Pseudomonas fluorescens contamination of a feline packed red blood cell unit and studies of canine units

Background: While screening programs have reduced the risk of infectious disease transmission by donors in human and veterinary blood banking, bacterial contamination of blood products has emerged as a major complication in human medicine. Objectives: To describe a Pseudomonas fluorescens (Pf)‐contaminated feline packed RBC (pRBC) unit and experimentally investigate Pf‐contaminated canine pRBCs. Methods: Canine pRBCs were inoculated with Pf‐rich pRBCs from the sentinel feline unit and stored at 4°C or 20°C for 72 hours. Aliquots from the pRBCs were serially evaluated by microscopy, culture, and a eubacterial 16S rRNA real‐time PCR assay. Results: One Pf‐contaminated feline unit turned black after 22 days of storage and was removed from the blood bank; a source was not found, and no other contaminated units were identified. Canine pRBCs spiked with 5 or 25 μL of the sentinel unit became culture‐ and/or 16S PCR‐positive at ≥8 hours at 20°C and 48 hours at 4°C and developed a color change at ≥24 hours. Sensitivity studies indicated that without incubation, inoculation of ≥100 μL Pf‐rich pRBCs was necessary for a positive 16S PCR test result. Conclusions: P. fluorescens grows in stored pRBCs slowly at 4°C and rapidly at 20°C. Screening of blood products for color change, estimating bacterial concentration with microscopy, and 16S PCR testing are simple and fast ways to detect bacteria in stored blood. Aseptic collection, temperature‐controlled storage, and regular visual monitoring of stored units is recommended. Discolored units should not be transfused, but examined for bacterial contamination or other blood product quality problems.     

Midwest (U.S.A.) reservoir water quality modification

The impact of a flood control, low flow argumentation reservoir in the Midwestern part of the United States on BOD, COD, and ammonia was evaluated in this paper. Fifteen years of weekly water quality data (9 yr before impoundment and 6 yr after impoundment) from four sampling stations upstream and downstream of the reservoir were available for analysis. The annual loading rates of these parameters (kg ha^?1 vr^?1) were found to correlate well with annual runoff (cm yr^?1). Besides, the reservoir was found to have had a significant and beneficial impact on the downstream loading rates of BOD and COD, which were reduced by 55 and 75%, respectively. As for ammonia, the results of this study indicate that its annual loadings at downstream locations were not significantly affected by the reservoir. Average non-point source contributions of BOD and ammonia loadings into the system were found to be about 80 and 55%, respectively.     

Method for the detection of synthetic cry3A in transgenic potatoes

All transgenic cultivars of potatoes registered in Canada and the United States have been modified to express a synthetic cry3A gene as a means of conferring resistance against the Colorado potato beetle, an important economic pest of potatoes. A PCR method was developed to amplify a 499 bp region of the synthetic cry3A gene. Using this method, synthetic cry3A could be detected in six different transgenic cultivars. Positive results could be confirmed with PvuII restriction digestion of the PCR-generated amplicon, which resulted in two fragments that were 283 and 216 bp in size. Of the 52 tuber extracts tested with this method, no false positive or false negative results were obtained, suggesting the method could be used with a high degree of accuracy. The absolute limit of detection was the number of cry3A copies present in one or perhaps two haploid copies of the potato genome. The practical limit of detection in tubers on a fresh weight basis was 0.02% for the NL 10-SUP and 0.01% for the remaining cultivars. Synthetic cry3A could also be detected in processed food products such as potato chips, shoestring potatoes, and frozen French fries. The method was suitable for screening potato tuber lots and some processed foods for the presence of synthetic cry3A.     

A comparison of constructed and natural habitat for frog conservation in an Australian agricultural landscape

Constructed ponds are an important consideration in the conservation of wetland biota in agricultural landscapes. Twenty-two natural ponds and 22 adjacent constructed ponds (farm dams) were surveyed on the Southern Tablelands of New South Wales to compare patterns of use by frogs and develop frog conservation recommendations. Farm dams supported similar numbers of frog species to natural ponds, although differences in frog assemblage were observed between the pond types. Limnodynastes tasmaniensis and Uperolia laevigata were significantly more likely to occur at farm dams while L. peronii was more likely to occur at natural ponds. Results suggest waterbodies with high levels of emergent vegetation cover that lack fish are likely to support a high number of frog species, regardless of origin (i.e. natural or constructed). However, it is important for landholders to conserve natural waterbodies as these environments appear likely to support frog species that do not use farm dams.     

Cocomposting of Cattle Manure and Hydrocarbon Contaminated Flare Pit Soils

The potential of using composting technology to remediate clayey soils with high levels of crude oil contamination was evaluated. An open air windrow comprised of flare pit soil, manure and wood chips was constructed at Olds College, Composting Technology Centre. Aeration and mixing were carried out by a skid steer loader and composting parameters were monitored for ten months. Temperature profile of this windrow gave cyclic patterns of high and low temperature recordings corresponded to the turning events. Most of the microbial metabolic activity occurred within the mesophilic temperature range and the hydrocarbon degrading microorganism populations remained high throughout the trial. Complete removal of BTEX compounds was achieved within six months and extractable carbons from C₅ to C₁₀ were reduced by 98.7% compared to the initial contaminated soil. Vegetative growth on the composted soil was also evaluated. Barley and timothy plants grown in the composted soil were compared to the control off-lease soil, contaminated soil, and other treatments of varying salinity and organic matter levels. Plant germination, survival, and biomass production was significantly better in the composted soil than in the contaminated soil. Furthermore, barley plants grown in the composted soil were more resilient than those grown in the control off-lease soil.     

The effect of immunosuppression on protective immunity of turkey poults against infection with turkey coronavirus

The objective of the present study was to evaluate the protective effect of humoral and cellular immunities on turkeys infected with turkey coronavirus (TCV). Two trials were conducted with two separate hatches of turkey poults. Turkey's were experimentally immunosuppressed with cyclosporin A (CsA) or cyclophosphamide (CY) and infected with TCV. Prior to infection, treatment with CsA selectively suppressed T cell activity as revealed by 2-3 fold decreased (p < 0.1) lymphocyte proliferation responses to a T cell mitogen, concanavalin A (Con A). Treatment with CY mainly induced B cell deficiency as indicated by significant reductions (p < 0.05) in antibody responses to sheep erythrocytes 7 days after injection. Body weight gain of turkeys treated with CY was significantly lower (p < 0.05) than that of untreated turkeys at 9 days post-infection (PI). Turkeys treated with CY had 1-2 fold higher immunofluorescent antibody assay (IFA) scores for TCV antigens (p < 0.05) in the intestine than untreated turkeys at 9 or 14 days PI. These results suggested that humoral immunity against TCV infection may be important in turkeys.     

Characterization of turkey coronavirus from turkey poults with acute enteritis.

The present study was to characterize turkey coronavirus associated with turkey poult enteritis and mortality. Intestinal contents or intestines from affected turkey poults and inoculated turkey embryos contained coronaviruses as revealed by electron microscopy or were positive for turkey coronavirus by immunofluorescent antibody assay. Sucrose density gradient ultracentrifugation of the virus-containing intestinal homogenate yielded two opalescent bands corresponding to the buoyant densities of 1.14-1.15 and 1.18-1.20 g/ml, respectively. Coronaviral particles from intestinal contents or the sucrose density gradient preparation were mainly spherical in shape and had envelope and central depression. They were surrounded by a fringe of regularly spaced petal-shaped projections attached to the particles by a short stalk. Purified viruses hemagglutinated rabbit erythrocytes with a titer of 16. Major protein bands of purified viruses analyzed by SDS-PAGE were located at 200, 100-110, 50-60, and 30-35 kDa. The patterns of protein bands were consistent with those of Minnesota or Quebec turkey coronavirus isolates. A 568 bp nucleotide fragment of turkey coronavirus spike protein gene was amplified from RNA of inoculated turkey embryo intestine or purified virus. Sequence analysis of the 568 bp PCR product revealed high degree of identity with the corresponding spike protein gene sequence of human and bovine coronaviruses. The results indicated that turkey coronavirus was associated with turkey poults with acute enteritis.     

Induction of hypericins and hyperforin in Hypericum perforatum L. in response to biotic and chemical elicitors

Hypericum perforatum L. produces hyperforins, a family of antimicrobial acylphloroglucinols; and hypericins, a family of phototoxic anthraquinones exhibiting anti-microbial, anti-viral, and anti-herbivore properties in vitro. To determine whether these secondary metabolites are part of the specific plant defense systems that are mediated by methyl jasmonate or salicylic acid, we used meristem cultures to assess the effects of exposure to exogenous application of these chemical elicitors. Levels of hypericins in plant tissue increased in response to both elicitor treatments; total hypericin levels increased as much as 3.3 times control levels when treated with 200 μ methyl jasmonate for 14 days. Increased hyperforin concentrations were detected when plantlets were treated with 1 m salicylic acid or 50 μ methyl jasmonate. For assessing responses to a biotic elicitor, greenhouse-grown plant materials were inoculated with the plant pathogen, Colletotrichum gloeosporioides. Levels of hypericins increased twice as much as the control when inoculated with 1 × 10⁴ spores per ml; higher doses of spores overwhelmed the plant defenses. The elevation of hypericins and hyperforin in response to chemical and biotic elicitors suggests that these secondary metabolites are components in the inducible plant defense responses of H. perforatum.