Antibacterial effect of five Zingiberaceae essential oils

Essential oil obtained by hydrodistillation and two different solvent extractions (petroleum ether and ethanol) from five Zingiberaceae species: ginger (Zingiber officinale Roscoe.), galanga (Alpinia galanga Sw.), turmeric (Curcuma longa L.), kaempferia (Boesenbergia pandurata Holtt.) and bastard cardamom (Amomum xanthioides Wall.) was characterized. Volatile components of all extracts were analyzed by gas chromatography-mass spectrometry (GC-MS). The major components of ginger, turmeric, galangal, bastard cardamom and kaempferia were zingiberene, turmerone, methyl chavicol, and gamma-terpinene, respectively. Their antibacterial effects towards Escherichia coli, Staphylococcus aureus, Bacillus cereus and Listeria monocytogenes were tested by a disc diffusion assay. Essential oil of kaempferia and bastard cardamom obtained by hydrodistillation extraction could inhibit growth of all tested bacteria. Essential oil of ginger extracted by hydrodistillation had the highest efficiency against three positive strains of bacteria (S. aureus, B. cereus and L. monocytogenes), with a minimum concentration to inhibit B. cereus and L. monocytogenes of 6.25 mg/mL.     

Polysaccharide differences between planktonic and biofilm-associated EPS from Pseudomonas fluorescens B52

The polysaccharides associated with free (planktonic) and surface-attached (biofilm) cells from cultures of Pseudomonas fluorescens strain B52 were compared. Variations in the attached matrix due to surface material (glass or stainless steel) were also analyzed. Two digestion methods were used to optimize the recoveries of sugars, uronic acids and acidic substituents. The yield of analyzable material after digestion reached 90% for the material associated to the biofilms, though only 20–30% for that bound to planktonic cells. The polysaccharide(s) in the biofilm had glucuronic and guluronic acids as main components, besides rhamnose, glucose and glucosamine. The proportion of glucuronic to guluronic acid was higher in the polysaccharide(s) found in biofilms formed on stainless steel than in those on glass.     

Architecture and high-resolution structure of Bacillus thuringiensis and Bacillus cereus spore coat surfaces

We have utilized atomic force microscopy (AFM) to visualize the native surface topography and ultrastructure of Bacillus thuringiensis and Bacillus cereus spores in water and in air. AFM was able to resolve the nanostructure of the exosporium and three distinctive classes of appendages. Removal of the exosporium exposed either a hexagonal honeycomb layer (B. thuringiensis) or a rodlet outer spore coat layer (B. cereus). Removal of the rodlet structure from B. cereus spores revealed an underlying honeycomb layer similar to that observed with B. thuringiensis spores. The periodicity of the rodlet structure on the outer spore coat of B. cereus was approximately 8 nm, and the length of the rodlets was limited to the cross-patched domain structure of this layer to approximately 200 nm. The lattice constant of the honeycomb structures was approximately 9 nm for both B. cereus and B. thuringiensis spores. Both honeycomb structures were composed of multiple, disoriented domains with distinct boundaries. Our results demonstrate that variations in storage and preparation procedures result in architectural changes in individual spore surfaces, which establish AFM as a useful tool for evaluation of preparation and processing "fingerprints" of bacterial spores. These results establish that high-resolution AFM has the capacity to reveal species-specific assembly and nanometer scale structure of spore surfaces. These species-specific spore surface structural variations are correlated with sequence divergences in a spore core structural protein SspE.     

Polysaccharide differences between planktonic and biofilm-associated EPS from Pseudomonas fluorescens B52

The polysaccharides associated with free (planktonic) and surface-attached (biofilm) cells from cultures of Pseudomonas fluorescens strain B52 were compared. Variations in the attached matrix due to surface material (glass or stainless steel) were also analyzed. Two digestion methods were used to optimize the recoveries of sugars, uronic acids and acidic substituents. The yield of analyzable material after digestion reached 90% for the material associated to the biofilms, though only 20–30% for that bound to planktonic cells. The polysaccharide(s) in the biofilm had glucuronic and guluronic acids as main components, besides rhamnose, glucose and glucosamine. The proportion of glucuronic to guluronic acid was higher in the polysaccharide(s) found in biofilms formed on stainless steel than in those on glass.     

EFFECT OF DIPICOLINIC ACID ON THE ULTRAVIOLET RADIATION RESISTANCE OF BACILLUS CEREUS SPORES

Abstract— The ultraviolet radiation (UV) resistance of B. cereus spores was shown to depend on their content of dipicolinic acid (DPA). Wild-type spores with decreasing amounts of DPA exhibited increased UV resistance. Similarly, spores devoid of DPA (DPA-minus), produced by a mutant strain of B. cereus unable to synthesize DPA, were more resistant to UV than mutant spores (DPA-plus) produced in the presence of exogenously supplied DPA. Resistance of both the wild type and mutant strains to ionizing radiation, however, was unaffected by DPA content. Comparison of the resistance of DPA-minus and DPA-plus mutant spores to UV of various wavelengths showed that the greater sensitivity of the latter DPA-plus spores appeared at wavelengths corresponding to the region of the first molecular absorption band of the calcium chelate of DPA. In the wild type and mutant, thymine photoproducts were produced at a greater rate and to a greater extent in spores with high levels of DPA than in spores with low DPA.
The data indicate that DPA transfers energy to DN A in vivo , which leads to the conclusion that DPA occurs in the spore protoplast.     

Fingerprinting species and strains of Bacilli spores by distinctive coat surface morphology

In this work, we applied high-resolution atomic force microscopy (AFM) to identify and characterize similarities and differences in the spore surface morphology of strains from four species of Bacilli: B. anthracis, B. cereus, B. pumilis, and B. subtilis. Common features of the examined spores in the dry state included ridges that spanned the long axis of each spore, and nanometer-scale fine rodlets that covered the entire spore surface. However, important differences in these features between species permitted them to be distinguished by AFM. Specifically, each species possessed significant variation in ridge architecture, and the rodlet width in B. anthracis was significantly less than that of the other species. To characterize similarities and differences within a species, we examined three B. subtilis strains. The ridge patterns among the three strains were largely the same; however, we detected significant differences in the ridge dimensions. Taken together, these experiments provide important information about natural variation in spore surface morphology, define structural features that can serve as species- and strain-specific signatures, and give insight into the dynamics of spore coat flexibility and its role during spore dormancy and germination.     

Detection of specific Bacillus anthracis spore biomarkers by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

Analysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) was applied for the characterization of Bacillus anthracis spore biomarkers. B. anthracis spores were extracted under a simple procedure, followed by linear mode analysis, using sinapinic acid as the matrix. Several markers with a mass range of 4-7 kDa were detected in three B. anthracis strains: Vollum, Sterne and V770-NP1-R. Similar spectra were also obtained for spore extracts of two members of the B. cereus group: B. thuringiensis and B. cereus, but not for B. mycoides, B. subtilis or B. licheniformis, suggesting that these markers are specific to closely related members of the B. cereus group. When alpha-cyano-4-hydroxycinnamic acid was used as the matrix, at least four additional new markers within a mass range of 2-4 kDa could be detected in all B. anthracis spore extracts. These markers, corresponding to a molecular weight of 2528.3, 2792.4, 3077.4, and 3590.7 Da, have not been observed in extracts of the three closely related Bacillus species - B. cereus, B. thuringiensis and B. mycoides. These unique B. anthracis biomarkers, which were isotopically resolved and reproducibly detected in the highly accurate MALDI-TOFMS reflectron mode, may be useful as a basis for rapid and specific identification of B. anthracis strains.     

Synthesis and antimicrobial activity of some new 1,3,4-thiadiazole and 1,2,4-triazole compounds having a D,L-methionine moiety

New 1,3,4-thiadiazole, 5a-e, and 1,2,4-triazolecompounds 6a-c, containing a D,L-methionine moiety were synthesized by intramolecular cyclization of 1,4-disubstituted thiosemicarbazides 4a-e in acid and alkaline media, respectively. The potential antimicrobial effects of the synthesized compounds were investigated using the Staphylococcus aureus ATCC 25923, Bacillus antracis ATCC 8705, Bacillus cereus ATCC 10987, Sarcina lutea ATCC 9341 and Escherichia coli ATCC 25922 strains. The newly synthesized compounds exhibited promising activities against Bacillus antracis and Bacillus cereus.     

3,4-二羟基噻吩交联壳聚糖的合成及其对白桃的抑菌保鲜性能

以氯乙酸乙酯、硫化钠、草酸二乙酯和壳聚糖为原料合成了3,4-二羟基噻吩交联壳聚糖(DTCC),测定了该交联聚合物对一系列菌种的抑菌能力;测定了以该交联聚合物为涂膜剂处理白桃后的失重率、腐烂率、叶绿素含量、膜透率和总糖含量随贮存时间的变化关系。 结果表明,部分交联的DTCC对革兰氏阳性、阴性菌有较强的抑菌效能,作用20 min后对铜绿假单胞菌、荧光假单胞菌、蜡样芽孢杆菌、枯草芽孢杆菌的抑菌率分别可达96.7％、95.8％、97.9％和98.2％,最小抑菌浓度分别为25.0、25.0、3.1和3.1 g/L;DTCC对黑曲霉菌和柑橘青霉菌亦有较强的抑菌效果,作用20 min后的抑菌率分别达到99.0％和28.4％,最小抑菌浓度分别为25.0和12.5 g/L。 以质量浓度为1.0 g/L的DTCC涂液处理白桃并在(27±0.5) ℃贮存12 d后,失重率仅有8.1％,腐烂率为40％,膜透率为34.1％,总糖含量可达75.1 mg/g。     

In vitro production of Clostridium difficile spores for use in the efficacy evaluation of disinfectants: a precollaborative investigation

Clostridium difficile is a strict anaerobic spore-forming bacterium, and an increasingly common nosocomial pathogen. The U.S. Environmental Protection Agency (EPA) is responsible for the registration of disinfectants, including products designed to treat environmental surfaces contaminated with spores of C. difficile. Product efficacy data are required for registration; however, there is a lack of methodology for generating high-quality spore suspensions for evaluating product performance. As such, a study was carried out to select a suitable C. difficile strain and to develop a stand-alone method to prepare a spore suspension that meets specific criteria necessary for quantitative testing of disinfectants. The criteria are: (1) a spore titer of > 8 log10/mL, (2) > or = 90% spores to vegetative cells, and (3) resistance of spores (determined by viability) to 2.5 M hydrochloric acid (HCl). Several strains of C. difficile (toxigenic and nontoxigenic) were grown on various media (solid and liquid) for varying lengths of time to determine the best combination of incubation conditions and media to optimize spore production and quality. Once the spore production procedure was optimized, a toxigenic strain of C. difficile [American Type Culture Collection (ATCC) 43598] was selected for use in trials to verify repeatability from one production run to the next. The spore suspension was initiated by spreading vegetative cells of C. difficile (ATCC 43598) on CDC anaerobic 5% sheep blood agar plates and incubating for 7-10 days at 36 +/- 1 degrees C under anaerobic conditions. Spores were harvested when > or = 90% of the cells converted to spores as determined by observation using phase-contrast microscopy. The spores were washed three times with saline-Tween-80, resuspended in cold deionized water, heated to 70 degrees C for 10 min, evaluated microscopically for quality, and enumerated on cycloserine-cefoxitin-fructose agar containing horse blood and taurocholate. The spore suspension was used to inoculate brushed stainless steel carriers (1 cm in diameter) with and without a soil load in accordance with the Standard Quantitative Carrier Disk Test Method (ASTM E-2197-02) to determine carrier load. Once it was determined that > 6 log10 spores/carrier could be recovered, spores were evaluated for resistance to HCI. The sporulation method presented in this report is simple and repeatable and results in spore suspension of high titer (> 8 log10/mL) and quality (> or = 90% spores to vegetative cells) that met acid resistance criteria (spores were resistant to 2.5 M HCI for 10 min). In addition, recovery from brushed stainless steel carriers with and without soil load was > 6 log10 spores/carrier. A 6 log10 performance standard was set forth in the EPA's interim guidance for generating data to support a label claim for effectiveness against C. difficile spores on hard, nonporous surfaces. This precollaborative investigation successfully demonstrated the use of a methodology for in vitro production of C. difficile spores (ATCC 43598) necessary for conducting efficacy tests. A proposal will be submitted to the AOAC INTERNATIONAL Methods Committee on Antimicrobial Efficacy Testing for a collaborative study; see Appendix.     

SYNTHESIS AND ANTIBACTERIAL ACTIVITY OF SOME METAL COMPLEXES OF BETA-LACTAMIC ANTIBIOTICS

Abstract

An overview of the results of the interaction of β-lactamic antibiotics with some transition metal ions is given. Several complexes have been synthesized and characterized by physicochemical and spectroscopic methods. Some exhibit very promising antibacterial activity. Clavulanic acid (L₁), penicillin (L₂), ampicillin (L₃), cephalexin (L₄), cefazolin (L₅) and cephalothin (L₆) were allowed to react with metal ions in methanol under nitrogen. IR spectra of clavulanic acid, penicillin and ampicillin complexes showed strong modifications of the carbonyl group located on the lactamic ring, indicating that this oxygen participates in the coordination to the metal ions along with the carboxylate group. Thus L₁ and L₂ behave as monoanionic bidentate ligands while L₃ behaves as a monoanionic tridentate ligand. The IR spectra of cephalexin, cefazolin and cephalotin chelates show that the beta lactamic carbonyl group does not participate in coordination to the metal ions. A relationship between the structure of the complexes and their antibacterial activity can be observed.

In vitro antibacterial activity of the antibiotics and the complexes were tested using the filter paper diffusion method and the chosen strains include Escherichia coli ATCC 10536, Pseudomonas aeruginosa ATCC 9027, Salmonella typhimorium ATCC 14028, Staphylococcus aureus ATCC 6538, Bacillus cereus ATCC 9634, Proteus mirabilis 35659, Proteus vulgaris ATCC 9920, Klebsiella pneumoniae ATCC 10031, Salmonella sp, Shigella sp ATCC 11126, Streptococcus viridans and Salmonella enteritidis ATCC 497.     

Biosurfactant production by Bacillus subtilis using cassava-processing effluent

A cassava flour-processing effluent (manipueira) was evaluated as a substrate for surfactant production by two Bacillus subtilis strains. B. subtilis ATCC 21332 reduced the surface tension of the medium to 25.9 mN/m, producing a crude biosurfactant concentration of 2.2 g/L. The wild-type strain, B. subtilis LB5a, reduced the surface tension of the medium to 26.6 mN/m, giving a crude biosurfactant concentration of 3.0 g/L. A decrease in surfactant concentration observed for B. subtilis ATCC 21332 seemed to be related to an increase in protease activity. The biosurfactant produced on cassava effluent medium by B. subtilis LB5a was similar to surfactin.     

Antimicrobial Activity of MgB2 Powders Produced via Reactive Liquid Infiltration Method

We report for the first time on the antimicrobial activity of MgB₂ powders produced via the Reactive Liquid Infiltration (RLI) process. Samples with MgB₂ wt.% ranging from 2% to 99% were obtained and characterized, observing different levels of grain aggregation and of impurity phases. Their antimicrobial activity was tested against Staphylococcus aureus ATCC BAA 1026, Enterococcus faecalis ATCC 29212, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, and Candida albicans ATCC 10231. A general correlation is observed between the antibacterial activity and the MgB₂ wt.%, but the sample microstructure also appears to be very important. RLI-MgB₂ powders show better performances compared to commercial powders against microbial strains in the planktonic form, and their activity against biofilms is also very similar.     

Antimicrobial activity of spices essential oils and its effectiveness on mature biofilms of human pathogens

Abstract

The antibacterial activity of Pimpinella anisum L., Cinnamomum zeylanicum, Syzygium aromaticum, and Cuminum cyminum L. essential oils (EOs) against some common pathogenic microorganisms (Staphylococcus aureus ATCC 6538, Staphylococcus epidermidis ATCC 14990, Enterococcus faecalis ATCC 29212, Streptococcus pyogenes ATCC 1915, Escherichia coli ATCC 8739, Pseudomonas aeruginosa ATCC 27853, Aeromonas hydrophila ATCC 7966, Proteus mirabilis ATCC 10005, Klebsiella pneumoniae ATCC 13883, and Candida albicans ATCC 10231) and their biofilms was studied. The EOs inhibitory effects were evaluated by both Agar Well Diffusion assay and Minimum Inhibitory Concentration (MIC) determination. The most active EOs, cinnamon and cloves, were also tested on 18, 24, 48, 72?hours mature biofilms. Cinnamon and cloves exhibited the best results showing a significant activity against all the tested bacteria. Concerning biofilm, results suggest that Cinnamomum zeylanicum oil may be a useful approach to impair the biofilm produced by the tested Gram-negative bacteria.      

Continuous nondestructive monitoring of <Emphasis Type="Italic">Bordetella pertussis</Emphasis> biofilms by Fourier transform infrared spectroscopy and other corroborative techniques

This work describes the application of several analytical techniques to characterize the development of Bordetella pertussis biofilms and to examine, in particular, the contribution of virulence factors in this development. Growth of surface-attached virulent and avirulent B. pertussis strains was monitored in continuous-flow chambers by techniques such as the crystal violet method, and nondestructive methodologies like fluorescence microscopy and Fourier transform (FT) IR spectroscopy. Additionally, B. pertussis virulent and avirulent strains expressing green fluorescent protein were grown adhered to the base of a glass chamber of 1-μm thickness. Three-dimensional images of mature biofilms, acquired by confocal laser scanning microscopy, were quantitatively analysed by means of the computer program COMSTAT. Our results indicate that only the virulent (Bvg⁺) phase of B. pertussis is able to attach to surfaces and develop a mature biofilm. In the virulent phase these bacteria are capable of producing a biofilm consisting of microcolonies of approximately 200 μm in diameter and 24 μm in depth. FTIR spectroscopy allowed us not only to follow the dynamics of biofilm growth through specific biomass and biofilm marker absorption bands, but also to monitor the maturation of the biofilm by means of the increase of the carbohydrate-to-protein ratio.     

Antibacterial activity of three medicinal Thai plants against Campylobacter jejuni and other foodborne pathogens

Leaves of Adenanthera pavonina, Moringa oleifera and Annona squamosa are used in traditional Thai medicine to treat dysentery and other diseases. This study investigated the antibacterial activity of these plants against six species of foodborne pathogen. Methods and solvents employed to extract active constituents were optimised using the disc diffusion assay. Phytochemical analysis of the optimised extracts was performed by thin layer chromatography (TLC). Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) were determined by broth microdilution. A. pavonina contained flavonoids, terpines and tannins, and was the most active extract against Campylobacter jejuni, inhibiting growth at 62.5-125 μg mL(-1). The A. squamosa extract contained flavonoids, terpines, tannins and alkaloids, and had the broadest spectrum of antibacterial activity, inhibiting Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus and C. jejuni between 62.5 and 500 μg mL(-1). MBCs were 2- to 4-fold higher than MICs against C. jejuni and B. cereus, suggesting the extracts are bactericidal against these species. Negligible activity was detected from M. oleifera. The data presented here show that A. pavonina and A. squamosa could potentially be used in modern applications aimed at the treatment or prevention of foodborne diseases.     

Antibacterial characteristics and activity of water-soluble chitosan derivatives prepared by the Maillard reaction

The antibacterial activity of water-soluble chitosan derivatives prepared by Maillard reactions against Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, Escherichia coli, Shigella dysenteriae, and Salmonella typhimurium was examined. Relatively high antibacterial activity against various microorganisms was noted for the chitosan-glucosamine derivative as compared to the acid-soluble chitosan. In addition, it was found that the susceptibility of the test organisms to the water-soluble chitosan derivative was higher in deionized water than in saline solution. Metal ions were also found to reduce the antibacterial activity of the water-soluble chitosan derivative on S. aureus. The marked increase in glucose level, protein content and lactate dehydrogenase (LDH) activity was observed in the cell supernatant of S. aureus exposed to the water-soluble chitosan derivative in deionized water. The results suggest that the water-soluble chitosan produced by Maillard reaction may be a promising commercial substitute for acid-soluble chitosan.     

Efficient isolation and identification of Bacillus cereus group

Bacillus cereus is a group of ubiquitous facultative anaerobic sporeforming Gram-positive rods commonly found in soil. The spores frequently contaminate a variety of foods, including produce, meat, eggs, and dairy products. Foodborne illnesses associated with toxins produced by B. cereus can result in self-limiting diarrhea or vomiting. Plate enumeration methods recommended by recognized food authorities to detect the presence of B. cereus in potentially contaminated food products do not inhibit other Gram-positive competitive bacteria. This study evaluated the use of Bacara, a new chromogenic agar, as an efficient method to identify and enumerate B. cereus group from food matrixes, even in the presence of background flora. Inclusivity and exclusivity testing was performed using four different selective and differential media for B. cereus, including Mannitol Egg Yolk Polymyxin (MYP), Polymyxin Pyruvate Egg-Yolk Mannitol Bromothymol Blue Agar, Bacillus Chromogenic Media, Brilliance, and Bacara. MYP and Bacara were also used in plate enumeration studies to isolate B. cereus from artificially contaminated foods.     

Antimicrobial activity and pollen composition of honey samples collected from different provinces in Turkey

The antibacterial activity of honey samples from different sources were collected and investigated against Bacillus cereus, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, Klebsiella pneumoniae ATCC 27736, Morganella morganii, Micrococcus luteus NRRL B-4375, Escherichia coli ATCC 35218, and Candida albicans. Pathogens exhibited different sensitivities towards the honey samples. The results showed that majority of the honey samples (75%) generally inhibitied the bacteria tested. The honey samples which were obtained from Izmir (samples 1 and 2) proved more effective as inhibitors against P. aeruginosa, E. coli, and S. aureus. The honey which was obtained from Mu?la (sample 5) exhibited high anticandidal activity on C. albicans. A comparison of the honey samples on the basis of pollen content revealed that they were heterofloral, and samples which had highest antibacterial activity against P. aeruginosa, E. coli, and S. aureus were dominated by pollen from Chenopodiaceae/Amaranthaceae (sample 1), and Trifolium, Trigonella, Cyperaceae, Zea mays and Anthemis taxa (sample 2). The honey proved more effective on bacteria than antibiotics.     

TRANSITORY UV RESISTANCE DURING GERMINATION OF UV-SENSITIVE SPORES PRODUCED BY A MUTANT OF Bacillus cereus 569

Abstract— A mutant of Bacillus cereus 569, isolated by us and designated 2422 is unable to excise cyclobutane-type dimers and spore-specific photoproducts from the DNA of UV-irradiated vegetative cells and dormant spores. The deficiency in the excision repair mechanism was found to be at the post-incision step in the exonuclease-mediated removal of the photoproducts. During germination, the mutant B. cereus 2422 exhibits UV-resistance and an efficient photoproduct removal which is followed by DNA repair synthesis. The data presented indicate the existence of germinative excision repair in B. cereus 569.