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Fisheries Science

, Volume 85, Issue 1, pp 187–197 | Cite as

Effect of Bacillus subtilis on intestinal apoptosis of grass carp Ctenopharyngodon idella orally challenged with Aeromonas hydrophila

  • Ding Zhang
  • Zhixin Wu
  • Xiaoxuan ChenEmail author
  • Huan Wang
  • Daoyuan Guo
Original Article Aquaculture

Abstract

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.

Keywords

Intestine Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling Real-time quantitative polymerase chain reaction Caspase Aquaculture 

Notes

Acknowledgment

This work was supported by the National Natural Science Foundation of China (grant nos. 31472310, 31672683).

References

  1. Banerjee G, Ray AK (2017) The advancement of probiotics research and its application in fish farming industries. Res Vet Sci 115:66–77CrossRefGoogle Scholar
  2. Banerjee C, Goswami R, Verma G, Datta M, Mazumder S (2012) Aeromonas hydrophila induced head kidney macrophage apoptosis in Clarias batrachus involves the activation of calpain and is caspase-3 mediated. Dev Comp Immunol 37:323–333CrossRefGoogle Scholar
  3. Banerjee G, Nandi A, Ray AK (2017) Assessment of hemolytic activity, enzyme production and bacteriocin characterization of Bacillus subtilis LR1 isolated from the gastrointestinal tract of fish. Arch Microbiol 199:115–124CrossRefGoogle Scholar
  4. Bialik S, Zalckvar E, Ber Y, Rubinstein AD, Kimchi A (2010) Systems biology analysis of programmed cell death. Trends Biochem Sci 35:556–564CrossRefGoogle Scholar
  5. Boatright KM, Salvesen GS (2003) Mechanisms of caspase activation. Curr Opin Cell Biol 15:725–731CrossRefGoogle Scholar
  6. Bröker LE, Kruyt FAE, Giaccone G (2005) Cell death independent of caspases: a review. Clin Cancer Res 11:3155–3162CrossRefGoogle Scholar
  7. Cao Y, He S, Zhou Z, Zhang M, Mao W, Zhang H, Yao B (2012) Orally administered thermostable N-acyl homoserine lactonase from Bacillus sp. strain AI96 attenuates Aeromonas hydrophila infection in zebrafish. Appl Environ Microbiol 78:1899–1908CrossRefGoogle Scholar
  8. Cerezuela R, Fumanal M, Tapia-Paniagua ST, Meseguer J, Morinigo MA, Esteban MA (2012) Histological alterations and microbial ecology of the intestine in gilthead seabream (Sparus aurata L.) fed dietary probiotics and microalgae. Cell Tissue Res 350:477–489CrossRefGoogle Scholar
  9. Chawla-Sarkar M, Lindner DJ, Liu Y-F, Williams BR, Sen GC, Silverman RH, Borden EC (2003) Apoptosis and interferons: role of interferon-stimulated genes as mediators of apoptosis. Apoptosis 8:237–249CrossRefGoogle Scholar
  10. Christofferson DE, Yuan J (2010) Necroptosis as an alternative form of programmed cell death. Curr Opin Cell Biol 22:263–268CrossRefGoogle Scholar
  11. 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
  12. Degterev A, Boyce M, Yuan J (2003) A decade of caspases. Oncogene 22:8543–8567CrossRefGoogle Scholar
  13. Fan T-J, Han L-H, Cong R-S, Liang J (2005) Caspase family proteases and apoptosis. Acta Biochim Biophys Sin 37:719–727CrossRefGoogle Scholar
  14. 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
  15. Florentin A, Arama E (2012) Caspase levels and execution efficiencies determine the apoptotic potential of the cell. J Cell Biol 196:513–527CrossRefGoogle Scholar
  16. 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
  17. Galluzzi L, Joza N, Tasdemir E, Maiuri MC, Hengartner M, Abrams JM, Tavernarakis N, Penninger J, Madeo F, Kroemer G (2008) No death without life: vital functions of apoptotic effectors. Cell Death Differ 15:1113–1123CrossRefGoogle Scholar
  18. Gao D, Xu Z, Zhang X, Zhu C, Wang Y, Min W (2013a) Cadmium triggers kidney cell apoptosis of pursered common carp (Cyprinus carpio) without caspase-8 activation. Dev Comp Immunol 41:728–737CrossRefGoogle Scholar
  19. 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
  20. Ghosh B, Cain KD, Nowak BF, Bridle AR (2016) Microencapsulation of a putative probiotic Enterobacter species, C6-6, to protect rainbow trout, Oncorhynchus mykiss (Walbaum), against bacterial coldwater disease. J Fish Dis 39:1–11CrossRefGoogle Scholar
  21. Gunther C, Neumann H, Neurath MF, Becker C (2013) Apoptosis, necrosis and necroptosis: cell death regulation in the intestinal epithelium. Gut 62:1062–1071CrossRefGoogle Scholar
  22. Gunther C, Buchen B, He GW, Hornef M, Torow N, Neumann H, Wittkopf N, Martini E, Basic M, Bleich A, Watson AJ, Neurath MF, Becker C (2015) Caspase-8 controls the gut response to microbial challenges by Tnf-alpha-dependent and independent pathways. Gut 64:601–610CrossRefGoogle Scholar
  23. Habil N, Al-Murrani W, Beal J, Foey AD (2011) Probiotic bacterial strains differentially modulate macrophage cytokine production in a strain-dependent and cell subset-specific manner. Benef Microbes 2:283–2937CrossRefGoogle Scholar
  24. Jia R, Cao LP, Du JL, Liu YJ, Wang JH, Jeney G, Yin GJ (2014) Grass carp reovirus induces apoptosis and oxidative stress in grass carp (Ctenopharyngodon idellus) kidney cell line. Virus Res 185:77–81CrossRefGoogle Scholar
  25. 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
  26. Kong W, Huang C, Tang Y, Zhang D, Wu Z, Chen X (2017) Effect of Bacillus subtilis on Aeromonas hydrophila-induced intestinal mucosal barrier function damage and inflammation in grass carp (Ctenopharyngodon idella). Sci Rep 7:1588CrossRefGoogle Scholar
  27. Kuranaga E (2011) Caspase signaling in animal development. Dev Growth Diff 53:137–148CrossRefGoogle Scholar
  28. Labbe K, Saleh M (2008) Cell death in the host response to infection. Cell Death Differ 15:1339–1349CrossRefGoogle Scholar
  29. Li H, Zhang X, Qiu Q, An Z, Qi Y, Huang D, Zhang Y (2013) 2,4-Dichlorophenol induces apoptosis in primary hepatocytes of grass carp (Ctenopharyngodon idella) through mitochondrial pathway. Aquat Toxicol.  https://doi.org/10.1016/j.aquatox.2013.05.015 Google Scholar
  30. Maloy KJ, Powrie F (2011) Intestinal homeostasis and its breakdown in inflammatory bowel disease. Nature 474:298–306CrossRefGoogle Scholar
  31. Marchiando AM, Graham WV, Turner JR (2010) Epithelial barriers in homeostasis and disease. Annu Rev Pathol 5:119–144CrossRefGoogle Scholar
  32. Nandi A, Banerjee G, Dan SK, Ghosh K, Ray AK (2017) Probiotic efficiency of Bacillus sp. in Labeo rohita challenged by Aeromonas hydrophila: assessment of stress profile, haemato-biochemical parameters and immune responses. Aquac Res 48:4334–4345CrossRefGoogle Scholar
  33. Nayak SK (2010) Probiotics and immunity: a fish perspective. Fish Shellfish Immunol 29:2–14CrossRefGoogle Scholar
  34. Newaj-Fyzul A, Adesiyun AA, Mutani A, Ramsubhag A, Brunt J, Austin B (2007) Bacillus subtilis AB1 controls Aeromonas infection in rainbow trout (Oncorhynchus mykiss, Walbaum). J Appl Microbiol 103:1699–1706CrossRefGoogle Scholar
  35. 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
  36. Olsen RE, Sundell K, Mayhew TM, Myklebust R, Ringø E (2005) Acute stress alters intestinal function of rainbow trout, Oncorhynchus mykiss (Walbaum). Aquaculture 250:480–495CrossRefGoogle Scholar
  37. Perez-Garijo A (2017) When dying is not the end: apoptotic caspases as drivers of proliferation. Semin Cell Dev Biol.  https://doi.org/10.1016/j.semcdb.2017.11.036 Google Scholar
  38. Perez-Garijo A, Steller H (2015) Spreading the word: non-autonomous effects of apoptosis during development, regeneration and disease. Development 142:3253–3262CrossRefGoogle Scholar
  39. Pirarat N, Pinpimai K, Endo M, Katagiri T, Ponpornpisit A, Chansue N, Maita M (2011) Modulation of intestinal morphology and immunity in Nile tilapia (Oreochromis niloticus) by Lactobacillus rhamnosus GG. Res Vet Sci.  https://doi.org/10.1016/j.rvsc.2011.02.014 Google Scholar
  40. Portt L, Norman G, Clapp C, Greenwood M, Greenwood MT (2011) Anti-apoptosis and cell survival: a review. Biochim Biophys Acta 1813:238–259CrossRefGoogle Scholar
  41. Ranger AM, Malynn BA, Korsmeyer SJ (2001) Mouse models of cell death. Nat Genet 28:113–118CrossRefGoogle Scholar
  42. Ren Y, Li S, Wu Z, Zhou C, Zhang D, Chen X (2017) The influences of Bacillus subtilis on the virulence of Aeromonas hydrophila and expression of luxS gene of both bacteria under co-cultivation. Curr Microbiol 74:718–724CrossRefGoogle Scholar
  43. Salinas I, Meseguer J, Esteban MA (2008) Antiproliferative effects and apoptosis induction by probiotic cytoplasmic extracts in fish cell lines. Vet Microbiol 126:287–294CrossRefGoogle Scholar
  44. Shao J-z, Liu J, L-x Xiang (2004) Aeromonas hydrophila induces apoptosis in Carassius auratus lymphocytes in vitro. Aquaculture 229:11–23CrossRefGoogle Scholar
  45. Song X, Zhao J, Bo Y, Liu Z, Wu K, Gong C (2014) Aeromonas hydrophila induces intestinal inflammation in grass carp (Ctenopharyngodon idella): an experimental model. Aquaculture 434:171–178CrossRefGoogle Scholar
  46. Strasser A, Pellegrini M (2004) T-lymphocyte death during shutdown of an immune response. Trends Immunol 25:610–615CrossRefGoogle Scholar
  47. Tang Y, Han L, Chen X, Xie M, Kong W, Wu Z (2018) Dietary supplementation of probiotic Bacillus subtilis affects antioxidant defenses and immune response in grass carp under Aeromonas hydrophila challenge. Probiotics Antimicrob Proteins.  https://doi.org/10.1007/s12602-018-9409-8 Google Scholar
  48. Thirabunyanon M, Thongwittaya N (2012) Protection activity of a novel probiotic strain of Bacillus subtilis against Salmonella Enteritidis infection. Res Vet Sci 93:74–81CrossRefGoogle Scholar
  49. 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
  50. Torrecillas S, Makol A, Betancor MB, Montero D, Caballero MJ, Sweetman J, Izquierdo M (2013) Enhanced intestinal epithelial barrier health status on European sea bass (Dicentrarchus labrax) fed mannan oligosaccharides. Fish Shellfish Immunol 34:1485–1495CrossRefGoogle Scholar
  51. Wang L, Ge C, Wang J, Dai J, Zhang P, Li Y (2017a) Effects of different combinations of Bacillus on immunity and antioxidant activities in common carp. Aquac Int 25:2091–2099CrossRefGoogle Scholar
  52. Wang X, Hu W, Zhu L, Yang Q (2017b) Bacillus subtilis and surfactin inhibit the transmissible gastroenteritis virus from entering the intestinal epithelial cells. Biosci Rep.  https://doi.org/10.1042/bsr20170082 Google Scholar
  53. 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
  54. 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

Copyright information

© Japanese Society of Fisheries Science 2018

Authors and Affiliations

  1. 1.Hubei Engineering Technology Research Center for Aquatic Animal Diseases Control and Prevention, Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Key Lab of Freshwater Animal Breeding, Ministry of Agriculture, College of FisheriesHuazhong Agricultural UniversityWuhanPeople’s Republic of China

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