Dietary supplementation of Streptococcus faecalis benefits the feed utilization, antioxidant capability, innate immunity, and disease resistance of blunt snout bream (Megalobrama amblycephala)

  • Xiao-Qun Zhong
  • Ming-Yang Liu
  • Chao Xu
  • Wen-Bin Liu
  • Kenneth-Prudence Abasubong
  • Xiang-Fei LiEmail author


This study aimed to investigate the effects of Streptococcus faecalis on the growth performance, intestinal histology, antioxidant capability, innate immunity, and disease resistance of blunt snout bream Megalobrama amblycephala. Fish were fed five experimental diets containing 0 (SF0, control), 1 × 105 (SF1), 1 × 106 (SF2), 1 × 107 (SF3), and 1 × 108 cfu/g (SF4) of Streptococcus faecalis, respectively, for 10 weeks. After the feeding trial, fish were challenged by Aeromonas hydrophila with the cumulative mortality recorded during a period of 96 h. The results showed that WG and FI of fish both showed no significant difference (P > 0.05) among all the treatments. However, the FCR was significantly (P < 0.05) affected by Streptococcus faecalis levels with the lowest value observed in the SF2 group, whereas the opposite was true for intestinal microvillus length (P < 0.05). Dietary supplementation of 1 × 106 cfu/g Streptococcus faecalis significantly (P < 0.05) increased the hepatic activities of SOD, CAT, and GPx; plasma activities of LZM, MPO, ACP, and AKP; and the levels of C3, C4, and IgM of fish, compared with the control group. Similar results were also observed in the tissue expressions of Leap-I, Leap-II, muc2, and muc5b (P < 0.05), whereas the opposite was true for liver MDA contents and plasma NO levels (P < 0.05). At 96 h after challenge, the cumulative mortality of the control was significantly (P < 0.05) higher than that of the SF2 group, but it showed no statistical difference (P > 0.05) with that of the other treatments. These results indicated that dietary supplementation of 1 × 106 cfu/g Streptococcus faecalis could not only improve the feed utilization of blunt snout bream but also enhance its antioxidant capability, innate immunity, and disease resistance.


Streptococcus faecalis Growth Innate immunity Disease resistance Blunt snout bream 



The authors also wanted to thank the following people for their technical assistance and help during fish husbandry and sampling: Xiufei Cao, Huajuan Shi, Jie Liu, Huihui Chen, Jiadai Liu, and Chenyuan Xu.

Funding information

The present study was funded by the National Technology System for Conventional Freshwater Fish Industries of China (CARS-45-14).


  1. Abid A, Davies SJ, Waines P, Emery M, Castex M, Gioacchini G, Carnevali O, Bickerdike R, Romero J, Merrifield DL (2013) Dietary synbiotic application modulates Atlantic salmon (Salmo salar) intestinal microbial communities and intestinal immunity. Fish Shellfish Immunol 35:1948–1956CrossRefGoogle Scholar
  2. Alexander JB, Ingram GA (1992) Noncellular nonspecific defence mechanisms of fish. Annu Rev Fish Dis 2:249–279CrossRefGoogle Scholar
  3. Allameh SK, Ringø E, Yusoff FM, Daud HM, Ideris A (2017) Dietary supplement of Enterococcus faecalis on digestive enzyme activities, short-chain fatty acid production, immune system response and disease resistance of Javanese carp (Puntius gonionotus, Bleeker 1850). Aquac Nutr 23:331–338CrossRefGoogle Scholar
  4. AOAC (1995) Agricultural chemicals; contaminants, drugs. In: official methods of analysis of AOAC International. AOAC International, Arlington, p 1298Google Scholar
  5. Buisine MP, Devisme L, Maunoury V, Deschodt E, Gosselin B, Copin MC et al (2000) Developmental mucin gene expression in the gastroduodenal tract and accessory digestive glands. I Stomach A relationship to gastric carcinoma. J Histochem Cytochem 48:1657–1666CrossRefGoogle Scholar
  6. Buisine MP, Desreumaux P, Leteurtre E, Copin MC, Colombel JF, Porcht NJ, Aubert P (2001) Mucin gene expression in intestinal epithelial cells in Crohn’s disease. Gut 49:544–551CrossRefGoogle Scholar
  7. Chang CI, Pleguezuelos O, Zhang YA, Zou J, Secombes CJ (2005) Identification of a novel cathelicidin gene in the rainbow trout, Oncorhynchus mykiss. Infect Immun 73:5053–5064CrossRefGoogle Scholar
  8. Cheng TC (1989) Immunology deficiency disease in marine mollusks: measurements of some variables. J Aquat Anim Health 1:209–216Google Scholar
  9. Chia TJ, Wu YC, Chen JY, Chi SC (2010) Antimicrobial peptides (AMP) with antiviral activity against fish nodavirus. Fish Shellfish Immunol 28:434–439CrossRefGoogle Scholar
  10. Cox CM, Sumners LH, Kim S, Mcelroy AP, Bedford MR, Dalloul RA (2010) Immune responses to dietary β-glucan in broiler chicks during an Eimeria challenge. Poult Sci 89:2597–2607CrossRefGoogle Scholar
  11. Dai ZL, Wu G, Zhu WY (2011) Amino acid metabolism in intestinal bacteria: links between gut ecology and host health. Front Biosci 16:1768–1786CrossRefGoogle Scholar
  12. Du Y, Yi M, Xiao P, Meng L, Li X, Sun G et al (2015) The impact of Aeromonas salmonicida, infection on innate immune parameters of Atlantic salmon (Salmo salar L.). Fish Shellfish Immunol 44:307–315CrossRefGoogle Scholar
  13. Ellis AE (2002) Immunity to bacteria in fish. Fish Shellfish Immunol 9:291–308CrossRefGoogle Scholar
  14. Eslamloo K, Akhavan SR, Henry MA (2013) Effects of dietary administration of Bacillus probiotics on the non-specific immune responses of tinfoil barb, Barbonymus schwanenfeldii (Actinopterygii: Cypriniformes: Cyprinidae). Acta Ichthyol Piscat 43:211–218CrossRefGoogle Scholar
  15. Esteban MA, Cordero H, Martínez-Tomé M, Jiménez-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 39:532–540CrossRefGoogle Scholar
  16. Gao Z, Luo W, Liu H, Zeng C, Liu X, Yi S et al (2012) Transcriptome analysis and SSR/SNP markers information of the blunt snout bream (Megalobrama amblycephala). PLoS One 7:e42637CrossRefGoogle Scholar
  17. González R, Brokordt K, Cárcamo CB, Tc DLP, Oyanedel D, Mercado L et al (2017) Molecular characterization and protein localization of the antimicrobial peptide big defensin from the scallop Argopecten purpuratus after Vibrio splendidus challenge. Fish Shellfish Immunol 68:173–179CrossRefGoogle Scholar
  18. Guardiola FA, Porcino C, Cerezuela R, Cuesta A, Faggio C, Esteban MA (2016) Impact of date palm fruits extracts and probiotic enriched diet on antioxidant status, innate immune response and immune-related gene expression of European seabass (Dicentrarchus labrax). Fish Shellfish Immunol 52:298–308CrossRefGoogle Scholar
  19. Han B, Long WQ, He JY, Liu YJ, Si YQ, Tian LX (2015) Effects of dietary Bacillus licheniformis on growth performance, immunological parameters, intestinal morphology and resistance of juvenile Nile tilapia (Oreochromis niloticus) to challenge infections. Fish Shellfish Immunol 46:225–231CrossRefGoogle Scholar
  20. Hara N (1975) Studies on the establishment of multi-drug-resistant strain BIO-4R of Streptococcus faecalis in the intestinal tract of germ-free mice: bacterial interaction and effect of antibiotics. Jpn J Microbiol 19:249–254CrossRefGoogle Scholar
  21. Hollingsworth MA, Swanson BJ (2004) Mucins in cancer: protection and control of the cell surface. Nat Rev Cancer 4:45–60CrossRefGoogle Scholar
  22. Hoseinifar SH, Sun Y, Caipang CM (2016) Short-chain fatty acids as feed supplements for sustainable aquaculture: an updated view. Aquac Res 48:1380–1391CrossRefGoogle Scholar
  23. Hoseinifar SH, Ahmadi A, Khalili M, Raeisi M, Doan HV, Caipang CM (2017a) The study of antioxidant enzymes and immune-related genes expression in common carp (Cyprinus carpio) fingerlings fed different prebiotics. Aquac Res 48(11):5447–5454CrossRefGoogle Scholar
  24. Hoseinifar SH, Dadar M, Ringø E (2017b) Modulation of nutrient digestibility and digestive enzyme activities in aquatic animals: the functional feed additives scenario. Aquac Res 48:3987–4000CrossRefGoogle Scholar
  25. Hoseinifar SH, Hoseini SM, Bagheri D (2017c) Effects of galactooligosaccharide and Pediococcus acidilactici on antioxidant defence and disease resistance of rainbow trout, Oncorhynchus mykiss. Ann Anim Sci 17(1):217–227CrossRefGoogle Scholar
  26. Hu CH, Xu Y, Xia MS, Xiong L, Xu ZR (2007) Effects of Cu2+-exchanged montmorillonite on growth performance, microbial ecology and intestinal morphology of Nile tilapia (Oreochromis niloticus). Aquaculture 270:200–206CrossRefGoogle Scholar
  27. Huang W, Li HQ, Luo L, Chen YJ, Bai FJ, Lin K (2017) Effect of Enterococcus faecalis on growth performance, body composition, physiological and biochemical indexes and digestive enzyme of GIFT tilapia (Oreochromis niloticus). J Fish China 41:1–9 (in Chinese with English abstract)Google Scholar
  28. Kim YR, Kim EY, Choi SY, Hossain MT, Oh R, Heo WS, Lee JM, Cho YC, Kong IS (2012) Effect of a probiotic strain, Enterococcus faecium, on the immune responses of olive flounder (Paralichthys olivaceus). J Microbiol Biotechnol 22:526–529CrossRefGoogle Scholar
  29. Lee S, Katya K, Park Y, Won S, Seong M, Hamidoghli A et al (2017) Comparative evaluation of dietary probiotics Bacillus subtilis, wb60 and lactobacillus plantarum, KCTC3928 on the growth performance, immunological parameters, gut morphology and disease resistance in Japanese eel, Anguilla japonica. Fish Shellfish Immunol 61:201–210CrossRefGoogle Scholar
  30. Li XF, Liu WB, Jiang YY, Zhu H, Ge XP (2010) Effects of dietary protein and lipid levels in practical diets on growth performance and body composition of blunt snout bream (Megalobrama amblycephala) fingerlings. Aquaculture 303:65–70CrossRefGoogle Scholar
  31. Li XF, Liu WB, Lu KL, Xu WN, Wang Y (2012) Dietary carbohydrate/lipid ratios affect stress, oxidative status and non-specific immune responses of fingerling blunt snout bream, Megalobrama amblycephala. Fish Shellfish Immunol 33:316–323CrossRefGoogle Scholar
  32. Li X, Ringø E, Hoseinifar SH, Lauzon HL, Birkbeck H, Yang D (2018) The adherence and colonization of microorganisms in fish gastrointestinal tract. Rev Aquac 10:1–45CrossRefGoogle Scholar
  33. Liang T, Ji W, Zhang GR, Wei KJ, Feng K, Wang WM, Zou GW (2013) Molecular cloning and expression analysis of liver-expressed antimicrobial peptide 1 (LEAP-1) and LEAP-2 genes in the blunt snout bream (Megalobrama amblycephala). Fish Shellfish Immunol 35:553–563CrossRefGoogle Scholar
  34. Liu S, Dong XF, Tong JM, Bao Y, Wang ZH (2017) Effects of dietary Enterococcus faecalis on performance, egg quality, lipid metabolism and intestinal microflora numbers of laying hens. Chin J Anim Nutr 29:202–213 (in Chinese with English abstract)Google Scholar
  35. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods 25:402–408CrossRefGoogle Scholar
  36. Marel MVD, Adamek M, Gonzalez SF, Frost P, Rombout JHWM, Wiegertjes GF et al (2012) Molecular cloning and expression of two β-defensin and two mucin genes in common carp (Cypinus carpio L.) and their up-regulation after β-glucan feeding. Fish Shellfish Immunol 32:494–501CrossRefGoogle Scholar
  37. Masso-Silva JA, Diamond G (2014) Antimicrobial peptides from fish. Pharmaceuticals 7:265–310CrossRefGoogle Scholar
  38. Merrifield DL, Dimitroglou A, Bradley G, Baker RT, Davies SJ (2009) Soybean meal alters autochthonous microbial populations, microvilli morphology and compromises intestinal enterocyte integrity of rainbow trout, Oncorhynchus mykiss (Walbaum). J Fish Dis 32:755–766CrossRefGoogle Scholar
  39. Mishra V, Shah C, Mokashe N, Chavan R, Yadav H, Prajapati J (2015) Probiotics as potential antioxidants: a systematic review. J Agric Food Chem 63:3615–3626CrossRefGoogle Scholar
  40. Moniaux N, Andrianifahanana M, Brand RE, Batra SK (2004) Multiple roles of mucins in pancreatic cancer, a lethal and challenging malignancy. Br J Cancer 91:1633–1638CrossRefGoogle Scholar
  41. Nawaz A, Bakhsh AJ, Irshad S, Hoseinifar SH, Xiong H (2018) The functionality of prebiotics as immunostimulant: evidences from trials on terrestrial and aquatic animals. Fish Shellfish Immunol 76:272–278CrossRefGoogle Scholar
  42. Nutten S, Sansonetti P, Huet G, Bisiaux CB, Meresse B, Colombel JF (2002) Desreumaux P, epithelial inflammation response induced by Shigella flexneri depends on mucin gene expression. Microbes Infect 4:1121–1124CrossRefGoogle Scholar
  43. Ozawa K (1985) Growth of Enterococcus faecalis (Streptococcus faecalis) BIO-4R in the intestinal tract of animals and humans. Biosci Microflora 4:15–22Google Scholar
  44. Ozawa K, Yabu-Uchi K, Yamanaka K, Yamashita Y, Nomura S, Oku I (1983) Effect of Streptococcus faecalis BIO-4R on intestinal flora of weanling piglets and calves. Appl Environ Microbiol 45:1513–1518PubMedPubMedCentralGoogle Scholar
  45. Panigrahi A, Kiron V, Satoh S, Hirono I, Kobayashi T, Sugita H, Puangkaew J, Aoki T (2007) Immune modulation and expression of cytokine genes in rainbow trout Oncorhynchus mykiss, upon probiotic feeding. Dev Comp Immunol 31:372–382CrossRefGoogle Scholar
  46. Rajanbabu V, Chen JY (2011) Applications of antimicrobial peptides from fish and perspectives for the future. Peptides 32:415–420CrossRefGoogle Scholar
  47. Rausch PG, Moore TG (1975) Granule enzymes of polymorphonuclear neutrophils: a phylogenetic comparison. Blood 46:913–919PubMedGoogle Scholar
  48. Ringø E, Olsen RE, Vecino JLG, Wadsworth S (2012) Use of immunostimulants and nucleotides in aquaculture: a review. J Mar Sci Res Dev 2:1–22Google Scholar
  49. Ringø E, Hoseinifar SH, Ghosh K, Doan HV, Beek BR, Song SK (2018) Lactic acid bacteria in finfish-an update. Front Microbiol 9:1–37CrossRefGoogle Scholar
  50. Sang YL, Nam YK (2017) Gene structure and expression characteristics of liver-expressed antimicrobial peptide-2 isoforms in mud loach (Misgurnus mizolepis, Cypriniformes). Fish Aquat Sci 20:31–41CrossRefGoogle Scholar
  51. Scharek L, Guth J, Reiter K, Weyrauch KD, Taras D, Schwerk P, Schierack P, Schmidt MFG, Wieler LH, Tedin K (2005) Influence of a probiotic Enterococcus faecium strain on development of the immune system of sows and piglets. Vet Immunol Immunopathol 105:151–161CrossRefGoogle Scholar
  52. Schlapbach C, Yawalkar N, Hunger RE (2009) Human β-defensin-2 and psoriasin are over expressed in lesions of acne inverse. J Am Acad Dermatol 61:58–65CrossRefGoogle Scholar
  53. Selim KM, Reda RM (2015) Improvement of immunity and disease resistance in the Nile tilapia, Oreochromis niloticus, by dietary supplementation with Bacillus amyloliquefaciens. Fish Shellfish Immunol 44:496–503CrossRefGoogle Scholar
  54. Shabir U, Ali S, Magray AR, Ganai BA, Firdous P, Hassan T, Nazir R (2018) Fish antimicrobial peptides (AMP's) as essential and promising molecular therapeutic agents: a review. Microb Pathog 114:50–56CrossRefGoogle Scholar
  55. Shaheen AA, Eissa N, Abouelgheit EN, Yao H, Wang HP (2014) Probiotic effect on molecular antioxidant profiles in yellow perch, Perca flavescens. Glob J Fish Aquacult Res 1:16–29Google Scholar
  56. Sheng YH, Hasnain SZ, Florin THJ, Mcguckin MA (2012) Mucins in inflammatory bowel diseases and colorectal cancer. J Gastroenterol Hepatol 27:28–38CrossRefGoogle Scholar
  57. Silphaduang U, Colorni A, Noga EJ (2006) Evidence for widespread distribution of piscidin antimicrobial peptides in teleost fish. Dis Aquat Org 72:241–252CrossRefGoogle Scholar
  58. Singh R, Kumar V, Kapoor V (2014) Isolation and screening of amylolytic and xylolytic actinomycetes from assorted habitats. Indo Am J Pharmaceut Res 4:2122–2132Google Scholar
  59. Smith AG, O'Doherty JV, Reilly P, Ryan MT, Bahar B, Sweeney T (2011) The effects of laminarin derived from Laminaria digitata on measurements of gut health: selected bacterial populations, intestinal fermentation, mucin gene expression and cytokine gene expression in the pig. Brit J Nutr 105:669–677CrossRefGoogle Scholar
  60. Srivastava A, Nigam AK, Mittal S, Mittal AK (2018) Role of aloin in the modulation of certain immune parameters in skin mucus of an Indian major carp, Labeo rohita. Fish Shellfish Immunol 73:252–261CrossRefGoogle Scholar
  61. Sun YZ, Yang HL, MA R-L, Song K, LI J-S (2012) Effect of Lactococcus lactis and Enterococcus faecium on growth performance, digestive enzymes and immune response of grouper Epinephelus coioides. Aquac Nutr 18:281–289CrossRefGoogle Scholar
  62. Talafantová M, Mandel L (1985) Protective activity of Streptococcus faecalis against pathogenic action of Escherichia coli O55 in gnotobiotic pigs. Folia Microbiol 30:329–333CrossRefGoogle Scholar
  63. Tian HY, Zhang DD, Li XF, Zhang CN, Qian Y, Liu WB (2015) Optimum feeding frequency of juvenile blunt snout bream (Megalobrama amblycephala). Aquaculture 437:60–66CrossRefGoogle Scholar
  64. Udayangani RMC, Dananjaya SHS, Nikapitiya C, Heo GJ, Lee J, Zoysa MD (2017) Metagenomics analysis of gut microbiota and immune modulation in zebrafish (Danio rerio) fed chitosan silver nanocomposites. Fish Shellfish Immunol 66:173–184CrossRefGoogle Scholar
  65. Uriel RE, Satoh S, Haga Y, Fushimi H, Sweetman J (2009) Effects of single and combined supplementation of Enterococcus faecalis, mannan oligosaccharide and polyhydroxybutyrate acid on growth performance and immune response of rainbow trout Oncorhynchus mykiss. Aquacult Sci 57:609–614Google Scholar
  66. Xu WN, Chen DH, Chen QQ, Liu WB (2017a) Growth performance, innate immune responses and disease resistance of fingerling blunt snout bream, Megalobrama amblycephala adapted to different berberine-dietary feeding modes. Fish Shellfish Immunol 68:458–465CrossRefGoogle Scholar
  67. Xu C, Liu WB, Zhang DD, Wang KZ, Xia SL, Li XF (2017b) Molecular characterization of AMP-activated protein kinase α2 from herbivorous fish Megalobrama amblycephala and responsiveness to glucose loading and dietary carbohydrate levels. Comp Biochem Physiol A Mol Integr Physiol 208:24–34CrossRefGoogle Scholar
  68. Xue CY, Xi BW, Ren M, Dong J, Xie J, Xu P (2015) Molecular cloning, tissue expression of gene Muc2 in blunt snout bream Megalobrama amblycephala and regulation after re-feeding. Chin J Oceanol Limnol 33:291–298CrossRefGoogle Scholar
  69. Yang HL, Xia HQ, Ye YD, Zou WC, Sun YZ (2014) Probiotic Bacillus pumilus SE5 shapes the intestinal microbiota and mucosal immunity in grouper Epinephelus coioides. Dis Aquat Org 111:119–127CrossRefGoogle Scholar
  70. Yuan XY, Liu WB, Liang C, Sun CX, Xue YF, Wan ZD, Jiang GZ (2017) Effects of partial replacement of fish meal by yeast hydrolysate on complement system and stress resistance in juvenile Jian carp (Cyprinus carpio var. Jian). Fish Shellfish Immunol 67:312–321CrossRefGoogle Scholar
  71. 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 35:1380–1386CrossRefGoogle Scholar
  72. Zhang CN, Li XF, Jiang GZ, Zhang DD, Tian HY, Li JY, Liu WB (2014a) Effects of dietary fructooligosaccharide levels and feeding modes on growth, immune responses, antioxidant capability and disease resistance of blunt snout bream (Megalobrama amblycephala). Fish Shellfish Immunol 41:560–569CrossRefGoogle Scholar
  73. Zhang YY, Liu B, Ge XP, Liu WB, Xie J, Ren M, Cui YT, Xia SL, Chen R, Zhou Q, Pan L, Yu Y (2014b) The influence of various feeding patterns of emodin on growth, non-specific immune responses, and disease resistance to Aeromonas hydrophila in juvenile Wuchang bream (Megalobrama amblycephala). Fish Shellfish Immunol 36:187–193CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Xiao-Qun Zhong
    • 1
  • Ming-Yang Liu
    • 2
  • Chao Xu
    • 1
  • Wen-Bin Liu
    • 1
  • Kenneth-Prudence Abasubong
    • 1
  • Xiang-Fei Li
    • 1
    Email author
  1. 1.Key Laboratory of Aquatic Nutrition and Feed Science of Jiangsu Province, College of Animal Science and TechnologyNanjing Agricultural UniversityNanjingChina
  2. 2.Wuxi Fisheries CollegeNanjing Agricultural UniversityWuxiChina

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