ABSTRACT
Globally, Ralstonia solanacearum (Smith) is ranked one of the most destructive bacterial pathogens inducing rapid and fatal wilting symptoms on tomatoes. Yield losses on tomatoes vary from 0 to 91% and most control measures are unaffordable to resource-poor farmers. This study investigated the antimicrobial activities of chitin and chitosan extracted from black soldier fly (BSF) pupal exuviae against R. solanacearum. Morphological, biochemical, and molecular techniques were used to isolate and characterize R. solanacearum for in vitro pathogenicity test using disc diffusion technique. Our results revealed that BSF chitosan significantly inhibited the growth of R. solanacearum when compared to treatments without chitosan. However, there was no significant difference in the antibacterial activities between BSF and commercial chitosan against R. solanacearum. Soil amended with BSF-chitin and chitosan demonstrated a reduction in bacterial wilt disease incidence by 30.31% and 34.95%, respectively. Whereas, disease severity was reduced by 22.57% and 23.66%, when inoculated tomato plants were subjected to soil amended with BSF chitin and chitosan, respectively. These findings have demonstrated that BSF pupal shells are an attractive renewable raw material for the recovery of valuable products (chitin and chitosan) with promising ability as a new type of eco-friendly control measure against bacterial wilt caused by R. solanacearum. Further studies should explore integrated pest management options that integrate multiple components including insect-based chitin and chitosan to manage bacterial wilt diseases, contributing significantly to increased tomato production worldwide.
ABSTRACT
Globally, the broad-spectrum antimicrobial activity of chitin and chitosan has been widely documented. However, very little research attention has focused on chitin and chitosan extracted from black soldier fly pupal exuviae, which are abundantly present as byproducts from insect-farming enterprises. This study presents the first comparative analysis of chemical and biological extraction of chitin and chitosan from BSF pupal exuviae. The antibacterial activity of chitosan was also evaluated. For chemical extraction, demineralization and deproteinization were carried out using 1 M hydrochloric acid at 100 °C for 2 h and 1 M NaOH for 4 h at 100 °C, respectively. Biological chitin extraction was carried out by protease-producing bacteria and lactic-acid-producing bacteria for protein and mineral removal, respectively. The extracted chitin was converted to chitosan via deacetylation using 40% NaOH for 8 h at 100 °C. Chitin characterization was done using FTIR spectroscopy, while the antimicrobial properties were determined using the disc diffusion method. Chemical and biological extraction gave a chitin yield of 10.18% and 11.85%, respectively. A maximum chitosan yield of 6.58% was achieved via chemical treatment. From the FTIR results, biological and chemical chitin showed characteristic chitin peaks at 1650 and 1550 cm-1-wavenumbers corresponding to amide I stretching and amide II bending, respectively. There was significant growth inhibition for Escherichia coli, Bacillus subtilis,Pseudomonas aeruginosa,Staphylococcus aureus, and Candida albicans when subjected to 2.5 and 5% concentrations of chitosan. Our findings demonstrate that chitosan from BSF pupal exuviae could be a promising and novel therapeutic agent for drug development against resistant strains of bacteria.
