The use of biomimetic surfaces to reduce single- and dual-species biofilms of Escherichia coli and Pseudomonas putida

The ability of bacteria to adhere to and form biofilms on food contact surfaces poses serious challenges, as these may lead to the cross-contamination of food products. Biomimetic topographic surface modifications have been explored to enhance the antifouling performance of materials. In this study, the topography of two plant leaves, Brassica oleracea var. botrytis (cauliflower, CF) and Brassica oleracea capitate (white cabbage, WC), was replicated through wax moulding, and their antibiofilm potential was tested against single- and dual-species biofilms of Escherichia coli and Pseudomonas putida. Biomimetic surfaces exhibited higher roughness values (SaWC = 4.0 ± 1.0 μm and SaCF = 3.3 ± 1.0 μm) than the flat control (SaF = 0.6 ± 0.2 μm), whilst the CF surface demonstrated a lower interfacial free energy (ΔGiwi) than the WC surface (−100.08 mJ m−2 and −71.98 mJ m−2, respectively). The CF and WC surfaces had similar antibiofilm effects against single-species biofilms, achieving cell reductions of approximately 50% and 60% for E. coli and P. putida, respectively, compared to the control. Additionally, the biomimetic surfaces led to reductions of up to 60% in biovolume, 45% in thickness, and 60% in the surface coverage of single-species biofilms. For dual-species biofilms, only the E. coli strain growing on the WC surface exhibited a significant decrease in the cell count. However, confocal microscopy analysis revealed a 60% reduction in the total biovolume and surface coverage of mixed biofilms developed on both biomimetic surfaces. Furthermore, dual-species biofilms were mainly composed of P. putida, which reduced E. coli growth. Altogether, these results demonstrate that the surface properties of CF and WC biomimetic surfaces have the potential for reducing biofilm formation

An electron microscopical study of the infection of the nematode Panagrellus redivivus by the endoparasitic fungus Verticillium balanoides

The infection of nematodes by the endoparasitic fungus Verticillium balanoides was studied by means of optical and electron microscopical techniques. Conidia of this fungus adhere randomly in high numbers to the entire cuticle surface of the nematode Panagrellus redivivus. Adhesion is accomplished by means of a tri-layered adhesive pad located at the apical end of the conidium, the outermost layer of which shows a radiated substructure. After attachment an appressorium which grows through the adhesive pad and establishes a firm contact between the fungal cell wall and the nematode cuticle is formed. Penetration of the cuticle by means of a hyphal outgrowth on the appressorium is followed by the formation of an infection bulb from which trophic hyphae develop that invade the nematode. Approximately 60 h after infection conidiophores develop outside the nematode, producing numerous conidia

Cellular distribution of COT1 kinase in Neurospora crassa

The Neurospora crassa cot-1 gene encodes a Ser/Thr protein kinase, which is involved in hyphal elongation. Many vacuoles, abnormally shaped mitochondria, and nuclei, along with differences in the structure of the cell wall and hyphal septa, were observed in hyphae of the cot-l mutant shortly after a shift to the restrictive temperature. Immunolocalization experiments indicated that COT1 was associated with the cytoplasmic membrane; COT1 was also detected in the cytoplasm, The membrane-associated COT1 was absent from the cot-1 mutant when shifted to the restrictive temperature, as was a lower molecular weight isoform of COT1, We propose that COT1 may be involved in several cellular processes, and the spatial and temporal regulation of COT1 activity involves trafficking of the kinase within the fungal cell and its possible interaction with additional proteins. (C) 2000 Academic Press

Thioalkalispira microaerophila gen. nov., sp nov., a novel lithoautotrophic, sulfur-oxidizing bacterium from a soda lake

An anaerobic enrichment medium (pH 10) with thiosulfate as electron donor and nitrate as electron acceptor was inoculated with sediment from soda lake Fazda (Wadi Natrun, Egypt); a novel strain, ALEN 1(T), was isolated from the subsequent enrichment culture. Cells of strain ALEN 1(T) had a spiral morphology (0.3-0.45 x 1-4 pm), were motile and had a single polar flagellum. Sphaeroplasts were formed by the cells and were rapidly lysed during prolonged aerobic incubation of cultures. Cells of strain ALEN 1(T) contained a membrane-associated yellow pigment. The metabolism of this novel organism was obligately chemolithoautotrophic, and thiosulfate or sulfide were utilized as electron donors. Washed cells of strain ALEN 1(T) oxidized thiosulfate, sulfide, polysulfide and elemental sulfur to sulfate. Best growth was observed when the strain was grown under micro-oxic conditions (1-2% O-2 in gas phase), whereas growth was inhibited under fully oxic conditions. Nitrate was reduced to nitrite without growth of the novel organism, but other nitrogen oxides were not utilized as electron acceptors. Strain ALEN 1(T) was alkaliphilic and moderately halophilic. it grew between pH 8 and 10.4 (optimum around pH 10) with a salt concentration of between 0.3 and 1.5 M Na+ (optimum 0.5 M). The maximum growth rate (0.08 h(-1)) of the organism was achieved in a thiosulfate-limited micro-oxic continuous culture (pH 10). Phylogenetic analyses of the 16S rDNA sequences of strain ALEN 1(T) and its closest relatives demonstrated that this strain formed a deep branch within the gamma-Proteobacteria, with no obvious association to any described cluster of species/genera. On the basis of its unique physiological properties and distinct phylogenetic position, it is proposed that strain ALEN 1(T) (= DSM 14786(T) = UNICEM 212(T)) represents a novel genus within the gamma-Proteobacteria, for which the name Thioalkalispira is proposed. It is also proposed that the type species of this novel genus be named Thioalkalispira microaerophila

The localization of chitin synthase in membranous vesicles (chitosomes) in Neurospora crassa

Polyclonal anti-chitin synthase antibodies raised against the Saccharomyces cerevisiae CHS2 gene product were used to identify and localize chitin synthase in the filamentous ascomycete Neurospora crassa. A single band of approximately 110 kDa was observed in Western blots of total protein extracts of N. crassa, probed with these antibodies. However, several additional bands were labelled when membrane fraction proteins (microsomes) were probed. Histo-immunochemical localization of chitin synthase confirmed that the polypeptide is compartmentalized in membranous vesicles (chitosomes), which are abundant in the vicinity of the hyphal tip. TEM analysis did not reveal chitin synthase in the plasma membrane. However, dense labelling of membrane-associated chitin synthase was observed by light-microscopic analysis of IV. crassa protoplasts and at young hyphal tips

In vivo crystal formation in Escherichia coli of an over-expressed soluble form of penicillin-binding protein 5

Accumulation of either native membrane-bound or soluble variants of PBP5 over-expressed in the cytoplasm was investigated by electron microscopy of ultra-thin sections. One of the soluble forms of PBP5 (PBP5s353) formed well-ordered crystals inside the cells. Cells sectioned perpendicular to their long axis showed a diamond-shaped crystal whereas cells cut parallel to their long axis contained a long, narrow crystal. In both sectioning directions an ordered ultra-structure was visible as shown by optical diffraction. Computer processing was used to enhance the crystal images. From this the unit cell parameters were calculated as a = 7.6 nm, b = 4 nm, c = 4.2 nm, gamma = 75-degrees. The calculated unit-cell volume of 120 nm3 is large enough to contain one protein molecule