Developmental stages of biofilm and characterization of extracellular matrix of manglicolous fungus Aspergillus niger BSC-1.

Fungal biofilm is ubiquitous in natural environment. The major constituent of fungal biofilm other than biomass is the extracellular matrix (ECM), in which fungal hyphae are embedded. Physical properties of biofilms such as attachment, mechanical strength, and antibiotic resistance can be attributed to ECM. The present work probes various stages of biofilm formation by filamentous manglicolous fungus Aspergillus niger BSC-1. The spectroscopic analysis revealed that with an increase in incubation time the biofilm formation was significantly increased (p < .0001) up to 36 h. Scanning electron micrograph and confocal micrograph depicted the development of fungal biofilm comprising of six stages, that is, (a) adsorption, (b) active attachment, (c) germling and monolayer formation, (d) hyphal development and formation of ECM, (e) maturation of ECM, and (f) dispersal of spores. At maturation stage, thickness of biofilm was observed upto approximately 15 µm. Approximately, 8.1 mg of ECM materials were extracted from 20 ml of broth culture using ethanol precipitation method. Furthermore, attenuated total reflectance Fourier-transformed infrared spectroscopic analysis exhibited peaks at 3,398, 2,930, 1,571, 1,391, 1,092, 977 cm-1 which confirmed the presence of protein, carbohydrate, and lipid in the biofilm-associated matrix.