16S rRNA gene microbial analysis of the skin of fleece rot resistant and susceptible sheep
T. J. Dixon A , S. I. Mortimer B and B. J. Norris A CA CSIRO Livestock Industries, 306 Carmody Rd, St Lucia, Qld 4067, Australia.
B NSW Department of Primary Industries Agricultural Research Centre, Trangie, NSW 2823, Australia.
C Corresponding author. Email: Belinda.Norris@csiro.au
Australian Journal of Agricultural Research 58(7) 739-747 https://doi.org/10.1071/AR06273
Submitted: 22 August 2006 Accepted: 29 March 2007 Published: 26 July 2007
Abstract
Fleece rot is a bacterial dermatitis that follows prolonged wetting of the sheep’s skin, and a major pre-disposing condition to body strike in the Australian Merino. Several studies have examined bacterial load of the fleece in relation to fleece rot using traditional culture-based techniques focussing on only a few bacterial species. We examined the natural bacterial diversity of the healthy sheep skin and changes that occurred in fleece-rot resistant and susceptible animals during fleece rot development. Presented is a preliminary molecular genetic analysis of the bacterial ecology of the sheep skin. Eight 16S rRNA gene libraries were constructed from susceptible and resistant sheep both before and after onset of the disease following induction by simulated rainfall. Approximately 75% of the sequences obtained in this study have not been previously identified in fleece-rot studies. Four operational taxonomic units (OTU; groups of >97% sequence similarity) of major interest were present on susceptible animals and absent from resistant animals. Data on these OTU expand current knowledge of bacteria involved in inflammation and wounding of sheep skin tissue, and provide direction for future research that may lead to new treatment options for fleece rot and body strike.
Additional keywords: Merino, fleece rot, 16S rRNA gene, Corynebacterium, Pseudomonas aeruginosa, inflammation, microbial, ulceration.
Acknowledgments
This work was partially supported by the Australian Sheep Industry Cooperative Research Centre. All animal trials were conducted at the NSW Department of Primary Industries’ Agricultural Research Centre, Trangie, NSW, and their staff are gratefully acknowledged for their role in sample collection and handling of the Merino Fleece Rot selection lines. We are also grateful for expertise in bioinformatics and data archiving from the CSIRO Livestock Industries Bioinformatics group, especially Mr Sean McWilliam and Dr Brian Dalrymple, technical advice from Ms Wendy Smith, and microbial and phylogenetic advice from Dr Stuart Denman.
Altschul SF,
Madden TL,
Schäffer AA,
Zhang J,
Zhang Z,
Miller W, Lipman DJ
(1997) Gapped BLASTN and PSI-BLASTN: a new generation of protein database search programs. Nucleic Acids Research 25, 3389–3402.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Batey RG
(1986) Pathogenesis of caseous lymphadenitis in sheep and goats. Australian Veterinary Journal 63, 269–272.
| PubMed |
Benson DA,
Karsch-Mizrachi I,
Lipman DJ,
Ostell J,
Rapp BA, Wheeler DL
(2002) GenBank. Nucleic Acids Research 30, 17–20.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Burrell DH, MacDiarmid JA
(1984) Characterisation of isolates of Pseudomonas aeruginosa from sheep. Australian Veterinary Journal 61, 277–279.
| PubMed |
Burrell DH,
Merritt GC,
Watts JW, Walker KH
(1982) The role of Pseudomonas aeruginosa in pathogenesis of fleece-rot and the effect of immunisation. Australian Veterinary Journal 58, 34–35.
| PubMed |
Chapman RE,
Hollis DE, Hemsley JA
(1984) How quickly does wetting affect the skin of Merino sheep? Proceedings of the Australian Society for Animal Production 15, 290–292.
Chin JC,
Dai Y, Watts JE
(1995) Antibody response against Pseudomonas aeruginosa membrane proteins in experimentally infected sheep. Veterinary Microbiology 43, 21–32.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Chin JC, Watts JE
(1991) Dermal and serological response against Pseudomonas aeruginosa in sheep bred for resistance and susceptibility to fleece-rot. Australian Veterinary Journal 68, 28–31.
| PubMed |
Colditz IG,
Lax J,
Mortimer SI,
Clarke RA, Beh KJ
(1994) Cellular inflammatory responses in skin of sheep selected for resistance or susceptibility to fleece rot and fly strike. Parasite Immunology 16, 289–296.
| PubMed |
Colditz IG,
Woolaston RR,
Lax J, Mortimer SI
(1992) Plasma leakage in skin of sheep selected for resistance or susceptibility to fleece rot and fly dtrike. Parasite Immunology 14, 587–594.
| PubMed |
Cole JR,
Chai B,
Farris RJ,
Wang Q,
Kulam SA,
McGarrell DM,
Garrity GM, Tiedje JM
(2005) The Ribosomal Database Project (RDP-II): sequences and tools for high-throughput rRNA analysis. Nucleic Acids Research 33, D294–D296.
| Crossref |
PubMed |
Collins MD,
(1982) Corynebacterium mycetoides sp. nov., nom review. 3, 399–400.
Dewinter LM,
Bernard KA, Romney MG
(2005) Human clinical isolates of Corynebacterium diphtheriae and Corynebacterium ulcerans collected in Canada from 1999 to 2003 but not fitting reporting criteria for cases of diphtheria. Journal of Clinical Microbiology 43, 3447–3449.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Engwerda CR,
Dale CJ, Sandeman RM
(1996) IgE, TNF alpha, IL1 beta, IL4 and IFN gamma gene polymorphisms in sheep selected for resistance to fleece rot and flystrike. International Journal for Parasitology 26, 787–791.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Felsenstein J
(1989) PHYLIP – Phylogeny Inference Package (Version 3.2). Cladistics 5, 164–166.
Frank DN,
Spiegelman GB,
Davis W,
Wagner E,
Lyons E, Pace NR
(2003) Culture-independent molecular analysis of microbial constituents of the healthy human outer ear. Journal of Clinical Microbiology 41, 295–303.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Funke G,
von Graevenitz A,
Clarridge JE, Bernard KA
(1997) Clinical microbiology of Coryneform bacteria. Clinical Microbiology Reviews 10, 125–159.
| PubMed |
Gilligan PH
(1991) Microbiology of airway disease in patients with cystic-fibrosis. Clinical Microbiology Reviews 4, 35–51.
| PubMed |
Hayman RH
(1953) Studies in fleece-rot of sheep. Australian Journal of Agricultural Research 4, 430–468.
| Crossref | GoogleScholarGoogle Scholar |
Hogg JC, Lehane MJ
(1999) Identification of bacterial species associated with the sheep scab mite (Psoroptes ovis) by using amplified genes coding for 16S rRNA. Applied and Environmental Microbiology 65, 4227–4229.
| PubMed |
Hogg JC, Lehane MJ
(2001) Microfloral diversity of cultured and wild strains of Psoroptes ovis infesting sheep. Parasitology 123, 441–446.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Hugenholtz P
(2002) Exploring prokaryotic diversity in the genomic era. Genome Biology
, reviews 3.1–3.8.
Jackson TA,
Pearson JF,
Young SD,
Armstrong J, O’Callaghan M
(2002) Abundance and distribution of microbial populations in sheep fleece. New Zealand Journal of Agricultural Research 45, 49–55.
James PJ,
Ponzoni RW,
Walkley JRW,
Smith DH, Stafford JE
(1984) Preliminary estimates of phenotypic and genetic-parameters for fleece rot susceptibility in the South Australian Merino. Wool Technology and Sheep Breeding 31, 152–157.
Kingsford NM, Raadsma HW
(1995) Detection of Pseudomonas aeruginosa from ovine fleece washings by PCR amplification of 16S ribosomal RNA. Veterinary Microbiology 47, 61–70.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Klappenbach JA,
Saxman PR,
Cole JR, Schmidt TM
(2001) rrndb: the ribosomal RNA operon copy number database. Nucleic Acids Research 29, 181–184.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
KÕljalg S,
Sults I,
Raukas E,
Truu J,
Ustav M, Mikelsaar M
(1999) Distribution of Acinetobacter baumannii in a neurointensive care unit. Scandanavian Journal of Infectious Diseases 31, 145–150.
| Crossref | GoogleScholarGoogle Scholar |
Lipson M,
Hilton RA,
Watts JE, Merritt GC
(1982) Factors influencing fleece rot in sheep. Australian Journal of Experimental Agriculture 22, 168–172.
| Crossref | GoogleScholarGoogle Scholar |
London CJ, Griffith IP
(1984) Characterization of Pseudomonads isolated from diseased fleece. Applied and Environmental Microbiology 47, 993–997.
| PubMed |
Lyness EW,
Pinnock DE, Cooper DJ
(1994) Microbial ecology of sheep fleece. Agriculture, Ecosystems & Environment 49, 103–112.
| Crossref | GoogleScholarGoogle Scholar |
MacDiarmid JA, Burrell DH
(1992) Degradation of the wool fiber by bacteria isolated from fleece rot. Wool Technology and Sheep Breeding 40, 123–126.
Mcguirk BJ, Atkins KD
(1984) Fleece rot in Merino sheep. 1: The heritability of fleece rot in unselected flocks of medium-wool Peppin Merinos. Australian Journal of Agricultural Research 35, 423–434.
| Crossref | GoogleScholarGoogle Scholar |
O’Meara TJ,
Nesa M,
Raadsma HW,
Saville DG, Sandeman RM
(1992) Variation in skin inflammatory responses between sheep bred for resistance or susceptibility to fleece rot and blowfly strike. Research in Veterinary Science 52, 205–210.
| PubMed |
O’Meara TJ,
Nesa M,
Seaton DS, Sandeman RN
(1995) A comparison of inflammatory exudates released from myiasis wounds on sheep bred for resistance or susceptibility to Lucilia cuprina. Veterinary Parasitology 56, 207–223.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Perričre GGM, Gouy M
(1996) WWW-query: An on-line retrieval system for biological sequence banks. Biochimie 78, 364–369.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Raadsma HW
(1991) The susceptibility to body strike under high rainfall conditions of flocks selected for and against fleece rot. Australian Journal of Experimental Agriculture 31, 757–759.
| Crossref | GoogleScholarGoogle Scholar |
Raadsma HW,
Gilmour AR, Paxton WJ
(1988) Fleece rot and body strike in merino sheep. 1: evaluation of liability to fleece rot and body strike under experimental conditions. Australian Journal of Agricultural Research 39, 917–934.
| Crossref | GoogleScholarGoogle Scholar |
Raadsma HW,
Kearins RD,
Bennett NW,
Coy J, Watts JE
(1987) An evaluation of classing merino sheep for fleece rot susceptibility. Australian Journal of Experimental Agriculture 27, 493–501.
| Crossref | GoogleScholarGoogle Scholar |
Saitou N, Nei M
(1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution 4, 406–425.
| PubMed |
Seddon HR
(1931) Conditions which predispose sheep to blowfly attack. Agricultural Gazette of New South Wales 42, 581–594.
Stackebrandt E, Göbel BM
(1994) Taxonomic note: a place for DNA-DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. Internationl Journal of Systematic Bacteriology 44, 846–849.
Stuart TPA
(1894) On green-producing chromogenic micro-organisms in wool. Journal of the Proceedings of the Royal Society of New South Wales 28, 320–322.
Van Tonder EM,
Colly PA,
Vermeulen SO,
Kellerman GE,
De Ruiter A, Whitehead CJ
(1990) Bolo disease: a bacteriological survey. Journal of the South African Veterinary Association 61, 96–101.
| PubMed |
Wang X,
Heazlewood SP,
Krause DO, Florin TH
(2003) Molecular characterization of the microbial species that colonize human ileal and colonic mucosa by using 16S rDNA sequence analysis. Journal of Applied Microbiology 95, 508–520.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Watts JE,
Nay T,
Merritt GC,
Coy JR,
Griffiths DA, Dennis JA
(1980) The significance of certain skin characters of sheep in resistance and susceptibility to fleece-rot and body strike. Australian Veterinary Journal 56, 57–63.
| PubMed |
Woo PC,
Tse H,
Lau SK,
Leung KW,
Woo GK,
Wong MK,
Ho CM, Yuen KY
(2005) Alkanindiges hongkongensis sp. nov. A novel Alkanindiges species isolated from a patient with parotid abscess. Systematic and Applied Microbiology 28, 316–322.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
