Molecular analysis of Dichelobacter nodosus isolated from footrot in sheep in Malaysia |
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| Institution: | 1. Unit of Models for Ecotoxicology and Toxicology (METO), INERIS, 60550 Verneuil en Halatte, France;2. UMR-I 02 SEBIO, INERIS, Parc ALATA, BP2, 60550 Verneuil-en-Halatte, France;1. Institute of Veterinary Bacteriology, Vetsuisse, University of Bern, Laenggasstrasse 122, 3001 Bern, Switzerland;2. Graduate School for Cellular and Biomedical Sciences, University of Bern, Bern CH-3012, Switzerland;1. Environmental Microbial and Food Safety Laboratory, Agricultural Research Service, United States Department of Agriculture, Building 173, BARC-East, 10300 Baltimore Avenue, Beltsville, MD 20705, USA;2. Universidade Federal de Uberlândia, Instituto de Ciências Biomédicas, Laboratório de Parasitologia, Av. Pará, 1720, Campus Umuarama, 38400-902 Uberlândia, Minas Gerais, Brazil;3. CAPES Foundation, Ministry of Education of Brazil Caixa, Postal 250, 70040-020 Brasília, DF, Brazil |
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| Abstract: | Pulsed field gel electrophoresis analysis of genomic DNA was used to investigate genetic diversity among Dichelobacter nodosus from footrot in sheep in Malaysia. Twelve Dichelobacter nodosus strains isolated from lesion materials from infected sheep were confirmed as Dichelobacter nodosus by polymerase chain reaction technique using the species-specific Dichelobacter nodosus 16S RNA sequence Ac and C as primers. Pulsed field gel electrophoresis banding profiles using restriction enzymes ApaI (5′GGGCCC3′), SfiI (5′GGCCNNNNNGGCC3′)and SmaI (′5CCCGGG3′) enabled the 12 Dichelobacter nodosus strains to be differentiated into eight different PFGE patterns and thus genome-types, with F (coefficient of similarity) values ranging from 0.17 to 1.0 (ApaI), 0.14 to 1.0 (SfiI) and 0.22 to 1.0 (SmaI). Strains with origin in different farms were shown to have different PFGE patterns (two strains, M7 and M8 were the only exception). On the basis of their PFGE, all field strains used in the study differed from the reference strains. Our data revealed that there are several clonal types of Dichelobacter nodosus isolates and indicated that there is probably more than one source of this pathogen on the farms studied. The study showed that strains of D. nodosus exhibited considerable genetic diversity using this method and that genomic analysis by pulsed field gel electrophoresis was useful in discriminating the D. nodosus strains. |
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