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Isolation and characterisation of Clostridia from different environments : project report
Lozič, Eva
Background: Clostridia are Gram-positive, anaerobic, sporogenic rods. Genus isvery diverse with its importance ranging from medical and veterinary relevance to the rising interest in industry. The ...main habitat of Clostridia is the gut from where they are distributed to different environments such as soil and water. They can survive extreme physical conditions as well as aerobic atmosphere in forms of spores. Certain species might also be transferred between different environments and survival in those environments can expand the spectrum of sources of human infections and-or might be of biotechnological importance. Some genera also contribute to degradation of organic matter or participate in nitrogen fixation. Although Clostridia have been well recognized in clinical and veterinary practice, little is known about the population structure of this genus in different habitats. Therefore we studied the population structure in four different environments and determined the Clostridia species with phenotypic (fatty acid profile) and molecular laboratory methods. Methods: Samples were collected from four different environments using classical culturing methods for isolation of Clostridia. Isolated strict anaerobes were Gram stained and Gram- positive rods with visible spores were further identified with MIDI system and by amplifying the ribosomal 16S gene, rpoB and cdd3 genes using PCR. Rsa\ and Mnl\ restrictions were used for screening and grouping of Clostridia isolates.Representative strains of different RFLP profiles were selected and used for sequencing and final identification. Results: Seventy nine confirmed Clostridia belonged to 18 different species among which Clostridium bifermentans, Clostridium metallolevans, and Clostridium sordellii predominated. The amplification of rpoB was achieved in 77.4% (82 out of 106 strains) while of 92 analysed strains, only 45 (48.9%) were identified as Clostridia species with MIDI system. Of all 4 environments, soil revealed the greatest diversity especially the compost sample. Conclusions: Our results were in the agreement with the proposed hypothesis. Clostridia were obtained from all the samples, although none of the species was isolated from all of the environments. The proposed "signature" species are: C. bifermentans, C. metallolevans and C. sporogenes for soil, C. sordellii for waste water samples, C. difficile, C. paraputrificum and C. sordellii for faeces of asymptomatic babies and C. celerecrescens / C. beijerinckii / C. spiriforme for faeces of less than one-week old chickens.
Type of material - master's thesis
; adult, serious
Publication and manufacture - London : [E. Lozič], 2011
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