Metagenomic Analysis of Uterine Microbiota in Postpartum Normal and Endometritic Water Buffaloes (Bubalus bubalis)
DOI:
https://doi.org/10.6000/1927-520X.2013.02.03.4Keywords:
Buffalo, endometritis, 16S rDNA cloning, bacterial populationAbstract
In Indian subcontinent the water buffalo (Bubalus bubalis) is one of the important livestock animals. As in cows, postpartum infection like endometritis in dairy buffaloes is major cause for the economic loss in the dairy industries. Till date, there is no study regarding metagenomic analysis of bacterial population of postpartum endometritic buffaloes. The purpose of this study was to identify and compare the uterine bacterial composition in normal and endometritic postpartum buffaloes using 16S rDNA cloning, which was a type of culture-independent methods. A total of 151 cloned plasmids for 16S rDNA from both normal and endometritic uterine samples were sequenced. Cloning library of 16S rDNA revealed clear cut difference between bacterial populations of normal and endometritic postpartum buffaloes. Cloned sequences were assigned to five major groups and one uncultured group. The five major groups include- Bacteroidetes, Firmicutes, Fusobacteria, Proteobacteria, and Tenericutes. Major cloned sequences from normal status endometrium were affiliated to phylum Proteobacteria, and most of the sequences showed high degree of similarity with bacteria Haemophilus felis. Most of the sequences from cloned library of endometritic status samples were affiliated to phylum Proteobacteria and Tenericutes. The most prevalent bacteria found in endometritic samples were Psychrobacter sp. PRwf-1, Psychrobacter pulmonis, Ureaplasma diversum strain T95 and Ureaplasma diversum strain A417. A major number of cloned sequences from both normal and endometritic samples were assigned to uncultured group. The present data showed bacterial population of postpartum normal and endometritic buffaloes and also described the presence of various types microbiota in uterine samples.
References
Olson JD, Bretzlaff KN, Mortimer RG, Ball L. The metritis-pyometra complex. In: Morrow DA, editor. Current therapy in theriogenology. Saunders WB, Co., Philadelphia, PA. USA 1986; pp. 227-36.
Jainudeen MR. Reproduction in water buffalo. In: Morrow DA, editor. Current therapy in theriogenology. Saunders WB, Philadelphia, PA, USA 1986; pp. 443-49.
Azawi OI. Clinical bacteriological and histopathological studies of uterine infections of Iraqi buffalo cows. Ph. D. Thesis, College of Veterinary Medicine, University of Baghdad, Baghdad, Iraq 2006.
Khan HM, Mohanty TK, Raina VS, Gupta AK, Bhakat M. Effect of peripartum disorders on reproduction performance traits in murrah buffaloes at an organized farm. Buffalo Bull 2009; 28: 176-83, 211.
El-Jakee JK, Ahmed WM, El-Seedy FR, Abd El-Moez SI. Bacterial profile of the genital tract in female-buffalo during the different reproductive stages. Global Veterinaria 2008; 2(1): 7-14.
Sheldon IM, Noakes DE, Rycroft AN, Pfeiffer DU, Dobson H. Influence of uterine bacterial contamination after parturition on ovarian dominant follicle selection and follicle growth and function in cattle. Reproduction 2002; 123: 837-45. http://dx.doi.org/10.1530/rep.0.1230837 DOI: https://doi.org/10.1530/rep.0.1230837
Williams EJ, Fischer DP, Pfeiffer DU, et al. Clinical evaluation of postpartum vaginal mucus reflects uterine bacterial infection and the immune response in cattle. Theriogenology 2005; 63: 102-17. http://dx.doi.org/10.1016/j.theriogenology.2004.03.017 DOI: https://doi.org/10.1016/j.theriogenology.2004.03.017
Miller AN, Williams EJ, Sibley K, et al. The effects of Arcanobacterium pyogenes on endometrial function in vitro, and on uterine and ovarian function in vivo. Theriogenology 2007; 68: 972-80. http://dx.doi.org/10.1016/j.theriogenology.2007.07.013 DOI: https://doi.org/10.1016/j.theriogenology.2007.07.013
Sheldon IM, Rycroft AN, Dogan B, et al. Specific strains of Escherichia coli are pathogenic for the endometrium of cattle and cause pelvic inflammatory disease in cattle and mice. PLoS One 2010; 5: e9192. http://dx.doi.org/10.1371/journal.pone.0009192 DOI: https://doi.org/10.1371/journal.pone.0009192
Azawi OI. Postpartum uterine infection in cattle. Anim Reprod Sci 2008; 105: 187-208. http://dx.doi.org/10.1016/j.anireprosci.2008.01.010 DOI: https://doi.org/10.1016/j.anireprosci.2008.01.010
Amann RI, Ludwig W, Schleifer KH. Phylogenetic identification and in situ detection of individual microbial cells without cultivation. Microbiol Rev 1995; 59: 143-69. DOI: https://doi.org/10.1128/mr.59.1.143-169.1995
Whitman WB, Coleman DC, Wiebe WJ. Prokaryotes: The unseen majority. Proc Natl Acad Sci USA 1998; 95: 6578-83. http://dx.doi.org/10.1073/pnas.95.12.6578 DOI: https://doi.org/10.1073/pnas.95.12.6578
Rondon MR, August PR, Bettermann AD, et al. Cloning the soil metagenome: A strategy for accessing the genetic and functional diversity of uncultured microorganisms. Appl Environ Microbiol 2000; 66: 2541-47. http://dx.doi.org/10.1128/AEM.66.6.2541-2547.2000 DOI: https://doi.org/10.1128/AEM.66.6.2541-2547.2000
Handelsman J. Metagenomics: Application of genomics to uncultured microorganisms. Microbiol Mol Biol Rev 2004; 68: 669-85. http://dx.doi.org/10.1128/MMBR.68.4.669-685.2004 DOI: https://doi.org/10.1128/MMBR.68.4.669-685.2004
Santos TMA, Gilbert RO, Bicalho RC. Metagenomic analysis of the uterine bacterial microbiota in healthy and metritic postpartum dairy cows. J Dairy Sci 2011; 94: 291-302. http://dx.doi.org/10.3168/jds.2010-3668 DOI: https://doi.org/10.3168/jds.2010-3668
Sheldon IM, Lewis GS, LeBlanc S, Gilbert RO. Defining postpartum uterine disease in cattle. Theriogenology 2006; 65: 1516-30. http://dx.doi.org/10.1016/j.theriogenology.2005.08.021 DOI: https://doi.org/10.1016/j.theriogenology.2005.08.021
Giovannoni SJ. The polymerase chain reaction. Sequencing and hybridization techniques in bacterial systematics. In: Stackebrandt E, Goodfellow M, editors. John Wiley and Sons, New York, NY: 1991; pp. 177-201.
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Biol 1990; 215: 403-10. DOI: https://doi.org/10.1016/S0022-2836(05)80360-2
Larkin MA, Blackshields G, Brown NP, et al. Clustal W and clustal X version 2.0. Bioinformatics 2007; 23: 2947-48. http://dx.doi.org/10.1093/bioinformatics/btm404 DOI: https://doi.org/10.1093/bioinformatics/btm404
Saitou N, Nei M. The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol Biol Evol 1987; 4: 406-25.
Jukes TH, Cantor CR. Evolution of protein molecules. Mammalian protein metabolism. Munro HN, editor. New York, NY: Academic Press 1969; Vol. 3: pp. 21-132. DOI: https://doi.org/10.1016/B978-1-4832-3211-9.50009-7
Machado VS, Oikonomou G, Bicalho MLS, Knauer WA, Gilbert R, Bicalho RC. Investigation of postpartum dairy cow’s uterine microbial diversity using metagenomic pyrosequencing of the 16S rRNA gene. Vet Microbiol 2012; 159: 460-69. http://dx.doi.org/10.1016/j.vetmic.2012.04.033 DOI: https://doi.org/10.1016/j.vetmic.2012.04.033
Santos TMA, Bicalho RC. Diversity and succession of bacterial communities in the uterine fluid of postpartum metritic, endometritic and healthy dairy cows. PLoS One 2012; 7(12): e53048. http://dx.doi.org/10.1371/journal.pone.0053048 DOI: https://doi.org/10.1371/journal.pone.0053048
Inzana TJ, Johnson JL, Shell L, Moller K, Kilian M. Isolation and characterization of a newly identified Haemophilus species from cats: Haemophilus felis. J Clin Microbiol 1992; 30: 2108-12. DOI: https://doi.org/10.1128/jcm.30.8.2108-2112.1992
Kwiecien JM, Little PB. Haemophilus somnus and reproductive disease in the cow: A review. Can Vet J 1991; 32: 595-01.
Bennett KW, Eley A, 1993. Fusobacteria: new taxonomy and related diseases. J Med Microbiol 1993; 39: 246-54. http://dx.doi.org/10.1099/00222615-39-4-246 DOI: https://doi.org/10.1099/00222615-39-4-246
Vela AI, Collins MD, Latre MV, et al. Psychrobacter pulmonis sp. nov., isolated from the lungs of lambs. Int J Syst Evol Microbiol 2003; 53: 415-19. http://dx.doi.org/10.1099/ijs.0.02413-0 DOI: https://doi.org/10.1099/ijs.0.02413-0
Mulira GL, Saunders R, Barth AD. Isolation of Ureaplasma diversum and Mycoplasmas from genital tracts of beef and dairy cattle in Saskatchewan. Can Vet J 1992; 33: 46-49.
Sanderson MW, Chenoweth PJ, Yeary T, Nietfeld JC. Prevalence and reproductive effects of Ureaplasma diversum in beef replacement heifers and the relationship to blood urea nitrogen level. Theriogenology 2000; 54: 401-408. http://dx.doi.org/10.1016/S0093-691X(00)00357-5 DOI: https://doi.org/10.1016/S0093-691X(00)00357-5
Doig PA, Ruhnke HL, Palmer NC. Experimental bovine genital ureaplasmosis. Can J Comp Med 1980; 44: 252-58.
Doig PA, Ruhnke HL, Waelchli-Suter R, Palmer NC, Miller RB. The role of Ureaplasma infection in bovine reproductive disease. Compend Contin Educ 1981; 3: S324-30.
Doig PA, Ruhnke HL, Palmer NC. Experimental bovine genital ureaplasmosis. II: granular vulvitis, endometritis and salpingitis following uterine inoculation. Can J Comp Med 1980; 44: 259-66.
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