Abstract
A 60-day feeding trial was conducted to investigate the effects of dietary Chinese herbal medicine (rhubarb, Rheum officinale Bail), probiotic (Bacillus cereus BC-01), prebiotic (yeast polysaccharide), and antibiotic (florfenicol) supplementation on the growth, intestinal morphology, and immune responses of juvenile sea cucumbers (Apostichopus japonicus Selenka; initial weight, 4.67 ± 0.06 g). The control treatment was fed only the basal diet (CT treatment), whereas four other treatments were fed the basal diet supplemented with rhubarb (RH treatment), B. cereus BC-01 (BC treatment), yeast polysaccharide (PO treatment), or florfenicol (FL treatment). Groups of ten sea cucumbers were housed in separate tanks, and each diet was fed to five randomly selected groups. The specific growth rates of the individuals in the RH and BC treatments were significantly higher than those in the CT, PO, and FL treatments. The apparent digestibility coefficient of crude protein in the RH and BC treatments was significantly higher than that of crude protein in the CT and PO treatments. The alkaline phosphatase activity of the coelomocytes in the RH and BC treatments was significantly higher than that in the CT and FL treatments. Coelomocytes in the PO treatment had the highest acid phosphatase activity. Higher than average lysozyme activity was observed in the PO and FL treatments. Fold and enterocyte heights in the mid-intestinal tract of sea cucumbers in the CT and RH treatments were lower than those in the BC, PO, and FL treatments. Microvillus height was greatest in the RH treatment and lowest in the FL treatment. Scanning electron microscopy revealed remarkable atrophy of microvilli and apoptosis of intestinal epithelial cells in the FL treatment. These results suggest that dietary rhubarb, B. cereus, and yeast polysaccharide supplementation can improve the health of sea cucumbers, whereas long-term florfenicol supplementation may have a negative impact on their intestinal healthy and immune response of sea cucumbers.
Similar content being viewed by others
References
Ai Q, Mai K, Zhang L, Tan B, Zhang W, Xu W, Li H (2007) Effects of dietary β-1,3 glucan on innate immune response of large yellow croaker, Pseudosciaena crocea. Fish Shellfish Immunol 22(4):394–402
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):155–161
Akinbowale OL, Peng H, Barton MD (2006) Antimicrobial resistance in bacteria isolated from aquaculture sources in Australia. J Appl Microbiol 100(5):1103–1113
Bagni M, Romano N, Finoia M, Abelli L, Scapigliati G, Tiscar P, Sarti M, Marino G (2005) Short- and long-term effects of a dietary yeast β-glucan (Macrogard) and alginic acid (Ergosan) preparation on immune response in sea bass (Dicentrarchus labrax). Fish Shellfish Immunol 18(4):311–325
Bai N, Zhang W, Mai K, Wang X, Xu W, Ma H (2010) Effects of discontinuous administration of β-glucan and glycyrrhizin on the growth and immunity of white shrimp Litopenaeus vannamei. Aquaculture 306(1):218–224
Cerezuela R, Fumanal M, Tapia-Paniagua ST, Meseguer J, Moriñigo MÁ, Esteban MÁ (2013) Changes in intestinal morphology and microbiota caused by dietary administration of inulin and Bacillus subtilis in gilthead sea bream (Sparus aurata L.) specimens. Fish Shellfish Immunol 34(5):1063–1070
Cha JH, Rahimnejad S, Yang SY, Kim KW, Lee KJ (2013) Evaluations of Bacillus spp. as dietary additives on growth performance, innate immunity and disease resistance of olive flounder (Paralichthys olivaceus) against Streptococcus iniae and as water additives. Aquaculture 402:50–57
Chang J, Zhang W, Mai K, Ma H, Xu W (2010) Effects of dietary β-glucan and glycyrrhizin on non-specific immunity and disease resistance of the sea cucumber (Apostichopus japonicus Selenka) challenged with Vibrio splendidus. J Ocean Univ China 9(4):389–394
Chia FS, Xing J (1996) Echinoderm coelomocytes. Zool Stud 35:231–254
Daniels CL, Merrifield DL, Boothroyd DP, Davies SJ, Factor JR, Arnold KE (2010) Effect of dietary Bacillus spp. and mannan oligosaccharides (MOS) on European lobster (Homarus gammarus L.) larvae growth performance, gut morphology and gut microbiota. Aquaculture 304(1):49–57
Dolmatova L, Eliseikina M, Romashina V (2004) Antioxidant enzymatic activity of coelomocytes of the Far East sea cucumber Eupentacta fraudatrix. J Evol Biochem Phys 40(2):126–135
Fink SL, Cookson BT (2007) Pyroptosis and host cell death responses during Salmonella infection. Cell Microbiol 9(11):2562–2570
Fisheries Bureau of Ministry of Agriculture (2013) China fishery statistical yearbook. China Agriculture Press, Beijing (in Chinese)
Fuchs VI, Schmidt J, Slater MJ, Zentek J, Buck BH, Steinhagen D (2015) The effect of supplementation with polysaccharides, nucleotides, acidifiers and Bacillus strains in fish meal and soy bean based diets on growth performance in juvenile turbot (Scophthalmus maximus). Aquaculture 437:243–251
Gaggìa F, Mattarelli P, Biavati B (2010) Probiotics and prebiotics in animal feeding for safe food production. Int J Food Microbiol 141:15–28
Gao QF, Wang Y, Dong S, Sun Z, Wang F (2011) Absorption of different food sources by sea cucumber Apostichopus japonicus (Selenka) (Echinodermata: Holothuroidea): evidence from carbon stable isotope. Aquaculture 319(1):272–276
Hao K, Liu JY, Ling F, Liu XL, Lu L, Xia L, Wang GX (2014) Effects of dietary administration of Shewanella haliotis D4, Bacillus cereus D7 and Aeromonas bivalvium D15, single or combined, on the growth, innate immunity and disease resistance of shrimp, Litopenaeus vannamei. Aquaculture 428:141–149
Hooton D, Lentle R, Monro J, Wickham M, Simpson R (2015) The secretion and action of brush border enzymes in the mammalian small intestine. Rev Physiol Biochem Pharmacol 168:59–118
Karin M, Lawrence T, Nizet V (2006) Innate immunity gone awry: linking microbial infections to chronic inflammation and cancer. Cell 124(4):823–835
Krishnappa L, Dreisbach A, Otto A, Goosens VJ, Cranenburgh RM, Harwood CR, Becher D, van Dijl JM (2013) Extracytoplasmic proteases determining the cleavage and release of secreted proteins, lipoproteins, and membrane proteins in Bacillus subtilis. J Proteome Res 12(9):4101–4110
Li Y, Li X, Sun Z (2011) Advance in detection techniques of florfenicol. Prog Vet Med 32(9):89–93 (in Chinese)
Liu CH, Chiu CS, Ho PL, Wang SW (2009) Improvement in the growth performance of white shrimp, Litopenaeus vannamei, by a protease-producing probiotic, Bacillus subtilis E20, from natto. J Appl Microbiol 107(3):1031–1041
Liu B, Xie J, Ge X, Xu P, Wang A, He Y, Zhou Q, Pan L, Chen R (2010) Effects of anthraquinone extract from Rheum officinale Bail on the growth performance and physiological responses of Macrobrachium rosenbergii under high temperature stress. Fish Shellfish Immunol 29(1):49–57
Liu B, Xie J, Ge X, Xu P, Miao L, Zhou Q, Pan L, Chen R (2012) Comparison study of the effects of anthraquinone extract and emodin from Rheum officinale bail on the physiological response, disease resistance of Megalobrama amblycephala under high temperature stress. Turk J Fish Aquat Sci 12:905–916
Liu ZM, Ma YX, Yang ZP, Li M, Liu J, Bao PY (2013) Immune responses and disease resistance of the juvenile sea cucumber Apostichopus japonicus induced by Metschnikowia sp. C14. Aquaculture 368–369:10–18
Ma YX, Liu ZM, Yang ZP, Li M, Liu J, Song J (2013) Effects of dietary live yeast Hanseniaspora opuntiae C21 on the immune and disease resistance against Vibrio splendidus infection in juvenile sea cucumber Apostichopus japonicus. Fish Shellfish Immunol 34(1):66–73
Merrifield DL, Burnard D, Bradley G, Davies SJ, Baker R (2009) Microbial community diversity associated with the intestinal mucosa of farmed rainbow trout (Oncoryhnchus mykiss Walbaum). Aquac Res 40(9):1064–1072
Ming J, Xie J, Xu P, Liu WB, Ge XP, Liu B, He YJ, Zhou QL, Xi BW, Pan LK (2010) Effects of emodin, vitamin C and their combination on growth, physiological and biochemical parameters, disease resistance and two HSP70 s mRNA expression of Wuchang bream (Megalobrama amblycephala). J Fish China 34(9):1447–1459 (in Chinese)
Misra CK, Das BK, Mukherjee SC, Pattnaik P (2006) Effect of long term administration of dietary β-glucan on immunity, growth and survival of Labeo rohita fingerlings. Aquaculture 255(1):82–94
Panigrahi A, Kiron V, Puangkaew J, Kobayashi T, Satoh S, Sugita H (2005) The viability of probiotic bacteria as a factor influencing the immune response in rainbow trout Oncorhynchus mykiss. Aquaculture 243(1):241–254
Parrado J, Rodriguez-Morgado B, Tejada M, Hernandez T, Garcia C (2014) Proteomic analysis of enzyme production by Bacillus licheniformis using different feather wastes as the sole fermentation media. Enzyme Microb Tech 57:1–7
Peng M, Xu W, Ai Q, Mai K, Liufu Z, Zhang K (2013) Effects of nucleotide supplementation on growth, immune responses and intestinal morphology in juvenile turbot fed diets with graded levels of soybean meal (Scophthalmus maximus L.). Aquaculture 392:51–58
Plotieau T, Lavitra T, Gillan DC, Eeckhaut I (2013) Bacterial diversity of the sediments transiting through the gut of Holothuria scabra (Holothuroidea; Echinodermata). Mar Biol 160(12):3087–3101
Reinus JF, Simon D (2014) Gastrointestinal anatomy and physiology: the essentials. Wiley, New York
Rescigno M (2011) The intestinal epithelial barrier in the control of homeostasis and immunity. Trends Immunol 32(6):256–264
Sarlin P, Philip R (2011) Efficacy of marine yeasts and baker’s yeast as immunostimulants in Fenneropenaeus indicus: a comparative study. Aquaculture 321(3):173–178
Serrano PH (2005) Responsible use of antibiotics in aquaculture. Food and Agriculture Org, Rome
Shah SQ, Cabello FC, L’Abée-Lund TM, Tomova A, Godfrey HP, Buschmann AH, Sørum H (2014) Antimicrobial resistance and antimicrobial resistance genes in marine bacteria from salmon aquaculture and non-aquaculture sites. Environ Microbiol 16(5):1310–1320
Tacchi L, Bickerdike R, Douglas A, Secombes CJ, Martin SA (2011) Transcriptomic responses to functional feeds in Atlantic salmon (Salmo salar). Fish Shellfish Immunol 31(5):704–715
Thitamadee S, Srisala J, Taengchaiyaphum S, Sritunyalucksana K (2014) Double-dose β-glucan treatment in WSSV-challenged shrimp reduces viral replication but causes mortality possibly due to excessive ROS production. Fish Shellfish Immunol 40(2):478–484
Tian XL, Zhao K, Wang J, Yang G, Yan FJ (2014) Effects of Bacillus cereus on the growth, activities of digestive enzyme and non-specific immune enzymes in Apostichopus japonicus: addition to water versus supplement in feed. J Ocean Univ China 45(1):18–25 (in Chinese)
Ubelmann F, Robine S, Louvard D (2011) Microvilli. In: Nabi IR (ed) Cellular domains, part III: cytoskeletal domains. Wiley, Hoboken, pp 213–228
Xie J, Liu B, Zhou Q, Su Y, He Y, Pan L, Ge X, Xu P (2008) Effects of anthraquinone extract from rhubarb Rheum officinale Bail on the crowding stress response and growth of common carp Cyprinus carpio var. Jian. Aquaculture 281(1):5–11
Yan FJ, Tian XL, Dong SL (2014) Growth performance, immune response, and disease resistance against Vibrio splendidus infection in juvenile sea cucumber Apostichopus japonicus fed a supplementary diet of the potential probiotic Paracoccus marcusii DB11. Aquaculture 420:105–111
Yang S, Wu Z, Jian J, Zhang X (2010) Effect of marine red yeast Rhodosporidium paludigenum on growth and antioxidant competence of Litopenaeus vannamei. Aquaculture 309(1):62–65
Yuan X, Yang H, Zhou Y, Mao Y, Zhang T, Liu Y (2006) The influence of diets containing dried bivalve feces and/or powdered algae on growth and energy distribution in sea cucumber Apostichopus japonicus (Selenka)(Echinodermata: Holothuroidea). Aquaculture 256(1):457–467
Zar JH (1999) Biostatistical analysis. Pearson Education, India
Zhang Q, Ma H, Mai K, Zhang W, Liufu Z, Xu W (2010) Interaction of dietary Bacillus subtilis and fructooligosaccharide on the growth performance, non-specific immunity of sea cucumber, Apostichopus japonicus. Fish Shellfish Immunol 29(2):204–211
Zhang Y, Liu B, Ge X, Liu W, Xie J, Ren M, Cui Y, Xia S, Chen R, Zhou Q (2014) 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(1):187–193
Zhao W, Liang M, Zhang P (2010) Effect of yeast polysaccharide on the immune function of juvenile sea cucumber, Apostichopus japonicus Selenka under pH stress. Aquac Int 18(5):777–786
Zhao W, Liang M, Xing Y, Wei J, Zhang P (2011a) The effect of yeast immunity polysaccharide (YSP) on the immunity function in sea cucumber, Apostichopus japonicus selenka. In: Remote sensing, environment and transportation engineering (RSETE), 2011 international conference on IEEE, pp 861–866
Zhao Y, Ma H, Zhang W, Ai Q, Mai K, Xu W, Wang X, Liufu Z (2011b) Effects of dietary β-glucan on the growth, immune responses and resistance of sea cucumber, Apostichopus japonicus against Vibrio splendidus infection. Aquaculture 315(3):269–274
Zhao Q, Jin S, Chen E, Qin F, Wang X, Wang G (2014) Pharmacokinetics of florfenicol in disease model of Portunus trituberculatus. Oceanol Limnol Sin 45(3):568–573 (in Chinese)
Zhou Y, Yang H, Liu S, Yuan X, Mao Y, Liu Y, Xu X, Zhang F (2006) Feeding and growth on bivalve biodeposits by the deposit feeder Stichopus japonicus Selenka (Echinodermata: Holothuroidea) co-cultured in lantern nets. Aquaculture 256:510–520
Zhu H, Liu H, Yan J, Wang R, Liu L (2012) Effect of yeast polysaccharide on some hematologic parameter and gut morphology in channel catfish (Ictalurus punctatus). Fish Physiol Biochem 38(5):1441–1447
Ziaei-Nejad S, Rezaei MH, Takami GA, Lovett DL, Mirvaghefi AR, Shakouri M (2006) The effect of Bacillus spp. bacteria used as probiotics on digestive enzyme activity, survival and growth in the Indian white shrimp Fenneropenaeus indicus. Aquaculture 252(2):516–524
Acknowledgments
This study was supported by the National Great Project of Scientific and Technical Supporting Programs (Grant No. 2011BAD13B03), the Program for Excellent Youth Foundation of Shandong Province (Grant No. JQ201009), and the Specialized Project of Regional demonstration for the Innovation and development of Marine economy in Guangdong Province (Grant No. GD2013-B003-005).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Yang, G., Tian, X., Dong, S. et al. Effects of dietary rhubarb, Bacillus cereus, yeast polysaccharide, and florfenicol supplementation on growth, intestinal morphology, and immune responses of sea cucumber (Apostichopus japonicus). Aquacult Int 24, 675–690 (2016). https://doi.org/10.1007/s10499-015-9957-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10499-015-9957-9