Aquaculture Department, Southeast Asian Fisheries Development Center, 5021 Tigbauan, Iloilo, Philippines; Laboratory of Aquaculture & Artemia Reference Center, Ghent University, Coupure Links 653, B-9000 Gent, Belgium.
Aquaculture Department, Southeast Asian Fisheries Development Center, 5021 Tigbauan, Iloilo, Philippines.
Fish Shellfish Immunol. 2017 Sep;68:202-210. doi: 10.1016/j.fsi.2017.07.023. Epub 2017 Jul 11.
In this study, the PHB-accumulating Bacillus sp. JL47 strain (capable of accumulating 55% PHB on cell dry weight) was investigated for its effects on the immune response of giant tiger shrimp (Penaeus monodon) postlarvae (PL) before and after the Vibrio campbellii challenge. Briefly, shrimp PL were cultured and fed with Artemia nauplii enriched with Bacillus sp. JL47. Shrimp receiving the Artemia nauplii without JL47 enrichment were used as control. After 15 days of feeding, the shrimp were challenged with pathogenic V. campbellii LMG 21363 at 10 cells mL by immersion. Relative expression of the immune related genes encoding for prophenoloxidase (proPO), transglutaminase (TGase) and heat shock protein 70 (Hsp70) in the shrimp were measured before (0 h) and after (3, 6, 9, 12, 24 h) the Vibrio challenge by quantitative real-time PCR using β-actin as the reference gene. The expressions of TGase and proPO were significantly up-regulated (p < 0.05) within 9 h and 12 h, respectively after challenge in shrimp receiving the Bacillus sp. JL47 as compared to the challenged and non-challenged controls. Hsp70 expression was significantly increased (p < 0.05) at 3 h post-challenge in all challenged shrimp. Interestingly, proPO and TGase genes were significantly up-regulated (p < 0.05) in Bacillus sp. JL47 treated shrimp even before the Vibrio challenge was applied. No up-regulation in the Hsp70 gene, however, was observed under these conditions. The data suggest that the protective effect of the PHB-accumulating Bacillus sp. JL47 in shrimp was due to its capacity to stimulate the innate immune related genes of the shrimp, specifically the proPO and TGase genes. The application of probiotic Bacillus species, capable of accumulating a significant amount of PHB, is suggested as potential immunostimulatory strategy for aquaculture.
在这项研究中,研究了聚羟基丁酸(PHB)积累芽孢杆菌 JL47 菌株(能够在细胞干重上积累 55%的 PHB)对凡纳滨对虾(Penaeus monodon)幼虾(PL)在弧菌攻毒前后免疫反应的影响。简而言之,用富含芽孢杆菌 JL47 的丰年虫培养和喂养虾 PL。用未富集 JL47 的丰年虫喂养的虾作为对照。喂养 15 天后,将虾用致病性弧菌 LMG 21363 通过浸泡以 10 个细胞 mL 进行攻毒。用定量实时 PCR 测量虾在弧菌攻毒前后(0 h)免疫相关基因编码的酚氧化酶原(proPO)、转谷氨酰胺酶(TGase)和热休克蛋白 70(Hsp70)的相对表达,β-肌动蛋白作为内参基因。与对照相比,在受到芽孢杆菌 JL47 处理的虾中,攻毒后 9 小时和 12 小时内 TGase 和 proPO 的表达分别显著上调(p < 0.05)。在所有攻毒的虾中,Hsp70 表达在攻毒后 3 小时显著增加(p < 0.05)。有趣的是,即使在施加弧菌攻毒之前,proPO 和 TGase 基因在 JL47 处理的虾中也显著上调(p < 0.05)。然而,在这些条件下,Hsp70 基因没有上调。数据表明,聚羟基丁酸积累芽孢杆菌 JL47 在虾中的保护作用是由于其刺激虾的固有免疫相关基因的能力,特别是 proPO 和 TGase 基因。建议应用能够积累大量 PHB 的益生菌芽孢杆菌作为水产养殖的潜在免疫刺激策略。