乳酸菌和噬菌体对中华鳖生理生化与肠道菌群的影响

doi:10.3969/j.issn.1004-1524.20240041

浙江农业学报 ›› 2025, Vol. 37 ›› Issue (1): 39-48.DOI: 10.3969/j.issn.1004-1524.20240041

乳酸菌和噬菌体对中华鳖生理生化与肠道菌群的影响

齐天鹏¹(), 刘莉², 夏美文¹, 吕孙建²^,^*(), 徐海圣¹^,³^,^*()

1.浙江大学动物科学学院,浙江杭州 310058
2.浙江省农业科学院水生生物研究所,浙江杭州 310021
3.湖州市南太湖现代农业科技推广中心,浙江湖州 313000

收稿日期:2024-01-08 出版日期:2025-01-25 发布日期:2025-02-14
作者简介:齐天鹏(1998—),男,浙江台州人,硕士研究生,从事水产动物养殖及其病害防治研究。E-mail:22117084@zju.edu.cn
通讯作者: *吕孙建,E-mail:lvsunjian@163.com;徐海圣,E-mail:hsxu@zju.edu.cn
基金资助:
浙江省重点研发计划(2022C02027);湖州市农业科技创新团队项目(2022HN02)

Effects of lactic acid bacteria and bacteriophage on physiology, biochemistry and gut microbiota of Chinese soft-shelled turtle (Pelodiscus sinensis)

QI Tianpeng¹(), LIU Li², XIA Meiwen¹, LYU Sunjian²^,^*(), XU Haisheng¹^,³^,^*()

1. College of Animal Sciences, Zhejiang University, Hangzhou 310058, China
2. Institute of Hydrobiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
3. Zhejiang University Huzhou South Taihu Lake Modern Agricultural Science and Technology Promotion Center, Huzhou 313000, Zhejiang, China

Received:2024-01-08 Online:2025-01-25 Published:2025-02-14

摘要/Abstract

摘要： 为探究饲料中添加乳酸菌和噬菌体对中华鳖生长性能、酶活力、免疫基因表达和肠道菌群的影响,以基础饲料为对照组,以在基础饲料中分别添加1×10⁷CFU·g^-1乳酸菌、1×10⁷CFU·g^-1乳酸菌+1×10⁹ PFU·g^-1噬菌体为试验组,以投喂方式养殖中华鳖90 d。对血清和肠道的酶活性,肝、脾、肾等器官的组织切片形态,免疫基因表达和肠道内容物的菌群组成等进行了检测。结果显示:与对照组相比,单独添加乳酸菌、联合添加乳酸菌和噬菌体的饲料均能显著(P<0.05)提高中华鳖的存活率,增加胰蛋白酶活性,乳酸菌+噬菌体组的增重率和特定生长率最高但没有显著性差异(P>0.05)。乳酸菌+噬菌体组的谷草转氨酶和酸性磷酸酶活性显著(P<0.05)降低,乳酸菌组的肝LYSC基因表达量显著(P<0.05)降低。然而,组织切片和肠道菌群的差异变化并不显著,各组织细胞均具有完整的形态与结构,肠道微生物的α多样性和门、属水平的微生物群落组成均无显著差异(P>0.05)。综上所述,饲粮中添加适量的乳酸菌和噬菌体能显著提高中华鳖的存活率,并在一定程度上提高中华鳖的健康水平。

关键词: 乳酸菌, 噬菌体, 中华鳖, 生化特性, 肠道菌群

Abstract:

This study investigated the effects of incorporating lactic acid bacteria and bacteriophages into the feed on the growth performance, enzyme activity, immune gene expression, and gut microbiota of Pelodiscus sinensis. The experimental groups included a lactic acid bacteria group, which received a basal feed supplemented with 1×10⁷CFU·g^-1 of lactic acid bacteria, and a lactic acid bacteria+bacteriophage group, which received a basal feed supplemented with 1×10⁷CFU·g^-1 of lactic acid bacteria and 1×10⁹ PFU·g^-1 of bacteriophages. The control group was provided with only the basal feed. After 90 days of feeding, the enzyme activity in serum and the intestine, the tissue morphology of the liver, spleen, kidney, and other organs, as well as the expression of immune-related genes and the microbiome composition in the intestinal contents were analyzed. The results revealed that, compared to the control group, diets supplemented with lactic acid bacteria alone and in combination with bacteriophages significantly (P<0.05) increased the survival rate of P. sinensis and trypsin activity. The lactic acid bacteria+bacteriophage group exhibited the highest weight gain rate and specific growth rate, although not significantly different (P>0.05). Activities of glutamine aminotransferase and acid phosphatase were significantly lower in the lactic acid bacteria+bacteriophage group, while hepatic LYSC gene expression was significantly (P<0.05) lower in the lactic acid bacteria group. However, there were no significant differential changes in tissue sections and gut microbiota. Tissue cells maintained intact morphology and structure, and the α diversity of intestinal microorganisms, as well as microbial community compositions at the phylum and genus levels, showed no significant differences (P>0.05). In conclusion, the addition of an appropriate amount of lactic acid bacteria and bacteriophages to the ration can significantly improve the survival rate of P. sinensis and improve the health level of P.sinensis to a certain extent.

Key words: lactic acid bacteria, bacteriophage, Pelodiscus sinensis, biochemical characteristic, gut microbiota

中图分类号:

S963

齐天鹏, 刘莉, 夏美文, 吕孙建, 徐海圣. 乳酸菌和噬菌体对中华鳖生理生化与肠道菌群的影响[J]. 浙江农业学报, 2025, 37(1): 39-48.

QI Tianpeng, LIU Li, XIA Meiwen, LYU Sunjian, XU Haisheng. Effects of lactic acid bacteria and bacteriophage on physiology, biochemistry and gut microbiota of Chinese soft-shelled turtle (Pelodiscus sinensis)[J]. Acta Agriculturae Zhejiangensis, 2025, 37(1): 39-48.

图/表 9

表1 目的基因引物序列

Table 1 Primer sequences of target genes

基因 Gene	上游引物 Forward primer(5'→3')	下游引物 Reverse primer(5'→3')
干扰素诱导的解螺旋酶C区域1 IFIH1	AAGCAAGCAATAGCCAGTA	CCTCACCTGACTCCTCTT
干扰因子IFN	CTACTACTCTATCCTGCTCAG	GCTTACCTCTGTCCAACTC
白细胞介素1 IL-1	CTCTAGGCTAGTGTAATGGTT	GGTTGGCTGAGACTTGAG
溶菌酶C LYSC	AAACCCGAGGGCAACTTT	GGCTTTCATAAATTCCCTACAC
肿瘤坏死因子TNF	CCATCATCCTCCATCCTTG	ACGGTCAGTGTGATATGTG

表2 乳酸菌和噬菌体对中华鳖生长性能的影响

Table 2 Effects of lactic acid bacteria and bacteriophages on growth performance of Pelodiscus sinensis

处理 Treatment	初体重 Initial weight/g	末体重 Final weight/g	存活率 Survival rate/%	增重率 Weight gain rate/%	特定生长率 Specific growth rate/(%·d^-1)
IF_CK	158.71±7.98	318.61±23.96 ab	80.00±3.50 b	102.00±18.79	0.76±0.10
IF_LL	152.63±7.68	291.78±19.02 b	91.43±3.50 a	92.39±17.04	0.71±0.09
IF_BP	162.33±6.06	369.43±23.96 a	85.71±0.01 a	125.50±26.62	0.87±0.14

表3 乳酸菌和噬菌体对中华鳖肠道消化酶活力的影响

Table 3 The effect of lactic acid bacteria and bacteriophages on the activity of digestive enzymes in the intestine of Pelodiscus sinensis

处理 Treatment	活性Activity
处理 Treatment	胰蛋白酶 Trypsin/(U·mg^-1)	α-淀粉酶 α-Amylase/(U·mg^-1)	糜蛋白酶 Chymotrypsin activity/(U·mg^-1)	脂肪酶 Lipase/(U·g^-1)
IF_CK	59.24±8.14 c	3.50±0.29	22.56±3.15	4.61±0.65
IF_LL	156.93±9.20 b	3.97±0.28	18.09±4.78	3.86±0.28
IF_BP	210.97±24.30 a	3.68±0.22	21.27±3.74	7.92±3.29

表4 乳酸菌和噬菌体对中华鳖血清生化指标的影响

Table 4 The effects of lactic acid bacteria and bacteriophages on blood biochemical indicators of Pelodiscus sinensis

处理 Treatment	含量Content/(mmol·L^-1)		活性Activity
处理 Treatment	甘油三酯 Triglyceride	总胆固醇 Total cholesterol	溶菌酶 Lysozyme/ (U·mL^-1)	超氧化物歧化酶 Superoxide dismutase/ (U·mL^-1)	谷丙转氨酶 Gammagluta- minase/ (U·L^-1)	谷草转氨酶 Glutamic transaminase/ (U·L^-1)	碱性磷酸酶 Alkaline phosphatase/ (U·L^-1)	酸性磷酸酶 Acid phosphatase/ (U·L^-1)
IF_CK	2.94±0.46	12.14±0.95	760.06±164.44	22.52±0.11	1.68±0.27	15.66±2.22 a	40.20±6.34 bc	278.54±44.66 a
IF_LL	2.63±0.70	13.49±1.19	754.84±90.51	21.83±0.34	2.58±0.66	11.76±0.82 a	47.90±47.27 ab	241.07±41.95 a
IF_BP	2.55±0.49	11.68±1.08	1 019.63±106.13	22.03±0.25	1.25±0.42	6.61±0.38 b	25.69±3.34 c	119.57±72.80 b

图1 乳酸菌和噬菌体对中华鳖组织的影响 H,肝细胞;HS,肝血窦;W,白髓;RP,红髓;AD,肺泡管;DCT,远曲小管;PCT,近曲小管;RC,肾小体;MF,心肌纤维;CL,中央乳糜管;CE,柱状上皮。IF_CK,对照组;IF_LL,乳酸菌组;IF_BP,乳酸菌+噬菌体组。下同。

Fig.1 The effects of lactic acid bacteria and bacteriophages on the tissue of Pelodiscus sinensis H,Hepatocyte; HS, Hepatic sinusoid; WP, White pulp; RP, Red pulp; AD, Alveolar duct; DCT, Distal convoluted tubule; PCT, Proximal convoluted tubule; RC, Renal corpuscle; MF, Myocardial fiber; CL, Central lacteal; CE, Columnar epithelium.IF_CK, Control group; IF_LL, Lactic acid bacteria group; IF_BP, Lactic acid bacteria + bacteriophage group.The same as bellow.

图2 乳酸菌和噬菌体对中华鳖免疫基因表达的影响柱状图上方无相同小写字母表示差异显著(P<0.05),无字母表示差异不显著(P>0.05)。

Fig.2 The effects of lactic acid bacteria and bacteriophages on the expression of immune genes in Pelodiscus sinensis No identical lowercase letters above the bar graphs indicate significant differences (P<0.05), while no letters indicate insignificant differences (P>0.05).

图3 不同处理中华鳖肠道菌群的韦恩图和主成分分析 A,韦恩图;B,主成分分析图。

Fig.3 Venn diagram and principal component analysis of gut microbiota of Pelodiscus sinensis in different treatment A, Venn diagram; B, Principal component analysis diagram.

表5 乳酸菌和噬菌体对中华鳖肠道内容物微生物α多样性的影响

Table 5 Effects of lactic acid bacteria and bacteriophages on the microbial α diversity of intestinal contents in Pelodiscus sinensis

处理 Treatment	香农指数 Shannon index	辛普森指数 Simpson index	Chao1指数 Chao1 index	OTU指数 Observed_OTUs index	覆盖率 Goods_coverage/%
IF_CK	4.74±0.34	0.88±0.04	179.38±18.90	168.70±17.76	100
IF_LL	4.37±0.27	0.87±0.26	154.45±29.68	145.80±26.68	100
IF_BP	4.20±0.21	0.87±0.16	133.31±13.04	129.20±12.81	100

图4 乳酸菌和噬菌体对中华鳖肠道菌群的影响 A,门水平;B,属水平。

Fig.4 The effects of lactic acid bacteria and bacteriophages on the gut microbiota of Pelodiscus sinensis A, Phylum level; B, Genus level.

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乳酸菌和噬菌体对中华鳖生理生化与肠道菌群的影响

Effects of lactic acid bacteria and bacteriophage on physiology, biochemistry and gut microbiota of Chinese soft-shelled turtle (Pelodiscus sinensis)

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