Dietary Supplementation of Probiotic Bacillus subtilis Affects Antioxidant Defenses and Immune Response in Grass Carp Under Aeromonas hydrophila Challenge

  • Ying Tang
  • Lei Han
  • Xiaoxuan Chen
  • Mengqi Xie
  • Weiguang Kong
  • Zhixin Wu
Article
  • 16 Downloads

Abstract

This study investigated whether Bacillus subtilis can provide protection for grass carp against oxidative stress damage induced by Aeromonas hydrophila. A total of 240 healthy grass carp (Ctenopharyngodon idellus) (average weight of 71.42 ± 4.36g) were randomly divided into four groups with three replicates: control group, A. hydrophila group, B. subtilis + A. hydrophila group, and A. hydrophila + B. subtilis group. After challenge with A. hydrophila, the lipid oxidative damage, antioxidant defenses, and the gene expression of inflammatory cytokines of the grass carp were investigated. Our results showed that A. hydrophila caused lipid oxidative damage, led to significant decreases in antioxidant defenses, and induced inflammatory responses of grass carp. However, the grass carp group fed the probiotic B. subtilis diet for 42 days before the challenge and the group fed the probiotic B. subtilis diet immediately after the challenge both showed (i) a reduced level of oxidative stress with a decrease in the level of MDA; (ii) an increase in antioxidant defenses, including an increase in total antioxidant capacity (T-AOC), increased activities of SOD and CAT, increased levels of GSH, and upregulated gene expression of antioxidant enzymes (SOD, CAT, and Gpx); and (iii) an improved immune response with the level of antiinflammatory cytokines IL-10 messenger RNA (mRNA) upregulated and the levels of pro-inflammatory cytokines TNF-α, IL-1β, and IL-8 mRNA downregulated. Based on this study, B. subtilis can provide effective protection of fish against oxidative stress damage induced by A. hydrophila infection.

Keywords

Bacillus subtilis Probiotic feed Oral administration Immune response Antioxidant defenses 

Notes

Acknowledgements

This work was supported by the National Natural Science Foundation of China (Nos. 31472310 and 31672683), the Innovation Training Plan of University Students (105042016047), and SRF (2016151).

Compliance with Ethical Standards

Conflict of Interest

The authors declare that there is no conflict of interest.

References

  1. 1.
    Song X, Li SF, Wang CH, Xu JW, Yang QL (2009) Grass carp (Ctenopharyngodon idellus) genetic structure analysis among native populations in China and introduced populations in USA, Europe and Japan based on mitochondrial sequence. Acta Hydrobiol Sin 33(4):709–716 (Chinese) CrossRefGoogle Scholar
  2. 2.
    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
  3. 3.
    Wu Z-Q, Jiang C, Ling F, Wang G-X (2015) Effects of dietary supplementation of intestinal autochthonous bacteria on the innate immunity and disease resistance of grass carp (Ctenopharyngodon idellus). Aquaculture 438:105–114CrossRefGoogle Scholar
  4. 4.
    Sahoo PK, Rauta PR, Mohanty BR, Mahapatra KD, Saha JN, Rye M, Eknath AE (2011) Selection for improved resistance to Aeromonas hydrophila in Indian major carp labeo rohita: survival and innate immune responses in first generation of resistant and susceptible lines. Fish Shellfish Immunol 31(3):432–438CrossRefGoogle Scholar
  5. 5.
    Chen CZ, Zhang XJ (1999) Detection and analysis of enteritis pathogen of grass carp. Chin J Vet Sci Technol 29(1):5–7 (Chinese) Google Scholar
  6. 6.
    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(3):921–971Google Scholar
  7. 7.
    Banerjee G, Ray AK (2017) The advancement of probiotics research and its application in fish farming industries. Res Vet Sci 115:66–77CrossRefGoogle Scholar
  8. 8.
    Seddik HA, Bendali F, Gancel F, Fliss I, Spano G, Drider D (2017) Lactobacillus plantarum and its probiotic and food potentialities. Probiotics Antimicro Prot 9(2):111–122CrossRefGoogle Scholar
  9. 9.
    Livingstone DR (2001) Contaminant-stimulated reactive oxygen species production and oxidative damage in aquatic organisms. Mar Pollut Bull 42(8):656–666CrossRefGoogle Scholar
  10. 10.
    Kassahn KS, Crozier RH, Portner HO, Caley MJ (2009) Animal performance and stress: responses and tolerance limits at different levels of biological organisation. Biol Rev Camb Philos Soc 84(2):277–292CrossRefGoogle Scholar
  11. 11.
    Moriarty DJW (1998) Control of luminous Vibrio species in penaeid aquaculture ponds. Aquaculture 164(1–4):351–358CrossRefGoogle Scholar
  12. 12.
    Liu H, Wang S, Cai Y, Guo X, Cao Z, Zhang Y, Liu S, Yuan W, Zhu W, Zheng Y, Xie Z, Guo W, Zhou Y (2017) Dietary administration of Bacillus subtilis HAINUP40 enhances growth, digestive enzyme activities, innate immune responses and disease resistance of tilapia, Oreochromis niloticus. Fish Shellfish Immunol 60:326–333CrossRefGoogle Scholar
  13. 13.
    Shen W-Y, Fu L-L, Li W-F, Zhu Y-R (2010) Effect of dietary supplementation with Bacillus subtilis on the growth, performance, immune response and antioxidant activities of the shrimp (Litopenaeus vannamei). Aquac Res 41(11):1691–1698CrossRefGoogle Scholar
  14. 14.
    Ai Q, Xu H, Mai K, Xu W, Wang J, Zhang W (2011) Effects of dietary supplementation of Bacillus subtilis and fructooligosaccharide on growth performance, survival, non-specific immune response and disease resistance of juvenile large yellow croaker, Larimichthys crocea. Aquaculture 317(1):155–161CrossRefGoogle Scholar
  15. 15.
    Nayak SK (2010) Probiotics and immunity: a fish perspective. Fish Shellfish Immunol 29(1):2–14CrossRefGoogle Scholar
  16. 16.
    Li WF, Zhang CN, Song WH, Deng B, Liang Q, Fu LQ et al (2012) Effects of Bacillus preparations on immunity and antioxidant activities in grass carp (Ctenopharyngodon idellus). Fish Physiol Biochem 38(6):1585–1592CrossRefGoogle Scholar
  17. 17.
    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(1):1588CrossRefGoogle Scholar
  18. 18.
    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(6):718–724CrossRefGoogle Scholar
  19. 19.
    Feng X, Wu ZX, Zhu DM, Wang Y, Pang SF, Yu YM, Chen XX (2008) Study on digestive enzyme-producing bacteria from the digestive tract of Ctenopharyngodon idellus and Carassius auratus gibelio. Freshw Fish 38:51–57 (Chinese) Google Scholar
  20. 20.
    Wu ZX, Feng X, Xie LL, Peng XY, Yuan J, Chen XX (2012) Effect of probiotic Bacillus subtilis Ch9 for grass carp, Ctenopharyngodon Idella (Valenciennes, 1844), on growth performance, digestive enzyme activities and intestinal microflora. J Appl Ichthyol 28(5):721–727CrossRefGoogle Scholar
  21. 21.
    Kong WG, Li SS, Chen XX, Huang YQ, Tang Y, Wu ZX (2017) A study of the damage of the intestinal mucosa barrier structure and function of Ctenopharyngodon idella with Aeromonas hydrophila. Fish Physiol Biochem 19:1–13Google Scholar
  22. 22.
    Dawood MA, Koshio S, Ishikawa M, Yokoyama S, El Basuini MF, Hossain MS, Moss AS (2016) Effects of dietary supplementation of Lactobacillus rhamnosus or/and Lactococcus lactis on the growth, gut microbiota and immune responses of red sea bream, Pagrus major. Fish Shellfish Immunol 49:275–285CrossRefGoogle Scholar
  23. 23.
    Söderhäll K, Cerenius L (1998) Role of the prophenoloxidase-activating system in invertebrate immunity. Curr Opin Immunol 10(1):23–28CrossRefGoogle Scholar
  24. 24.
    Livingstone DR, Lips F, Martinez PG, Pipe RK (1992) Antioxidant enzymes in the digestive gland of the common mussel Mytilus edulis. Mar Biol 112(2):265–276CrossRefGoogle Scholar
  25. 25.
    Wang HL, Wen HS, Zhang XY (2016) Effects of salinity stress on antioxidant enzyme and non-specific immunity activities in the intestine of juvenile lateolabrax maculatus. Mod Agric Sci Technol 4:236–265 (Chinese) Google Scholar
  26. 26.
    Adeyemi JA (2014) Oxidative stress and antioxidant enzymes activities in the African catfish, Clarias gariepinus, experimentally challenged with Escherichia coli and Vibrio fischeri. Fish Physiol Biochem 40(2):347–354CrossRefGoogle Scholar
  27. 27.
    Aiassa V, Barnes AI, Albesa I (2010) Resistance to ciprofloxacin by enhancement of antioxidant defenses in biofilm and planktonic Proteus mirabilis. Biochem Biophys Res Commun 393(1):84–88CrossRefGoogle Scholar
  28. 28.
    Krzyminska S, Tanska A, Kaznowski A (2011) Aeromonas spp. induce apoptosis of epithelial cells through an oxidant-dependent activation of the mitochondrial pathway. J Med Microbiol 60(Pt 7):889–898CrossRefGoogle Scholar
  29. 29.
    Mathew S, Kumar KA, Anandan R, Viswanathan Nair PG, Devadasan K (2007) Changes in tissue defence system in white spot syndrome virus (WSSV) infected Penaeus monodon. Comp Biochem Physiol C Toxicol Pharmacol 145(3):315–320CrossRefGoogle Scholar
  30. 30.
    Sarathi M, Ahmed VPI, Venkatesan C, Balasubramanian G, Prabavathy J, Hameed ASS (2007) Comparative study on immune response of Fenneropenaeus indicus to Vibrio alginolyticus and white spot syndrome virus. Aquaculture 271(1–4):8–20CrossRefGoogle Scholar
  31. 31.
    Banerjee G, Ray AK, Pandey S, Kumar R (2016) An alternative approach of toxic heavy metal removal by Arthrobacter phenanthrenivorans: assessment of surfactant production and oxidative stress. Curr Sci 110:1–5CrossRefGoogle Scholar
  32. 32.
    Nogueira CW, Quinhones EB, Jung EAC, Zeni G, Rocha JBT (2003) Anti-inflammatory and antinociceptive activity of diphenyl diselenide. Inflamm Res 52(2):56–63CrossRefGoogle Scholar
  33. 33.
    Falcón RM, Carvalho HF, Joazeiro PP, Gatti MSV, Yano T (2001) Induction of apoptosis in HT29 human intestinal epithelial cells by the cytotoxic enterotoxin of Aeromonas hydrophila. Biochem Cell Biol 79(4):525–531CrossRefGoogle Scholar
  34. 34.
    Esteban MA, Cordero H, Martinez-Tome M, Jimenez-Monreal AM, Bakhrouf A, Mahdhi A (2014) Effect of dietary supplementation of probiotics and palm fruits extracts on the antioxidant enzyme gene expression in the mucosae of gilthead seabream (Sparus aurata L.) Fish Shellfish Immunol 39(2):532–540CrossRefGoogle Scholar
  35. 35.
    Cascon A, Yugueros J, Temprano A, Sanchez M, Hernanz C, Luengo JM, Naharro G (2000) A major screted elastase is essential for pathogenicity of Aeromonas hydrophila. Infect Immun 68(6):3233–3241CrossRefGoogle Scholar
  36. 36.
    Geng X, Dong XH, Tan BP, Yang QH, Chi SY, Liu HY, Liu XQ (2011) Effects of dietary chitosan and Bacillus subtilis on the growth performance, non-specific immunity and disease resistance of cobia, Rachycentron canadum. Fish Shellfish Immunol 31(3):400–406CrossRefGoogle Scholar
  37. 37.
    Giri SS, Sukumaran V, Sen SS, Jena PK (2013) Effects of dietary supplementation of potential probiotic Bacillus subtilis VSG1 singularly or in combination with Lactobacillus plantarum VSG3 or/and Pseudomonas aeruginosa VSG2 on the growth, immunity and disease resistance of Labeo rohita. Aquac Nutr 20(2):163–171CrossRefGoogle Scholar
  38. 38.
    Zhang CN, Li XF, Xu WN, Jiang GZ, Lu KL, Wang LN, Liu WB (2013) Combined effects of dietary fructooligosaccharide and Bacillus licheniformis on innate immunity, antioxidant capability and disease resistance of triangular bream (Megalobrama terminalis). Fish Shellfish Immunol 35(5):1380–1386CrossRefGoogle Scholar
  39. 39.
    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
  40. 40.
    Chiu CH, Guu YK, Liu CH, Pan TM, Cheng W (2007) Immune responses and gene expression in white shrimp, Litopenaeus vannamei, induced by Lactobacillus plantarum. Fish Shellfish Immunol 23(2):364–377CrossRefGoogle Scholar
  41. 41.
    Gullian M, Thompson F, Rodriguez J (2004) Selection of probiotic bacteria and study of their immunostimulatory effect in Penaeus vannamei. Aquaculture 233(1–4):1–14CrossRefGoogle Scholar
  42. 42.
    Zhou JM, Wu ZX, Zeng LB, Chen XX, Yang L, Zhao Y (2012) Selection and characterization of pathogen antagonised probiotics from intestinal tract of yellow catfish (Pelteobagrus fulvidraco). Acta Hydrobiol Sin 36:78–84 (Chinese)Google Scholar
  43. 43.
    Aly SM, Abdel-Galil Ahmed Y, Abdel-Aziz Ghareeb A, Mohamed MF (2008) Studies on Bacillus subtilis and Lactobacillus acidophilus, as potential probiotics, on the immune response and resistance of Tilapia nilotica (Oreochromis niloticus) to challenge infections. Fish Shellfish Immunol 25(1–2):128–136CrossRefGoogle Scholar
  44. 44.
    Giri SS, Sukumaran V, Dangi NK (2012) Characteristics of bacterial isolates from the gut of freshwater fish, Labeo rohita, that may be useful as potential probiotic bacteria. Probiotics Antimicro Prot 4(4):238–242CrossRefGoogle Scholar
  45. 45.
    Ramesh D, Vinothkanna A, Rai AK, Vignesh VS (2015) Isolation of potential probiotic Bacillus spp. and assessment of their subcellular components to induce immune responses in Labeo rohita against Aeromonas hydrophila. Fish Shellfish Immunol 45(2):268–276CrossRefGoogle Scholar
  46. 46.
    Sugita H, Hirose Y, Matsuo N, Deguchi Y (1998) Production of the antibacterial substance by Bacillus sp. strain NM 12, an intestinal bacterium of Japanese coastal fish. Aquaculture 165(3–4):269–280CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.College of FisheriesHuazhong Agricultural UniversityWuhanPeople’s Republic of China
  2. 2.Hubei Engineering Technology Research Center for Aquatic Animal Diseases Control and PreventionWuhanPeople’s Republic of China
  3. 3.Hubei Provincial Engineering Laboratory for Pond AquacultureWuhanPeople’s Republic of China
  4. 4.Key Lab of Freshwater Animal BreedingMinistry of AgricultureWuhanPeople’s Republic of China

Personalised recommendations