Effects of feeding Candida ethanolica GXU01 on growth, immunity and intestinal flora of tilapia
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摘要: 为探究乙醇假丝酵母 (Candida ethanolica GXU01) 在罗非鱼养殖中的益生潜能，为罗非鱼可持续养殖寻找生态友好的饲用添加菌，该研究将乙醇假丝酵母作为饲料添加菌饲喂尼罗罗非鱼 (Oreochroms niloticus)，通过测定罗非鱼的生长性能、肠道消化酶、血清非特异性免疫指标、肠道微生物群落结构以及用无乳链球菌 (Streptococcus agalactiae) 对罗非鱼进行攻毒试验，全面评价了乙醇假丝酵母对罗非鱼生长及其免疫力的影响。结果表明，摄食乙醇假丝酵母可以使罗非鱼的生长性能、消化酶活性、血清溶菌酶活性和补体C3含量显著提高 (P<0.05)。投喂乙醇假丝酵母后，罗非鱼肠道中梭杆菌门、鲸杆菌属 (Cetobacterium) 和艾克曼菌属 (Akkermansia) 等有益菌群的丰度显著上调，蓝细菌门大幅减少。攻毒试验中，摄食酵母饲料的罗非鱼成活率增加26.66%。研究表明乙醇假丝酵母对罗非鱼肠道消化能力和免疫抗病能力均有促进作用。Abstract: To explore the probiotic potential of Candida ethanolica GXU01 in tilapia culture and to find ecological and friendly feeding bacteria for tilapia sustainable cultivation, we used C. ethanolica GXU01 as feed-additive to feed tilapia. Then we determined the growth performance, intestinal digestive enzymes, serum non-specific immune indexes and intestinal microbial community structure, and challenged the tilapia with Streptococcus agalactiae, so as to comprehensively evaluate the effects of C. ethanolica GXU01 on the growth and immunity of tilapia. The results show that the growth performance, digestive enzyme activity, serum lysozyme activity and complement C3 content of tilapia could be significantly improved by feeding C. ethanolica GXU01 (P<0.05). The abundance of Fusobacteria, Cetobacterium and Akkermansia in the intestinal tract of tilapia increased significantly and those of Cyanobacteria decreased significantly after feed of C. ethanolica GXU01. In the challenge test, the survival rate of tilapia fed with diet containing C. ethanolica GXU01 increased by 26.66%. The study shows that C. ethanolica GXU01 promotes the intestinal digestive ability and immune resistance of tilapia.
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Key words:
- Tilapia /
- Candida ethanolica GXU01 /
- Intestinal flora /
- Probiotic
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图 1 GXU01对罗非鱼肠道蛋白酶、淀粉酶、木聚糖酶和纤维素酶活性的影响
CK和T5分别为对照组和GXU01实验组；Day 30、Day 60分别表示饲养30和60 d取样；柱上不同字母表示差异性显著 (P<0.05)；图2同此。
Figure 1. Effects of GXU01 on activity of gut proteinase, amylase, xylanase and cellulase in tilapias
CK and T5 indicate control and GXU01 treated group, respectively; samples were marked as Day 30 on 30th day and Day 60 on 60th day of feeding; different letters on the bar indicate significant difference (P<0.05). The same case in Figure 2.
图 2 GXU01对罗非鱼血清超氧化物歧化酶活性、溶菌酶活性、补体C3和补体C4质量浓度的影响
Figure 2. Effects of GXU01 on serum SOD activity, lysozyme activity, mass concentrations of Complement 3 and Complement 4 in tilapia
图 3 基于16S rDNA基因序列的罗非鱼肠道在门水平和属水平菌群结构及相对丰度图
CK和T5分别代表对照组和GXU01的实验组；投喂30 d取样标为CK-1、T5-1，投喂60 d取样标为CK-2、T5-2；表2、图4同此。
Figure 3. Structure and relative abundance of intestinal flora at phylum and genus of tilapia based on 16s rDNA sequence
CK and T5 indicate control and GXU01 treated group, respectively; samples were marked as CK-1, T5-1 at 30th day and CK-2, T5-2 at 60th day of feeding. The same case in Table 2 and Figure 4.
图 4 基于属水平上的物种丰度聚类热图
Figure 4. Clustering heat map based on species abundance at genus level
图 5 基于binary_chisq距离算法的属水平PCoA分析图
Figure 5. Genus level PCoA analysis diagram based on binary_chisq distance algorithm
图 6 用无乳链球菌进行攻毒试验后7 d成活率
Figure 6. Survival rate of 7 d after challenge test with S. agalactiae
表 1 GXU01饲料对罗非鱼生长性能的影响
Table 1. Effects of GXU01 feed on growth performance of tilapia
指标
Index对照组
CK Group实验组
T5 Group初始体质量 Initial mass/g 108.96±14.29 106.07±13.53 最后体质量 Final mass/g 287.41±25.98 297.69±27.73 总增质量 Total mass gain/g 178.45±18.23 196.42±25.62 饲料转化率 Feed conversion rate/% 1.78±0.20b 1.56±0.30a 饲料效率 Feed efficiency 0.57±0.06 0.66±0.13 成活率 Survival rate/% 100 100 注：同行数据上标不同字母表示差异性显著 (P<0.05)。 Note: Values in the same row with different superscripts are significantly different (P<0.05). 
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表 2 Alpha多样性指数分析
Table 2. Analysis of Alpha diversity index
组别
GroupOUT数
OTUsChao1指数
Chao1辛普森指数
Simpson香浓指数
ShannonCK-1 590 601.90 0.045 2b 4.251 7a CK-2 628 649.48 0.085 1b 4.001 7a T5-1 557 615.44 0.150 3a 3.138 6b T5-2 563 605.90 0.168 3a 3.268 6b 注：同列数值不同小写字母表示差异性显著 (P<0.05)。 Note: Values within the same column with different superscripts are significantly different (P<0.05).
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