Immune-Related Differentially Expressed Genes of Spleen in Broilers Fed with Probiotic Bacillus cereus Screened by SSH Library and DGGE Technique

doi:10.7668/hbnxb.20190909

Abstract

Abstract: To explore the molecular mechanism of the effect of Bacillus cereus PAS38 on the immunity in broilers. 60 7-day-old broilers were divided into two groups,the control group was fed with basal diet,and the treatment group was fed with basal diet containing Bacillus cereus PAS38 1×10⁶ cfu/g. Six broilers were randomly selected from each group at the age of 42 days,spleens were taken and total RNA was extracted,then,and reversed transcription to cDNA. SSH and ACSSH techniques were used to construct splenic differential gene library and screen immune-related genes respectively. The results showed that 119 differentially expressed genes were screened by SSH,including 63 up-regulated genes and 56 down-regulated genes,and including 9 immunoregulatory genes,while five differentially expressed genes were screened by ACSSH,all of which were down-regulated genes,and no immunoregulatory genes were found. The results indicated that feeding Bacillus cereus PAS38 could affect the expression of immune genes such as JCHAIN, FTH1 and P2RX7 in the spleen of broilers,SSH library was superior to ACSSH library,and ACSSH technology was defective.

Key words: Broilers, Bacillus cereus, SSH technology, ACSSH technology, Differential gene

CLC Number:

Q78
S432.4

LI Jiajun, LI Jianzhen, LI Wanqiang, WANG Zhenhua, WANG Junrui, XIAO Dan, PAN Kangcheng, YANG Miao. Immune-Related Differentially Expressed Genes of Spleen in Broilers Fed with Probiotic Bacillus cereus Screened by SSH Library and DGGE Technique[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(S1): 392-398. doi: 10.7668/hbnxb.20190909.

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References

[1] Yan F,Polk D B. Probiotics:progress toward novel therapies for intestinal diseases[J]. Current Opinion in Gastroenterology,2010,26(2):95-101. doi:10.1097/MOG.0b013e328335239a.
[2] 张李荣. 三种益生菌对维鸡生长、小肠发育及盲肠微生物区系的影响[D]. 扬州:扬州大学,2018.
Zhang L R. Effects of three probiotics on growth,small intestine development and cecum microorganism of chicks[D]. Yangzhou:Yangzhou University,2018.
[3] Narayanan G,Baskaralingam V,Jiann C C,Rekha R,Vijayakumar S,Anjugam M,Iswarya A. Dietary supplementation of probiotic Bacillus licheniformis Dahb1 improves growth performance,mucus and serum immune parameters,antioxidant enzyme activity as well as resistance against Aeromonas hydrophila in tilapia Oreochromis mossambicus[J]. Fish & Shellfish Immunology,2018(74):501-508. doi:10.1016/j.fsi.2017.12.066.
[4] Madej J P,Stefaniak T,Bednarczyk M. Effect of in ovo-delivered prebiotics and synbiotics on lymphoid-organs' morphology in chickens[J]. Poultry Science,2015,94(6):1209-1219. doi:10.3382/ps/pev076.
[5] Zhang J C,Yu J F,Hong H L,Liu J,Lu H,Yan C. Identification of heavy metal pollutant tolerance-associated genes in Avicennia marina(Forsk.)by suppression subtractive hybridization[J]. Marine Pollution Bulletin,2017,119(1):81-91. doi:10.1016/j.marpolbul.2017.03.023.
[6] Yu Z B,Mu C K,Song W W,Li R H,Chen Y E,Wang C L. Screening of genes related to ovarian development in the swimming crab, Portunus trituberculatus,by suppression subtractive hybridization[J]. Genetics and Molecular Research,2015,14(4):18675-18686. doi:10.4238/2015.December.28.16.
[7] Kang X L,Liu Y F,Zhang J B,Xu Q,Liu C,Fang M. Characteristics and expression profile of KRT71 screened by suppression subtractive hybridization cDNA library in curly fleece Chinese Tan Sheep[J]. DNA and Cell Biology,2017,36(7):1-13. doi:10.1089/dna.2017.3718.
[8] Muyzer G,Smalla K. Application of denaturing gradient gelelectrophoresis(DGGE)and temperature gradient gelelectrophoresis(TGGE)in microbialecology[J]. Antonie Van Leeuwenhoek,1998,73(1):127-141. doi:10.1023/a:1000669317571.
[9] 王雪寒,夏冬双,武志华,任兴波,丁一秀,马强,刘惠荣. 内蒙古西部地区土壤放线菌的多样性分析[J]. 基因组学与应用生物学,2018,37(10):4340-4348. doi:10.13417/j.gab.037.004340.
Wang X H,Xia D S,Wu Z H,Ren X B,Ding Y X,Ma Q,Liu H R. Analysis on the diversity of soil actinomycetes in the western region of Inner Mongolia[J]. Genomics and Applied Biology,2018,37(10):4340-4348.
[10] Sevcan A,Aiyoub S E,Gozde O. Use of PCR-DGGE based molecular methods to assessment of microbial diversity during anaerobic treatment of antibiotic combinations[J]. Bioresource Technology,2015(192):735-740. doi:10.1016/j.biortech.2015.05.086.
[11] Garofalo C,Bancalari E,Milanović V,Cardinali F,Osimani A,Sardaro M L S,Bottari B,Bernini V,Aquilanti L,Clementi F,Neviani E,Gatti M. Study of the bacterial diversity of foods:PCR-DGGE versus LH-PCR[J]. International Journal of Food Microbiology,2017(242):24-36. doi:10.1016/j.ijfoodmicro.2016.11.008.
[12] Karina E M,Christian M,Marco V,Valencia F E,Ruas-Madiedo P,Delgado S. Bacterial diversity of the colombian fermented milk "Suero Costen o" assessed by culturing and high-throughput sequencing and DGGE analysis of 16S rRNA gene amplicons[J]. Food Microbiology,2017(68):129-136. doi:10.1016/j.fm.2017.07.011.
[13] Andrea O,Vesna M,Cristiana G,Cardinali F,Roncolini A,Sabbatini R,De Filippis F,Ercolini D,Gabucci C,Petruzzelli A,Tonucci F,Clementi F,Aquilanti L. Revealing the microbiota of marketed edible insects through PCR-DGGE,metagenomic sequencing and real-time PCR[J]. International Journal of Food Microbiology,2018(276):54-62. doi:10.1016/j.ijfoodmicro.2018.04.013.
[14] Gadde U,Oh S T,Lee Y S,Davis E,Zimmerman N,Rehberger T,Lillehoj H S. The effects of direct-fed microbial supplementation,as an alternative to antibiotics,on growth performance,intestinal immune status,and epithelial barrier gene expression in broiler chickens[J]. Probiotics & Antimicro,2017(9):397-405. doi:10.1007/s12602-017-9275-9.
[15] Gong Y,Li H,Li Y. Effects of Bacillus subtilis on epithelial tight junctions of mice with inflammatory bowel disease[J]. Journal of Interferon & Cytokine Research,2016,36(2):1-11. doi:10.1089/jir.2015.0030.
[16] 朱沛霁.枯草芽孢杆菌对雪山鸡生产性能、肠道健康和免疫机能的影响及机制[D].扬州:扬州大学,2017.
Zhu P Q. Effect of dietary supplementation of Bacillus Subtilis on growth performance,intestinal health and immune function responses of Xueshan Broiler[D]. Yangzhou:Yangzhou Universuty,2017.
[17] He Y,Wang A Q,Chen S,Wu Z,Zhang J Y,Wang M S,Jia R Y,Zhu D K,Liu M F,Yang Q,Wu Y,Sun K F,Chen X G,Cheng A C. Differential immune-related gene expression in the spleens of duck Tembusu virus-infected goslings[J]. Veterinary Microbiology,2017(212):39-47. doi:10.1016/j.vetmic.2017.08.002.
[18] Byadgi O,Chen Y C,Maekawa S,Wang P C,Chen S C. Immune-Related functional differential gene expression in Koi Carp(Cyprinus carpio)after challenge with Aeromonas sobria[J]. International Journal of Molecular Sciences,2018,19(7):2107. doi:10.3390/ijms19072107.
[19] Gao L,Yang Y,Yao Q,Chen K. Differentially expressed genes in the midguts of BmNPV-susceptible and resistant silkworm strains determined using suppression subtractive hybridization[J]. Invertebrate Survival Journal,2018,15(1):256-264.
[20] Liu Q N,Xin Z Z,Chai X Y,Jiang S H,Li C F,Zhang H B,Ge B M,Zhang D Z,Zhou C L,Tang B P. Characterization of immune-related genes in the yellow catfish Pelteobagrus fulvidraco in response to LPS challenge[J]. Fish & Shellfish Immunology,2016(56):248-254. doi:10.1016/j.fsi.2016.05.019.
[21] Fujita T,Kitaura F,Fujii H. A critical role of the Thy28-MYH9 axis in b cell-specific expression of the Pax5 gene in chicken B cells[J]. PLoS One,2015,10(1):e0116579. doi:10.1371/journal.pone.0116579.
[22] Jamrog L,Chemin G,Fregona V,Coster L,Pasquet M,Oudinet C,Rouqui N,Prade N,Lagarde S,Cresson C,H brard S,Nguyen H N S,Bousquet M,Quelen C,Brousset P,Mancini S J C,Delabesse E,Khamlichi A A,Gerby B,Broccardo C. PAX5-ELN oncoprotein promotes multistep B-cell acute lymphoblastic leukemia in mice[J]. PNAS,2018,115(41):10357-10362. doi:10.1073/pnas.1721678115.
[23] Danilo E. PCR-DGGE fingerprinting:novel strategies for detection of microbes in food[J]. Journal of Microbiological Methods,2004,56(3):297-314. doi:10.1016/j.mimet.2003.11.006.
[24] Giusy F,Giovina R. Detection of polymorphisms within the FAS cDNA gene sequence by GC-clamp denaturing gradient gel electrophoresis[J]. Immunogenetics,1994,39(6):437-439. doi:10.1007/bf00176163.
[25] Luo H F,Qi H Y,Xue K,Wang X Y,Wang C,Zhang H X. Influence of application of GC-clamp on study of soil microbial diversity by PCR-DGGE[J]. Acta Ecologica Sinica,2003,23(10):2170-2175.

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