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Among various organic wastes, shrimp shell powder (SSP) was the most suitable carbon/nitrogen source for producing antinematode compounds (ANCs) via Bacillus veleznesis RB.EK7 fermentation. The fermentation process for the enhancement of antinematode activity was investigated. B. veleznesis RB.EK7 produced the highest antinematode activity in the medium containing 0.8% SSP with an initial pH of 6.5–7.0, and fermentation was performed at 35–37 °C with a saking speed of 150 rpm for 72 h. Targeting ANCs were purified from the fermented culture broth and identified as thymine (1) and hexahydropyrrolo [1,2-a]pyrazine-1,4-dione (2) based on nuclear magnetic resonance (NMR) and mass spectra analysis and were compared to those of the reported compounds. Notably, for the first time, these compounds were found as novel ANCs. Thymine (1) demonstrated a potential nematicidal effect with near 100% mortality of second-stage juvenile (J2) nematodes and anti-egg hatching effects of 70.1%, while hexahydropyrrolo [1,2-a]pyrazine-1,4-dione showed moderate antinematode activities with 64.2% mortality of J2 nematodes and anti-egg hatching effects of 57.9%. The docking study coupled with experimental enzyme inhibition results indicated that the potent nematicidal effect of these compounds may be possibly due to the inhibition of the targeting enzyme acetylcholinesterase. The data of this study suggest that SSP can be potentially reused for the eco-friendly production of ANCs for the management of black pepper nematodes.

This study aimed to reuse groundnut oil processing by-product, groundnut cake (GNC) for the low-cost production of prodigiosin (PG) via microbial technology and to assess its novel potential application for the management of black pepper nematodes. Serratia marcescens TUN02 was found as the most active PG-producing strain. Various small-scale experiments conducted in flasks indicated that GNC at 1% may be used as the sole carbon/nitrogen source for cost-effective PG production by fermentation. Notably, no further commercial ingredients and salts are required to supplement into the culture medium of this fermentation. PG was further investigated for scale-up production in a 14-L bioreactor system and PG was produced at high yield (6886 mg/L) with large-scale volume (4 L) in a short cultivation time (10 h). PG was then purified and its nematicidal activity was evaluated and showed effective inhibition of juveniles and egg hatching of Meloidogyne incognita species, harmful on black pepper, with low IC50 values of 0.2 and 0.32 mg/mL, respectively. The simple medium containing 1% GNC is the first report of cost-effective biosynthesis of PG, as well as potential in vitro anti-egg hatching activity of PG. These results indicated the potential application of GNC for low-cost bioproduction of PG for promising and novel use in the management of black pepper nematodes.

Robusta coffee is a major commercial crop in the Central Highland of Vietnam with high economic and export value. However, this crop is adversely affected by various pathogens, particularly nematodes. This study aimed to screen active anti-nematode rhizobacterial strains for sustainable coffee production. Among more than 200 isolates, the isolate TUN03 demonstrated efficient biocontrol with nearly 100% mortality of J2 coffee nematodes Meloidogyne spp. and 84% inhibition of nematode egg hatching. This active strain was identified as Pseudomonas aeruginosa TUN03 based on its 16S rRNA gene sequence and phylogenetic analysis. In greenhouse tests, the strain TUN03 significantly reduced the coffee nematode population in the rhizome-soil with an 83.23% inhibition rate and showed plant growth-promoting effects. Notably, this is the first report of the nematicidal effect of P. aeruginosa against coffee nematodes. This potent strain further showed an antifungal effect against various crop-pathogenic fungi and was found to be the most effective against Fusarium solani F04 (isolated from coffee roots) with a 70.51% inhibition rate. In addition, high-performance liquid chromatography analysis revealed that this bacterial strain also secretes plant growth regulators including indole acetic acid (IAA), gibberellic acid (GA3), kinetin, and zeatin in significant amounts of 100, 2700, 37, and 9.5 µg/mL, respectively. The data from this study suggest that P. aeruginosa TUN03 may be a potential biocontrol agent and biofertilizer for the sustainable production of Robusta coffee and other crops.