羊源Lactobacillus plantarum对腹泻羔羊空肠菌群及肠道黏膜屏障的调控作用

doi:10.11843/j.issn.0366-6964.2024.08.027

Abstract

Abstract:

The aim of the experiment was to investigate the mitigatingregulating effect of Lactobacillus plantarum on diarrhea lambs in indexes including the damage of anti-oxidative capacity in serum, jejunal microbiota, and jejunal mucosal barrier damagein diarrhea lambs. Twenty-four newly weaned lambs were selected and distributed into four groups using a completely randomized experimental design: the control group (C), the diarrhea group (D), the group of antibiotic-treated under diarrhea (DA), and the group of Lactobacillus plantarum-treated under diarrhea (DL), with six replicates in each group. Lambs in groups D, DA and DL were dosed with 20 mL of ETEC K99 starting on the first day of the formal trial for 8 d consecutively. On the morning of the ninth day, lambs in group DA were injected regulated with florfenicol, and lambs in group DL were regulated by dosinggavaged with Lactobacillus plantarum, and the regulationabove treatment lasted for 4 consecutive days. The blood was collected fasting on days 1, 9 and 13 of the trial for the detection of serum antioxidant indexes as well as diamine oxidase (DAO) and D-lactic acid (D-LA) concentration in lambs. After slaughter the jejunal content was collected to determine the composition of bacterial community. The jejunal mucosal were collected to explore the expression of intestinal mucosal barrier-associated proteins and mucus proteins. Alisin Blue staining was uesd to study the changes of acidic mucus. The results showed that: 1) After 8 d of ETEC K99 treatment, the feces of lambs in D, DA and DL were all shapeless, and the concentrations of DAO and D-LA in the serum of lambs in the three groups were significantly higher than those in group C (P < 0.05), which indicated that the model of diarrhea had been successfully constructed; 2) On the 9^th day, compared with that of group C, the T-AOC and SOD in serum of lambs in D, DA and DL groups decreased significantly (P < 0.05), and the MDA content increased significantly (P < 0.05). Compared to on the 9^th day, the antioxidant capacity of lambs in DA and DL groups increased significantly on the 13^th day (P < 0.05), but there were significant decrease on the concentration of DAO, D-LA and MDA in serum of lambs in these two groups (P < 0.05);3) Compared to group C, the expression of Occludin, Claudin1 and MUC2 protein in group D decreased significantly (P < 0.05). Compared with group D, the expression of Occludin protein in jejunal tissues of DA lambs was significantly higher (P < 0.05), but the expression of Claudin1 and MUC2 proteins reduced slightly (P>0.05). The expression of Occludin, Claudin1 and MUC2 protein in DL group were significantly higher than that of DA group (P < 0.05). 4) Compared with group C, the Chao and Shannon values of the bacterial community in the jejunal content of lambs in D and DA decreased significantly (P < 0.05), and while increased in DL group (P>0.05).The abundances of Firmicutes in the jejunum of lambs from C, D and DL groups were significantly higher than in DA group (P>0.05). The abundances of Actinobacteriota in the jejunum of lambs was significantly higher in C, D and DA than in DL group (P < 0.05). The abundances of Proteobacteria in the jejunum of lambs was a little higher in D group than in C and DL groups (P>0.05), while the abundances of Proteobacteria in the jejunum of lambs in group DA was significantly higher than in C, D and DL groups(P < 0.05). Correlation analysis revealed that more microorganisms associated with MUC2 expression and the phylum Proteobacteria abundance was significantly negatively correlated with the expression of MUC2 protein (P < 0.05), while the phylaabundance of Firmicutes and Verrucomicrobiota were significantly positively correlated with the expression of MUC2 protein (P < 0.05). NK4A214_group had a significant positive correlation with MUC2 expression (P < 0.05) and Escherichia-Shigella was significantly negatively correlated with MUC2 expression (P < 0.05). In conclusion, ETEC K99 induced diarrhea in lambs, decreased serum antioxidant capacity and microbial diversity of jejunal content, and inhibited the expression of jejunal mucus barrier protein. Compared to antibiotic therapy, gavaging Lactobacillus plantarum increases the diversity of jejunal contents flora and promotes the expression of jejunal acidic mucus and mucus tissue barrier proteins. Above all, Lactobacillus plantarum had a better effect in improving lamb diarrhea.

Key words: Lactobacillus plantarum, lambs, anti-oxidative capacity, mucosal barrier of jejunum, bacterial community

CLC Number:

S858.26

Bibo LI, Ke WU, Xiaolong SHI, Yining YAN, Jiahao LI, Guoqing DUAN, Xiong LI, Yanpeng REN, Jianing DONG, Chunxiang ZHANG, Youshe REN. Sheep-derived Lactobacillus plantarum Regulates the Bacterial Community and Mucosal Barrier in Jejunum of Diarrheic Lambs[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(8): 3552-3569.

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组别 Group	绵羊编号 Sheep number	粪便得分 Fecal score
C	C1	0
	C2	0
	C3	0
	C4	0
	C5	0
	C6	0
D	D1	1
	D2	2
	D3	0
	D4	1
	D5	2
	D6	0
DA	DA1	0
	DA2	1
	DA3	1
	DA4	1
	DA5	0
	DA6	2
DL	DL1	0
	DL2	1
	DL3	1
	DL4	2
	DL5	1
	DL6	1分

项目Item	时间/d Time	组别Group				SEM	P值(处理) P-value (Treatment)
项目Item	时间/d Time	C	D	DA	DL	SEM	P值(处理) P-value (Treatment)
二胺氧化酶/(pg·mL^-1)	1	243.67	237.68^B	239.12^B	234.78^B	2.97	0.814
DAO	9	237.04^b	318.62^aA	323.98^aA	280.81^aA	11.89	0.005
	13	238.19	—	216.62^B	228.66^B	5.71	0.345
SEM		2.57	18.59	17.65	10.19
P		0.596	0.001	0.003	0.042
D-乳酸/(pg·mL^-1)	1	81.42	93.57^B	96.32^A	90.05^AB	2.41	0.125
D-LA	9	82.56^c	123.96^aA	99.22^bA	99.96^bA	4.65	< 0.001
	13	81.68	—	74.40^B	78.40^B	1.63	0.199
SEM		0.85	7.05	3.73	3.70
P		0.882	0.002	0.002	0.038

项目Item	时间/d Time	组别Group				SEM	P值(处理) P-value (Treatment)
项目Item	时间/d Time	C	D	DA	DL	SEM	P值(处理) P-value (Treatment)
谷胱甘肽过氧化物酶/(U·mL^-1)	1	195.88	186.39	196.29	189.00	7.05	0.962
GSHPx	9	193.54	144.60	191.07	167.56	9.04	0.181
	13	189.94	—	199.05	203.43	5.49	0.657
SEM		7.44	16.49	3.31	9.20
P		0.960	0.241	0.669	0.313
总抗氧化能力/(mmol·mL^-1)	1	0.51	0.47^A	0.49^B	0.48^B	0.01	0.191
TAOC	9	0.51^a	0.39^cB	0.43^bcB	0.40^cC	0.02	0.049
	13	0.56	—	0.56^A	0.57^A	0.02	0.989
SEM		0.02	0.02	0.02	0.03
P		0.446	0.034	0.024	0.002
超氧化物歧化酶/(U·mL^-1)	1	13.17	13.38^A	12.30^A	11.71^B	0.37	0.399
SOD	9	11.42^a	8.55^bB	7.53^bB	8.30^bC	0.54	0.020
	13	13.31	—	14.18^A	15.47^A	1.02	0.266
SEM		0.46	1.33	1.05	1.05
P		0.284	0.042	0.002	< 0.001
丙二醛/(nmol·mL^-1)	1	1.55	1.06^B	1.41^B	1.67^B	0.11	0.211
MDA	9	1.67^b	2.15^abA	2.41^aA	2.47^aA	0.13	0.069
	13	1.52	—	1.43^B	1.61^B	0.11	0.069
SEM		0.10	0.27	0.19	0.17
P		0.840	0.009	0.017	0.023

项目Item	组别Group				SEM	P值(处理) P-value (Treatment)
项目Item	C	D	DA	DL	SEM	P值(处理) P-value (Treatment)
ACE	685.89^a	382.09^b	175.05^b	681.25^a	63.29	< 0.001
Chao	675.28^a	378.60^b	191.56^c	669.09^a	60.25	< 0.001
Shannon	4.01^a	3.78^a	2.97^b	4.01^a	0.14	0.010
Simpson	0.080^b	0.118^a	0.127^a	0.045^b	0.012	0.048
Coverage	0.998	0.999	0.999	0.998	0.000	0.079

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Sheep-derived Lactobacillus plantarum Regulates the Bacterial Community and Mucosal Barrier in Jejunum of Diarrheic Lambs

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