Expression, purification and characterisation of recombinant Escherichia coli derived chicken interleukin-12

Zoonotic viruses, such as H5N1 Avian Influenza, pose major threats to both animals and humans, and with this in mind there is a need for the development of new anti-viral strategies. The cytokine interleukin-12 (IL-12) is known to play a pivotal regulatory role in the anti-viral response due to its role in the induction of the key anti-viral cytokine IFN-gamma. Therefore, strategies which provide a means for the production of therapeutic quantities of IL-12 may be of major benefit. Here we describe the development of biologically active Escherichia coli (E. coli) derived chicken IL-12 (ChIL-12). The single chain ChIL-12 gene was cloned into the pET32b expression vector, transformed into the BL-21 E. coli strain and expression induced with IPTG. Over expressed protein was solubilised with zwittergent detergent and isolated utilising Nickel ion affinity chromatography. Biological activity was determined as ChIL-12 stimulated proliferation of pre-treated T-cells in vitro. This study is the first example of a biologically active E. coli derived IL-12 from a non-mammalian vertebrate subsequently providing a means for testing the anti-viral therapeutic potential of ChIL-12 in an in vivo model.     

Development and evaluation of an online computer-aided learning (CAL) package to promote small-animal welfare

RATIONALE FOR THE STUDY: The aims of the study, conducted as a student research project in the 2004-5 session, were to develop an interactive, online computer-aided learning (CAL) package on the topic of small-animal husbandry; to validate the resource as a suitable lecture replacement for first-year veterinary students; and to raise awareness of current guidelines and legislation relating to small-animal housing among local catteries and kennels and in the wider community. METHODOLOGY: Quantitative feedback was collected from student and teaching staff using paper-based questionnaires. Qualitative feedback was gathered from open questionnaire responses and through focus-group discussions with students. Student examination marks were compared for 2004 and 2005, allowing a comparison of student performance before and after the replacement of the traditional lecture with the CAL package. Ethical approval for the study was granted by the faculty's Ethics Committee. RESULTS AND CONCLUSIONS: The CAL package on small-animal housing was well received by teaching staff and students; student performance in examinations improved after the introduction of the CAL program, suggesting that it provides a suitable alternative to didactic teaching. The creation and distribution of the CAL package on CD-ROM and its availability via the Internet are intended to contribute to small-animal welfare education in local catteries and kennels and further afield. The package sets a precedent for the development of more Internet-based, student-authored CAL packages in the future, providing additional resources for independent learning.     

Review of research methodologies for tigers: Telemetry

Over the past half century, wildlife research has relied on technological advances to gain additional insight into the secretive lives of animals. This revolution started in the 1960s with the development of radio telemetry and continues today with the use of Global Positioning System (GPS)-based research techniques. In the present paper we review the history of radio telemetry from its origins with grizzly bears in Yellowstone to its early applications in tiger research and conservation in Asia. We address the different types of data that are available using radio telemetry as opposed to using other research techniques, such as behavioral observations, camera trapping, DNA analysis and scat analysis. In the late 1990s, the rapid development of GPS collar technology revolutionized wildlife research. This new technology has enabled researchers to dramatically improve their ability to gather data on animal movements and ecology. Despite the ecological and conservation benefits of radio telemetry, there have been few telemetry studies of tigers in the wild, and most have been on the Bengal or Amur subspecies. We close with an assessment of the current tiger conservation efforts using GPS technology and discuss how this new information can help to preserve tigers for future generations.     

Clinical,pathologic, and toxicologic characterization of Salvia reflexa (lance-leaf sage) poisoning in cattle fed contaminated hay

Salvia reflexa (lance-leaf sage)-contaminated alfalfa hay was fed to ~500 mixed-breed beef cattle. Within hours of exposure, nearly half of the cattle developed lethargy, anorexia, depression, and recumbency, followed by bellowing, colic, and death. Even though the uneaten contaminated hay was removed the first day, nearly 100 animals died within the first 48 h. Three of these cattle were examined postmortem, and tissues and hay samples were collected for microscopic and chemical analysis. Several days later, a smaller number of the clinically poisoned cattle developed neurologic disease with aberrant behavior, aggression, icterus, blindness, exhaustion, and death. A total of 165 cattle were fatally poisoned. Poisoned cattle had swollen, dark, mottled livers that had a prominent nutmeg-like lobular pattern on cut section. Histologically, there was severe centrilobular-to-panlobular hepatic necrosis with marked hepatocellular swelling, degeneration, and necrosis. The surviving cattle developed liver disease characterized by altered serum biochemical analyses and microscopic hepatocellular degeneration and necrosis. In subsequent biopsies and analysis, these lesions resolved within 6–7 mo. After confirming toxicity of the hay in cattle, goats, and mice, followed by a mouse bioassay–guided chemical fractionation process, Salvia reflexa was identified as the contaminant in the hay responsible for the hepatotoxicity. S. reflexa has not been reported previously to cause fatal hepatotoxicity in livestock in North America, to our knowledge.     

Surgical removal of a mammary adenocarcinoma and a granulosa cell tumor in an African pygmy hedgehog

A 3-year-old, female African pygmy hedgehog (Atelerix albiventris) was referred with a history of hematuria. Hyperglycemia and glucosuria were found at presentation. Mammary adenocarcinoma and a granulosa cell tumor were found and removed surgically. Glucosuria and hematuria resolved, and the hedgehog has done well for 10 mo postoperatively.     

Influence of Prescribed Fire on Ecosystem Biomass,Carbon, and Nitrogen in a Pinyon Juniper Woodland

Increases in pinyon and juniper woodland cover associated with land-use history are suggested to provide offsets for carbon emissions in arid regions. However, the largest pools of carbon in arid landscapes are typically found in soils, and aboveground biomass cannot be considered long-term storage in fire-prone ecosystems. Also, the objectives of carbon storage may conflict with management for other ecosystem services and fuels reduction. Before appropriate decisions can be made it is necessary to understand the interactions between woodland expansion, management treatments, and carbon retention. We quantified effects of prescribed fire as a fuels reduction and ecosystem maintenance treatment on fuel loads, ecosystem carbon, and nitrogen in a pinyon–juniper woodland in the central Great Basin. We found that plots containing 30% tree cover averaged nearly 40 000 kg · ha^?1 in total aboveground biomass, 80 000 kg · ha^?1 in ecosystem carbon (C), and 5 000 kg · ha^?1 in ecosystem nitrogen (N). Only 25% of ecosystem C and 5% of ecosystem N resided in aboveground biomass pools. Prescribed burning resulted in a 65% reduction in aboveground biomass, a 68% reduction in aboveground C, and a 78% reduction in aboveground N. No statistically significant change in soil or total ecosystem C or N occurred. Prescribed fire was effective at reducing fuels on the landscape and resulted in losses of C and N from aboveground biomass. However, the immediate and long-term effects of burning on soil and total ecosystem C and N is still unclear.     

Development of an assay to determine single nucleotide polymorphisms in the prion gene for the genetic diagnosis of relative susceptibility to classical scrapie in sheep.

The objective of this study was to develop a reliable Taqman 5' Nuclease Assay for genotyping sheep for scrapie susceptibility. The sheep prion gene contains 2 single nucleotide polymorphisms (SNPs) that may mediate resistance to classical scrapie, one at codon 136, alanine (A) or valine (V), and another at codon 171, arginine (R) or glutamine (Q). The R allele appears to confer resistance to classical scrapie, with the AA(136) RR(171) genotype the most resistant to scrapie and QR(171) only rarely infected in the US sheep population. The Assays by Design protocol was used for development of probes and primers for codon 136 and Primer Express for codon 171. Commercially available kits were used to isolate genomic DNA from blood or muscle. For validation, 70 SNP determinations for each codon were compared to commercial testing with an error rate of less than 1%. Then, 935 samples from blood (n = 818) and muscle (n = 117) were tested for both codons with 928 successful determinations and only 7 samples (<1% of total samples) that needed repeating. Genotypes were AA QQ (n = 102; 11.0%), AV QQ (n = 28; 3.0%), AA QR (n = 396; 42.7%), AV QR (n = 54; 5.8%), and AA RR (n = 348; 37.5%). Thus, 86% of the sheep tested (n = 798) contained R at codon 171 and were expected to be scrapie-resistant. This new Taqman 5' Nuclease SNP genotyping assay is accurate, easy to perform, and useful in the study of classical scrapie in sheep and its prevention through selective breeding programs to eliminate highly susceptible animals.