The accumulation of petrochemical plastic waste is detrimental to the environment. Polyhydroxyalkanoates (PHAs) are bacterial-derived polymers utilized for the production of bioplastics. PHA-plastics exhibit mechanical and thermal properties similar to conventional plastics. However, high production cost and obtaining high PHA yield and productivity impedes the widespread use of bioplastics. This study demonstrates the concept of cyclic fed-batch fermentation (CFBF) for enhanced PHA productivity by Bacillus thuringiensis using a glucose-rich hydrolyzate as the sole carbon source. The statistically optimized fermentation conditions used to obtain high cell density biomass (OD600 of 2.4175) were: 8.77 g L−1 yeast extract; 66.63% hydrolyzate (v/v); a fermentation pH of 7.18; and an incubation time of 27.22 h. The CFBF comprised three cycles of 29 h, 52 h, and 65 h, respectively. After the third cyclic event, cell biomass of 20.99 g L−1, PHA concentration of 14.28 g L−1, PHA yield of 68.03%, and PHA productivity of 0.219 g L−1 h−1 was achieved. This cyclic strategy yielded an almost threefold increase in biomass concentration and a fourfold increase in PHA concentration compared with batch fermentation. FTIR spectra of the extracted PHAs display prominent peaks at the wavelengths unique to PHAs. A copolymer was elucidated after the first cyclic event, whereas, after cycles CFBF 2–4, a terpolymer was noted. The PHAs obtained after CFBF cycle 3 have a slightly higher thermal stability compared with commercial PHB. The cyclic events decreased the melting temperature and degree of crystallinity of the PHAs. The approach used in this study demonstrates the possibility of coupling fermentation strategies with hydrolyzate derived from lignocellulosic waste as an alternative feedstock to obtain high cell density biomass and enhanced PHA productivity.

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Phytochemicals are being explored as therapeutic alternatives in aquaculture since they have destressing, growth-promoting, appetite-increasing, immune-stimulating, and antimicrobial properties. The susceptibility of 28 Flavobacterium johnsoniae-like isolates and nine selected Flavobacterium spp. isolates to three phytochemicals, viz.: cinnamaldehyde (10-250 µg/ml), vanillin (5-500 µg/ml) and four crude Kigelia africana extracts (4-10 mg/ml ethyl acetate, dichloromethane, methanol and hexane), were assessed using disk diffusion assays and compared to standard antimicrobial agents, ampicillin and tetracycline using activity indices. Cinnamaldehyde (250 µg/ml) was more effective than 250 µg/ml vanillin, which was ineffective even at higher concentrations. K. africana extract (4 mg/ml) antibacterial efficacy decreased in the following order: Ethyl acetate, methanol, dichloromethane and hexane. The 10 mg/ml methanolic K. africana extract was most effective, with 100% of isolates displaying susceptibility, irrespective of the isolation source. Methanolic extract (10 mg/ml) activity indices ≥ 1 were obtained for 67.9 and 71.4% of isolates, respectively, relative to AMP10 and TE30. Cinnamaldehyde and the K. africana methanol extract are promising candidates to be tested for their efficacy in the treatment of Flavobacterium-associated fish infections. These phytochemicals might be environmentally-friendly, cost-effective alternatives to antimicrobial agent use in aquaculture, with a lesser potential of resistance development.