Effects of Dietary Bacillus subtilis on Growth, Intestinal Tissue Structure, Antioxidant Capacity, Immunity and Enteritis of Juvenile Largemouth Bass (Micropterus salmoides)

ZHANG Dongmei; YAN Haoxiao; LUO Maolin; YANG Yi; HU Yifan; GONG Chenxin; LI Zhiqiong; YANG Song; ZHAO Liulan

doi:10.3969/j.issn.1006-267x.2022.01.053

2022 , Vol. 34 >Issue 1: 575 - 588

DOI: https://doi.org/10.3969/j.issn.1006-267x.2022.01.053

RESEARCH PAPER

Effects of Dietary Bacillus subtilis on Growth, Intestinal Tissue Structure, Antioxidant Capacity, Immunity and Enteritis of Juvenile Largemouth Bass (Micropterus salmoides)

ZHANG Dongmei ,
YAN Haoxiao ,
LUO Maolin ,
YANG Yi ,
HU Yifan ,
GONG Chenxin ,
LI Zhiqiong ,
YANG Song ,
ZHAO Liulan

Expand

1. College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, China;
2. Sichuan Tequ Investment Group Co., Ltd., Chengdu 611430, China

Received date: 2021-06-18

Online published: 2022-01-18

Fold

Abstract

This experiment was conducted to investigate the effects of dietary Bacillus subtilis on the growth, intestinal tissue structure, antioxidant capacity, immunity and enteritis of juvenile largemouth bass (Micropterus salmoides). Three groups were set up to carry the feeding trial for 6 weeks, and they were control group (a basal diet), 0.5% Bacillus subtilis group (the basal diet+0.5% Bacillus subtilis preparation) and 1.0% Bacillus subtilis group (the basal diet+1.0% Bacillus subtilis preparation). Each group had three replicates and each replicate had 20 fish with the initial body weight of (16.0±0.5) g. The results showed as follows:1) compared with control group, the weight gain rate (WGR), specific growth rate (SGR)and body weight after 6weeks in Bacillus subtilis groups were increased, but the differences were not significant (P>0.05). 2) After injection of trinitro-benzene-sulfonic acid (TNBS) to induce enteritis, experimental fish in all group showed a certain mortality, but the addition of Bacillus subtilis reduced the cumulative mortality. 3) After the addition of Bacillus subtilis, the midgut villi of largemouth bass were slender, closely arranged, and the height and number of intestinal villi and the thickness of muscle layer were significantly increased compared with the control group (P<0.05). TNBS induced enteritis damaged the intestinal structure characterized by villous fusion, submucosa widening and muscle layer, but the addition of Bacillus subtilis obviously reduced the damage. 4) With the increase of dietary Bacillus subtilis supplemental level, the activities of antioxidant enzymes in the intestine showed an upward trend. The activities of total superoxide dismutase (T-SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) in the intestine in 1.0% Bacillus subtilis group were significantly higher than those in control group and 0.5% Bacillus subtilis group (P<0.05), and the malondialdehyde (MDA) content was significantly lower than that in control group (P<0.05). Enteritis could improve the activities of antioxidant enzymes in the intestine of juvenile largemouth bass, and the intestinal antioxidant capacity of juvenile largemouth bass was stronger after adding Bacillus subtilis. 5) The activities of acid phosphatase (ACP) and lysozyme (LZM) in the intestine were significantly increased with the increase of Bacillus subtilis supplemental level (P<0.05). The activity of alkaline phosphatase (AKP) in the intestine in 1.0% Bacillus subtilis group was significantly higher than that in control group and 0.5% Bacillus subtilis group (P<0.05). After enteritis was induced by TNBS, the intestinal immune enzyme activities of the three groups were significantly increased (P<0.05), and the intestinal ACP, AKP and LZM activities in 1.0% Bacillus subtilis group were significantly higher than those in control group (P<0.05). 6) Compared with the control group, the addition of 0.5% or 1.0% Bacillus subtilis could significantly reduce the mRNA relative expression levels of intestinal proinflammatory factors such as interleukin-1β (IL-1β), interleukin-15 (IL-15), tumor necrosis factor-α (TNF-α) and interleukin-8 (IL-8), and significantly increase the mRNA relative expression levels of anti-inflammatory factors such as interleukin-10 (IL-10) and transforming growth factor-β1 (TGF-β1) (P<0.05). To sum up, the addition of Bacillus subtilis in the diet can improve the intestinal antioxidant capacity and immunity of juvenile largemouth bass, and alleviate the intestinal injury caused by TNBS. The effect is better with the addition of 1.0%.

Key words： Bacillus subtilis; largemouth bass; growth; intestinal tissue structure; antioxidant enzyme; immune enzyme; enteritis

Cite this article

ZHANG Dongmei , YAN Haoxiao , LUO Maolin , YANG Yi , HU Yifan , GONG Chenxin , LI Zhiqiong , YANG Song , ZHAO Liulan . Effects of Dietary Bacillus subtilis on Growth, Intestinal Tissue Structure, Antioxidant Capacity, Immunity and Enteritis of Juvenile Largemouth Bass (Micropterus salmoides)[J]. Chinese Journal of Animal Nutrition, 2022 , 34(1) : 575 -588 . DOI: 10.3969/j.issn.1006-267x.2022.01.053

References

[1] 韩刚,许玉艳,刘琪,等.科学制定水产养殖业绿色发展标准的思考与建议[J].中国渔业质量与标准,2019,9(5):55-60. HAN G,XU Y Y,LIU Q,et al.Thoughts and suggestions on the scientific establishment for the green development of aquaculture[J].Chinese Fishery Quality and Standards,2019,9(5):55-60.(in Chinese)
[2] PANDIYAN P,BALARAMAN D,THIRUNAVUKKARASU R,et al.Probiotics in aquaculture[J].Drug Invention Today,2013,5(1):55-59.

[3] YARAHMADI P,KOLANGI MIANDARE H,HOSEINIFAR S H.Haemato-immunological and serum biochemical parameters,intestinal histomorphology and growth performance of rainbow trout (Oncorhynchus mykiss) fed dietary fermentable fibre (Vitacel)[J].Aquaculture Nutrition,2016,22(5):1134-1142.

[4] FAN Y,PEDERSEN O.Gut microbiota in human metabolic health and disease[J].Nature Reviews Microbiology,2021,19(1):55-71.

[5] ZHAO L L,TANG G,XIONG C,et al.Chronic chlorpyrifos exposure induces oxidative stress,apoptosis and immune dysfunction in largemouth bass (Micropterus salmoides)[J].Environmental Pollution,2021,282:117010.
[6] CHAUHAN A,SINGH R.Probiotics in aquaculture:a promising emerging alternative approach[J].Symbiosis,2018,77(2):99-113.
[7] 惠明,窦丽娜,田青,等.枯草芽孢杆菌的应用研究进展[J].安徽农业科学,2008,36(27):11623-11624,11627. HUI M,DOU L N,TIAN Q,et al.Advances in application research of Bacillus subilis[J].Journal of Anhui Agricultural Sciences,2008,36(27):11623-11624,11627.(in Chinese)
[8] ZHOU Y H,ZENG Z H,XU Y B,et al.Application of Bacillus coagulans in animal husbandry and its underlying mechanisms[J].Animals,2020,10(3):454.
[9] 翟继鹏,张金枝.枯草芽孢杆菌在养殖业中的应用研究进展[J].浙江畜牧兽医,2010,35(3):7-9. ZHAI J P,ZHANG J Z.Recent advance on application of Bacillus subtilis in the animal industry[J].Zhejiang Journal Animal Science and Veterinary Medicine,2010,35(3):7-9.(in Chinese)
[10] GOBI N,VASEEHARAN B,CHEN J C,et al.Dietary supplementation of probiotic Bacillus licheniformis Dahb1 improves growth performance,mucus and serum immune parameters,antioxidant enzyme activity as well as resistance against Aeromonas hydrophila in tilapia Oreochromis mossambicus[J].Fish & Shellfish Immunology,2018,74:501-508.
[11] LEE S,KATYA K,PARK Y,et al.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[J].Fish & Shellfish Immunology,2017,61:201-210.
[12] STANDEN B T,RAWLING M D,DAVIES S J,et al.Probiotic Pediococcus acidilactici modulates both localised intestinal- and peripheral-immunity in tilapia (Oreochromis niloticus)[J].Fish & Shellfish Immunology,2013,35(4):1097-1104.

[13] FENG J C,CAI Z L,CHEN Y Y,et al.Effects of an exopolysaccharide from Lactococcus lactis Z-2 on innate immune response,antioxidant activity,and disease resistance against Aeromonas hydrophila in Cyprinus carpio L.[J].Fish & Shellfish Immunology,2020,98:324-333.
[14] 张冬梅,赵柳兰,刘巧,等.大口黑鲈leptin A基因的组织表达及其对急性低氧胁迫的响应[J].水产学报,2020,44(10):1609-1618. ZHANG D M,ZHAO L L,LIU Q,et al.Tissue expression of leptin A gene in Micropterus salmoides and its response to acute hypoxia stress[J].Journal of Fisheries of China,2020,44(10):1609-1618.(in Chinese)
[15] 农业农村部渔业渔政管理局,全国水产技术推广总站,中国水产学会.2020中国渔业统计年鉴[M].北京:中国农业出版社,2020. Fishery Administration of the Ministry Agriculture and Rural Affairs,National Fisheries Technology Extension Center,China Society of Fisheries.2020 China fishery statistical yearbook[M].Beijing:China Agriculture Press,2020.(in Chinese)
[16] 黄太寿,白俊杰,李胜杰,等.我国加州鲈养殖现状和绿色发展问题探讨[J].中国水产,2017(12):44-47. HUANG T S,BAI J J,LI S J,et al.Present situation and green development of California perch culture in China[J].China Fisheries,2017(12):44-47.(in Chinese)
[17] ZHOU Y L,HE G L,JIN T,et al.High dietary starch impairs intestinal health and microbiota of largemouth bass,Micropterus salmoides[J].Aquaculture,2021,534:736261.
[18] SUN J L,ZHAO L L,LIAO L,et al.Interactive effect of thermal and hypoxia on largemouth bass (Micropterus salmoides) gill and liver:aggravation of oxidative stress,inhibition of immunity and promotion of cell apoptosis[J].Fish & Shellfish Immunology,2020,98:923-936.
[19] 王成强,李宝山,王际英,等.饲料中添加枯草芽孢杆菌和酵母培养物对珍珠龙胆石斑鱼幼鱼生长性能、体组成及养殖水质的影响[J].浙江大学学报(农业与生命科学版),2019,45(4):490-499. WANG C Q,LI B S,WANG J Y,et al.Effects of dietary Bacillus subtilis and yeast culture on growth performance,body composition and farming water quality of juvenile hybrid grouper (Epinephelus fuscoguttatus ♀×Epinephelus lanceolatus ♂)[J].Journal of Zhejiang University (Agriculture & Life Sciences),2019,45(4):490-499.(in Chinese)
[20] MORRIS G P,BECK P L,HERRIDGE M S,et al.Hapten-induced model of chronic inflammation and ulceration in the rat colon[J].Gastroenterology,1989,96(3):795-803.

[21] Scheiffele F,Fuss I J.Induction of TNBS colitis in mice.[J].Current Protocols in Immunology,2002,49(1).
[22] Cheng W,Chiu C S,Guu Y K,et al.Expression of recombinant phytase of Bacillus subtilis E20 in Escherichia coli HMS 174 and improving the growth performance of white shrimp, Litopenaeus vannamei, juveniles by using phytase-pretreated soybean meal-containing diet[J].Aquaculture Nutrition,2013,19(2):117-127.

[23] Liu C H,Wu K,Chu T W,et al.Dietary supplementation of probiotic,Bacillus subtilis E20, enhances the growth performance and disease resistance against Vibrio alginolyticus in parrot fish (Oplegnathus fasciatus)[J].Aquaculture International,2017,26(1):63-74
[24] DANIELS C L,MERRIFIELD D L,BOOTHROYD D P,et al.Effect of dietary Bacillus spp. and mannan oligosaccharides (MOS) on European lobster (Homarus gammarus L.) larvae growth performance,gut morphology and gut microbiota[J].Aquaculture,2010,304(1/2/3/4):49-57.
[25] 黄灵,李小梅,舒琥,等.两种微生态制剂对虎龙斑体成分、肠道消化酶及组织学结构的影响[J].饲料工业,2018,39(12):24-32. HUANG L,LI X M,SHU H,et al.The effects of two kinds of microecologics on body composition,intestinal digestive enzyme and histological structure of hybrid grouper (Epinephelus fuscoguttatus ♀×E. lanceolatus ♂)[J].Feed Industry,2018,39(12):24-32.(in Chinese)
[26] BAI K W,HUANG Q,ZHANG J F,et al.Supplemental effects of probiotic Bacillus subtilis fmbJ on growth performance,antioxidant capacity,and meat quality of broiler chickens[J].Poultry Science,2017,96(1):74-82.

[27] 从光雷,王强,肖蕴祺,等.饲粮添加橡椀单宁对肉鸡生长性能、屠宰性能、肉品质、抗氧化功能和肠道发育的影响[J].动物营养学报,2020,32(12):5948-5957. CONG G L,WANG Q,XIAO Y Q,et al.Effects of dietary Quercus acutissima Carruth tannin on growth performance,slaughter performance,meat quality,antioxidant function and intestinal development of broilers[J].Chinese Journal of Animal Nutrition,2020,32(12):5948-5957.(in Chinese)
[28] 张克烽,张子平,陈芸,等.动物抗氧化系统中主要抗氧化酶基因的研究进展[J].动物学杂志,2007,42(2):153-160. ZHANG K F,ZHANG Z P,CHEN Y,et al.Antioxidant defense system in animals[J].Chinese Journal of Zoology,2007,42(2):153-160.(in Chinese)
[29] 吴益星,叶坤,王志勇,等.低盐养殖对大黄鱼生长、肌肉营养成分及抗氧化能力的影响[J].集美大学学报(自然科学版),2020,25(3):171-179. WU Y X,YE K,WANG Z Y,et al.Effects of low salinity culture on growth,muscle nutritional composition and antioxidant capacity of Larimichthys crocea[J].Journal of Jimei University (Natural Science),2020,25(3):171-179.(in Chinese)
[30] TOVAR-RAMÍREZ D,MAZURAIS D,GATESOUPE J F,et al.Dietary probiotic live yeast modulates antioxidant enzyme activities and gene expression of sea bass (Dicentrarchus labrax) larvae[J].Aquaculture,2010,300(1/4):142-147.
[31] CELIK H,CELIK N,KOCYIGIT A,et al.The relationship between plasma aluminum content,lymphocyte DNA damage,and oxidative status in persons using aluminum containers and utensils daily[J].Clinical Biochemistry,2012,45(18):1629-1633.

[32] 刘慧玲,黄翔鹄,李长玲,等.不同浓度的枯草芽孢杆菌对罗非鱼鱼苗的养殖水体水质及其抗病力的影响[J].水产养殖,2009,30(10):5-9. LIU H L,HUANG X H,LI C L,et al.Effect of different concentrations of Bacillus substilis on water quality and disease resistence of tilapia fry[J].Journal of Aquaculture,2009,30(10):5-9.(in Chinese)
[33] 谢佳磊,肖丹,殷蝶,等.枯草芽孢杆菌对克氏原螯虾免疫机能的影响[J].淡水渔业,2007,37(6):24-27. XIE J L,XIAO D,YIN D,et al.Effects of Bacillus subtilis on immune functions of Procambarus clarkii[J].Freshwater Fisheries,2007,37(6):24-27.(in Chinese)
[34] 张永安,聂品.鱼类体液免疫因子研究进展[J].水产学报,2000,24(4):376-381. ZHANG Y A,NIE P.Humoral immune factors of fish:a review[J].Journal of Fisheries of China,2000,24(4):376-381.(in Chinese)
[35] 陈剑杰,曹谨玲,贺鑫晋,等.苯并芘(BaP)对鲤鱼肝、肾抗氧化、非特异性免疫能力及组织结构的影响[J].核农学报,2019,33(3):623-630. CHEN J J,CAO J L,HE X J,et al.Effects of benzopyrene on the capacity of antioxidases,non-specific immunity and organization structure in liver and kidney of carp (Cyprinus carpio)[J].Journal of Nuclear Agricultural Sciences,2019,33(3):623-630.(in Chinese)
[36] GENG X,DONG X H,TAN B P,et al.Effects of dietary chitosan and Bacillus subtilis on the growth performance,non-specific immunity and disease resistance of cobia,Rachycentron canadum[J].Fish & Shellfish Immunology,2011,31(3):400-406.

[37] MOURIÑO J L P,DO NASCIMENTO VIEIRA F,JATOBÁ A B,et al.Effect of dietary supplementation of inulin and W. cibaria on haemato-immunological parameters of hybrid surubim (Pseudoplatystoma sp)[J].Aquaculture Nutrition,2012,18(1):73-80.

[38] AI Q H,XU H G,MAI K S,et al.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[J].Aquaculture,2011,317(1/2/3/4):155-161.
[39] 胡金娥.益生菌对炎症和免疫系统影响的研究进展[J].实用心脑肺血管病杂志,2013,21(12):5-7. HU J E.Effect of probiotics on inflammatory and immune system[J].Practical Journal of Cardiac Cerebral Pneumal and Vascular Disease,2013,21(12):5-7.(in Chinese)
[40] BOUMA G,STROBER W.The immunological and genetic basis of inflammatory bowel disease[J].Nature Reviews Immunology,2003,3(7):521-533.

[41] PIZARRO T T,ARSENEAU K O,BAMIAS G,et al.Mouse models for the study of Crohn's disease[J].Trends in Molecular Medicine,2003,9(5):218-222.

[42] TE VELDE A A,VERSTEGE M I,HOMMES D W.Critical appraisal of the current practice in murine TNBS-induced colitis[J].Inflammatory Bowel Diseases,2006,12(10):995-999.

[43] ASADUZZAMAN M,IEHATA S,AKTER S,et al.Effects of host gut-derived probiotic bacteria on gut morphology,microbiota composition and volatile short chain fatty acids production of Malaysian Mahseer Tor tambroides[J].Aquaculture Reports,2018,9:53-61.
[44] RINGØ E,OLSEN R E,GIFSTAD T Ø,et al.Prebiotics in aquaculture:a review[J].Aquaculture Nutrition,2010,16(2):117-136.

[45] 宫秀燕,韦明,蒋秋斐,等.凝结芽孢杆菌对肠炎沙门氏菌感染肉鸡生产性能和抗氧化功能的影响[J].中国畜牧杂志,2015,51(17):74-79,98. GONG X Y,WEI M,JIANG Q F,et al.Effect of Bacillus coagulans on growth performance and antioxidant fuction of broilers challenged with Salmonella enteritis[J].Chinese Journal of Animal Science,2015,51(17):74-79,98.(in Chinese)
[46] LOWRY S F.Cytokine mediators of immunity and inflammation[J].Archives of Surgery,1993,128(11):1235-1241.

[47] 丰培金.鱼类细胞因子研究进展[J].中国兽医杂志,2005,41(8):44-46. FENG P J.Research advances of fish cytokines[J].Chinese Journal of Veterinary Medicine,2005,41(8):44-46.(in Chinese)
[48] SONG X H,ZHAO J,BO Y X,et al.Aeromonas hydrophila induces intestinal inflammation in grass carp (Ctenopharyngodon idella):an experimental model[J].Aquaculture,2014,434:171-178.
[49] FEHRMANN-CARTES K,CORONADO M,HERNÁNDEZ A J,et al.Anti-inflammatory effects of aloe vera on soy meal-induced intestinal inflammation in zebrafish[J].Fish & Shellfish Immunology,2019,95:564-573.
[50] 刘晓勇,张颖,齐茜,等.枯草芽孢杆菌对杂交鲟幼鱼生长性能、消化酶活性及非特异性免疫的影响[J].中国水产科学,2011,18(6):1315-1320. LIU X R,ZHANG Y,QI Q,et al.Effects of Bacillus subtilis on growth,digestive enzyme activity,and non-specific immunity in hybrid sturgeon (Acipenser baeri ♂×Acipenser schrenkii ♀) juveniles[J].Journal of Fishery Sciences of China,2011,18(6):1315-1320.(in Chinese)
[51] MIDHUN S J,NEETHU S,ARUN D,et al.Dietary supplementation of Bacillus licheniformis HGA8B improves growth parameters,enzymatic profile and gene expression of Oreochromis niloticus[J].Aquaculture,2019,505:289-296.
[52] SINGH V,SINGH K,AMDEKAR S,et al.Innate and specific gut-associated immunity and microbial interference[J].FEMS Immunology and Medical Microbiology,2009,55(1):6-12.

Options

Outlines

模态框（Modal）标题

Abstract

Cite this article

References