Catechin as the Most Efficient Bioactive Compound from Azadirachta indica with Antibiofilm and Anti-quorum Sensing Activities Against Dental Biofilm: an In Vitro and In Silico Study.

Neem (Azadirachta indica [AI]) is a unique and traditional source of antioxidant and antibacterial compounds. The GC-MS studies revealed that phytoextract of Azadirachta indica comprises a large number of phytocompounds that possess the efficacy of inhibiting the biofilm. It was observed that phytocompounds like catechin showed maximum eradication of biofilm along with the degradation of EPS structural components like carbohydrates and proteins compared to quercetin, nimbolide, nimbin, and azardirachtin, and hence, catechin was proved to be the best against dental plaque-forming bacteria. It was also observed that catechin was able to bring about a marked reduction in quorum sensing (QS) both in Alcaligenes faecalis and Pseudomonas gingivalis dental biofilm-forming strains. The extent of such reduction was maximum for catechin (94.56±2.56% in P. gingivalis & 96.56±2.5 in A. faecalis) in comparison to other bioactive compounds. It was further observed that the bioactive compounds possess the ability to quickly pass across the membrane and bring about inhibition in the DNA and RNA content of the sessile cells. This was further validated by microscopic and in silico studies. Thus, this study revealed that catechin obtained from the phytoextract of AI showed a marked ability to inhibit the dental biofilm and can be used as a natural drug-like compound in treating biofilm-associated chronic infections.

Upregulation of a marine fungal biosynthetic gene cluster by an endobacterial symbiont.

Fungal-bacterial associations are present in nature, playing important roles in ecological, evolutionary and medicinal processes. Here we report a fungus-bacterial symbiont from marine sediment. The bacterium lives inside the fungal mycelium yet is robust enough to survive independent of its host; the independently grown bacterium can infect the fungal host in vitro and continue to grow progenitively. The bacterial symbiont modulates the fungal host to biosynthesize a polyketide antimicrobial, spiromarmycin. Spiromarmycin appears to endow upon the symbiont pair a protective/defensive means of warding off competitor organisms, be they prokaryotic or eukaryotic microorganisms. Genomic analyses revealed the spiromarmycin biosynthetic machinery to be encoded, not by the bacterium, but rather the fungal host. This unique fungal-bacterial symbiotic relationship and the molecule/s resulting from it dramatically expand our knowledge of marine microbial diversity and shed important insights into endosymbionts and fungal-bacterial relationships.

Characterization of Alcaligenes faecalis strain AD15 indicating biocontrol activity against plant pathogens.

Bacterial strain possessing both bacteriostatic and fungistatic activity (biocontrol activity) against pathogens of cyclamen (Cyclamen sp.) was isolated from the soil in Gifu Prefecture, Japan, and characterized with respect to its taxonomic and biocontrol properties. The sequence of its 16S rRNA gene, morphology, biochemistry, and fatty acid composition demonstrated that it is a strain most closely related to Alcaligenes faecalis subsp. faecalis LMG 1229(T). The isolate was named A. faecalis strain AD15. A. faecalis AD15 produced hydroxylamine at maximum yields of 33.3±1.7 mg/L after 16 h cultivation in LB medium and 19.0±0.44 mg/L after 19 h cultivation in synthetic medium. Moreover, minimum inhibitory concentrations of hydroxylamine against the cyclamen pathogens Pantoea agglomerans and Colletotrichum gloeosporioides were 4.20±0.98 and 16.5±0.67 mg/L. These results indicated that the biocontrol activity of strain AD15 might be attributed to hydroxylamine, a metabolite in the culture medium, and it had the potential for biopesticide application.

Isolation and characterization of a heterotrophic nitrifier from coke plant wastewater.

This study investigated some factors affecting ammonium removal and nitrite accumulation by Alcaligenes faecalis C16, which was isolated from the activated sludge of a coking wastewater treatment plant. Nitrite was produced from ammonium only in the presence of citrate, acetate, meat extract, peptone or ethanol. The highest amount of nitrite was found with citrate as carbon source. A. faecalis C16 could not use glucose, fructose, sucrose and methanol. Under the optimum conditions of initial pH 6.0, C/N 14, 30 °C and 120 rpm, a maximum nitrite accumulation of 28.29 mg/L NO(2)(-)-N was achieved when the organism grew with citrate in four days. Nitrite accumulation increased with the increase of NH(4)(+)-N. Furthermore, A. faecalis C16 was shown to have phenol-degrading capacity during ammonium removal. Metabolism of phenol resulted in acidification of the media, which is not favorable for nitrification, whereas many other carbon sources made the medium more alkaline. However, no inhibitory effect by phenol was observed when phenol and acetate were used as mixed carbon source at different phenol/sodium acetate (P/S) ratios and their pH values were all controlled above 9.2 or P/S ratios below 5:5. These results suggested that A. faecalis C16 has some potential application in industrial wastewater treatment systems.

Surface properties of bacteria from activated sludge in relation to bioflocculation.

Two bacterial stains were isolated from the activated sludge and identified as Leucobacter sp. and Alcaligenesfaecalis by 16S rDNA sequencing. Pure cultures of these two strains, representing well or poorly settled bacteria, were used to investigate the mechanism of bioflocculation in activated sludge. Based on the analyses of the characteristics of cells hydrophobicity, zeta-potential, flocculation ability and extracellular polymeric substance (EPS) composition under different growth stages, it was found that the ratio of cell EPS protein had the highly influence on zeta-potential and hydrophobicity, which were important factors to bioflocculation. Cellulase and Proteinase K could destroy the extracellular biopolymer and resulted in a decrease in the hydrophobicity and zeta-potential. However, in our study, the flocculation characteristics exhibited differently in relation to cellulase and Proteinase K. Flocculation of cells treated with cellulase and Proteinase K decreased sharply, and then recovered quickly in cellulase treatment, while cells treated with Proteinase K showed no sign of recovery. This reveals that the presence of protein in extracellular biopolymer plays an important role to the bioflocculation of cells.

Impact of antifungals producing rhizobacteria on the performance of Vigna radiata in the presence of arbuscular mycorrhizal fungi.

Plant growth-promoting rhizobacteria (PGPR) that produce antifungal metabolites are potential threats for the arbuscular mycorrhizal (AM) fungi known for their beneficial symbiosis with plants that is crucially important for low-input sustainable agriculture. To address this issue, we used a compartmented container system where test plants, Vigna radiata, could only reach a separate nutrient-rich compartment indirectly via the hyphae of AM fungi associated with their roots. In this system, where plants depended on nutrient uptake via AM symbiosis, we explored the impact of various PGPR. Plants were inoculated with or without a consortium of four species of AM fungi (Glomus coronatum, Glomus etunicatum, Glomus constrictum, and Glomus intraradices), and one or more of the following PGPR strains: phenazine producing (P(+)) and phenazine-less mutant (P(-)), diacetylphloroglucinol (DAPG) producing (G(+)) and DAPG-less mutant (G(-)) strains of Pseudomonas fluorescens, and an unknown antifungal metabolite-producing Alcaligenes faecalis strain, SLHRE425 (D). PGPR exerted only a small if any effect on the performance of AM symbiosis. G(+) enhanced AM root colonization and had positive effects on shoot growth and nitrogen content when added alone, but not in combination with P(+). D negatively influenced AM root colonization, but did not affect nutrient acquisition. Principal component analysis of all treatments indicated correlation between root weight, shoot weight, and nutrient uptake by AM fungus. The results indicate that antifungal metabolites producing PGPR do not necessarily interfere with AM symbiosis and may even promote it thus carefully chosen combinations of such bioinoculants could lead to better plant growth.