemental analysis. The results of in vitro antibacterial activity of all the synthesized compounds were screened against Gram-negative (S. typhi, E. coli, P. aeruginosa) and Gram-positive (B. subtilis, S. aureus) bacteria and were found to be decent inhibitors. Amongst the synthesized molecules, 3e showed lowest minimum inhibitory concentration MIC = 7.52±0.μg/mL against S. Typhi, credibly due to the presence of 2-bromobenzyl group, relative to the reference standard, ciprofloxacin, having MIC = 7.45±0.58μg/mL.
Matched MeSH terms: Gram-Negative Bacteria/growth & development
Antibacterial activity of cell-free supernatant from Escherichia coli E against selected pathogenic bacteria in food and aquaculture was the highest against Edwardsiella tarda 3, a significant aquaculture pathogen. Biochemical properties of the bacteriocins were studied and bacteriocin was found to be sensitive to proteinase K, demonstrating its proteinaceous nature. In addition, pH and temperature affected bacteriocin activity and stability. The bacteriocins were partially purified by ammonium sulfate precipitation. The antibacterial activity was only detected in 20% ammonium sulfate fraction and direct detection of its activity was performed by overlaying on the indicator strains. The inhibition zone associated with the antibacterial activity was detected in the sample overlaid by E. tarda 3 and Staphylococcus aureus DMST8840 with the relative molecular mass of about 27 kDa and 10 kDa, respectively. Bacteriocin showed no cytotoxic effect on NIH-3T3 cell line; however, two virulence genes, aer and sfa, were detected in the genome of E. coli E by PCR. The characteristics of bacteriocins produced by E. coli E exhibited the antibacterial activity against both Gram-positive and Gram-negative pathogenic bacteria and the safe use determined by cytotoxicity test which may have interesting biotechnological applications.
Matched MeSH terms: Gram-Negative Bacteria/growth & development
Brown algae of genus Sargassum are known to produce relatively higher amount of alginic acid. Optimal extraction of this algalcolloid for local consumption requires in-depth studies on post-harvest treatment of the algal fronds. Present investigation endeavors to establish the dynamics and inter-relationship of moisture content and bacteria found on the surface of the alga and alginic acid content during post-harvest desiccation of Sargassum stolonifolium Phang et Yoshida. Harvested fronds were subjected to desiccation for 31 days and bacterial dynamics were monitored with relation to moisture content and water activity index (a(w)). There was 85% decrease in moisture content, however, a(w) showed a more gradual decrease. Total bacterial count increased during the first week and attained maximal value on day 7. Thereafter, a drastic decrease was seen until day 14, followed by a gradual decline. Six species of bacteria were isolated and identified, i.e. Azomonas punctata, Azomonas sp., Escherichia coli, Micrococcus sp., Proteus vulgaris and Vibrio alginolyticus. Calculated ratios for increase in alginic acid content and decrease in moisture content were almost the same throughout the desiccation process, implying that extracellular alginase-producing bacteria did not use the alginic acid produced by the algae as its carbon source. It became apparent that drastic decrease in bacterial count after day 7 could not be attributed to salinity, moisture content, a(w) or lack of carbon source for the bacteria. The possible exposure of these bacteria to algal cell sap which is formed due to the rupture of algal cells was seen as the most likely reason for the drop in bacterial population. Scanning electron microscope (SEM) micrograph taken on day 10 of desiccation showed the presence of cracks and localities where bacteria were exposed to algal cell sap. In vitro antibacterial tests were carried out to verify the effect of algal extracts. Separation and purification of crude algal extracts via bioassay guided separation methodology revealed the identity of active compounds (i.e. gylcolipids and free fatty acids) involved in this inherently available antibacterial defense mechanism during algal desiccation.
Matched MeSH terms: Gram-Negative Bacteria/growth & development
Orthosiphon stamineus is considered an important traditional folk medicine. In this study ethanol and aqueous extracts of O. stamineus were evaluated in vitro for their antioxidant, antimicrobial as well as for their immunomodulatory properties on human peripheral blood mononuclear cells (PBMCs). The DPPH radical scavenging method was used for the determination of antioxidant activity, while the antibacterial efficacy was investigated by both disc diffusion method and Minimum Inhibitory Concentration (MIC) against four bacterial strains (Gram-positive and Gram-negative). Furthermore, the immunomodulatory potential of the extracts was investigated through the MTT assay. Aqueous extract of O. stamineus exhibited significant free radical scavenging activity with IC₅₀ 50 9.6 µg/mL, whereas the IC₅₀ for the ethanol extract was 21.4 µg/mL. The best antimicrobial activity was shown by the aqueous extract of O. stamineus against Staphylococcus aureus, with inhibition zone of 10.5 mm and MIC value 1.56 mg/mL. Moreover, the results observed from the MTT assay showed that both plant extracts stimulated the PBMCs proliferation in vitro in a concentration-dependent manner, but the aqueous extract has remarkable activity against PBMCs. These findings indicate that O. stamineus showed high antioxidant activity and may be considered as an immunomodulatory agent.
Matched MeSH terms: Gram-Negative Bacteria/growth & development
The objective of this study was to evaluate the effects of ultraviolet (UV) radiation (UVB; 90 J/m²) on growth, bioconversion of isoflavones and probiotic properties of parent and subsequent passages of L. casei FTDC 2113. UV radiation significantly enhanced (P < 0.05) the growth of parent cells in mannitol-soymilk fermented at 37°C for 24 h. This had led to an enhanced intracellular and extracellular β-glucosidase activity with a subsequent increase in bioconversion of isoflavones in mannitol-soymilk (P < 0.05). UV radiation also promoted (P < 0.05) the tolerance of parent cells towards acidic condition (pH 2 and 3) and intestinal bile salts (oxgall, taurocholic and cholic acid). In addition, parent treated cells also exhibited better (P < 0.05) adhesion ability to mucin and antimicrobial activity compared to that of the control. All these positive effects of UV radiation were only prevalent in the parent cells without inheritance by first, second and third passage of cells. Although temporary, our results suggested that UV radiation could enhance the bioactive and probiotic potentials of L. casei FTDC 2113, and thus could be applied for the production of probiotic products with enhanced bioactivity.
Matched MeSH terms: Gram-Negative Bacteria/growth & development
Non-healing wounds have placed an enormous stress on both patients and healthcare systems worldwide. Severe complications induced by these wounds can lead to limb amputation or even death and urgently require more effective treatments. Electrospun scaffolds have great potential for improving wound healing treatments by providing controlled drug delivery. Previously, we developed fibrous scaffolds from complex carbohydrate polymers [i.e. chitin-lignin (CL) gels]. However, their application was limited by solubility and undesirable burst drug release. Here, a coaxial electrospinning is applied to encapsulate the CL gels with polycaprolactone (PCL). Presence of a PCL shell layer thus provides longer shelf-life for the CL gels in a wet environment and sustainable drug release. Antibiotics loaded into core-shell fibrous platform effectively inhibit both gram-positive and -negative bacteria without inducting observable cytotoxicity. Therefore, PCL coated CL fibrous gel platforms appear to be good candidates for controlled drug release based wound dressing applications.
Matched MeSH terms: Gram-Negative Bacteria/growth & development
Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) have reached epidemic proportions globally. Therefore, there is an urgent need for a continuous supply of antibiotics to combat the problem. In this study, bacteria initially identified as species belonging to the Bacillus amyloliquefaciens operational group were re-identified based on the housekeeping gene, gyrB. Cell-free supernatants (CFS) from the strains were used for antimicrobial tests using the agar well diffusion assay against MRSA and various types of pathogenic bacteria. The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and physicochemical characteristics of the CFS were determined. Based on gyrB sequence analysis, five strains (PD9, B7, PU1, BP1 and L9) were identified as Bacillus velezensis. The CFS of all B. velezensis strains showed broad inhibitory activities against Gram-negative and -positive as well as MRSA strains. Strain PD9 against MRSA ATCC 33742 was chosen for further analysis as it showed the biggest zone of inhibition (21.0 ± 0.4 mm). The MIC and MBC values obtained were 125 μl/ml. The crude antimicrobial extract showed bactericidal activity and was stable at various temperatures (40-80°C), pH (4-12), surfactants (Tween 20, Tween 80, SDS and Triton X-100) and metal ions (MgCI2, NaCI2, ZnNO3 and CuSO4) when tested. However, the crude extract was not stable when treated with proteinase K. All these properties resembled the characteristics of peptides. The antimicrobial compound from the selected strain was purified by using solvent extraction method and silica gel column chromatography. The purified compound was subjected to High Performance Liquid Chromatography which resulted in a single peak of the anti-MRSA compound being detected. The molecular weight of the anti-MRSA compound was determined by using SDS-PAGE and zymogram. The size of the purified antimicrobial peptide was approximately ~ 5 kDa. The antimicrobial peptide produced from B. velezensis strain PD9 is a promising alternative to combat the spread of MRSA infections in the future.
Matched MeSH terms: Gram-Negative Bacteria/growth & development
The urgent need to treat multi-drug resistant pathogenic microorganisms in chronically infected patients has given rise to the development of new antimicrobials from natural resources. We have tested Elaeis guineensis Jacq (Arecaceae) methanol extract against a variety of bacterial, fungal and yeast strains associated with infections. Our studies have demonstrated that E. guineensis exhibits excellent antimicrobial activity in vitro and in vivo against the bacterial and fungal strains tested. A marked inhibitory effect of the E. guineensis extracts was observed against C. albicans whereby E. guineensis extract at ½, 1, or 2 times the MIC significantly inhibited C. albicans growth with a noticeable drop in optical density (OD) of the bacterial culture. This finding confirmed the anticandidal activity of the extract on C. albicans. Imaging using scanning (SEM) and transmission (TEM) electron microscopy was done to determine the major alterations in the microstructure of the extract-treated C. albicans. The main abnormalities noted via SEM and TEM studies were the alteration in morphology of the yeast cells. In vivo antimicrobial activity was studies in mice that had been inoculated with C. albicans and exhibited good anticandidal activity. The authors conclude that the extract may be used as a candidate for the development of anticandidal agent.
Matched MeSH terms: Gram-Negative Bacteria/growth & development
Defatted Jatropha curcas L. (J. curcas) seed kernels contained a high percentage of crude protein (61.8%) and relatively little acid detergent fiber (4.8%) and neutral detergent fiber (9.7%). Spectrophotometric analysis of the methanolic extract showed the presence of phenolics, flavonoids and saponins with values of 3.9, 0.4 and 19.0 mg/g DM, respectively. High performance liquid chromatography (HPLC) analyses showed the presence of gallic acid and pyrogallol (phenolics), rutin and myricetin (flavonoids) and daidzein (isoflavonoid). The amount of phorbol esters in the methanolic extract estimated by HPLC was 3.0 ± 0.1 mg/g DM. Other metabolites detected by GC-MS include: 2-(hydroxymethyl)-2 nitro-1,3-propanediol, β-sitosterol, 2-furancarboxaldehyde, 5-(hydroxymethy) and acetic acid in the methanolic extract; 2-furancarboxaldehyde, 5-(hydroxymethy), acetic acid and furfural (2-furancarboxaldehyde) in the hot water extract. Methanolic and hot water extracts of kernel meal showed antimicrobial activity against both Gram positive and Gram negative pathogenic bacteria (inhibition range: 0-1.63 cm) at the concentrations of 1 and 1.5 mg/disc. Methanolic extract exhibited antioxidant activities that are higher than hot water extract and comparable to β-carotene. The extracts tended to scavenge the free radicals in the reduction of ferric ion (Fe(3+)) to ferrous ion (Fe(2+)). Cytotoxicity assay results indicated the potential of methanolic extract as a source of anticancer therapeutic agents toward breast cancer cells.
Matched MeSH terms: Gram-Negative Bacteria/growth & development