3 Biotech

, 8:176 | Cite as

Bacterial lactase genes diversity in intestinal mucosa of mice with dysbacterial diarrhea induced by antibiotics

Original Article

Abstract

The current study aimed at exploring the diversity of bacterial lactase genes in the intestinal mucosa of mice with dysbacterial diarrhea induced by antibiotics and to provide experimental basis for antibiotics-induced diarrhea. Mice model of dysbacterial diarrhea was established by gastric perfusion with mixture of cephradine capsules and gentamicin sulfate (23.33 mL kg−1 d−1), twice a day and continuously for 5 days. Intestinal mucosa from jejunum to ileum was collected, and bacterial metagenomic DNA was extracted for Miseq metagenome sequencing to carry out diversity analysis. The results showed that specific operational taxonomic units (OTUs) were 45 in the control group and 159 in the model group. The Chao1, ACE, Shannon and Simpson indices in model group were significantly higher (P < 0.01 or P < 0.05) than control group. Principal component analysis (PCA) and box chart of the control group were relatively intensive, while in the model group, they were widely dispersed. Furthermore, the inter-group box area was higher than that in the intra-group. Compared with the model group, the abundance of bacterial lactase genes in Proteobacteria from the intestinal mucosa of the control group was higher, but lower in Actinobacteria and unclassified bacteria. At the genus level, the relative abundance of bacterial species and taxon units in model group was obviously increased (P < 0.05). Our results indicate that antibiotics increased the diversity and abundance of bacterial lactase genes in the intestinal mucosa, as the abundance of Betaproteobacteria, Cupriavidus, Ewingella, Methyloversatilis, Rhodocyclaceae and Rhodocyclales. In addition, antibiotics become an additional source for lactase genes of Ewingella, Methyloversatilis, Mycobacterium, Microbacterium, Beutenberqia and Actinomyces.

Keywords

Dysbacterial diarrhea Lactase gene Intestinal mucosa Gene diversity High-throughput sequencing Antibiotics 

Notes

Acknowledgements

Thanks are extended to the National Natural Science Foundation of China for research funding (no. 81573951).

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest related to this article.

References

  1. Amann RI, Ludwig W, Schleifer KH (1995) Phylogenetic identification and in situ detection of individual microbial-cells without cultivation. Microbiol Res 59:143–169Google Scholar
  2. Blaxter M, Mann J, Chapman T, Thomas F, Whitton C, Floyd R, Abebe E (2005) Defining operational taxonomic units using DNA barcode data. Philos Trans R Soc Lond B Biol Sci 360:1935–1943CrossRefGoogle Scholar
  3. Cao R, Wang H, Guo KX, Peng MJ, He YS, Zhang QL, Peng CY, Tan ZJ (2014) Effects of ultra-micro Qiweibaizhusan on disaccharides metabolism of intestinal microbiotia in diarrheal mice with dysbacteriosis. Int J Curr Microbiol Appl Sci 3:446–457Google Scholar
  4. Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK, Fierer N, Peṅa AG, Goodrich JK, Gordon JI, Hutley GA, Kelley ST, Knights D, Koeniq JE, Ley RE, Lozupone CA, McDonald D, Muegge BD, Pirrung M, Reeder J, Sevinsky JR, Turnbaugh PJ, Walters WA, Widmann J, Yatsunenko T, Zaneveld J, Knignt R (2010) QIIME allows analysis of high-throughput community sequencing data. Nat Methods 7:335–336CrossRefGoogle Scholar
  5. Chen XQ, Huang XJ, Shi DY, Guo SN (2014) Research progress on interaction of Chinese materia medica with intestinal flora. Chin Tradit Herbal Drugs 45:1031–1036Google Scholar
  6. Edgar RC (2010) Search and clustering orders of magnitude faster than BLAST. Bioinformatics 26:2460–2461CrossRefGoogle Scholar
  7. Fukuda S, Toh H, Hase K, Oshima K, Nakanishi Y, Yoshimura K, Tobe T, Clarke JM, Topping DL, Suzuki T, Taylor TD, Itoh K, Kikuchi J, Morita H, Hattori M, Ohno H (2011) Bifidobacteria can protect from enteropathogenic infection through production of acetate. Nature 469:543–547CrossRefGoogle Scholar
  8. Guo BH (2001) Physiological function of intestinal flora. J Chin Dairy Ind 29:4Google Scholar
  9. Hong N, Zhan XB (2014) Correlation of gut microflora and intestinal mucosal immunity. J Med Postgra 27:444–446Google Scholar
  10. Jiang XD, Guo GG, Zhang J (2014) Association of genetic diversity for Amy6-4 gene with α-amylase activity in germplasm of barley. Acta Agron Sin 40:205–213CrossRefGoogle Scholar
  11. Jin L, Yang XH, Ren JL, Li JL, Guo XY, Cao P, Wang Z (2012) Effect of dietary compound probiotics on disaccharidase in small intestine mucosa of layer breeders. China Poultry 34:14–17Google Scholar
  12. Juajun O, Nguyen TH, Maischberger T, Iqbal S, Haltrich D, Yammabhai M (2011) Cloning, purification, and characterization of β-galactosidase from Bacillus licheniformis DSM 13. Appl Microbiol Biotechnol 89:645–654CrossRefGoogle Scholar
  13. Li G, Xiao XH, Jin C, Li ZL, Luo Y (2007) Chinese medicinal compound and modulation of intestinal microecology. Chin J Integr Tradit West Med 27:466–469Google Scholar
  14. Liu QS, Liu H, Peng W, She Y, Tan ZJ (2015) The influence of antibiotic modeling on intestinal mucosa in dysbacteriotic diarrhea mice. Chin J Microecol 27:501–504Google Scholar
  15. Long CX, He L, Guo YF, Liu YW, Xiao NQ, Tan ZJ (2017a) Diversity of bacterial lactase genes in intestinal contents of mice with antibiotics-induced diarrhea. World J Gastroenterol 23:7584–7593CrossRefGoogle Scholar
  16. Long CX, He L, Liu YJ, Hui HY, Tan ZJ, Li DD (2017b) Universal primer for analysis of the diversity of intestinal bacterial lactase gene. Chin J Appl Environ Biol 23:758–763Google Scholar
  17. Lu DY (2001) Medical microbiology. People’s Medical Publishing House, Beijing, p 89Google Scholar
  18. Luo WX, Xie M, Gao LW (2016) Observation on the therapeutic effect of lactase on lactose intolerance in infants with diarrhea. Strait Pharm J 28:153–154Google Scholar
  19. Mahaffee WF, Kloepper JW (1997) Temporal changes in the bacterial communities of soil, rhizosphere, and endorhiza associated with field-grown cucumber (Cucumis sativus L). Microb Ecol 34:210–223CrossRefGoogle Scholar
  20. Merga Y, Campbell BJ, Rhodes JM (2014) Mucosal barrier, bacteria and inflammatory bowel disease: possibilities for therapy. Dig Dis 32:475–483CrossRefGoogle Scholar
  21. Parvin R, Louie T, Pitchumoni CS (2013) Infectious complications of acute pancreatitis. Infect Dis Clin Pract 21:94–104CrossRefGoogle Scholar
  22. Peng HZ, Ren LH (2011) Relationship between antibiotic associated diarrhea and lactose intolerance. Chin Gen Pract 14:2999–3006Google Scholar
  23. Pitta DW, Pinchak E, Dowd SE, Osterstock J, Gontcharova V, Youn E, Dorton K, Yoon I, Min BR, Fulford JD, Wickersham TA, Malinowski DP (2010) Rumen bacterial diversity dynamics associated with changing from bermudagrass hay to grazed winter wheat diets. Microb Ecol 59:511–522CrossRefGoogle Scholar
  24. Pitta DW, Parmar N, Patel AK, Induqu N, Kumar S, Prajapathi KB, Patel AB, Reddy B, Joshi C (2014) Bacterial diversity dynamics associated with different diets and different primer pairs in the rumen of Kankrej cattle. PLoS One 9:e111710CrossRefGoogle Scholar
  25. Ramette A (2007) Multivariate analyses in microbial ecology. FEMS Microbiol Ecol 62:142–160CrossRefGoogle Scholar
  26. Rhimi M, Aghajari N, Jaouadi B, Juy M, Boudebbouze S, Maquin E, Haser R, Bejar S (2009) Exploring the acidotolerance of beta-galactosidase from Lactobacillus delbrueckii subsp. bulgaricus: an attractive enzyme for lactose bioconversion. Res Microbiol 160:775–784CrossRefGoogle Scholar
  27. Segata N, Izard J, Waldron L, Gevers D, Miropolsky L, Garrett WS, Huttenhower C (2011) Metagenomic biomarker discovery and explanation. Genome Biol 12:1–18CrossRefGoogle Scholar
  28. Shannon CE (1997) The mathematical theory of communication. 1963. MD Comput Comput Med Pract 14:306–317Google Scholar
  29. Tan CC, Shi L (2008) Diversity of Helicobacter pylori clinical isolates by the method of polymerase chain reaction-restriction fragment length polymorphism. Mod Dig Int 13:18–21Google Scholar
  30. Tan ZY, Peng GX, Xu PZ, Ai SY, Tang SH, Zhang GX, Zeng FY (2009) Diversity and nitrogenase activity of endogenous azotobacteria in Oryza rufipogon. Chin Sci Bull 54:1885–1893CrossRefGoogle Scholar
  31. Tan ZJ, Zhang HL, Zhou SN, Yu WY, Zeng A, Cai Y, Cai GX (2013) Change of intestinal microbes in dysbacteriosis-modeled mice treated with ultra-micro powder of Qiweibaizhusan. Chin J Appl Environ Biol 19:449–453CrossRefGoogle Scholar
  32. Wang JZ, Liang JR, Qiu HY, Duan R, Xiao YC, Wang X, Jing HQ (2014) Analysis on polymorphism of Yersinia enterocolitica urease gene and urease activity. Chin J Zoon 30:140–145Google Scholar
  33. Wu H, Zhou SN, Guo C, Tan ZJ, Cai GX, Zeng A, Zhang HL (2012) A metagenome DNA extracting method of intestinal flora in mice for molecular diversity analysis based on PCR technology. Chin J Microecol 24:648–651Google Scholar
  34. Yang Y, Huang L, Yang L, Xie D, Wang T (2015) Reduction characteristics and differential expression of Acidiphilium cryptum XTS Cr(VI)-reduced related gene. Microbiol China 42:64–73Google Scholar
  35. Zeng A, Zhang HL, Tan ZJ, Cai Y, Cai GX, Zhou SN (2012) The construction of mice diarrhea model due to dysbacteriosis and curative effect of ultra-micro Qiweibaizhusan. Microbiol China 39:1341–1348Google Scholar
  36. Zhang J, Zhang HW, Li XY, Su ZC, Zhang CG (2006) Soil microbial ecological process and microbial functional gene diversity. Chin J Appl Ecol 17:1129–1132Google Scholar
  37. Zhang HL, Cai Y, Tan ZJ, Zhou SN, Guo KX, She Y, Cai GX (2014) Effects of ultra-micro powder Qiweibaizhusan on metabolism diversity of intestinal microflora in diarrhea mice with dysbacteriosis. Chin J Appl Environ Biol 1:93–100Google Scholar
  38. Zhao WJ, Liu SY, Ding JM, Dai RH, Meng H (2016) Metagenomic sequencing of gut microbiota along the intestinal tracts and feces in mice. J Shanghai Jiaotong Univ (Agric Sci) 34:15–21Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Hunan University of Chinese MedicineChangshaChina
  2. 2.College of Mathematics and FinanceHunan University of Humanities, Science and TechnologyLoudiChina
  3. 3.School of Animal Science and TechnologyHuazhong Agricultural UniversityWuhanChina
  4. 4.Rongjun Hospital of Hunan ProvinceChangshaChina

Personalised recommendations