title: Bacterial community profiling of milk samples as a means to understand culture-negative bovine clinical mastitis.
abstract: Inflammation and infection of bovine mammary glands, commonly known as mastitis, imposes significant losses each year in the dairy industry worldwide. While several different bacterial species have been identified as causative agents of mastitis, many clinical mastitis cases remain culture negative, even after enrichment for bacterial growth. To understand the basis for this increasingly common phenomenon, the composition of bacterial communities from milk samples was analyzed using culture independent pyrosequencing of amplicons of 16S ribosomal RNA genes (16S rDNA). Comparisons were made of the microbial community composition of culture negative milk samples from mastitic quarters with that of non-mastitic quarters from the same animals. Genomic DNA from culture-negative clinical and healthy quarter sample pairs was isolated, and amplicon libraries were prepared using indexed primers specific to the V1-V2 region of bacterial 16S rRNA genes and sequenced using the Roche 454 GS FLX with titanium chemistry. Evaluation of the taxonomic composition of these samples revealed significant differences in the microbiota in milk from mastitic and healthy quarters. Statistical analysis identified seven bacterial genera that may be mainly responsible for the observed microbial community differences between mastitic and healthy quarters. Collectively, these results provide evidence that cases of culture negative mastitis can be associated with bacterial species that may be present below culture detection thresholds used here. The application of culture-independent bacterial community profiling represents a powerful approach to understand long-standing questions in animal health and disease.
authors: Department of Veterinary Microbiology and Preventive Medicine, College of Veterinary Medicine, Iowa State University, Ames, Iowa, United States of America.
authors: Joanna S
authors: JS
authors: Kuehn
authors: Patrick J
authors: PJ
authors: Gorden
authors: Daniel
authors: D
authors: Munro
authors: Ruichen
authors: R
authors: Rong
authors: Qunfeng
authors: Q
authors: Dong
authors: Paul J
authors: PJ
authors: Plummer
authors: Chong
authors: C
authors: Wang
authors: Gregory J
authors: GJ
authors: Phillips
mesh_terms: D000818:Animals; D001419:Bacteria / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification*; D002417:Cattle; D046508:Culture Techniques; D004275:DNA, Ribosomal / Q000235:genetics; D005260:Female; D008414:Mastitis, Bovine / Q000382:microbiology*; D008892:Milk / Q000382:microbiology*; D017422:Sequence Analysis, DNA
chemical_list: D004275:DNA, Ribosomal
references: Bioinformatics. 2010 Mar 15;26(6):715-21
references: 20130030
references: N Z Vet J. 2009 Feb;57(1):44-9
references: 19252542
references: Vet Clin North Am Food Anim Pract. 2003 Mar;19(1):199-221
references: 12682943
references: J Clin Microbiol. 1991 Sep;29(9):2041-2
references: 1774332
references: Vet Rec. 1986 Oct 4;119(14):350-1
references: 3787992
references: J Dairy Sci. 2011 Sep;94(9):4476-87
references: 21854920
references: Appl Environ Microbiol. 2002 Apr;68(4):1548-55
references: 11916667
references: BMC Microbiol. 2011 Aug 30;11:194
references: 21878094
references: J Appl Microbiol. 2007 Oct;103(4):754-64
references: 17897177
references: PLoS One. 2010 May 06;5(5):e10518
references: 20463896
references: PLoS Genet. 2008 Nov;4(11):e1000255
references: 19023400
references: Lett Appl Microbiol. 2012 Jun;54(6):572-6
references: 22443137
references: Res Vet Sci. 2003 Apr;74(2):105-12
references: 12589733
references: J Dairy Sci. 2011 Apr;94(4):1893-9
references: 21426978
references: Appl Environ Microbiol. 2007 Nov;73(22):7162-8
references: 17890340
references: Clin Microbiol Infect. 2003 Apr;9(4):263-73
references: 12667235
references: J Hosp Infect. 2006 Mar;62(3):278-84
references: 16337309
references: J Hosp Infect. 2010 Jul;75(3):153-7
references: 20434794
references: Appl Environ Microbiol. 2012 Jan;78(2):326-33
references: 22081572
references: Appl Environ Microbiol. 2009 Dec;75(23):7537-41
references: 19801464
references: Int J Food Microbiol. 2011 Apr 29;146(3):253-62
references: 21429612
references: J Clin Microbiol. 2001 Mar;39(3):990-4
references: 11230416
references: Appl Environ Microbiol. 2007 Mar;73(6):1882-91
references: 17259356
references: Vet Microbiol. 2009 Feb 16;134(1-2):100-5
references: 18930612
references: J Vet Diagn Invest. 2010 Nov;22(6):995-7
references: 21088192
references: N Engl J Med. 2001 Apr 12;344(15):1171-2
references: 11302149
references: BMC Bioinformatics. 2006 Aug 07;7:371
references: 16893466
references: Nat Methods. 2010 Dec;7(12):943-4
references: 21116242
references: Appl Environ Microbiol. 2007 Aug;73(16):5261-7
references: 17586664
references: ISME J. 2010 Jan;4(1):17-27
references: 19710709
references: Pediatr Infect Dis J. 2004 Dec;23(12):1169-71
references: 15626961
references: BMC Genomics. 2006 Mar 20;7:57
references: 16549033
references: Biochem Biophys Res Commun. 2002 Jun 28;294(5):1138-43
references: 12074595
references: J Dairy Sci. 1994 Jul;77(7):2103-12
references: 7929968
references: Appl Environ Microbiol. 2004 Jul;70(7):4187-92
references: 15240300
references: Nucleic Acids Res. 2007;35(21):7188-96
references: 17947321
references: PLoS Pathog. 2008 Feb 8;4(2):e20
references: 18248093
references: FEBS Lett. 1993 Feb 8;317(1-2):96-100
references: 8428640
references: Vet Microbiol. 2012 Jun 15;157(3-4):363-8
references: 22306036
references: PLoS Biol. 2012;10(8):e1001377
references: 22904687
references: Appl Environ Microbiol. 2003 Jan;69(1):130-8
references: 12513987
references: Foodborne Pathog Dis. 2012 Feb;9(2):132-7
references: 22283638
references: J Dairy Sci. 2009 May;92(5):2020-6
references: 19389959
references: Infect Control Hosp Epidemiol. 2008 Oct;29(10):990-2
references: 18808348
references: Int J Syst Evol Microbiol. 2002 Jul;52(Pt 4):1291-1297
references: 12148642
references: BMC Genomics. 2006 Aug 23;7:216
references: 16928277
references: J Dairy Sci. 2003 Nov;86(11):3466-72
references: 14672176
references: BMC Microbiol. 2007 Nov 30;7:108
references: 18047683
references: Appl Environ Microbiol. 2005 Dec;71(12):8228-35
references: 16332807
references: PLoS One. 2012;7(10):e47671
references: 23082192
references: Nature. 2010 Jul 15;466(7304):334-8
references: 20631792
references: FEMS Microbiol Rev. 2003 Jun;27(2-3):385-410
references: 12829276
references: J Vet Med B Infect Dis Vet Public Health. 2005 May;52(4):153-60
references: 16000109
references: Nature. 2008 Apr 3;452(7187):629-32
references: 18337718
references: J Mammary Gland Biol Neoplasia. 2011 Dec;16(4):339-56
references: 21947764
references: J Am Vet Med Assoc. 2006 Jul 15;229(2):259-62
references: 16842049
references: J Dairy Sci. 2011 Jul;94(7):3411-9
references: 21700026
references: Rev Environ Contam Toxicol. 2009;201:71-115
references: 19484589
references: Appl Environ Microbiol. 2007 Mar;73(5):1576-85
references: 17220268
references: Environ Microbiol. 2012 Jan;14(1):240-53
references: 22004107
references: J Am Vet Med Assoc. 1984 Mar 15;184(6):671-3
references: 6427159
references: FEMS Microbiol Lett. 2003 Nov 7;228(1):57-62
references: 14612237
references: J Appl Microbiol. 2005;98(1):96-105
references: 15610421
references: PLoS One. 2011;6(6):e21313
references: 21695057
references: Infect Immun. 1995 Aug;63(8):2899-905
references: 7542635
references: Bioinformatics. 2011 Aug 15;27(16):2194-200
references: 21700674