In vitro method to assess the antimicrobial activity and potential efficacy of novel types of wound dressings

AIMS: To develop a simple, reproducible in vitro static diffusion method using cellulose disks and defined species to test antimicrobial efficacy of wound dressings. METHODS AND RESULTS: Cellulose disks were inoculated by immersion in cell suspensions of target species Staphylococcus epidermidis, Candida albicans and Fusobacterium nucleatum. Test and control wound dressings were cut into equal sized squares (25 x 25 mm) and applied to the surface of 10-mm thick tryptone yeast extract agar on test beds. Following a 2-h equilibration period, inoculated cellulose disks were inserted (one per dressing) at the interface between dressing and agar surface and a small weight applied over each square. At various sampling times, disks were removed and surviving cells enumerated by viable counts. Disk to disk variation for microbial loading was assessed using S. epidermidis for both initial (n = 16) and standard treatment (n = 16) conditions. The coefficient of variation was low (<5%) indicating good reproducibility for cell loading and treatment position on the test bed. Replicate assays (n = 6) using S. epidermidis and oxyzyme gels produced similar kill rates with low scatter (R2 > 0.9) indicating good reproducibility between assays. Significant differences (P < 0.05) in kill rates were observed for different target species, types of dressing and test bed conditions (+/-blood and nutrients). CONCLUSIONS: The method is reproducible and useful in tracking the death kinetics of test species, enabling the comparison of different types of dressing. SIGNIFICANCE AND IMPACT OF THE STUDY: The reported method has significant advantages over established test procedures; it can be applied equally across a wide range of target species (including anaerobes and yeasts), a wide range of conditions, and different types of surface dressings, including those relying upon oxygen diffusion.     

一株代谢煤油兼性厌氧菌的筛选鉴定和特性

从大港油田的油层水样中分离得到一株能以煤油为碳代谢生成表面活性剂、产酸和产气的菌株DG7,该菌为革兰氏阴性杆菌,兼性厌氧,运动,能在NaCl浓度为18．5％的培养基中生长,经鉴定,可能为气单杆菌属（Aeromonas sp.）中的一个新种,但学需进一步确定。     

The presence of black-pigmented Gram-negative anaerobes in the oral cavity of edentolous subjects

Abstract The aim of this study was to investigate whether black-pigmented Gram-negative anaerobes are part of the indigenous oral flora of edentulous subjects with or without dentures. Group I consisted of 11 subjects with dentures (mean age 63.6 years, range 52–75) and Group II consisted of 39 subjects with complete dentures in both jaws (mean age 59.3 years, range 37–80). Two microbial samples for microbiological examination were taken from each subject. One of them was from the dorsum of the tongue and the other was from saliva. Black-pigmented Gram-negative anaerobes > 10⁷ cfu/ml were found in both samples. In 50 edentulous subjects, they were found more commonly from tongue (40%) than from saliva (26%). None of the subjects had Porphyromonas gingivalis . There was no statistically significant difference ( P > 0.05) between the presence of black-pigmented Gram-negative anaerobes on tongue and in saliva in the two groups. Our results suggested that high levels of black-pigmented Gram-negative anaerobes may belong to the indigenous oral flora in edentulous mouths with or without dentures.     

Isolation and physiological characterisation of Thiobacillus aquaesulis sp. nov., a novel facultatively autotrophic moderate thermophile

A moderately thermophilic, facultatively chemolithoautotrophic thiobacillus isolated from a thermal sulphur spring is described. It differs from all other species currently known to be in culture. It grows lithoautotrophically on thiosulphate, trithionate or tetrathionate, which are oxidized to sulphate. Batch cultures on thiosulphate do not produce tetrathionate, but do precipitate elemental sulphur during growth. In autotrophic chemostat cultures the organism produces yields on thiosulphate, trithionate and tetrathionate that are among the highest observed for a Thiobacillus. Autotrophic cultures contain ribulose bisphosphate carboxylase. Heterotrophic growth has been observed only on complex media such as yeast extract and nutrient broth. It is capable of autotrophic growth and denitrification under anaerobic conditions with thiosulphate and nitrate. It grows between 30 to 55° C, and pH 7 to 9, with best growth at about 43°C and pH 7.6. It contains ubiquinone Q-8, and its DNA contains 65.7 mol% G+C. The organism is formally described and named as Thiobacillus aquaesulis.Now the Department of Biological Sciences     

Temperature range variants of Bacillus megaterium

Facultatively and obligately thermophilic variants were isolated from 3 out of 12 tested mesophilic Bacillus megaterium strains. The variants occurred at a frequency of 10^-8–10^-9. The ability to grow at elevated temperatures was cured by means of treatment with acridine orange. Stable revertants were isolated from facultatively and obligately thermophilic variants. An unknown type of megacin was produced by the facultative thermophiles. This megacin attacked mesophilic and obligately thermophilic strains. The thermophiles displayed a few divergent taxonomic characteristics but a close relationship between the strains was indicated by the megacin spectrum and sensitivity to phage. Arrhenius plots revealed that the strains could be considered as temperature range variants and that the temperature characteristic increased with growth at a higher temperature range. The case for a plasmid involvement in the phenomenon is discussed.Abbreviations M Mesophilic - Fp facultatively psychrophilic - Ft facultatively thermophilic - Ot obligately thermophilic     

Vesicular arbuscular mycorrhizae in floating wetland mat communities dominated by Typha

Low levels of vesicular arbuscular mycorrhizae were present in floating mats dominated by clones of Typha angustifolia L., T. x glauca Godr., and T. latifolia L. Floating mats composed of rhizomes (underwater-ground stems with high starch accumulation), roots, decaying organic matter, and wind deposited soil, easily supported human activities. The majority of roots isolated from the root cores were connected to Typha rhizomes. Tests employing the gridline intersect method, intensity, spore counts and most probable number (MPN) were used to define the level of colonization. Mycorrhizal colonization from the T. angustifolia and T. x glauca clones averaged 4 to 5%, while the T. latifolia clone averaged 13%. When colonization was encountered, intensities varied from a high of 3.0 to a low of 0.4 on a sclae of 0 to 4. Although arbuscules were not found, abundant hyphae, vesicles and spores indicated that presumed facultative associations occurred between the vesicular arbuscular fungi and the indicated that presumed facultative associations plant communities found on floating mats. The mycorrhizal fungi identified from these communities in cluded Glomus albidum Walker & Rhodes, G. caledonium (Nicol. & Gerd.) Trappe & Gerdemann, G. etunicatum Becker & Gerdemann, and G. microcarpum Tul. & Tul. Spore counts ranged from 16 to 76 spores per gram dried organic soil. The recolonization ability of VAM propagules by way of a most probable number bioassay with maize yielded numbers that ranged from zero to 96 propagules per gram soil, with G. etunicatum the only species recovered.     

Rewilding traditional grazing areas affects scavenger assemblages and carcass consumption patterns

The abandonment of traditional livestock farming systems in Mediterranean countries is triggering a large-scale habitat transformation, which, in general, consists of the replacement of open grazing areas by woodlands through non-managed regeneration. As a consequence, wild ungulates are occupying rapidly the empty niche left by domestic ungulates. Both types of ungulates represent the main trophic resource for large vertebrate scavengers. However, a comparison of how vertebrate scavengers consume ungulate carcasses in different habitats with different ungulate species composition is lacking. This knowledge is essential to forecast the possible consequences of the current farmland abandonment on scavenger species. Here, we compared the scavenging patterns of 24 wild and 24 domestic ungulate carcasses in a mountainous region of southern Spain monitored through camera trapping. Our results show that carcasses of domestic ungulates, which concentrate in large numbers in open pasturelands, were detected and consumed earlier than those of wild ungulate carcasses, which frequently occur in much lower densities at more heterogenous habitats such as shrublands and forest. Richness and abundance of scavengers were also higher at domestic ungulate carcasses in open habitats. Vultures, mainly griffons (Gyps fulvus), consumed most of the carcasses, although mammalian facultative scavengers, mainly wild boar (Sus scrofa) and red fox (Vulpes vulpes), also contributed importantly to the consumption of wild ungulate carcasses in areas with higher vegetation cover. Our findings evidence that the abandonment of traditional grazing may entail consequences for the scavenger community, which should be considered by ecologists and wildlife managers.     

Factors affecting proton translocation by facultative methylotrophs

The general properties of respiration-driven proton translocation by the two facultative methylotrophs, Pseudomonas AM1 and Pseudomonas extorquens, were similar to those of other bacteria. The stoichiometry of H⁺ extruded/O atom consumed ($\text{H}\text{←}{}^{\mathrm{+}}\text{O}$) during respiration with a particular substrate depended, however, on the concentration of the permeant anion SCN^? used to abolish the membrane potential and on the physiological state of the organism. This variability makes the use of proton translocation data of dubious value in the elucidation of electron-transport pathways, at least in these species, unless the physiological condition of the organisms can be accurately described and reproduced. Methanol oxidation was inhibited by SCN^? but substitution of valinomycin for most of the SCN^? during pulse oxidant experiments allowed measurement of proton translocation when methanol was the substrate. Starved organisms were used to eliminate ambiquity as to whether added test substrates or endogenous reserve materials were being oxidised. Viability remained high during starvation and endogenous O₂ uptake remained detectable long after endogenously driven proton translocation was undetectable. In the absence of endogenously driven proton translocation, measured $\text{H}\text{←}{}^{\mathrm{+}}\text{O}$ stoichiometries differed substantially from those when it was present, suggesting that the physiological state of the organisms is an essential parameter in assessing proton translocation data.     

Induction of proteins during phage reactivation induced by UV irradiation, oxygen and peroxide in Bacteroides fragilis

Abstract Phage reactivation systems in Bacteroides fragilis were induced by far-UV irradiation, O₂ and H₂O₂. These three treatments also induced the synthesis of 3, 6, and 4 protein bands, respectively, which were easily detectable by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Two proteins with apparent M _r s of approx. 90 000 and 70 000 were induced by all three treatments. Caffeine completely inhibited UV- and O₂-induced phage reactivation and prevented the synthesis of the M _r 90 000 and M _r 70 000 proteins. The results suggest that these two proteins may be involved in phage reactivation processes induced by UV, O₂ and H₂O₂ in B. fragilis .