Given that morphological identification of willow is difficult, willow lines being investigated for their suitability for use as short rotation crops for biomass production in Saskatchewan were investigated with various molecular techniques as possible tools for DNA fingerprinting. Flow cytometry was used to assess variation in nuclear DNA content and thus ploidy level of the lines of the five species (<i>Salix purpurea, Salix eriocephala, Salix sachalinensis, and Salix dasyclados</i>) and three hybrids (<i>S. purpurea x S. miyabeana, S. sachalinensis x S. miyabeana, S. viminalis x S. miyabeana</i>). The DNA content varied between 1.14 and 3.00pg. Ploidy levels of the species varied from triploid to hexaploid while all hybrids were tetraploid. RAPD and ISSR marker systems were used to assess genetic and taxonomic relationships among all willow lines. Of 90 RAPD primers tested, 60 were selected and 99 polymorphic bands scored. Of 35 ISSR primers tested, 19 were selected and 35 polymorphic bands scored. Both RAPD and ISSR dendrograms clustered together lines belonging to the same species and same hybrid combination. A combination of strong and reproducible RAPD and ISSR bands was used to develop identification keys for lines belonging to the same species.<p> The ribosomal RNA gene region, including the entire 5.8S RNA gene and the internal transcribed spacers (ITS1 and ITS2) was amplified and sequenced to assess sequence homology between the five species. The total length of the amplified region was 601bp, with the ITS1, 5.8 S and ITS2 being 223, 163, and 215bp respectively. Intra- and inter-species SNPs were observed, 6 within ITS1, and 3 within ITS2. No polymorphisms were found in the 5.8S gene. The low rate of variation within the sequenced ITS fragment between species supports the monophyly of the five species involved in this study, and confirms their belonging to the subgenus Caprisalix. SCAR primers were designed from species-specific polymorphic nucleotides and applied to the willow collection to test their use for species identification. A species identification key based on SNPs is proposed.

Previous studies examining the canine intestinal microflora have focused on cultivation of bacteria from intestinal content. Recently, it has been recognized that the majority of bacteria cannot be identified using standard culture techniques. The aim of this study was to describe the composition and dynamics of the canine intestinal microflora using molecular methods based on identification of the 16S ribosomal DNA (16S rDNA) and to evaluate the clinical use of a 13C-glycocholic acid blood test (13CGCBT) as a serum marker for small intestinal bacterial biomass. Intestinal content was obtained from healthy dogs and the microflora was characterized in different compartments of each dog by denaturing gradient gel electrophoresis (DGGE) and comparative 16S rDNA analysis. A 13C-glycocholic acid blood test (13C-GCBT) was developed as a marker for small intestinal bacterial biomass and the influence of tylosin administration on the 13C-GCBT, serum concentrations of cobalamin, folate, and unconjugated cholic acid (SUCA) was evaluated. There was marked variation in DGGE profiles between individual dogs and also between different intestinal compartments within dogs. DGGE profiles from duodenal juice samples collected endoscopically at different time-points varied within individuals, possibly due to variations over time or a slight variation in sampling location. Direct sequencing revealed 106 individual 16S rDNA sequences. Forty-two sequences showed less than 98% similarity to described sequences in public databases and may constitute previously uncharacterized bacterial species. Serum folate concentrations, SUCA, and the cumulative percent dose/min of 13C administered as 13C-glycocholic acid (CUMPCD) increased significantly following tylosin administration (p<0.01). The results indicate that dogs have a complex intestinal microflora with marked differences between individual dogs. Different intestinal compartments appear to host a unique microflora and the assessment of a fecal sample does not yield accurate information about the composition of the microflora in proximal compartments of the gut. The intestine harbors many previously uncharacterized bacterial species. The clinical significance of these uncharacterized intestinal bacterial species needs to be further investigated in dogs with gastrointestinal disease. Increased serum folate, SUCA, and CUMPCD in the 13C-GCBT suggest that, in the dogs described here, tylosin administration increased the biomass of organisms carrying out these metabolic functions.

Thesis (M. Phil.)--Hong Kong University of Science and Technology, 2003. / Includes bibliographical references (leaves 103-111). Also available in electronic version. Access restricted to campus users.

Thesis (MTech. degree in Policing) -- Tshwane University of Technology, 2010. / Looks at the causes for poor investigation of murder cases in Loate police station and seek ways which would assist in improving the investigation of murder cases.

published_or_final_version / Zoology / Master / Master of Philosophy

In order to establish a method of assessing the degree of inbreeding within herds of cattle, we constructed a calibration index relating kinship and the degree of DNA band sharing in DNA fingerprints. Firstly, chickens were used as a model system to test the possibility of using microsatellite DNA as a probe for DNA fingerprinting in inbreeding analysis. Six genetic groups of chickens with estimated coefficients of inbreeding ranging from 0.026 to $>$ 0.98 (pedigree analysis) were fingerprinted using the minisatellite probe derived from M13 and the microsatellite probe (CAC)$ sb5$. The degree of band sharing using either probe increased in concert with the known amount of inbreeding and was described by the equation Y = 0.56X ($ pm$0.06) + 0.42 ($ pm$0.03); r = 0.998. Since in-gel hybridization using the microsatellite probes was faster and less labour intensive than using the minisatellite probe, it was used in the subsequent studies. Pedigree analysis in Holstein dairy cattle allowed for the empirical calibration of the association of band sharing with the coefficient of relatedness, (r), defined as the expected proportion of genes in 2 individuals that are identical by descent (i.e. for monozygous twins r = 1; for first order relatives r = 0.5; for half sibs r = 0.25 etc.). The average band sharing between pairs (6 pairs at each r value) of individuals within each class formed the basis for calibration. DNA was digested using RsaI. The relationship between band sharing and relatedness was well represented by a linear approximation Y = 0.51X ($ pm$0.09) + 0.50 ($ pm$0.04); r = 0.992. Using this calibration curve, random samples of animals within herds can be tested to establish the herd variability and to minimize inbreeding.

A concern in transgenic crops is the potential risk posed by unintended effects which could result from genetic transformation. The objective of this work was to develop an untargeted approach that could characterize transgenic crops, as well as conventional crops, at the molecular level. An experimental approach was designed and used to compare conventional and transgenic soybean varieties. Varieties were compared using their metabolite fingerprints obtained by reverse-phase high performance liquid chromatography (HPLC) and both the analytical and biological variability were assessed. Multivariate and univariate statistical analyses were applied to the data to detect significant differences between the varieties. It was found that transgenic variety PS 46 RR was the most different variety analyzed and that it differed most from Mandarin (Ottawa) and AC Dundas. The statistical analyses also determined that PS 46 RR differed more from the conventional varieties tested than 2601R did.

Thesis (MSc)--University of Stellenbosch, 2007. / Bibliography. Also available via the Internet

Thesis (Ph. D.)--Washington State University, August 2008. / Includes bibliographical references.

Thesis (M.S.)--Michigan State University. School of Criminal Justice, 2008. / Title from PDF t.p. (viewed on Aug. 7, 2009) Includes bibliographical references (p. 52-54). Also issued in print.