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.     

Assessing obesity in adult dogs and cats presenting for routine vaccination appointments in the North Island of New Zealand using electronic medical records data

Aims: To assess the prevalence of obesity in adult dogs and cats presented to first-opinion veterinary clinics in the North Island of New Zealand for routine vaccination appointments, using electronic medical records.

Methods: Ten first-opinion veterinary clinics across the North Island of New Zealand provided electronic medical records for all routine vaccination appointments for adult (>1 year old) dogs and cats between 1 January 2011 and 30 June 2016. Animals with a body condition score (BCS) of 6 or 7 on a 9-point scale and 4 on a 5-point scale were classified as overweight; those with a BCS of 8 or 9 on a 9-point scale and 5 on a 5-point scale were classified as obese. A total of 106,144 records were available over the study period, of which 48,041 (45.2%) had both a recorded weight and BCS.

Results: Of the 24,247 records for dogs with both BCS and weight, 6,324 (26.1%) were classified as overweight, and 551 (2.3%) as obese. The prevalence of dogs classified as overweight or obese was highest in dogs aged between 5–13 years. The odds of desexed dogs being classified as overweight or obese was greater than the odds for intact dogs (OR=1.42 (95% CI=1.29–1.57), p<0.001) adjusting for the effects of age. Of the 23,794 records for cats with a recorded weight and BCS, 5,222 (21.9%) were classified as overweight, and 622 (2.6%) as obese. The prevalence of cats classified as overweight or obese was highest in cats aged between 5–11 years. The odds of desexed cats being classified as overweight or obese tended to be greater than the odds for intact cats (OR=1.14 (95% CI=0.98–1.31); p=0.075), adjusting for the effects of age.

Conclusions: Although there are limitations with using electronic medical records to estimate the prevalence of obesity in companion animal populations, the results highlight that a significant number of animals presenting for routine vaccination appointments were classified as overweight or obese.

Clinical Relevance: It is important for veterinarians to record both patient body condition and weight during routine preventative care appointments to allow accurate ongoing monitoring of trends in obesity at both the patient and population levels.

Abbreviations: BCS: Body condition score     

Indicators of success for smart law enforcement in protected areas: A case study for Russian Amur tiger (Panthera tigris altaica) reserves

Although considerable conservation resources have been committed to develop and use law enforcement monitoring and management tools such as SMART, measures of success are ill‐defined and, to date, few reports detail results post‐implementation. Here, we present 4 case studies from protected areas with Amur tigers (Panthera tigris altaica) in Russia, in which indicators of success were defined and evaluated at each. The ultimate goal was an increase in tiger numbers to 1 individual/100 km² at each site. We predicted that improvements in law enforcement effectiveness would be followed by increases in prey numbers and, subsequently, tiger numbers. We used short‐term and long‐term indicators of success, including: (i) patrol team effort and effectiveness; (ii) catch per unit effort indicators (to measure reductions in threats); and (iii) changes in target species numbers. In addition to implementing a monitoring system, we focused on improving law enforcement management using an adaptive management process. Over 4 years, we noted clear increases in patrol effort and a partial reduction in threats. Although we did not detect clear trends in ungulate numbers, tiger populations remained stable or increased, suggesting that poaching of tigers may be more limiting than prey depletion. Increased effectiveness is needed before a clear reduction in threats can be noted, and more time is needed before detecting responses in target populations. Nonetheless, delineation of concrete goals and indicators of success provide a means of evaluating progress and weaknesses. Such monitoring should be a central component of law enforcement strategies for protected areas.     

Application of unmanned aerial systems for high throughput phenotyping of large wheat breeding nurseries

Background

Low cost unmanned aerial systems (UAS) have great potential for rapid proximal measurements of plants in agriculture. In the context of plant breeding and genetics, current approaches for phenotyping a large number of breeding lines under field conditions require substantial investments in time, cost, and labor. For field-based high-throughput phenotyping (HTP), UAS platforms can provide high-resolution measurements for small plot research, while enabling the rapid assessment of tens-of-thousands of field plots. The objective of this study was to complete a baseline assessment of the utility of UAS in assessment field trials as commonly implemented in wheat breeding programs. We developed a semi-automated image-processing pipeline to extract plot level data from UAS imagery. The image dataset was processed using a photogrammetric pipeline based on image orientation and radiometric calibration to produce orthomosaic images. We also examined the relationships between vegetation indices (VIs) extracted from high spatial resolution multispectral imagery collected with two different UAS systems (eBee Ag carrying MultiSpec 4C camera, and IRIS+ quadcopter carrying modified NIR Canon S100) and ground truth spectral data from hand-held spectroradiometer.

Results

We found good correlation between the VIs obtained from UAS platforms and ground-truth measurements and observed high broad-sense heritability for VIs. We determined radiometric calibration methods developed for satellite imagery significantly improved the precision of VIs from the UAS. We observed VIs extracted from calibrated images of Canon S100 had a significantly higher correlation to the spectroradiometer (r = 0.76) than VIs from the MultiSpec 4C camera (r = 0.64). Their correlation to spectroradiometer readings was as high as or higher than repeated measurements with the spectroradiometer per se.

Conclusion

The approaches described here for UAS imaging and extraction of proximal sensing data enable collection of HTP measurements on the scale and with the precision needed for powerful selection tools in plant breeding. Low-cost UAS platforms have great potential for use as a selection tool in plant breeding programs. In the scope of tools development, the pipeline developed in this study can be effectively employed for other UAS and also other crops planted in breeding nurseries.

     

Validation of a new dental plaque quantification method in dogs

Current methods for the quantification of plaque accumulation in cats and dogs are well-accepted adaptations of traditional human models, but have required substantial modifications in order to compensate for the inherent differences in compliance, cooperation, and temperament between animals and humans. While these modifications have sought to maintain or improve upon the accuracy and reproducibility of the original methods, they also have increased the complexity of the technique and have required additional scorer time and animal cooperation, which leads to increased cost of trials. Therefore, research was directed toward the development and validation of a new substrate scoring system that reduces resources while maintaining or increasing the reproducibility attributed to the more traditional methods. This new gingival contour plaque index was shown to be accurate and reproducible, but used fewer animals, required less time, and eliminated the need for many of the specialized procedures required by traditional methods.     

ULTRASOUND AS AN AID FOR DIAGNOSIS OF OVARIAN ABSCESSES IN TWO MARES

This report describes two mares presented for evaluation of anorexia, fever of unknown origin, and weight loss. Clinical examination, laboratory findings, and transrectal ultrasonographic images suggested ovarian abscessation. One mare was successfully treated medically. Because of financial considerations, the second mare was euthanatized and a postmortem examination was performed. At necropsy, there was an enlarged right ovary with an adhesion to the large colon. Microscopic findings were characteristic of an ovarian abscess.     

Pulmonary Thromboembolism in 29 Dogs: 1985–1995

Pulmonary thromboembolism (PTE) occurs as a complication to a number of commonly encountered clinical diseases. Antemortem recognition of this life-threatening disorder is hampered by nonspecificity of clinical signs. This retrospective study was performed to analyze clinical features, laboratory findings, imaging abnormalities, and concurrent postmortem diagnoses in 29 dogs with confirmed pulmonary embolism. A variety of clinicopathologic and radiographic abnormalities were noted but there were no pathognomonic findings for PTE. Arterial blood gas analyses were performed in 15 (52%) of 29 dogs; 12 (80%) of 15 exhibited hypoxemia and 15 (100%) of 15 had increased alveolar-arterial oxygen gradients. Response to supplemental O2 was variable and did not correlate with the presence or absence of additional pulmonary pathology on postmortem. At postmortem, 25 (86%) of 29 dogs had grossly visible emboli, 17 (59%) of 29 dogs had multiple disease processes, and 16 (55%) of 29 dogs had additional pulmonary pathology. PTE was suspected antemortem in 11 (38%) of 29 dogs. In dogs with respiratory signs consistent with PTE, the condition was a differential diagnosis in 11 of 17 animals; all had diseases previously reported to be associated with PTE. Neoplasia, systemic bacterial disease, and immune-mediated hemolytic anemia were diagnosed most frequently.     

Ascorbate Oxidase,Protein Disulfide Isomerase,Ascorbic Acid,Dehydroascorbic Acid and Protein Levels in Developing Wheat Kernels and Their Relationship to Protein Disulfide Bond Formation

Developing wheat kernels of three New Zealand wheat cultivars at one to nine weeks postanthesis (WPA) were analyzed for kernel weight, salt‐soluble protein, salt‐insoluble protein, ascorbate oxidase (AOX), protein disulfide isomerase (PDI), ascorbic acid (AA), and dehydroascorbic acid (DHA). Kernel weight and salt‐soluble protein increased from one to five or six WPA then declined. The amount of salt‐insoluble protein per kernel rapidly increased from two to five or six WPA, then remained constant. AOX activity increased from one to two or three WPA then declined to almost zero at seven WPA. PDI activity increased from one to three WPA, stayed constant to six WPA, then declined about 25%. DHA increased from one to two or three WPA then declined to zero at seven WPA. AA increased from one to three or five WPA then declined to zero at seven WPA. When DHA, at similar concentrations (≈1 mM) to that found in one to six WPA kernels, was mixed with reduced‐gluten proteins, it was reduced to AA. The results are interpreted as a scheme showing that, during the formation of protein‐disulfide bonds, electrons are transferred from nascent protein thiols, ultimately to oxygen, through a series of electron carriers including PDI, AA‐DHA, and AOX.     

Influence of harvest date on seed yield and quality in forage kochia

Forage kochia (Bassia prostrata) is used for rangeland reclamation and livestock and wildlife forage, but limited research has been conducted on its seed production. Therefore, this research evaluated the effect of harvest date on seed weight, germination, and seed yield of forage kochia subspecies virescens and grisea. Seed was harvested from individual plants for 3 years during October, November, and December. October harvest had the lightest 100-seed weights, with the November harvest slightly heavier than December, for most accessions. Cultivar Snowstorm and breeding line Sahsel, both subsp. grisea, had the greatest 100-seed weights in November, 155 and 143 mg, respectively, whereas, cv. Immigrant (subsp. virescens), the standard for forage kochia, ranked among the least for 100-seed weight. For most accessions, germination was lowest from the October harvest (11%–43%), with greater germination with November and December harvested seeds (43%–64%). Viable seed yields were greatest in November with the exception of two accessions, which peaked in October, indicating earlier maturity. Results indicate that forage kochia usually reaches optimum seed maturity by early November, after plants are exposed to freezing temperatures; however, earlier maturing accessions exist in both subspecies virescens and grisea.