TY - JOUR
T1 - Feed supplementation with quorum quenching probiotics with anti-virulence potential improved innate immune responses, antioxidant capacity and disease resistance in Asian seabass (Lates calcarifer)
AU - Ghanei-Motlagh, Reza
AU - Gharibi, Darioush
AU - Mohammadian, Takavar
AU - Khosravi, Mohammad
AU - Mahmoudi, Esmaeil
AU - Zarea, Mojtaba
AU - Menanteau-Ledouble, Simon
AU - El-Matbouli, Mansour
N1 - Funding Information:
This study was supported by a grant from Shahid Chamran University of Ahvaz Research Council (Grant No. vC98.299). The funding body played no role in the design of the study or analysis of the data. We gratefully thank Prof. Dr. Masoud Ghorbanpoor for providing the specific primers. The authors appreciate Dr. Sakineh Mashjoor for her assistance in assessment of the antioxidant parameters, Mojtaba Emam and Javad Seyedi for their assistance during in vivo trials, and Dr. M Shokouhmand and Mag. M. Srivastava for the valuable help in bacteriological experiments.
Publisher Copyright:
© 2021 Elsevier B.V.
PY - 2021/3/30
Y1 - 2021/3/30
N2 - Application of multifunctional probiotics is considered to be a practical strategy for the control of infections caused by Gram-negative bacteria. Consequently, the present study was designed to investigate the possible inhibitory effects of quorum quenching (QQ) probiotics on several N-acyl-homoserine lactone (AHL)-associated virulence factors in Vibrio alginolyticus and V. harveyi in vitro, and to evaluate the impacts of their administration in the diet on non-specific immune responses, antioxidant capacity and resistance to V. alginolyticus in Asian seabass. For the in vitro assay, the effect of exogenous AHL was assayed on the regulation of several virulence factors including lytic enzymes (amylase, gelatinase, hemolysin, lecithinase, lipase and protease productions), swarming and swimming motilities, and biofilm formation in V. alginolyticus and V. harveyi. Exogenous 3-oxo-C10-HSL positively regulated the production of amylase, gelatinase, protease and biofilm in V. alginolyticus. Exogenous 3-oh-C4-HSL positively regulated swarming, swimming and biofilm formation in V. harveyi. To determine the virulence-inhibitory effects of QQ probiotics on the AHL-dependent virulence factors, cell-free lysates were obtained from Bacillus thuringiensis QQ1 and B. cereus QQ2. Both probionts could reduce the production of amylase, gelatinase and protease in V. alginolyticus. Bacillus thuringiensis QQ1 was also able to decrease the biofilm formed by V. alginolyticus. Bacillus thuringiensis QQ1 and B. cereus QQ2 decreased swarming and swimming motilities, and biofilm formation in V. harveyi. In order to assay the effectiveness of the QQ probiotics in vivo, Asian seabass were fed with basal diet (control) and basal diet supplemented with 109 CFU/g feed of B. thuringiensis QQ1 or B. cereus QQ2 for 8 weeks. At the end of pre-infection trial (day 42), catalase (CAT), superoxide dismutase (SOD), lysozyme, antiprotease, myeloperoxidase (MPO), respiratory burst (RB) and bactericidal activities were significantly (p < 0.05) higher in the probiotic-treated fish than un-treated fish. After challenge with V. alginolyticus, significantly (p < 0.05) higher levels of glutathione (GSH) in the QQ2 treatment (day 49) and lower levels of malondialdehyde (MDA) in both probiotic groups were observed compared to the control group. Moreover, CAT, MPO, RB, alternative pathway of complement (APC), lysozyme, antiprotease and bactericidal activities were significantly (p < 0.05) higher in the probiotic treatments compared to the control group after 7 and 14 days post-infection. Both probionts significantly (p < 0.05) improved the cumulative survival in Asian seabass against V. alginolyticus. Our results indicate that QQ probiotics can modulate immune and antioxidant responses, and provide high protection against Vibrio infection in Asian seabass.
AB - Application of multifunctional probiotics is considered to be a practical strategy for the control of infections caused by Gram-negative bacteria. Consequently, the present study was designed to investigate the possible inhibitory effects of quorum quenching (QQ) probiotics on several N-acyl-homoserine lactone (AHL)-associated virulence factors in Vibrio alginolyticus and V. harveyi in vitro, and to evaluate the impacts of their administration in the diet on non-specific immune responses, antioxidant capacity and resistance to V. alginolyticus in Asian seabass. For the in vitro assay, the effect of exogenous AHL was assayed on the regulation of several virulence factors including lytic enzymes (amylase, gelatinase, hemolysin, lecithinase, lipase and protease productions), swarming and swimming motilities, and biofilm formation in V. alginolyticus and V. harveyi. Exogenous 3-oxo-C10-HSL positively regulated the production of amylase, gelatinase, protease and biofilm in V. alginolyticus. Exogenous 3-oh-C4-HSL positively regulated swarming, swimming and biofilm formation in V. harveyi. To determine the virulence-inhibitory effects of QQ probiotics on the AHL-dependent virulence factors, cell-free lysates were obtained from Bacillus thuringiensis QQ1 and B. cereus QQ2. Both probionts could reduce the production of amylase, gelatinase and protease in V. alginolyticus. Bacillus thuringiensis QQ1 was also able to decrease the biofilm formed by V. alginolyticus. Bacillus thuringiensis QQ1 and B. cereus QQ2 decreased swarming and swimming motilities, and biofilm formation in V. harveyi. In order to assay the effectiveness of the QQ probiotics in vivo, Asian seabass were fed with basal diet (control) and basal diet supplemented with 109 CFU/g feed of B. thuringiensis QQ1 or B. cereus QQ2 for 8 weeks. At the end of pre-infection trial (day 42), catalase (CAT), superoxide dismutase (SOD), lysozyme, antiprotease, myeloperoxidase (MPO), respiratory burst (RB) and bactericidal activities were significantly (p < 0.05) higher in the probiotic-treated fish than un-treated fish. After challenge with V. alginolyticus, significantly (p < 0.05) higher levels of glutathione (GSH) in the QQ2 treatment (day 49) and lower levels of malondialdehyde (MDA) in both probiotic groups were observed compared to the control group. Moreover, CAT, MPO, RB, alternative pathway of complement (APC), lysozyme, antiprotease and bactericidal activities were significantly (p < 0.05) higher in the probiotic treatments compared to the control group after 7 and 14 days post-infection. Both probionts significantly (p < 0.05) improved the cumulative survival in Asian seabass against V. alginolyticus. Our results indicate that QQ probiotics can modulate immune and antioxidant responses, and provide high protection against Vibrio infection in Asian seabass.
KW - Antioxidant capacity
KW - Asian seabass
KW - Innate immune response
KW - Quorum quenching probiotics
KW - Vibrio infection
KW - Virulence factor
UR - http://www.scopus.com/inward/record.url?scp=85099613598&partnerID=8YFLogxK
U2 - 10.1016/j.aquaculture.2021.736345
DO - 10.1016/j.aquaculture.2021.736345
M3 - Journal article
AN - SCOPUS:85099613598
SN - 0044-8486
VL - 535
JO - Aquaculture
JF - Aquaculture
M1 - 736345
ER -