Major Cellular and Physiological Impacts of Ocean Acidification on a Reef Building Coral

As atmospheric levels of CO2 increase, reef-building corals are under greater stress from both increased sea surface temperatures and declining sea water pH. To date, most studies have focused on either coral bleaching due to warming oceans or declining calcification due to decreasing oceanic carbonate ion concentrations. Here, through the use of physiology measurements and cDNA microarrays, we show that changes in pH and ocean chemistry consistent with two scenarios put forward by the Intergovernmental Panel on Climate Change (IPCC) drive major changes in gene expression, respiration, photosynthesis and symbiosis of the coral, Acropora millepora, before affects on biomineralisation are apparent at the phenotype level. Under high CO2 conditions corals at the phenotype level lost over half their Symbiodinium populations, and had a decrease in both photosynthesis and respiration. Changes in gene expression were consistent with metabolic suppression, an increase in oxidative stress, apoptosis and symbiont loss. Other expression patterns demonstrate upregulation of membrane transporters, as well as the regulation of genes involved in membrane cytoskeletal interactions and cytoskeletal remodeling. These widespread changes in gene expression emphasize the need to expand future studies of ocean acidification to include a wider spectrum of cellular processes, many of which may occur before impacts on calcification.

Physiological and behavioral responses of sheep to gaseous ammonia

Ammonia can accumulate in highly stocked sheep accommodation, for example during live export shipments, and could affect sheep health and welfare. Thus, the objective of this experiment was to test the effects of 4 NH3 concentrations, 4 (control), 12, 21, and 34 mg/m(3), on the physiology and behavior of wether sheep. Sheep were held for 12 d under a micro-climate and stocking density similar to shipboard conditions recorded on voyages from Australia to the Middle East during the northern hemispheric summer. Ammonia increased macrophage activity in transtracheal aspirations, indicating active pulmonary infl ammation; however, it had no effect (P > 0.05) on hematological variables. Feed intake decreased (P = 0.002) in proportion to ammonia concentration, and BW gain decreased (P < 0.001) at the 2 greatest concentrations. Exposure to ammonia increased (P = 0.03) the frequency of sneezing, and at the greatest ammonia concentration, sheep were less active, with less locomotion, pawing, and panting. Twenty-eight days after exposure to NH3, the pulmonary macrophage activity and BW of the sheep returned to that of sheep exposed to only 4 mg/m(3). It was concluded that NH3 induced a temporary inflammatory response of the respiratory system and reduced BW gain, which together indicated a transitory adverse effect on the welfare of sheep.

Acute and chronic effects of dietary nitrate supplementation on blood pressure and the physiological responses to moderate-intensity and incremental exercise

Dietary nitrate (NO3−) supplementation with beetroot juice (BR) over 4–6 days has been shown to reduce the O2 cost of submaximal exercise and to improve exercise tolerance. However, it is not known whether shorter (or longer) periods of supplementation have similar (or greater) effects. We therefore investigated the effects of acute and chronic NO3− supplementation on resting blood pressure (BP) and the physiological responses to moderate-intensity exercise and ramp incremental cycle exercise in eight healthy subjects. Following baseline tests, the subjects were assigned in a balanced crossover design to receive BR (0.5 l/day; 5.2 mmol of NO3−/day) and placebo (PL; 0.5 l/day low-calorie juice cordial) treatments. The exercise protocol (two moderate-intensity step tests followed by a ramp test) was repeated 2.5 h following first ingestion (0.5 liter) and after 5 and 15 days of BR and PL. Plasma nitrite concentration (baseline: 454 ± 81 nM) was significantly elevated (+39% at 2.5 h postingestion; +25% at 5 days; +46% at 15 days; P < 0.05) and systolic and diastolic BP (baseline: 127 ± 6 and 72 ± 5 mmHg, respectively) were reduced by ∼4% throughout the BR supplementation period (P < 0.05). Compared with PL, the steady-state V̇o2 during moderate exercise was reduced by ∼4% after 2.5 h and remained similarly reduced after 5 and 15 days of BR (P < 0.05). The ramp test peak power and the work rate at the gas exchange threshold (baseline: 322 ± 67 W and 89 ± 15 W, respectively) were elevated after 15 days of BR (331 ± 68 W and 105 ± 28 W; P < 0.05) but not PL (323 ± 68 W and 84 ± 18 W). These results indicate that dietary NO3− supplementation acutely reduces BP and the O2 cost of submaximal exercise and that these effects are maintained for at least 15 days if supplementation is continued.

How weed control and fertilisation influence tree physiological processes and growth at early establishment in an exotic F1 hybrid pine plantation of subtropical Australia

Purpose This study investigated how nitrogen (N) nutrition and key physiological processes varied under changed water and nitrogen competition resulting from different weed control and fertilisation treatments in a 2-year-old F1 hybrid (Pinus elliottii Engelm var. elliottii × P. caribaea var. hondurensis Barr. ex Golf.) plantation on a grey podzolic soil type, in Southeast Queensland. Materials and methods The study integrated a range of measures including growth variables (diameter at ground level (DGL), diameter at breast height (DBH) and height (H)), foliar variables (including foliar N concentration, foliar δ13C and δ15N) and physiological variables (including photosynthesis (An), stomatal conductance (gs), transpiration (E), intrinsic water use efficiency (WUEi) (A/gs) and xylem pressure potential (ΨXPP)) to better understand the mechanisms influencing growth under different weed control and fertilisation treatments. Five levels of weed control were applied: standard (routine), luxury, intermediate, mechanical and nil weed control, all with routine fertilisation plus an additional treatment, routine weed control and luxury fertilisation. Relative weed cover was assessed at 0.8, 1.1 and 1.6 years after plantation establishment to monitor the effectiveness of weed control treatments. Soil investigation included soil ammonium (NH4 +-N), nitrate (NO3 −-N), potentially mineralizable N (PMN), gravimetric soil moisture content (MC), hot water extractable organic carbon (HWETC), hot water extractable total N (HWETN), total C, total N, stable C isotope composition (δ13C), stable N isotope composition (δ15N), total P and extractable K. Results and discussion There were significant relationships between foliar N concentrations and relative weed cover and between tree growth and foliar N concentration or foliar δ15N, but initial site preparation practices also increased soil N transformations in the planting rows reducing the observable effects of weed control on foliar δ15N. A positive relationship between foliar N concentration and foliar δ13C or photosynthesis indicated that increased N availability to trees positively influenced non-stomatal limitations to photosynthesis. However, trees with increased foliar N concentrations and photosynthesis were negatively related to xylem pressure potential in the afternoons which enhanced stomatal limitations to photosynthesis and WUEi. Conclusions Luxury and intermediate weed control and luxury fertilisation positively influenced growth at early establishment by reducing the competition for water and N resources. This influenced fundamental key physiological processes such as the relationships between foliar N concentration, A n, E, gs and ΨXPP. Results also confirmed that time from cultivation is an important factor influencing the effectiveness of using foliar δ15N as an indicator of soil N transformations.

A physiological framework to explain genetic and environmental regulation of tillering in sorghum

Tillering determines the plant size of sorghum (Sorghum bicolor) and an understanding of its regulation is important to match genotypes to prevalent growing conditions in target production environments. The aim of this study was to determine the physiological and environmental regulation of variability in tillering among sorghum genotypes, and to develop a framework for this regulation. * Diverse sorghum genotypes were grown in three experiments with contrasting temperature, radiation and plant density to create variation in tillering. Data on phenology, tillering, and leaf and plant size were collected. A carbohydrate supply/demand (S/D) index that incorporated environmental and genotypic parameters was developed to represent the effects of assimilate availability on tillering. Genotypic differences in tillering not explained by this index were defined as propensity to tiller (PTT) and probably represented hormonal effects. * Genotypic variation in tillering was associated with differences in leaf width, stem diameter and PTT. The S/D index captured most of the environmental effects on tillering and PTT most of the genotypic effects. * A framework that captures genetic and environmental regulation of tillering through assimilate availability and PTT was developed, and provides a basis for the development of a model that connects genetic control of tillering to its phenotypic consequences.

Synthesis, Reactivity and Biological Activity of 17β-HSD1 Inhibitors Based on a Thieno[2,3-d]pryrimidin-4(3H)-one Core

Breast cancer is the most common cancer in women in Western countries. In the early stages of development most breast cancers are hormone-dependent, and estrogens, especially estradiol, have a pivotal role in their development and progression. One approach to the treatment of hormone-dependent breast cancers is to block the formation of the active estrogens by inhibiting the action of the steroid metabolising enzymes. 17beta-Hydroxysteroid dehydrogenase type 1 (17beta-HSD1) is a key enzyme in the biosynthesis of estradiol, the most potent female sex hormone. The 17beta-HSD1 enzyme catalyses the final step and converts estrone into the biologically active estradiol. Blocking 17beta-HSD1 activity with a specific enzyme inhibitor could provide a means to reduce circulating and tumour estradiol levels and thus promote tumour regression. In recent years 17beta-HSD1 has been recognised as an important drug target. Some inhibitors of 17beta-HSD1 have been reported, however, there are no inhibitors on the market nor have clinical trials been announced. The majority of known 17beta-HSD1 inhibitors are based on steroidal structures, while relatively little has been reported on non-steroidal inhibitors. As compared with 17beta-HSD1 inhibitors based on steroidal structures, non-steroidal compounds could have advantages of synthetic accessibility, drug-likeness, selectivity and non-estrogenicity. This study describes the synthesis of large group of novel 17beta-HSD1 inhibitors based on a non-steroidal thieno[2,3-d]pyrimidin-4(3H)-one core. An efficient synthesis route was developed for the lead compound and subsequently employed in the synthesis of thieno[2,3-d]pyrimidin-4(3H)-one based molecule library. The biological activities and binding of these inhibitors to 17beta-HSD1 and, finally, the quantitative structure activity relationship (QSAR) model are also reported. In this study, several potent and selective 17beta-HSD1 inhibitors without estrogenic activity were identified. This establishment of a novel class of inhibitors is a progressive achievement in 17beta-HSD1 inhibitor development. Furthermore, the 3D-QSAR model, constructed on the basis of this study, offers a powerful tool for future 17beta-HSD1 inhibitor development. As part of the fundamental science underpinning this research, the chemical reactivity of fused (di)cycloalkeno thieno[2,3-d]pyrimidin-4(3H)-ones with electrophilic reagents, i.e. Vilsmeier reagent and dimethylformamide dimethylacetal, was investigated. These findings resulted in a revision of the reaction mechanism of Vilsmeier haloformylation and further contributed to understanding the chemical reactivity of this compound class. This study revealed that the reactivity is dependent upon a stereoelectronic effect arising from different ring conformations.

Physiological Factors Influencing Toughness in Cooked Saddletail Snapper (Lutjanus malabaricus)

Saddletail snapper (Lutjanus malabaricus) is a commercially significant tropical species in Australia and has been the subject of consumer complaints of extreme toughness in cooked fillets. Textural and biochemical analyses including collagen and hydroxylysyl pyridinoline (PYD) cross-links concentrations were conducted on 101 commercially harvested Saddletail snapper to identify causes of toughness. Fish age was found to account for 75.6% of observed variation in cooked muscle texture (work done) of Saddletail snapper. A significant linear relationship (P < 0.001) between PYD content and cooked muscle texture was also identified accounting for 50.3% of observed variation. The concentration ratio of PYD to total collagen (TC) ranged from 0.04 to 0.38 mol PYD per mol of TC. Fish size was also found to be a poor indicator of fish age and therefore a poor indicator of the potential risk of toughness of the cooked muscle.

QTL analysis in multiple sorghum populations facilitates the dissection of the genetic and physiological control of tillering

Tillering in sorghum can be associated with either the carbon supply–demand (S/D) balance of the plant or an intrinsic propensity to tiller (PTT). Knowledge of the genetic control of tillering could assist breeders in selecting germplasm with tillering characteristics appropriate for their target environments. The aims of this study were to identify QTL for tillering and component traits associated with the S/D balance or PTT, to develop a framework model for the genetic control of tillering in sorghum. Four mapping populations were grown in a number of experiments in south east Queensland, Australia. The QTL analysis suggested that the contribution of traits associated with either the S/D balance or PTT to the genotypic differences in tillering differed among populations. Thirty-four tillering QTL were identified across the populations, of which 15 were novel to this study. Additionally, half of the tillering QTL co-located with QTL for component traits. A comparison of tillering QTL and candidate gene locations identified numerous coincident QTL and gene locations across populations, including the identification of common non-synonymous SNPs in the parental genotypes of two mapping populations in a sorghum homologue of MAX1, a gene involved in the control of tiller bud outgrowth through the production of strigolactones. Combined with a framework for crop physiological processes that underpin genotypic differences in tillering, the co-location of QTL for tillering and component traits and candidate genes allowed the development of a framework QTL model for the genetic control of tillering in sorghum.

Measuring Physiological Stress in Australian Flying-Fox Populations

Flying-foxes (pteropid bats) are the natural host of Hendra virus, a recently emerged zoonotic virus responsible for mortality or morbidity in horses and humans in Australia since 1994. Previous studies have suggested physiological and ecological risk factors for infection in flying-foxes, including physiological stress. However, little work has been done measuring and interpreting stress hormones in flying-foxes. Over a 12-month period, we collected pooled urine samples from underneath roosting flying-foxes, and urine and blood samples from captured individuals. Urine and plasma samples were assayed for cortisol using a commercially available enzyme immunoassay. We demonstrated a typical post-capture stress response in flying-foxes, established urine specific gravity as an attractive alternative to creatinine to correct urine concentration, and established population-level urinary cortisol ranges (and geometric means) for the four Australian species: Pteropus alecto 0.5–305.1 ng/mL (20.1 ng/mL); Pteropus conspicillatus 0.3–370.9 ng/mL (18.9 ng/mL); Pteropus poliocephalus 0.3–311.3 ng/mL (10.1 ng/mL); Pteropus scapulatus 5.2–205.4 ng/mL (40.7 ng/mL). Geometric means differed significantly except for P. alecto and P. conspicillatus. Our approach is methodologically robust, and has application both as a research or clinical tool for flying-foxes, and for other free-living colonial wildlife species

Rhipicephalus microplus serine protease inhibitor family: annotation, expression and functional characterisation assessment

Background: Rhipicephalus (Boophilus) microplus evades the host's haemostatic system through a complex protein array secreted into tick saliva. Serine protease inhibitors (serpins) conform an important component of saliva which are represented by a large protease inhibitor family in Ixodidae. These secreted and non-secreted inhibitors modulate diverse and essential proteases involved in different physiological processes. Methods: The identification of R. microplus serpin sequences was performed through a web-based bioinformatics environment called Yabi. The database search was conducted on BmiGi V1, BmiGi V2.1, five SSH libraries, Australian tick transcriptome libraries and RmiTR V1 using bioinformatics methods. Semi quantitative PCR was carried out using different adult tissues and tick development stages. The cDNA of four identified R. microplus serpins were cloned and expressed in Pichia pastoris in order to determine biological targets of these serpins utilising protease inhibition assays. Results: A total of four out of twenty-two serpins identified in our analysis are new R. microplus serpins which were named as RmS-19 to RmS-22. The analyses of DNA and predicted amino acid sequences showed high conservation of the R. microplus serpin sequences. The expression data suggested ubiquitous expression of RmS except for RmS-6 and RmS-14 that were expressed only in nymphs and adult female ovaries, respectively. RmS-19, and -20 were expressed in all tissues samples analysed showing their important role in both parasitic and non-parasitic stages of R. microplus development. RmS-21 was not detected in ovaries and RmS-22 was not identified in ovary and nymph samples but were expressed in the rest of the samples analysed. A total of four expressed recombinant serpins showed protease specific inhibition for Chymotrypsin (RmS-1 and RmS-6), Chymotrypsin / Elastase (RmS-3) and Thrombin (RmS-15). Conclusion: This study constitutes an important contribution and improvement to the knowledge about the physiologic role of R. microplus serpins during the host-tick interaction.

Performance and physiological responses of combined t-bar and PIT tagged giant mud crabs (Scylla serrata)

Mud crabs (Scylla spp.) are intensively caught throughout South-East Asia and support a very substantial commercial, recreational fishing and aquaculture industry. Identification of individual animals is important to improve understanding and management of this species. However, tagging of crustaceans is difficult as they frequently molt and internal tags can pose a hazard to consumers. In this pilot study we tested a new method combining passive integrated transponder tags and t-bar tags externally. 45 giant mud crabs (Scylla serrata) were captured from the wild and kept in tanks for a maximum of 10 months. We inserted tags into the abdomen of 35 giant mud crabs and tested a modified method where the combined t-bar/PIT-tag was inserted into the muscle tissue of the rear leg between the dorsal carapace plate and the top of the abdominal flap. Tagged crabs with the modified method showed 85% tag retention for molting crabs. We tested the same method in the field where 852 individuals were tagged with combined t-bar/PIT-tags of which 82 were recaptured showing 100% tag retention but without any evidence of molting having occurred. The tested method of combined t-bar/PIT-tags in giant mud crabs can further improve monitoring for wild and aquaculture populations and can be deployed widely with low cost.

Performance and physiological responses of combined t-bar and PIT tagged giant mud crabs (Scylla serrata)

Mud crabs (Scylla spp.) are intensively caught throughout South-East Asia and support a very substantial commercial, recreational fishing and aquaculture industry. Identification of individual animals is important to improve understanding and management of this species. However, tagging of crustaceans is difficult as they frequently molt and internal tags can pose a hazard to consumers. In this pilot study we tested a new method combining passive integrated transponder tags and t-bar tags externally. 45 giant mud crabs (Scylla serrata) were captured from the wild and kept in tanks for a maximum of 10 months. We inserted tags into the abdomen of 35 giant mud crabs and tested a modified method where the combined t-bar/PIT-tag was inserted into the muscle tissue of the rear leg between the dorsal carapace plate and the top of the abdominal flap. Tagged crabs with the modified method showed 85% tag retention for molting crabs. We tested the same method in the field where 852 individuals were tagged with combined t-bar/PIT-tags of which 82 were recaptured showing 100% tag retention but without any evidence of molting having occurred. The tested method of combined t-bar/PIT-tags in giant mud crabs can further improve monitoring for wild and aquaculture populations and can be deployed widely with low cost.

Physiological basis of yield variation in response to row spacing and plant density of mungbean grown in subtropical environments

In this study, we investigated the extent and physiological bases of yield variation due to row spacing and plant density configuration in the mungbean Vigna radiata (L.) Wilczek variety “Crystal” grown in different subtropical environments. Field trials were conducted in six production environments; one rain-fed and one irrigated trial each at Biloela and Emerald, and one rain-fed trial each at Hermitage and Kingaroy sites in Queensland, Australia. In each trial, six combinations of spatial arrangement of plants, achieved through two inter-row spacings of 1 m or 0.9 m (wide row), 0.5 m or 0.3 m (narrow row), with three plant densities, 20, 30 and 40 plants/m2, were compared. The narrow row spacing resulted in 22% higher shoot dry matter and 14% more yield compared to the wide rows. The yield advantage of narrow rows ranged from 10% to 36% in the two irrigated and three rain-fed trials. However, yield loss of up to 10% was also recorded from narrow rows at Emerald where the crop suffered severe drought. Neither the effects of plant density, nor the interaction between plant density and row spacing, however, were significant in any trial. The yield advantage of narrow rows was related to 22% more intercepted radiation. In addition, simulations by the Agricultural Production Systems Simulator model, using site-specific agronomy, soil and weather information, suggested that narrow rows had proportionately greater use of soil water through transpiration, compared to evaporation resulting in higher yield per mm of soil water. The long-term simulation of yield probabilities over 123 years for the two row configurations showed that the mungbean crop planted in narrow rows could produce up to 30% higher grain yield compared to wide rows in 95% of the seasons.

Preliminary investigation of some physiological responses of Bos indicus heifers to surgical spaying

Objective To determine the value of peripheral blood concentrations of cortisol, creatine phosphokinase (CPK), aspartate aminotransferase (AST), non-esterified fatty acids (NEFAs) and haptoglobin as indicators of welfare in Brahman heifers spayed by either the Willis dropped ovary technique (WDOT) or the flank laparotomy method. Design A total of 24, 2-year-old Brahman heifers were allocated to: crush (head-bail) restraint alone (Control, n = 5); crush restraint and ear-punch (Ear-punch, n = 5); crush restraint, WDOT spay and ear-punch (WDOT, n = 9); or crush restraint, elecrtoimmobilisation, flank spay and ear-punch (Flank; n = 5). Cattle were blood sampled frequently to 8 h, and then daily to day 4 and were monitored to 42 days post-procedure. Peripheral blood concentrations of bound and unbound cortisol, CPK, AST, NEFAs and haptoglobin were determined. Results Concentrations of plasma bound cortisol peaked in the spayed heifers 3-4 h post-procedure; values in the Flank (1603 nmol/L) and WDOT (1290 nmol/L) groups were similar and significantly greater (P < 0.05) than in the Controls (519 nmol/L). Flank heifers had elevated plasma haptoglobin levels to day 4 postprocedure. Liveweights were significantly lower in the spayed compared with the Control heifers at 21 and 42 days post-procedure, with liveweight gains also significantly reduced at day 21. Conclusions Bound cortisol responses in spayed heifers were elevated to 6 h post-procedure and similar in WDOT- and flank-spayed animals, indicating comparable levels of pain and stress. An inflammatory response, indicated by haptoglobin concentrations, was sustained for longer in Flank than in WDOT spayed heifers, suggesting longer-lasting adverse effects on welfare from flank spaying than WDOT spaying. © 2011 The State of Queensland (Department of Employment, Economic Development and Innovation). Australian Veterinary Journal © 2011 Australian Veterinary Association.