Effect of Bacillus subtilis on intestinal apoptosis of grass carp Ctenopharyngodon idella orally challenged with Aeromonas hydrophila
The influence of Bacillus subtilis Ch9 on intestinal apoptosis of grass carp Ctenopharyngodon idella challenged with Aeromonas hydrophila was studied. Groups of grass carp were orally intubated with B. subtilis or phosphate buffered saline for 3 days and subjected to oral A. hydrophila challenge. Intestinal tissues were collected and prepared for terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) assay, real-time quantitative polymerase chain reaction analysis of caspase-3 and -8 and b-cell lymphoma–2 protein (bcl-2), histopathological analyses and measurement of caspase-3 and -8 activities at 0, 12, 24, 48, 72 and 96 h post-challenge. TUNEL analyses demonstrated that B. subtilis induced intestinal cell apoptosis, and caspase-3 and -8 activities increased significantly compared to in the control and A. hydrophila groups. The intestinal damage due to A. hydrophila was also reduced. However, messenger RNA transcript levels of caspase-3 and -8 and bcl-2 were not markedly elevated. These results suggested that B. subtilis Ch9 increased intestinal apoptosis in fish challenged with A. hydrophila, and that B. subtilis Ch9-induced caspase-dependent apoptosis may play an important role in reducing intestinal damage following A. hydrophila challenge.
KeywordsIntestine Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling Real-time quantitative polymerase chain reaction Caspase Aquaculture
This work was supported by the National Natural Science Foundation of China (grant nos. 31472310, 31672683).
- De BC, Meena DK, Behera BK, Das P, Das Mohapatra PK, Sharma AP (2014) Probiotics in fish and shellfish culture: immunomodulatory and ecophysiological responses. Fish Physiol Biochem 40:921–971Google Scholar
- Feng L, Luo JB, Jiang WD, Liu Y, Wu P, Jiang J, Kuang SY, Tang L, Zhang YA, Zhou XQ (2015) Changes in barrier health status of the gill for grass carp (Ctenopharyngodon idella) during valine deficiency: regulation of tight junction protein transcript, antioxidant status and apoptosis-related gene expression. Fish Shellfish Immunol 45:239–249CrossRefGoogle Scholar
- Galindo CL, Fadl AA, Sha J, Gutierrez C Jr, Popov VL, Boldogh I, Aggarwal BB, Chopra AK (2004) Aeromonas hydrophila cytotoxic enterotoxin activates mitogen-activated protein kinases and induces apoptosis in murine macrophages and human intestinal epithelial cells. J Biol Chem 279:37597–37612CrossRefGoogle Scholar
- Gao D, Ze Xu, Qiao P, Liu S, Zhang L, He P, Zhang X, Wang Y, Min W (2013b) Cadmium induces liver cell apoptosis through caspase-3A activation in purse red common carp (Cyprinus carpio). PLoS One 8:1–11Google Scholar
- Kanaly ST, Nashleanas M, Hondowicz B, Scott P (1999) TNF receptor p55 is required for elimination of inflammatory cells following control of intracellular pathogens. J Immunol 163:3883–3889Google Scholar
- Ngamkala S, Futami K, Endo M, Maita M, Katagiri T (2010) Immunological effects of glucan and Lactobacillus rhamnosus GG, a probiotic bacterium, on Nile tilapia Oreochromis niloticus intestine with oral Aeromonas challenges. Fish Sci 76:833–840. https://doi.org/10.1007/s12562-010-0280-0 CrossRefGoogle Scholar
- Tong N, Yin XY, Li DP, Tang R (2014) Effects of acute thermal stress on kidney cell of grass carp (Ctenopharyngodon idella). J Huazhong Agric Univ 33:88–92 (in Chinese with English abstract) Google Scholar
- Wu P, Liu Y, Jiang WD, Jiang J, Zhang YA, Zhou XQ, Feng L (2017) Intestinal immune responses of Jian carp against Aeromonas hydrophila depressed by choline deficiency: varied change patterns of mRNA levels of cytokines, tight junction proteins and related signaling molecules among three intestinal segments. Fish Shellfish Immunol 65:34–41CrossRefGoogle Scholar
- Xu HJ, Jiang WD, Feng L, Liu Y, Wu P, Jiang J, Kuang SY, Tang L, Tang WN, Zhang YA, Zhou XQ (2016) Dietary vitamin C deficiency depresses the growth, head kidney and spleen immunity and structural integrity by regulating NF-kappaB, TOR, Nrf2, apoptosis and MLCK signaling in young grass carp (Ctenopharyngodon idella). Fish Shellfish Immunol 52:111–138CrossRefGoogle Scholar