Aims of the study
1) Examination and quantification of environmental samples from piggeries for the presence of hmycobacteria. To develop and apply DNA and rRNA-based methods for the detection of potentially hazardous mycobacteria in the piggery environment.
2) Sequencing (16S rDNA) and typing (RFLP, MIRU-VNTR) of the mycobacterial isolates from porcine origin and compared them to isolates of humans for examination the similarity of the M. avium strains originating from slaughter pigs and human cases in regards of public health aspects in Finland.
3) To investigate the possibilities of parallel application of Restricted Fragment Length Polymorphism (RFLP) patterns and Variable-Number Tandem Repeat (VNTR) typing of genetic interspersed repetitive units of Mycobacteria (MIRUs).
4) Quantification of Mycobacterium avium subspecies in pig tissues by real-time quantitative PCR.
3.Materials and methods
3.1. Samples and experimental design
3.1.1.Piggery environmental samples and experimental design
Piggery environmental samples were collected from birth to slaughter farms with high condemnation rates for environmental mycobacteria . The total viable mycobacteria contents were analyzed from environmental samples taken from five piggeries, about 15-20 samples per piggery, total 94 samples, 2 parallels. The prevalence of tuberculous lesions in meat control, selected 5 piggeries was more than 4% during years 2002 to 2004
(Table XX). TABLE XX,PAKARINEN 2007, PAPER I OLI TÄSSÄ VÄLISSÄ, JÄTETÄÄNKÖ POIS ?
Experimental design for piggery environmental samples is showing in Fig.6. The results were confirmed by 16S rRNA sandwich hybridization.
KUVA 6 TULEE TÄHÄN
Fig 6. Experimental design for piggery environmental samples.
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