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Biomedical subjects

K P Pruess

Publications and source records attributed to K P Pruess.

6 recordsLinked to original sources
PubMed2004

Molecular differentiation of two sibling species of the black fly Simulium vittatum (Diptera: Simuliidae) based on random amplified polymorphic DNA.

Larvae of the black fly morphospecies Simulium vittatum from Colorado, Montana, Nebraska, and New Hampshire were cytologically identified as either the IS-7 or the IIIL-1 cytospecies. DNA was PCR amplified from cytotyped larvae using eight 10-mer primers, labeled with 33P, and run on polyacrylamide gels. The entire data set of 96 amplicons produced incomplete separation of the two cytospecies when subjected to neighbor-joining and maximum parsimony analyses. However, when analyzed within geographical regions, separate species status was supported. Bootstrap support for distinctness of the two cytospecies was best in Colorado where they were collected in true sympatry. The IS-7 cytospecies was more polymorphic in the western states, where it differed most from IIIL-1, which was most variable in the eastern states. The frequencies of the 17 most common amplicons in the two cytospecies were inversely correlated. A model of speciation derived from the molecular evidence suggests that IS-7 evolved in the west and spread eastward, whereas IIIL-1 later originated in the east and spread westward.

Animals↗
PubMed2000

Utility of the mitochondrial cytochrome oxidase II gene for resolving relationships among black flies (Diptera: simuliidae).

The complete mitochondrial cytochrome oxidase II gene was sequenced from 17 black flies, representing 13 putative species, and used to infer phylogenetic relationships. A midge (Paratanytarsus sp.) and three mosquitoes (Aedes aegypti, Anopheles quadrimaculatus, and Culex quinquefasciatus) were used as outgroup taxa. All outgroup taxa were highly divergent from black flies. Phylogenetic trees based on weighted parsimony (a priori and a posteriori), maximum likelihood, and neighbor-joining (log-determinant distances) differed topologically, with deeper nodes being the least well-supported. All analyses supported current classification into species groups but relationships among those groups were poorly resolved. The majority of phylogenetic signal came from closely related sister taxa. The CO-II gene may be useful for exploring relationships at or below the subgeneric level, but is of questionable value at higher taxonomic levels. The weighting method employed gave phylogenetic results similar to those reported by other authors for other insect CO-II data sets. A best estimate of phylogenetic relationships based on the CO-II gene is presented and discussed in relation to current black fly classification.

Amino Acid Sequence↗
PubMed1998

Greenbug (Homoptera: Aphididae) biotypes characterized using random amplified polymorphic DNA.

Genomic DNA was extracted from seven greenbug, Schizaphis graminum, biotypes (B, C, E, F, G, H and I) obtained from laboratory colonies maintained by USDA-ARS, Stillwater, Oklahoma. DNA was amplified using single 10-base primers. Of 100 primers tested, four were found which either alone, or in combination, distinguished all biotypes by distinct size differences in amplified fragments. Results were repeatable using aphids obtained from the same colonies 2 years later. These diagnostic primers produced unvarying banding patterns for all biotype E greenbugs collected in the field in Nebraska, Kansas, Oklahoma, and Texas.

Animals↗
PubMed1998

Effects of preservation methods, parasites, and gut contents of black flies (Diptera: Simuliidae) on polymerase chain reaction products.

Molecular analysis of biological specimens usually requires extraction of high-molecular weight DNA free of foreign DNA contaminants. DNA was extracted from black flies at different life stages that had been preserved by 4 methods: larvae and adults in ethanol, larvae in Carnoy's solution, adults on card-points, and adults hand-swatted and sun-dried. Using specific primers for the mitochondrial ND4 gene, a 257-bp amplicon was obtained from specimens preserved by ethanol, card-point mounting, and sun-drying. Successful amplification often required DNA dilutions > or = 1:20 (< 1-10 ng). DNA from specimens preserved in Carnoy's solution (ethanol: acetic acid, 3:1) yielded degraded DNA, resulting in fewer successful amplications. Parasitic nematodes and, to a lesser extent, gut contents resulted in extra products when amplified with randomly amplified polymorphic DNA (RAPD) primers. Sufficient DNA was extracted from the head of a larva for a successful polymerase chain reaction (PCR), eliminating the need to remove the contaminating gut and parasites.

Animals↗
PubMed1992

Mitochondrial transfer RNA genes in a black fly, Simulium vittatum (Diptera: Simuliidae), indicate long divergence from mosquito (Diptera: Culicidae) and fruit fly (Diptera: Drosophilidae).

Sequences are given for nine complete genes and one partial mitochondrial tRNA gene of the black fly, Simulium vittatum (Zetterstedt). Sequenced tRNA genes were for alanine, arginine, asparagine, aspartic acid, glutamic acid, glycine, leucine(CUN), lysine, serine(AGN), and valine. Nucleotides were aligned with the same previously sequenced genes in Aedes albopictus Skuse and Drosophila yakuba Burla. A cluster of six tRNA genes, which differ in arrangement in Ae. albopictus and D. yakuba, was amplified by PCR and found to have the same position and orientation in S. vittatum as in D. yakuba. Overall, similarity with either D. yakuba or Ae. albopictus was 86%. Sequences that were common to the three insects suggest that black flies and mosquitoes are as divergent from each other as either is from Drosophila. Sequences for nine species of black flies were obtained for tRNA leucine(CUN) from DNA amplified with another primer set. Little variation occurred within the tRNA gene but, by including the flanking regions to provide 175 base pairs, a phylogeny of the nine species was obtained that was largely consistent with current classification.

Animals↗