The Antibiofilm Effects of Antimony Tin Oxide Nanoparticles against Polymicrobial Biofilms of Uropathogenic Escherichia coli and Staphylococcus aureus.

Biofilms are responsible for persistent or recurring microbial infections. Polymicrobial biofilms are prevalent in environmental and medical niches. Dual-species biofilms formed by Gram-negative uropathogenic Escherichia coli (UPEC) and Gram-positive Staphylococcus aureus are commonly found in urinary tract infection sites. Metal oxide nanoparticles (NPs) are widely studied for their antimicrobial and antibiofilm properties. We hypothesized that antimony-doped tin (IV) oxide (ATO) NPs, which contain a combination of antimony (Sb) and tin (Sn) oxides, are good antimicrobial candidates due to their large surface area. Thus, we investigated the antibiofilm and antivirulence properties of ATO NPs against single- and dual-species biofilms formed by UPEC and S. aureus. ATO NPs at 1 mg/mL significantly inhibited biofilm formation by UPEC, S. aureus, and dual-species biofilms and reduced their main virulence attributes, such as the cell surface hydrophobicity of UPEC and hemolysis of S. aureus and dual-species biofilms. Gene expression studies showed ATO NPs downregulated the hla gene in S. aureus, which is essential for hemolysin production and biofilm formation. Furthermore, toxicity assays with seed germination and Caenorhabditis elegans models confirmed the non-toxic nature of ATO NPs. These results suggest that ATO nanoparticles and their composites could be used to control persistent UPEC and S. aureus infections.

Inhibition of growth, biofilm formation, virulence, and surface attachment of Agrobacterium tumefaciens by cinnamaldehyde derivatives.

Agrobacterium tumefaciens, a soil-borne, saprophytic plant pathogen that colonizes plant surfaces and induces tumors in a wide range of dicotyledonous plants by transferring and expressing its T-DNA genes. The limited availabilities and efficacies of current treatments necessitate the exploration of new anti-Agrobacterium agents. We examined the effects of trans-cinnamaldehyde (t-CNMA) and its derivatives on the cell surface hydrophobicity, exopolysaccharide and exo-protease production, swimming motility on agar, and biofilm forming ability of A. tumefaciens. Based on initial biofilm inhibition results and minimum inhibitory concentration (MIC) data, 4-nitro, 4-chloro, and 4-fluoro CNMAs were further tested. 4-Nitro, 4-chloro, and 4-fluoro CNMA at ≥150 µg/ml significantly inhibited biofilm formation by 94-99%. Similarly, biofilm formation on polystyrene or nylon was substantially reduced by 4-nitro and 4-chloro CNMAs as determined by optical microscopy and scanning electron microscopy (SEM) and 3-D spectrum plots. 4-Nitro and 4-chloro CNMAs induced cell shortening and concentration- and time-dependently reduced cell growth. Virulence factors were significantly and dose-dependently suppressed by 4-nitro and 4-chloro CNMAs (P ≤ 0.05). Gene expressional changes were greater after 4-nitro CNMA than t-CNMA treatment, as determined by qRT-PCR. Furthermore, some genes essential for biofilm formation, motility, and virulence genes significantly downregulated by 4-nitro CNMA. Seed germination of Raphanus sativus was not hindered by 4-nitro or 4-fluoro CNMA at concentrations ≤200 µg/ml, but root surface biofilm formation was severely inhibited. This study is the first to report the anti-Agrobacterium biofilm and anti-virulence effects of 4-nitro, 4-chloro, and 4-fluoro CNMAs and t-CNMA and indicates that they should be considered starting points for the development of anti-Agrobacterium agents.

Trans-Cinnamaldehyde Eluting Porous Silicon Microparticles Mitigate Cariogenic Biofilms.

Dental caries, a preventable disease, is caused by highly-adherent, acid-producing biofilms composed of bacteria and yeasts. Current caries-preventive approaches are ineffective in controlling biofilm development. Recent studies demonstrate definite advantages in using natural compounds such as trans-cinnamaldehyde in thwarting biofilm assembly, and yet, the remarkable difficulty in delivering such hydrophobic bioactive molecules prevents further development. To address this critical challenge, we have developed an innovative platform composed of components with a proven track record of safety. We fabricated and thoroughly characterised porous silicon (pSi) microparticles to carry and deliver the natural phenyl propanoid trans-cinnamaldehyde (TC). We investigated its effects on preventing the development of cross-kingdom biofilms (Streptococcus mutans and Candida albicans), typical of dental caries found in children. The prepared pSi microparticles were roughly cubic in structure with 70-75% porosity, to which the TC (pSi-TC) was loaded with about 45% efficiency. The pSi-TC particles exhibited a controlled release of the cargo over a 14-day period. Notably, pSi-TC significantly inhibited biofilms, specifically downregulating the glucan synthesis pathways, leading to reduced adhesion to the substrate. Acid production, a vital virulent trait for caries development, was also hindered by pSi-TC. This pioneering study highlights the potential to develop the novel pSi-TC as a dental caries-preventive material.

Inhibition of Agrobacterium tumefaciens Growth and Biofilm Formation by Tannic Acid.

Agrobacterium tumefaciens underlies the pathogenesis of crown gall disease and is characterized by tumor-like gall formation on the stems and roots of a wide variety of economically important plant species. The bacterium initiates infection by colonizing and forming biofilms on plant surfaces, and thus, novel compounds are required to prevent its growth and biofilm formation. In this study, we investigated the ability of tannic acid, which is ubiquitously present in woody plants, to specifically inhibit the growth and biofilm formation of A. tumefaciens. Tannic acid showed antibacterial activity and significantly reduced the biofilm formation on polystyrene and on the roots of Raphanus sativus as determined by 3D bright-field and scanning electron microscopy (SEM) images. Furthermore, tannic acid dose-dependently reduced the virulence features of A. tumefaciens, which are swimming motility, exopolysaccharide production, protease production, and cell surface hydrophobicity. Transcriptional analysis of cells (Abs600 nm = 1.0) incubated with tannic acid for 24 h at 30 °C showed tannic acid most significantly downregulated the exoR gene, which is required for adhesion to surfaces. Tannic acid at 100 or 200 µg/mL limited the iron supply to A. tumefaciens and similarly reduced the biofilm formation to that performed by 0.1 mM EDTA. Notably, tannic acid did not significantly affect R. sativus germination even at 400 µg/mL. The findings of this study suggest that tannic acid has the potential to prevent growth and biofilm formation by A. tumefaciens and thus infections resulting from A. tumefaciens colonization.

Effect of halogenated indoles on biofilm formation, virulence, and root surface colonization by Agrobacterium tumefaciens.

Agrobacterium tumefaciens is a plant pathogen that causes crown gall disease in several plant species by transferring its T-DNA to the host genome. Its chemotactic response to a range of chemical compounds released by hosts facilitates its colonization to host surfaces, and thus, novel anti-agrobacterium compounds are needed to prevent its biofilm formation. Here, we investigated 83 indole derivatives against A. tumefaciens, and based on the screening, 4-chloroindole, 6-iodoindole, and 5-chloro-2-methyl indole were selected as candidates that at 50 µg mL-1 significantly inhibited the adherence and biofilm formation of A. tumefaciens to abiotic (nitrocellulose and polystyrene) and biotic (roots of Brassica juncea) surfaces. Furthermore, they reduced bacterial growth in a time and concentration-dependent manner and significantly reduced log CFU mL-1 and survival (%). Changes in biofilm morphologies and biomasses, thicknesses, and substratum coverages were determined, and 2-D and 3-D analyses were performed using a crystal violet assay and bright field, CLSM, and SEM microscopies. Virulence factors such as swimming motility, exopolysaccharide, and exo-protease production, and cell surface hydrophobicity were markedly inhibited by the three compounds. Transcriptional analysis showed multi-fold downregulation of biofilm, virulence, motility, and stress-related genes; however, the degrees of these downregulations were variably affected. B. juncea seed germination was only severely affected by 4-chloroindole. This study demonstrates the promising antibiofilm and antivirulence activities of the three indole derivatives tested and their potentials for targeting and curbing A. tumefaciens infections.

Differential bioaccumulations and ecotoxicological impacts of metal-oxide nanoparticles, bulk materials, and metal-ions in cucumbers grown in sandy clay loam soil.

Expanding applications of metal-oxide nanoparticles (NPs) and increased environmental deposition of NPs followed by their interactions with edible crops threaten yields. This study demonstrates the effects of aging (45 days in soil) of four NPs (ZnO, CuO, Al2O3, TiO2; 3.9-34 nm) and their corresponding metal oxide bulk particles (BPs; 144-586 nm) on cucumbers (Cucumis sativus L.) cultivated in sandy-clay-loam field soil and compares these with the phytotoxic effects of readily soluble metal salts (Zn2+, Cu2+, and Al3+). Data revealed the cell-to-cell translocations of NPs, their attachments to outer and inner cell surfaces, nuclear membranes, and vacuoles, and their upward movements to aerial parts. Metal bioaccumulations in cucumbers were found in the order: (i) ZnO-NPs > ZnO-BPs > Zn2+, (ii) CuO-NPs > CuO-BPs > Cu2+, (iii) Al3+> Al2O3-NPs > Al2O3-BPs and (iv) TiO2-NPs > TiO2-BPs. Aging of NPs in soil for 45 days significantly enhanced metal uptake (P ≤ 0.05), for instance aged ZnO-NPs at 1 g kg-1 increased the uptake by 20.7 % over non-aged ZnO-NPs. Metal uptakes inhibited root (RDW) and shoot (SDW) dry weight accumulations. For Cu species, maximum negative impact (%) was exhibited by Cu2+ (RDW:SDW = 94:65) followed by CuO-NPs (RDW:SDW = 78:34) and CuO-BPs (RDW:SDW = 27:22). Aging of NPs/BPs at 1-4 g kg-1 further enhanced the toxic impact of tested materials on biomass accumulations and chlorophyll formation. NPs also induced membrane damage of root tissues and enhanced levels of antioxidant enzymes. The results of this study suggest that care is required when aged metal-oxide NPs of both essential (Zn and Cu) and non-essential (Al and Ti) metals interact with cucumber plants, especially, when they are used for agricultural purposes.