Universal PCR primers for ribosomal protein gene introns of fish

Human ribosomal protein (RP) gene sequences with respect to intron/exon structures and corresponding cDNA or genomic data of fish species were obtained from the GenBank database. Based on conserved exon sequences, 128 primer pairs for 41 genes were designed for exon-primed intron-crossing (EPIC) polymerase chain reaction (PCR). In reference to the draft genome sequences of the Pacific bluefin tuna (Thunnus orientalis), 12 primer pairs expected to amplify introns of the bluefin tuna with lengths of 500–1000 bp were selected and applied to six distantly related fish species belonging to the Orders Clupeiformes, Tetraodontiformes, Pleuronectiformes, Perciformes, Scorpaeniformes, and Anguilliformes. PCR amplification was observed for at least four species in each primer pair, and all fragments were larger than those expected for intronless amplification. Single fragment amplification was observed for at least seven primer pairs per species. Fragment sizes of the bluefin tuna for nine primer pairs corresponded to those expected from the genomic data. Thus, our primer pairs are potentially applicable to a wide variety of fish species and serve as an initial step for isolating single-copy nuclear DNA sequences.     

Comparison of bacterial culture, polymerase chain reaction, and a mix-enzyme-linked immunosorbent assay for the detection of Salmonella status in grow-to-finish pigs in western Canada with a Bayesian approach

Among grow-to-finish pigs from 10 herds in Alberta and Saskatchewan, 23 (16%) of 144 fecal samples were culture-positive and 40 (28%) of 144 pigs were seropositive for Salmonella. With a Bayesian model specifying dependence between the 2 tests, the sensitivity (Se) of culture and real-time polymerase chain reaction (RT-PCR) was 79% to 86%, depending on the cut-off value for the enzyme-linked immunosorbent assay (ELISA). Culture specificity (Sp) was assumed to be 100%; RT-PCR Sp was found to be 94%. The ELISA Se was 76% and 51% at optical density cut-off values ≥ 20% and ≥ 40%, respectively; the Sp was 94% at each cut-off value. The model showed some sensitivity to ELISA prior information, the ELISA Se being approximately 8% lower when informative prior information was specified in the model. When there was no adjustment for dependence between culture and RT-PCR, the posterior estimates for both culture and RT-PCR Se were 11% higher than with the conditional-dependence model and had considerably narrower probability intervals, which suggests that correlation between culture and PCR is important and should be adjusted for in future studies.     

Unilateral ventral strabismus in an aquarium Silver Arowana (Osteoglossum bicirrhosum)

An adult silver arowana (Osteoglossum bicirrhosum) presented with a 2‐month history of acute onset ventral strabismus of the right eye associated with a cystic structure overlying the dorsal aspect of the globe. Aspiration of the cyst, removal of redundant conjunctival tissue, and apposition of the surgical edges of the conjunctiva did not correct the strabismus. Magnetic resonance imaging of the head and histopathology of the eye did not reveal significant differences between affected and unaffected eyes except for a more posterior dorsal rectus muscle insertion site in the affected eye. This case report documents and describes a syndrome well known among arowana hobbyists as ‘drop eye’.     

Infrared spectral characteristics and surface inactivation of wood at high temperatures

Summary Spectra resulting from chemical changes in white spruce [Picea glauca (Moench) Voss] microsections heated in air or nitrogen at high temperatures (100 to 240°C) were continuously recorded on an infrared spectrophotometer.A significant change occurred in the intensity of the 1730 cm^-1 band which indicates a carbonyl absorption of carboxyl and ester groups of wood. This intensity initially decreased and the increased at a greater rate. The time periods to reach the minimum inflection point, termed the times to initiate a significant oxidative carboxylation or oxidation, showed a good curvilinear relationship with heating temperatures.Quantification of this time-temperature relationship required to reach a significant level of oxidation was achieved, using wood microsections that had extractives removed to varying degrees. It was concluded that the extractives served only as catalysts for oxidation. When drying wood at temperatures over 180°C, in addition to oxidation, pyrolytic degradation occurred.Chemical evidence was further confirmed by tests of plywood panels bonded with phenolformaldehyde glue. Three separate types of veneers were investigated—non-extracted, acetone extracted, and veneer with the surface chemically stabilized by treatment with sodium borohydride. The results suggest that the time period to reach a significant level of oxidative carboxylation is also the time period to initiate a wood surface inactivated to polymer adhesion.The author wishes to thank Miss D. Bouchard and Mr. H. N. Mukai for their assistance in the experiments. Appreciation also is due to Mr. H. MacLean for a critical review of this paper.     

A method for routine measurements of total sugar and starch content in woody plant tissues

Several extraction and measurement methods currently employed in the determination of total sugar and starch contents in plant tissues were investigated with the view to streamline the process of total sugar and starch determination. Depending on the type and source of tissue, total sugar and starch contents estimated from samples extracted with 80% hot ethanol were significantly greater than from samples extracted with a methanol:chloroform:water solution. The residual ethanol did not interfere with the sugar and starch determination, rendering the removal of ethanol from samples unnecessary. The use of phenol-sulfuric acid with a phenol concentration of 2% provided a relatively simple and reliable colorimetric method to quantify the total soluble-sugar concentration. Performing parallel sugar assays with and without phenol was more useful for accounting for the interfering effects of other substances present in plant tissue than using chloroform. For starch determination, an enzyme mixture of 1000 U alpha-amylase and 5 U amyloglucosidase digested starch in plant tissue samples more rapidly and completely than previously recommended enzyme doses. Dilute sulfuric acid (0.005 N) was less suitable for starch digestion than enzymatic hydrolysis because the acid also broke down structural carbohydrates, resulting in overestimates of starch content. After the enzymatic digestion of starch, the glucose hydrolyzate obtained was measured with a peroxidase-glucose oxidase/o-dianisidine reagent; absorbance being read at 525 nm after the addition of sulfuric acid. With the help of this series of studies, we developed a refined and shortened method suitable for the rapid measurement of total sugar and starch contents in woody plant tissues.     

Thermo-directional movement of wood and wood composites

Summary Thermal movement of wood and wood composites was studied and compared with that of random and oriented phenolic foams. Cell orientation was a dominate factor determining the thermal response of these structures. In dried wood, thermal movement in the direction parallel to the cells (longitudinal) decreased in dimension during heating while the radial and tangential directions expanded under similar heating condition. Oriented foams showed more restraint in thermal movement in the parallel to orientation direction. These findings indicate constraining forces act on the direction parallel to the cells while more flexibility exists in perpendicular directions. Wood element size and orientation and the manufacturing process also influence the thermal response in wood composites. The experimental results also revealed the potential fire hazard of waferboard type of composites.     

Wettability of four Asian meranti species

Summary The wettability of four Asian meranti species (dark red, light red, white, and yellow) in various liquids was studied by a cone-immersion technique. Wettability was expressed in terms of the area under the force-immersion curve and referred to as the wettability index. Wettability was strongly influenced by the pH of the solution, generally increasing with increasing pH. Surfactant type affected wettability, with the cationic type yielding the greatest wettability followed by anionic and nonionic types. Among the liquids used, distilled water gave the lowest, and acetone-isopropyl alcohol solution gave the greatest wettability. Of the meranti species, yellow meranti showed the greatest wettability, followed by white, light red, and dark red meranti.A plot of the relative amount of liquid absorbed into the wood sample versus wettability index yielded two regions, one attributed mainly to liquid trapped in crevices on the wood surface, and another which may be more indicative of wood-liquid interactions. A highly significant linear correlation between wettability index and liquid surface tension was found for those species with low and medium wettability, while that with high wettability showed no correlation. A new empirical approach of determining the critical surface tension of wood is discussed. Limited data for the present method indicated close agreement with result from the literature obtained by the use of the well-known Zisman's contact angle approach.