Enterococci as Dominant Xylose Utilizing Lactic Acid Bacteria in Eri Silkworm Midgut and the Potential Use of Enterococcus hirae as Probiotic for Eri Culture

SIMPLE SUMMARY: This study focused on isolation and identification of xylose utilizing lactic acid bacteria from the midgut of Eri silkworm to understand their characteristics such as tannin tolerance, production of cellulolytic enzymes, and antimicrobial activity against insect pathogenic bacteria. The Enterococcus was found as the dominant genus among xylose utilizing lactic acid bacteria. Within this genus, Enterococcus hirae SX2 showed the potential to be used as a probiotic in Eri silkworm culture due to its tannin tolerance and antimicrobial activity against insect pathogenic bacteria. The trial experiment for applying live E. hirae SX2 supplemented to castor leaves in Eri silkworm rearing showed a positive effect for improving larval weight and survival. These findings led to the development of a new probiotic for Eri culture and also could be the experimental model for screening of the potential probiotic from mulberry silkworm (Bombyx mori). ABSTRACT: A total of 51 pentose utilizing lactic acid bacteria (LAB) were isolated from acid-forming bacteria in the midgut of healthy mature Eri silkworm using de Man, Rogosa and Sharpe (MRS) agar containing 10 g/L xylose (MRS-xylose) as the carbon source supplemented with 0.04% (w/v) bromocresol purple. Further analysis of 16S rRNA gene sequences revealed the highest prevalence of up to 35 enterococci isolates, which included 20 isolates of Enterococcus mundtii, followed by Entercoccus faecalis (eight isolates), Weissella cibaria (four isolates), Enterococcus hirae (two isolates), Enterococcus lactis (one isolate), and Enterococcus faecium (one isolate). All 51 LAB isolates showed positive growth on MRS containing a range of polysaccharides as the sole carbon source. All isolates were able to grow and form clear zones on MRS supplemented with 1 g/L xylose, while E. faecalis SC1, E. faecalis SCT2, and E. hirae SX2 showed tannin tolerance ability up to 5 g/L. Moreover, five isolates showed antimicrobial activity against Eri silkworm pathogens, including Bacillus cereus, Staphylococcus aureus, and Proteus vulgaris, with E. hirae SX2 having the highest inhibitory effect. Supplementation of live E. hirae SX2 on castor leaves significantly improved the weight and reduced the silkworm mortality when compared with the control group (p < 0.05). This cocci LAB can be considered as the new probiotic for Eri culture. Additionally, this finding presented the perspective of non-mulberry silkworm that could also be used as the model for further applying to new trends of the sericulture industry.