ALBERT

Description: Close-to-market weight grain fed cattle experience high heat loads during summer. There are health, welfare and production impacts on these high value animals. Two cohorts of 600 kg Black Angus steers (n = 12) were subjected to heatwave conditions during a thermal challenge in climate chambers. Frequent blood sampling enabled a detailed description of the metabolic and endocrine trajectories during high heat load and recovery in feedlot cattle. In high heat load ruminants, blood flow is diverted from the major organs impacting metabolic rate and cellular functions. The metabolic rate will slow with falls in the thyroid hormone plasma concentrations. Insulin and the adipokines gave an indifferent response. The high heat load cattle were hypoglycaemic and oxidising fatty acids. Liver involvement was evidenced by the build-up of bilirubin in plasma, and reduced release of cholesterol and ALP. Thermal challenge saw markedly increased plasma creatinine and urea implicating reduced glomerular filtration; although the kidneys were working to retain chloride ions to balance the loss of bicarbonate from the increased respiration rate. As heat load reduced during recovery, rumen temperature and respiration rate normalised and feed intake gradually returned. Plasma glucose levels increased also. With increased blood supply to the organs, there was a rise in liver enzymes into the blood, although liver function had not fully restored during the recovery period; plasma bilirubin concentrations were still high, and ALP and cholesterol levels low. Twelve days after the thermal challenge, most blood parameters had returned to normal and the steers had gained weight.

Description: Introduction: Unnatural amino acids (UAAs) share the same basic structure as proteinogenic amino acids. However, UAAs permit additional functions and applications to proteins due to their different side chains. Recent UAA applications include using fluorescent UAAs to label proteins. The UAA system provides an alternative method to traditional protein labeling mechanisms (antibodies, GFP, and tags, such as HA and HIS), which can affect protein functionality and topology. The purpose of this study was to visualize the hepatitis C virus (HCV) core protein using the fluorescent UAA Anap (3-[(6-acetyl-2-naphthalenyl)amino]-L-alanine). Methods: Huh-7.5 cells were co-transfected with HCV core plasmids containing amber stop codons at various positions throughout the coding sequence and a second plasmid encoding the orthogonal tRNA/synthetase pair that facilitates Anap incorporation. Three days post transfection, cells were stained for core protein and lipid droplets (LDs) and visualized using immunofluorescence or confocal microscopy. Results: We have optimized transfection protocols for the efficient expression of the tRNA/synthetase pair required for Anap incorporation and are able to visualize our core mutant proteins containing Anap. We have successfully substituted Anap into 11 different positions within the core, including substitutions for tryptophan, tyrosine, and phenylalanine residues. In addition, we have shown that our core mutants associate with cellular LDs, suggesting that the incorporation of the UAA did not disrupt core protein expression, stability, or cellular localization. Conclusions: We have demonstrated the establishment of a UAA incorporation system in an HCV protein without any obvious impact on core protein function. The ability to label viral proteins using fluorescent UAAs eliminates the requirement of antibodies or tags for protein visualization. In conclusion, the UAA system is a useful method to study HCV proteins and can potentially be used to label viruses for live cell and animal studies.

Description: Introduction: Pyroptosis (inflammatory programmed cell death) is induced after the activation of an inflammasome, ultimately resulting in pore formation and cell lysis. One factor in the pathology associated with chronic hepatitis C virus (HCV) infection is non-inflammatory caspase-3-mediated apoptosis. Our lab has found both apoptosis and pyroptosis occurring in HCV-infected Huh-7.5 cells. In the context of some viral infections, pyroptosis is beneficial to the virus; for others, pyroptosis is believed to represent an innate antiviral response. This study aimed to test the effects of knocking out components of the inflammasome pathway on caspase activation in HCV-infected cells. Methods: FAM-FLICA (Carboxyfluorescein - Fluorochrome Inhibitor of Caspases) probes or antibodies were used to visualize active caspase-1 and active caspase-3 in vitro. Huh-7.5 cells with components of the pyroptotic or apoptotic pathways knocked out (NLRP3, GSDM-D or caspase-3) were used to determine the effects of their absence on the virus and caspase activation using confocal microscopy and flow cytometry. Results: Increased levels of caspase-1 were consistently observed in HCV-infected cells compared to those in uninfected cells, and these levels increased with subsequent days post-infection. The inhibition of inflammasome activation using knock out cell lines induced the differential activation of caspase-1 and caspase-3, with the inhibition of pyroptosis, resulting in a trend towards greater expression of caspase-3, indicative of apoptosis. The inhibition of NLRP3 did not fully stop caspase-1 activation, but it was decreased. The flow cytometry results revealed a small sub-set of cells positive for both caspase-1 and caspase-3. Conclusions: These data confirm the occurrence of pyroptosis in HCV-infected cells and demonstrate the involvement of the NLRP3 inflammasome, although other inflammasome sensors might be involved. Since the inhibition of one cell death pathway resulted in the increased activation of the other, along with the presence of double-positive cells, there may be cross-talk between apoptotic and pyroptotic pathways; the role of this cross-talk during infection remains to be elucidated.

Description: There is limited research that evaluates the effect of farmer involvement in agrichemical exposure surveillance on their attitudes and behaviour towards pesticide handling and use of personal protective equipment. This limited follow-up study aimed to (i) evaluate attitudes/behaviours towards the use of personal protective equipment (PPE) among farmers who participated in the In-Field Personalised Cholinesterase Assessment Project (PCAP) (2016/17); and (ii) qualitatively assess the effect of monthly presentation of acetylcholinesterase (AChE) testing results on farmer agrichemical safety practices and behaviours prior to, and following participation in PCAP. This study surveyed 42 farming men and women, asking questions about agrichemical usage and hygiene practices. The majority of surveyed farmers’ self-apply agrichemicals on their farm (97.6%), with 81% reporting that involvement in PCAP research changed the way they handled Organophosphates (OPs)—a widely used insecticide in agriculture. By enabling people to think critically about their exposure, there was a 66% increase in frequency of respirator usage post-PCAP. Following this, participants were invited to take part in one-on-one interviews to further discuss their involvement in PCAP. Many responses were positive, with participants stating they were more aware and cautious of their own practices. This study determined that research participation and point-of-care testing and education can result in effective engagement of farmers and farm workers, increase health literacy and change farming practice—highlighting the importance of an interactive, participatory model in order to bring about change, to reduce possible pesticide exposures.

Description: COVID-19 is a wicked problem for policy makers internationally as the complexity of the pandemic transcends health, environment, social and economic boundaries. Many countries are focusing on two key responses, namely virus containment and financial measures, but fail to recognise other aspects. The systems approach, however, enables policy makers to design the most effective strategies and reduce the unintended consequences. To achieve fundamental change, it is imperative to firstly identify the “right” interventions (leverage points) and implement additional measures to reduce negative consequences. To do so, a preliminary causal loop diagram of the COVID-19 pandemic was designed to explore its influence on socio-economic systems. In order to transcend the “wait and see” approach, and create an adaptive and resilient system, governments need to consider “deep” leverage points that can be realistically maintained over the long-term and cause a fundamental change, rather than focusing on “shallow” leverage points that are relatively easy to implement but do not result in significant systemic change.

Description: A multiple sensor payload for a multi-rotor based UAV platform was developed and tested for measuring land surface albedo and spectral measurements at user-defined spatial, temporal, and spectral resolutions. The system includes a Matrice 600 UAV with an RGB camera and a set of four downward pointing radiation sensors including a pyranometer, quantum sensor, and VIS and NIR spectrometers, measuring surface reflected radiation. A companion ground unit consisting of a second set of identical sensors simultaneously measure downwelling radiation. The reflected and downwelling radiation measured by the four sensors are used for calculating albedo for the total shortwave broadband, visible band and any narrowband at a 1.5 nm spectral resolution within the range of 350–1100 nm. The UAV-derived albedo was compared with those derived from Landsat 8 and Sentinel-2 satellite observations. Results show the agreement between total shortwave albedo from UAV pyranometer and Landsat 8 (R2 = 0.73) and Sentinel-2 (R2 = 0.68). Further, total shortwave albedo was estimated from spectral measurements and compared with the satellite-derived albedo. This UAV-based sensor system promises to provide high-resolution multi-sensors data acquisition. It also provides maximal flexibility for data collection at low cost with minimal atmosphere influence, minimal site disturbance, flexibility in measurement planning, and ease of access to study sites (e.g., wetlands) in contrast with traditional data collection methods.

Description: Rice is the principal source of calories for more than half of the world’s population, and while an excellent source of energy is a relatively poor source of micronutrients. [...]

Description: In beef cattle, horn management is practiced to physically or surgically remove horns for the safety of animals and workers. However, invasive practices of dehorning and disbudding are a great threat to animal welfare, health, production and human safety, as well as labour intensive and costly. The most effective way to limit the impacts and costs of horns is to prevent their occurrences by breeding naturally polled (hornless) herds. Horn development is complex, although two mutually exclusive genetic variants (Celtic and Friesian) have been found prevalent on each copy of chromosome 1 in most polled cattle. Predicting genotypes in an animal is challenging. Available genetic testing assays were often limited in tropically adapted beef cattle. In this study we present a new optimized poll testing (OPT) assay, which has been bundled with SNP genotyping arrays being used for genomic evaluation in cattle. Breeding schemes can profile future parents for pure-polled stock based on the OPT results. We also evaluated the factors causing complexity in horn conditions. Thus, we coupled OPT predictions with head-status and sex distributions, by modelling genetic and non-genetic impacts, revealing that genetics, sex and sex hormones control horn ontology. Finally, concerns of polledness adversely affecting production and reproduction were investigated by using estimated breeding values of several beef traits. We found no detrimental effects of polledness on production or reproduction. Overall, this research concludes that genetically polled cattle will minimize issues about animal welfare and management costs without reducing production potentials in the tropically adapted beef cattle.