Effect of Seasonal Variation on Intestinal Microflora of Free-range Luhua Chicken Analyzed by DGGE

doi:10.11924/j.issn.1000-6850.casb2021-0237

Abstract

Abstract:

To explore the effect of seasonal variation on the intestinal microflora of adult chicken, the PCR-DGGE method was used in this study to determine the microbial community in caecum and small intestine, which were collected from free-range Luhua chicken in winter, spring and summer. The results showed that the intestinal microflora of adult chicken changed greatly with season. There were only 66.7% and 62.5% similarity between microbial community in caecum and small intestine in summer and those in winter, respectively. The diversity of microbial community in caecum was much higher than that in small intestine. However, seasonal variation had different effects on the microflora of caecum and small intestine. The microflora diversity of caecum was lower in summer and winter than that in spring, while that of small intestine in summer and winter were higher. The common strains in caecum included Megamonas, Bacteroides and Clostridium, whose amount did not change with season; while the common strain in small intestine was mainly Lactobacillus, and the amount of its different species changed with season. These results provide a theoretical basis for adding distinct probiotics to forage according the seasonal variation.

Key words: intestinal microflora, seasonal variation, diversity, Luhua chicken

CLC Number:

S831.5

ZHANG Tengshuai, ZHANG Yanying, LIU Jingguo. Effect of Seasonal Variation on Intestinal Microflora of Free-range Luhua Chicken Analyzed by DGGE[J]. Chinese Agricultural Science Bulletin, 2022, 38(8): 129-134.

Figures/Tables 4

References 32

[1]	KAU A L, AHERN P P, GRIFFIN N W, et al. Human nutrition, the gut microbiome and the immune system[J]. Nature: International weekly journal of science, 2011, 474(7351):
[2]	SHANG Y, KUMAR S, OAKLEY B, et al. Chicken Gut Microbiota: Importance and Detection Technology[J]. Frontiers in Veterinary Science, 2018, 5.
[3]	DIAZ CARRASCO J M, CASANOVA N A, fernÁNdez MIYAKAWA M E. Microbiota, gut health and chicken productivity: what is the connection?[J]. Microorganisms, 2019, 7(10):374. doi: 10.3390/microorganisms7100374 URL
[4]	张亚楠. 蛋鸡肠道微生物菌群结构特征与产蛋水平关联性研究[D]. 郑州:河南农业大学, 2016.
[5]	KUMAR S, CHEN C, INDUGU N, et al. Effect of antibiotic withdrawal in feed on chicken gut microbial dynamics, immunity, growth performance and prevalence of foodborne pathogens[J]. PloS ONE, 2018, 13.
[6]	BALLOU A L, ALI R A, MENDOZA M A, et al. Development of the chick microbiome: how early exposure influences future microbial diversity[J]. Frontiers in Veterinary Science, 2016, 3:2.
[7]	STANLEY D, GEIER M S, CHEN H, et al. Comparison of fecal and cecal microbiotas reveals qualitative similarities but quantitative differences[J]. BMC Microbiology, 2015, 15:51. doi: 10.1186/s12866-015-0388-6 URL
[8]	WILLSON N L, NATTRASS G S, HUGHES R J, et al. Correlations between intestinal innate immune genes and cecal microbiota highlight potential for probiotic development for immune modulation in poultry[J]. Applied Microbiology and Biotechnology, 2018, 102:9317-9329. doi: 10.1007/s00253-018-9281-1 URL
[9]	FARAG M R, ALAGAWANY M. Physiological alterations of poultry to the high environmental temperature[J]. Journal of Thermal Biology, 2018, 76:101-106. doi: 10.1016/j.jtherbio.2018.07.012 URL
[10]	LARA L J, ROSTAGNO M H. Impact of heat stress on poultry production[J]. Animals, 2013, 3:356-369. doi: 10.3390/ani3020356 URL
[11]	BURKHOLDER K M, THOMPSON K L, EINSTEIN M E, et al. Influence of stressors on normal intestinal microbiota, intestinal morphology, and susceptibility to Salmonella enteritidis colonization in broilers[J], Poultry Science, 2008, 87(9):1734-1741. doi: 10.3382/ps.2008-00107 URL
[12]	彭骞骞, 王雪敏, 张敏红, 等. 持续偏热环境对肉鸡盲肠菌群多样性的影响[J]. 中国农业科学, 2016, 49(1):186-194.
[13]	OAKLEY B B, VASCONCELOS E J R, DINIZ P P V P, et al. The cecal microbiome of commercial broiler chickens varies significantly by season[J]. Poultry Science, 2018, 10;3635-3644.
[14]	TONG Q, LIU X N, HU Z F, et al. Effects of captivity and season on the gut microbiota of the brown frog (Rana dybowskii)[J]. Frontiers in Microbiology, 2019, 10:1912. doi: 10.3389/fmicb.2019.01912 URL
[15]	郭森, 孙全友, 魏凤仙, 等. 抗菌肽和地衣芽孢杆菌对肉仔鸡肠道微生物及免疫器官指数的影响[J]. 中国家禽, 2016, 38(18):26-31.
[16]	行浩. 饲粮中添加益生菌对蛋鸡早期生长、养分利用及肠道菌群的影响[D]. 扬州:扬州大学, 2016.
[17]	WEI S, MORRISON M, YU A. Bacterial census of poultry intestinal microbiome[J]. Poultry Science, 2013, 92:671-683. doi: 10.3382/ps.2012-02822 URL
[18]	MUYZER G, DE WAAL E C, UITTERLINDEN A G. Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA[J]. Applied and Environmental Microbiology, 1993, 59(3):695-700. doi: 10.1128/aem.59.3.695-700.1993 URL
[19]	STALEY J T, KONOPKA A. Measurement of in situ activities of nonphotosynthetic microorganisms in aquatic and terrestrial habitats[J]. Annual Review of Microbiology, 1985, 39:321-346. doi: 10.1146/micro.1985.39.issue-1 URL
[20]	RAPPE M S, GIOVANNONI S J. The uncultured microbial majority[J]. Annual Review of Microbiology, 2003, 57:369-394. doi: 10.1146/micro.2003.57.issue-1 URL
[21]	JOHNSTON-MONJE D, MEJIA J L. Botanical microbiomes on the cheap: inexpensive molecular fingerprinting methods to study plant-associated communities of bacteria and fungi[J]. Applications in Plant Sciences, 2020, 8(4):e11334.
[22]	MORGAN H H, TOIT M D, SETATI M E. The grapevine and wine microbiome: insights from high-throughput amplicon sequencing[J]. Frontiers in Microbiology, 2017, 8:820. doi: 10.3389/fmicb.2017.00820 URL
[23]	夏围围, 贾仲君. 高通量测序和DGGE分析土壤微生物群落的技术评价[J]. 微生物学报, 2014, 54(12):1489-1499.
[24]	ZHOU Y, YAO Q, ZHU H H. Soil organic carbon attenuates the influence of plants on root-associated bacterial community[J]. Frontiers in Microbiology, 2020, 11:594890. doi: 10.3389/fmicb.2020.594890 URL
[25]	MARGIOTTA E, MIRAGOLI F, CALLEGARI M L, et al. Gut microbiota composition and frailty in elderly patients with Chronic Kidney Disease[J]. PLoS One, 2020, 15(4):e0228530. doi: 10.1371/journal.pone.0228530 URL
[26]	王雪洁, 冯京海. 热应激对家禽肠道黏膜结构的影响及可能原因[J]. 动物营养学, 2018, 30(4):1224-1229.
[27]	SIEGERSTETTER S C, SCHMITZ-ESSER S, MAGOWAN E, et al. Intestinal microbiota profiles associated with low and high residual feed intake chickens across two geographical locations[J]. PLoS ONE, 2017, 12,e0187766. doi: 10.1371/journal.pone.0187766 URL
[28]	ZHOU X, JIANG X, YANG C, et al. Cecal microbiota of Tibetan Chickens from five geographic regions were determined by 16S rRNA sequencing[J]. MicrobiologyOpen, 2016, 5:753-762. doi: 10.1002/mbo3.2016.5.issue-5 URL
[29]	KNAPP O, BENZ R, POPOFF M R. Pore-forming activity of clostridial binary toxins[J]. Biochimica et Biophysica Acta, 2016, 1858(3):512-525.
[30]	FORTE C, MANUALI E, ABBATE Y, et al. Dietary Lactobacillus acidophilus positively influences growth performance, gut morphology, and gut microbiology in rurally reared chickens[J]. Poultry Science, 2018, 97(3):930-936. doi: 10.3382/ps/pex396 URL
[31]	ALAQIL A A, ABBAS A O, El-BELTAGI H S, et al. Dietary supplementation of probiotic Lactobacillus acidophilus modulates cholesterol levels, immune Response, and productive performance of laying hens[J]. Animals (Basel), 2020, 10(9):1588.
[32]	WANG S, MARTINS R, SULLIVAN M C, et al. Diet-induced remission in chronic enteropathy is associated with altered microbial community structure and synjournal of secondary bile acids[J]. Microbiome, 2019, 7:126. doi: 10.1186/s40168-019-0740-4 URL

引物	序列
GC357F	5′-CGCCCGCCGCGCGCGGCGGGCGGGGCGGGGGCACGGGGGGCCTACGGGAGGCAGCAG-3′
357F	5′-CCTACGGGAGGCAGCAG-3′
518R	5′-ATTACCGCGGCTGCTGG-3′

引物	序列
GC357F	5′-CGCCCGCCGCGCGCGGCGGGCGGGGCGGGGGCACGGGGGGCCTACGGGAGGCAGCAG-3′
357F	5′-CCTACGGGAGGCAGCAG-3′
518R	5′-ATTACCGCGGCTGCTGG-3′

	冬盲	春盲	夏盲	冬小	春小	夏小
相似系数	100%	82.3%	66.7%	100%	77.8%	62.5%
香浓指数(H)	3.02	3.49	3.19	2.82	2.07	2.49
均匀度(E)	0.95	0.94	0.96	0.89	0.80	0.89

	冬盲	春盲	夏盲	冬小	春小	夏小
相似系数	100%	82.3%	66.7%	100%	77.8%	62.5%
香浓指数(H)	3.02	3.49	3.19	2.82	2.07	2.49
均匀度(E)	0.95	0.94	0.96	0.89	0.80	0.89

条带编号	序列长度	数据库中最相近的菌种名称	登陆号	相似性%
1	189	Bacteroides barnesiae	NR_041446.1	100
2	172	Clostridium viride	NR_026204.1	95
3	196	Megamonas funiformis	NR_041590.1	97
4	194	Lactobacillus johnsonii	NR_117574.1	100
5	195	Lactobacillus amylovorus	NR_117064.1	99
6	189	Bacteroides salanitronis	NR_074616.1	95
7	169	Kineothrix alysoides	NR_156081.1	100
8	194	Clostridium spiroforme	NR_119030.1	100
9	170	Eubacterium coprostanoligenes	NR_104907.1	94
10	171	Colidextribacter massiliensis	NR_147375.1	96
11	169	Paraclostridium benzoelyticum	NR_148815.1	99
a	195	Lactobacillus aviaries	NR_112692.1	99
b	170	Clostridium hiranonis	NR_028611.1	98
c	211	Lactobacillus acidophilus	NR_113638.1	96
d	194	Lactobacillus gallinarum	NR_042111.1	100
e	195	Lactobacillus satsumensis	NR_028658.1	95
f	169	Uncultured bacterium	FJ471300.1	100
g	169	Paraclostridium benzoelyticum^*	NR_148815.1	98
h	169	Paraclostridium benzoelyticum^**	NR_148815.1	98