Browse > Article
http://dx.doi.org/10.14405/kjvr.2017.57.2.117

Identification of microbiome with 16S rRNA gene pyrosequencing and antimicrobial effect of egg white in bovine mastitis  

Kim, Danil (Department of Farm Animal Medicine, College of Veterinary Medicine, Seoul National University)
Kim, Eun-Kyung (Department of Farm Animal Medicine, College of Veterinary Medicine, Seoul National University)
Seong, Won-Jin (Department of Farm Animal Medicine, College of Veterinary Medicine, Seoul National University)
Ro, Younghye (Department of Farm Animal Medicine, College of Veterinary Medicine, Seoul National University)
Ko, Dae-Sung (Department of Farm Animal Medicine, College of Veterinary Medicine, Seoul National University)
Kim, Nam-Hyung (Laboratory of Avian Diseases, College of Veterinary Medicine, Seoul National University)
Kim, Jae-Hong (Laboratory of Avian Diseases, College of Veterinary Medicine, Seoul National University)
Kwon, Hyuk-Joon (Department of Farm Animal Medicine, College of Veterinary Medicine, Seoul National University)
Publication Information
Korean Journal of Veterinary Research / v.57, no.2, 2017 , pp. 117-126 More about this Journal
Abstract
Bovine mastitis is an important microbial disease in the dairy industry. We investigated the frequencies of bacterial pathogens in 62 farms and pathogen antibiotic resistance from mastitis samples (n = 748). We tested the antimicrobial activity of chicken and duck egg white and lysozyme purified from chicken egg white. Moreover, we compared the microbiomes of normal and mastitic raw milk obtained by 16S rRNA gene pyrosequencing and culture methods. The results showed that the frequencies of Gram-positive pathogens (Enterococcus faecalis 37% and Staphylococcus aureus 36%) were higher than that of a Gram-negative pathogen (Escherichia coli 15%). Resistance frequencies to ampicillin and norfloxacin were lowest in Staphylococcus aureus (21%), Enterococcus faecalis (23%), and Escherichia coli (33%), and the antimicrobial activity of chicken egg white was higher than those of lysozyme and duck egg white. Pyrosequencing results revealed clear differences between the microbiomes of mastitic and normal raw milk samples and revealed a slightly similar, but clearly different, composition of pathogens compared to that from the culture method. Thus, pyrosequencing may be useful for elucidating changes in microbiomes during mastitis progression and treatment. A chicken egg white and antibiotic combination may help with mastitis treatment; however, further studies are needed.
Keywords
antibiotic resistance; bovine mastitis; microbial pathogens; microbiome; raw milk;
Citations & Related Records
Times Cited By KSCI : 2  (Citation Analysis)
연도 인용수 순위
1 Huber T, Faulkner G, Hugenholtz P. Bellerophon: a program to detect chimeric sequences in multiple sequence alignments. Bioinformatics 2004, 20, 2317-2319.   DOI
2 Hur M, Kim Y, Song HR, Kim JM, Choi YI, Yi H. Effect of genetically modified Poplars on soil microbial communities during the phytoremediation of waste mine tailings. Appl Environ Microbiol 2011, 77, 7611-7619.   DOI
3 Kim JH, Ko MJ, Kim KH, Lee SH, Choi SS. Antimicrobial susceptibility of Staphylococci sp. isolated from bovine milk. Korean J Microbiol 2010, 46, 341-345.
4 Kim OS, Cho YJ, Lee K, Yoon SH, Kim M, Na H, Park SC, Jeon YS, Lee JH, Yi H, Won S, Chun J. Introducing EzTaxon-e: a prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured species. Int J Syst Evol Microbiol 2012, 62, 716-721.   DOI
5 Kojima H, Fukui M. Sulfuritalea hydrogenivorans gen. nov., sp. nov., a facultative autotroph isolated from a freshwater lake. Int J Syst Evol Microbiol 2011, 61, 1651-1655.   DOI
6 Kovacs-Nolan J, Zhang JW, Hayakawa S, Mine Y. Immunochemical and structural analysis of pepsin-digested egg white ovomucoid. J Agric Food Chem 2000, 48, 6261-6266.   DOI
7 Lee ES, Kang HM, Chung C, Moon JS. Antimicrobial susceptibility and prevalence of gram-negative bacteria isolated from bovine mastitis. Korean J Vet Res 2007, 47, 67-75.
8 Masoud W, Vogensen FK, Lillevang S, Abu Al-Soud W, Sorensen SJ, Jakobsen M. The fate of indigenous microbiota, starter cultures, Escherichia coli, Listeria innocua and Staphylococcus aureus in Danish raw milk and cheeses determined by pyrosequencing and quantitative real time (qRT)-PCR. Int J Food Microbiol 2012, 153, 192-202.   DOI
9 Nam HM. Current situation of mastitis and relative frequency of pathogens isolated from subclinical mastitis in dairy cattle in Korea. Korean J Vet Public Health 2010, 34, 265-272.
10 Nam HM, Lim SK, Jang GC, Joung DY, Kim H, Lee CS, Jung SC. Culture results from quarter milk samples submitted to veterinary diagnostic laboratories during January-November 2012 in Korea. J Prev Vet Med 2013, 37, 111-119.   DOI
11 Oikonomou G, Machado VS, Santisteban C, Schukken YH, Bicalho RC. Microbial diversity of bovine mastitic milk as described by pyrosequencing of metagenomic 16s rDNA. PLoS One 2012, 7, e47671.   DOI
12 Rodrigues MX, Lima SF, Higgins CH, Canniatti-Brazaca SG, Bicalho RC. The Lactococcus genus as a potential emerging mastitis pathogen group: A report on an outbreak investigation. J Dairy Sci 2016, 99, 9864-9874.   DOI
13 Osborne AD, Armstrong K, Catrysse NH, Butler G, Versavel L. An outbreak of pseudomonas mastitis in dairy cows. Can Vet J 1981, 22, 215-216.
14 Quigley L, O'Sullivan O, Beresford TP, Ross RP, Fitzgerald GF, Cotter PD. Molecular approaches to analysing the microbial composition of raw milk and raw milk cheese. Int J Food Microbiol 2011, 150, 81-94.   DOI
15 Quigley L, O'Sullivan O, Stanton C, Beresford TP, Ross RP, Fitzgerald GF, Cotter PD. The complex microbiota of raw milk. FEMS Microbiol Rev 2013, 37, 664-698.   DOI
16 Tarlera S, Denner EBM. Sterolibacterium denitrificans gen. nov., sp. nov., a novel cholesterol-oxidizing, denitrifying member of the ${\beta}$-Proteobacteria. Int J Syst Evol Microbiol 2003, 53, 1085-1091.   DOI
17 Varahan S, Iyer VS, Moore WT, Hancock LE. Eep confers lysozyme resistance to Enterococcus faecalis via the activation of the extracytoplasmic function sigma factor SigV. J Bacteriol 2013, 195, 3125-3134.   DOI
18 Vernon CA. The mechanisms of hydrolysis of glycosides and their relevance to enzyme-catalysed reactions. Proc R Soc Lond B Biol Sci 1967, 167, 389-401.   DOI
19 Watts JL. Etiological agents of bovine mastitis. Vet Microbiol 1988, 16, 41-66.   DOI
20 Zadoks RN, Watts JL. Species identification of coagulasenegative staphylococci: genotyping is superior to phenotyping. Vet Microbiol 2009, 134, 20-28.   DOI
21 Delgado S, Rachid CTCC, Fernandez E, Rychlik T, Alegria A, Peixoto RS, Mayo B. Diversity of thermophilic bacteria in raw, pasteurized and selectively-cultured milk, as assessed by culturing, PCR-DGGE and pyrosequencing. Food Microbiol 2013, 36, 103-111.   DOI
22 Zhen YH, Jin LJ, Li XY, Guo J, Li Z, Zhang BJ, Fang R, Xu YP. Efficacy of specific egg yolk immunoglobulin (IgY) to bovine mastitis caused by Staphylococcus aureus. Vet Microbiol 2009, 133, 317-322.   DOI
23 Abeyrathne EDNS, Lee HY, Ahn DU. Egg white proteins and their potential use in food processing or as nutraceutical and pharmaceutical agents-a review. Poult Sci 2013, 92, 3292-3299.   DOI
24 Ben-Ami R, Navon-Venezia S, Schwartz D, Schlezinger Y, Mekuzas Y, Carmeli Y. Erroneous reporting of coagulasenegative staphylococci as Kocuria spp. by the Vitek 2 system. J Clin Microbiol 2005, 43, 1448-1450.   DOI
25 Bera A, Herbert S, Jakob A, Vollmer W, Gotz F. Why are pathogenic staphylococci so lysozyme resistant? The peptidoglycan O-acetyltransferase OatA is the major determinant for lysozyme resistance of Staphylococcus aureus. Mol Microbiol 2005, 55, 778-787.
26 Chun J, Kim KY, Lee JH, Choi Y. The analysis of oral microbial communities of wild-type and toll-like receptor 2-deficient mice using a 454 GS FLX Titanium pyrosequencer. BMC Microbiol 2010, 10, 101.   DOI
27 CLSI. Performance Standards for Antimicrobial Disk Susceptibility Tests; Approved Standard-Ninth Edition. Clinical and Laboratory Standards Institute document M2-A9. Clinical and Laboratory Standards Institute, Wayne, 2006.
28 de Oliveira GB, Favarin L, Luchese RH, McIntosh D. Psychrotrophic bacteria in milk: how much do we really know? Braz J Microbiol 2015, 46, 313-321.   DOI
29 Gong D, Wilson PW, Bain MM, McDade K, Kalina J, Herve-Grepinet V, Nys Y, Dunn IC. Gallin; an antimicrobial peptide member of a new avian defensin family, the ovodefensins, has been subject to recent gene duplication. BMC Immunol 2010, 11, 12.   DOI
30 Glynn AA. Lysozyme: antigen, enzyme and antibacterial agent. Sci Basis Med Annu Rev 1968, 31-52.
31 Hamady M, Lozupone C, Knight R. Fast UniFrac: facilitating high-throughput phylogenetic analyses of microbial communities including analysis of pyrosequencing and PhyloChip data. ISME J 2010, 4, 17-27.   DOI
32 Hogeveen H, Huijps K, Lam TJ. Economic aspects of mastitis: new developments. N Z Vet J 2011, 59, 16-23.   DOI
33 Bhatt VD, Ahir VB, Koringa PG, Jakhesara SJ, Rank DN, Nauriyal DS, Kunjadia AP, Joshi CG. Milk microbiome signatures of subclinical mastitis-affected cattle analysed by shotgun sequencing. J Appl Microbiol 2012, 112, 639-650.   DOI