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Mechanism of Alginate Oligosaccharides Fermented with Gut Microbiota Inoculum against Salmonella enterica by Transcriptomic Analysis

CHENG Jiaying, XIAO Mengshi, REN Xinmiao, YU Ying, FU Xiaodan, MOU Haijin

Journal of Henan Agricultural Sciences 2023, 52 (6): 139-149. DOI: 10.15933/j.cnki.1004-3268.2023.06.015

Abstract （391）

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In order to explore the effect of alginate oligosaccharides（AOS）on S.enterica in the intestinal environment，AOS was prepared by enzymatic degradation，furthermore the simulated digestion characteristics of AOS were evaluated in vitro.The total number of bacteria and S.enterica in the process of AOS fermentation by chicken gut microbiota were quantified by in vitro simulated fermentation model，and the response of S.enterica to AOS in the intestinal environment was analyzed by transcriptomics.The results showed that the content of reducing sugar and molecular weight of AOS did not change significantly，indicating that AOS were not digested.When AOS was fermented by gut microbiota，the relative abundance of S.enterica in the flora decreased from 4.6% to 1.1%.Transcriptomics and reverse transcription⁃quantitative PCR analysis showed that the expression of key genes of S.enterica flagella assembly，two⁃component system，infection and virulence was down⁃regulated.Therefore，the gut microbiota metabolites of AOS could inhibit the growth and virulence of S.enterica.

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Effects of Clostridium butyricum on Growth，Intestinal Morphology and Barrier Function of Broilers

LI Wenjia, XU Bin, WEI Fengxian, DENG Wen, MA Huihui, WANG Linyi, BAI Jie, SUN Quanyou, XI Yanyan, LI Shaoyu

Journal of Henan Agricultural Sciences 2023, 52 (1): 144-153. DOI: 10.15933/j.cnki.1004-3268.2023.01.015

Abstract （505）

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The experiment was conducted to investigate the effects of Clostridium butyricum in diet on growth performance，intestinal morphology and intestinal barrier function of broilers. A total of 324 healthy one‑day‑old Arbor Acres male broilers with the same specifications were randomly divided into 3 treatments with 6 replicates per treatment and 18 broilers per replicate. Control group（CON）was fed with a basal diet，antibiotic group（ANT）and Clostridium butyricum group（CB）were fed with the basal diet containing 10 mg/kg virginiamycin and 1×10 ⁹ cfu/kg Clostridium butyricum，respectively.The experiment period was 42 days. At the age of 21 and 42 days，the growth performance，intestinal tissue morphology，intestinal mucosal pyroptosis and intestinal barrier related gene mRNA expression abundance of each group were statistically analyzed. The results showed that compared with the CON group，the average daily gain（ADG）and final body weight（FBW）in ANT group and CB group were significantly increased by 6.31% and 8.43%，and 6.26% and 8.31%，respectively.At 42 days，the levels of Caspase‑1，IL‑18 and IL‑1β in CB group were significantly decreased by 24.91%，42.96% and 19.91% compared with the CON group，respectively. The jejunum villus height and villus height/crypt depth（V/C）ratio were significantly increased by 13. 40% and 54.20%，respectively；and the ileum crypt depth was significantly decreased by 32.62%，and the V/C ratio of ileum was significantly increased by 60.76%.At 42 days，the mRNA relative expression levels of IL‑1β， T NF‑α， Caspase‑1 and IL‑18 in jejunum mucosa in CB and ANT groups were significantly lower than those in CON group and the expression levels of ZO‑1 and Claudin‑1 in CB group were significantly up‑regulated，respectively. In conclusion，dietary supplementation of 1×10 ⁹ cfu/kg C.butyricum can improve the growth performance and feed utilization of broilers，improve the intestinal morphology，enhance the intestinal immune and barrier functions，and alleviate the intestinal cell pyroptosis of broilers to a certain extent.

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Effect of Fermented Traditional Chinese Medicine on Intestinal Microflora of Broilers with Avian Colibacillosis

GU Yanli, CAO Yanzi, WANG Xiaoyu, SHAN Chunqiao, LIU Yan, JIANG Guotuo

Journal of Henan Agricultural Sciences 2022, 51 (7): 145-153. DOI: 10.15933/j.cnki.1004-3268.2022.07.015

Abstract （460）

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This study aimed to evaluate the effect of fermented traditional Chinese medicine on intestinal microflora of broilers with avian colibacillosis.270 healthy 1‑day‑old AA white‑feathered broilers were divided into three groups，three replicates for each treatment and 30 broilers for each repetition.Broilers in the control group（D group）were routinely fed.The fermented traditional Chinese medicine group （PEG group）and the antibiotic group（K group）were modeled with E.coli O78 and treated.The samples were analyzed through 16S rRNA sequencing technology.The result showed that after treatment with the fermented traditional Chinese medicine composed of Andrographis herba， Pulsatillae Radix， Patrinia， Rhizoma Coptidis，etc，there was no significant difference in the richness and diversity of microflora with the control group，and the content of Akkermansia was significantly increased.The fermented traditional Chinese medicine can regulate intestinal microflora and substance transport and metabolism，increase the abundance of beneficial bacteria and inhibit harmful bacteria in broilers.

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Research Progress on the Relationship between Intestinal Barrier Damage and Intestinal Inflammation Development in Fish

CHEN Xiumei, WANG Guiqin, SHAN Xiaofeng, QIAN Aidong

Journal of Henan Agricultural Sciences 2022, 51 (5): 1-9. DOI: 10.15933/j.cnki.1004-3268.2022.05.001

Abstract （559）

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Fish intestinal barrier mainly includes biological，chemical，mechanical and immune barriers.Its barrier function plays an important role in maintaining the intestinal homeostasis，and will affect the intestinal health once the damage occurs. A large number of studies have shown that intestinal barrier damage was closely related to intestinal inflammation，and intestinal barrier damage induced by various factors could lead to intestinal inflammation. Therefore，domestic and foreign research progress on the relationship of fish intestinal barrier damage and intestinal inflammation development is reviewed.Based on understanding of the structure and function of intestinal barrier in fish，this paper analyzes the relationship between intestinal barrier injury and intestinal inflammation in fish，and explores the control strategy of intestinal inflammation in fish based on the pathway of stablizing intestinal barrier，to provide
new methods for intestinal inflammation prevention in fish and other aquatic animals.

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Effect of Fermented Traditional Chinese Medicine on Intestinal Flora Structure of Broilers under Heat Stress

HAO He, ZHONG Cuihong, LI Xinze, WANG Haiying, WU Ya’nan, WANG Fengshen, LIU Guanhui, SHI Yuxiang, ZHANG Yongying

Journal of Henan Agricultural Sciences 2021, 50 (9): 135-142. DOI: 10.15933/j.cnki.1004-3268.2021.09.016

Abstract （536）

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The purpose of this study was to explore the effect of fermented Chinese medicine on the intestinal flora structure of broilers under heat stress，and to provide theoretical support for the prevention and control of heat stress and the research on the mechanism of heat stress.180 Ross 308 broilers were randomly divided into six groups with three replicates per group and 10 broilers per replicate. Control group was routine feeding；Broilers in each heat stress group were fed at 34 ℃. The heat stress control group was fed with basal diet under heat stress.In the heat stress experimental group，0.5 g/kg，1.0 g/kg and 1.5 g/kg fermented Chinese medicine and 1.5 g/kg unfermented Chinese medicine were added to the diet of broilers.Feeding lasted for 14 d.The composition of cecum flora of broilers in each group was detected by high⁃throughput detection technology based on 16S rDNA amplicon. The number of Lactobacillus，Salmonella and Escherichia coli in cecum of broilers in each group was counted by plate colony counting method. The results showed that the effect of adding 1.5 g/kg fermented Chinese medicine in broiler diet was the best. Compared with the heat stress control group，the average daily weight gain and average daily feed intake of broilers under heat stress were significantly increased by 36.30% and 26.42%，respectively，and the feed to meat ratio was significantly decreased by 8. 82%. Blautia abundance increased in cecum，the number of Lactobacillus increased by 25. 04%，and the number of Escherichia coli and Salmonella decreased by 11.25% and 13.74%，respectively. The above results showed that fermented traditional Chinese medicine could improve the growth performance of broilers under heat stress，regulate the cecum flora of broilers，increase the number of beneficial bacteria and reduce the number of harmful bacteria.

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Effects of Lactobacillus plantarum LR-39 on Growth and Intestinal Microbiota Regulation of Dogs

CHEN Zedong, ZHU Jianping, GUO Shuyu, LU Wei, LIU Lei

Journal of Henan Agricultural Sciences 2021, 50 (3): 148-156. DOI: 10.15933/j.cnki.1004-3268.2021.03.020

Abstract （526）

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To evaluate the growth-promoting and intestinal flora regulation effects of Lactobacillus plantarum LR-39 isolated from fermented pickles,ten 3.5-month-old beagle dogs were divided into control group and experimental group. The control group was fed with basic dog food,while the experimental group was daily given LR-39 with the basic dog food for 28 days.The average daily gain,feed weight ratio,nutrient digestibility and other indicators were measured at the end of the test. Based on highthroughput sequencing,the relative abundance of intestinal flora at the phylum level and the differences in the characteristic microbes of each group of intestinal flora were analyzed. PICRUSt was performed to further analyze the expression difference of microbial functional genes in the intestinal tract.The whole genome sequencing of LR-39 and subsequent COG and CAZy annotation analysis were performed.The results of the growth-promoting test showed that the average daily gain of the experimental group was significantly( P<0. 01) higher than that of the control group;the feed weight ratio of the experimental group was significantly( P<0.05) lower than that of the control group.The digestibility of crude protein,crude fat and calcium in the experimental group was significantly( P<0.05) higher than those of the control group.High-throughput sequencing results showed that at the phylum level,the relative abundance of Fusobacteria in the experimental group was significantly( P<0.05) higher than that in the control group,while the relative abundance of Actinobacteria was significantly( P<0.05) lower than that in the control group.LDA effect size(LEfSe) analysis showed that the characteristic bacterial generus of the intestinal flora in the experimental group was Fusobacterium,while the characteristic bacterial genera of the control group were Bulleidia, Catenibacterium,Eubacterium,Blautia,and Faecalibacterium.At the species level,the relative abundance of Lactobacillus helveticus in the experimental group was significantly( P<0.01)increased.Intestinal functional gene difference analysis showed that the relative expression abundance of intracellular transport and extracellular structure genes in the experimental group was significantly( P<0.05) higher than that in the control group,and the relative expression abundance of secondary metabolite gene was significantly( P<0.05) lower than that in the control group.Whole genome sequencing showed that the strain had a full length of 247 554 bp, no plasmid, with GC content of 44.26%.The abundance of genes related to carbohydrate metabolism was the highest,followed by amino acid metabolism related genes,the abundance of genes related to lipid metabolism was lower,and the genes related to antagonistic mechanism had a certain proportion.The enzyme system mainly included glycoside hydrolases, glycosyltransferases, carbohydrate esterases, carbohydrate binding molecules and auxiliary enzymes.In summary,feeding LR-39 can promote the growth of dogs,change the structure of the intestinal flora,regulate the expression of functional genes in intestinal flora,and it has the potential to develop into probiotic products.

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