High temperature stress (HTS) adversely affects reproduction in most plant species studied to date. HTS during flowering may result in an almost total inhibition of seed production in crop plants. Increasing our knowledge of the effects of HTS on seed production will aid the breeding of more thermotolerant crop plants and improve our understanding of the effects of stress on plants. An investigation of the effects of both drought and high temperature stress on the yields of barley, canola, flax, durum and spring wheat in five locations in Saskatchewan over a 25-year period was performed using multivariate analysis. Higher temperatures during June and July, when the plants were flowering, were correlated with reductions in yields of all the crops studied (except barley in June). A positive correlation between yields and precipitation during May and the winter preceding the growing season was observed.<p>In growth chambers, <i>Brassica napus</i> silique and seed production were inhibited during a ramping HTS treatment. This was due to a decrease in pollen germinability rather than a reduction in the number of flowers produced. HTS also caused reductions in megagametophyte fertility and disrupted embryo and/or seed development.<p>Transgenic plants were developed to overcome the effects of HTS on seed production. Two DNA constructs, one with the <i>Arabidopsis thaliana LEAFY</i> (<i>AtLFY</i>) promoter controlling <i>A. thaliana HEAT SHOCK PROTEIN 101</i> (<i>AtHSP101</i>) ORF expression and another with the <i>AtHSP101</i> promoter controlling <i>AtLFY</i> ORF expression, were inserted into <i>B. napus</i>. Other DNA constructs were made, using the constitutively expressed Cauliflower Mosaic Virus <i>35S</i> or the synthetic <i>EntCup4</i> promoters to control expression of the <i>AtHSP101</i> or <i>A. thaliana HEAT SHOCK TRANSCRIPTION FACTOR 3</i> (<i>AtHSF3</i>) ORFs. These constructs were inserted into both <i>B. napus</i> and <i>A. thaliana</i>. Transgenic plants were tested using a ramping temperature regime but were found not to have increased flower thermotolerance. During the manufacture of the DNA constructs it was determined that, in <i>A. thaliana</i>, 573 bp of <i>AtHSP101</i> had been copied between Terminal Inverted Repeats of a <i>Mu-Like Element</i> (<i>MULE</i>). This fragment was named <i>HSP101B</i>. In some transgenic <i>B. napus</i> and <i>A. thaliana</i> lines, containing 2046 bp of the <i>HSP101B</i> upstream regulatory region controlling <i>B</i>-glucuronidase (GUS) expression, cold-inducible GUS expression was observed. Methylation may have a role in control of endogenous <i>HSP101B</i> transcription.

Current knowledge indicates that nonthermal muscle metaboreflex activity plays a critical role in the modulation of skin vasodilation and sweating. However, the mechanisms of control have primarily been studied during isometric handgrip exercise in which muscle metaboreceptor activation is induced by a brief post-exercise ischemia of the upper limb. While the reflex increase in mean arterial pressure associated with this period of ischemia is consistent with the activation of muscle metaboreceptors, the change in baroreflex activity may in itself modulate the response. Thus, we sought to understand how these nonthermal stimuli interact in modulating the control of skin perfusion and sweating under conditions of elevated hyperthermia. Furthermore, we examined the mechanisms responsible for the maintenance of arterial blood pressure under varying levels of heat stress during isometric handgrip exercise. Our study findings indicate that the parallel activation of muscle metaboreceptors and baroreceptors during post-exercise ischemia causes divergent influences on the control of skin blood flow and sweating; and these nonthermal stimuli are dependent on the level of hyperthermia. Moreover, we report that heat stress reduces the increase in arterial blood pressure during isometric handgrip exercise and this attenuation is attributed to a blunted increase in peripheral resistance, since cardiac output increased to similar levels for all heat stress conditions. These results provide important insight and understanding into the role of muscle metabo- and baroreflex activity on the control of skin blood flow and sweating; along with further knowledge into the cardiovascular mechanisms responsible for the regulation of arterial blood pressure during hyperthermia.

Carbon flow into developing ovaries has been reported to be important in seed retention and seed size. Invertase, which cleaves sucrose into glucose and fructose has been shown to be important in rapidly expanding tissue, such as early root growth or during tiller expansion. The manipulation of invertase activity with over-the-top applications of agrochemicals may prevent the detrimental effects of abiotic stress by altering the source/sink relationship. These experiments examined economically important tissues in rice production during critical developmental stages under abiotic stress. Field and greenhouse studies were conducted under normal growing conditions using local management practices. Plants were treated with exogenous chemicals that affect the activity of invertase during the early-grain fill stage on field grown plants. Other plants were exposed to elevated nighttime temperature of 30°C for 4 d using a free-air, infrared heating device in the greenhouse. Rice was also treated at mid- to late-grain fill stage of the main crop to identify the impact of the exogenous chemicals on developing ratoon tiller buds. The activity of soluble acid invertase (SAI), concentrations of glucose, fructose, sucrose, and starch were determined in penultimate leaves, panicles, and main-crop stem segments during ratoon tiller bud expansion, using the enzyme-coupled stoichiometric production of NADH measured spectrophotometrically at 340nm. The results suggest SAI, carbohydrates, and agronomic characters are influenced by exogenous chemicals at the applied rates. The thidiazuron treatment caused an unidentified stress event. The stress was confirmed by increased hexose concentration and the proportion of hexose concentration to sucrose concentration. This stress reduced the main-crop grain yield, but not the ratoon yield or total grain yield. An interaction between the ammonium molybdate treatment and high nighttime temperature was seen in the panicle. The ranked difference was reduced by the high nighttime temperature from the ambient nighttime temperature for the SAI activity, TSC content, starch content, and TNC content. The tested chemicals and rates are not appropriate for commercial rice production because the effect of the exogenous chemicals do not appear to consistently aid rice plants to counteract the detrimental effects of abiotic stress.

It was hypothesized that technical factors associated with in vitro production (IVP) of embryos may influence rate of blastocyst development of oocytes matured at 38.5 or 41.0 C. To test this hypothesis, a retrospective meta-analysis was performed. Simple linear regression was performed to analyze continuous variables and ANOVA for categorical variables. Interactions among factors and maturation temperature on blastocyst development were analyzed using dummy regression for continuous variables, and using a factorial treatment design and ANOVA for categorical variables. Month of collection was the only variable that impacted responsiveness of ova to heat stress. Independent of maturation temperature, variables that explained most variation in blastocyst development included technician, total number of sliced ovaries per collection, ova number placed per well of oocyte maturation media, oocyte collection time, bull ID, sperm concentration added to ova, and ova age at IVF. The proportion of 8 to 16-cell embryos at time of cleavage assessment was the best predictor of blastocyst development. Results of model selection showed that development of ova matured at 38.5 C was associated with size of the collection, while development of ova matured at 41.0 C was mainly associated with ova age at fertilization. When data for ova matured at 38.5 and 41.0 C were combined, the effect of number of PZ per well on blastocyst development became evident. Use of these findings for optimizing efficiency of IVP procedures would effectively reduce experimental costs related to embryo production and increase laboratory productivity.

The cardiovascular effects of whole body heat stress during exercise are well established. However the independent contribution of elevated skin temperature (Tsk) or core temperature (Tc) on these responses remains unclear. The purpose of this study was to determine how increases in Tsk and Tc alone and in combination, impact cardiovascular responses during moderate intensity exercise. To accomplish this goal, eight healthy, recreationally active males were immersed to the neck in a cold (14 - 17°C) or hot (40 - 42.5°C) water bath for 20 to 25 min to alter Tc immediately prior to exercise with either cool Tsk (i.e. fans) or warm Tsk (i.e. heaters). Conditions during exercise were cool skin and cool core (CC), warm skin and cool core (WC), cool skin and warm core (CW), and warm skin and warm core (WW), and were conducted in a randomized crossover design. When data was combined (n=16), warm core conditions (CW and WW) were associated with significantly higher average heart rate (HR) and lower stroke volume (SV) during exercise compared to cool core conditions (CC and WC); 168.1 ± 3.2 vs. 152.2 ± 4.0 beats/min and 139.2 ± 7.3 vs. 147.7 ± 9.4 mL/beat, respectively. The approximate 9 mL/beat decline in SV and 16 beat/min increase in HR in warm core conditions tended to increase cardiac output (Q), 23.2 ± 0.6 vs. 22.2 ± 0.7 L/min, P=0.078. Similarly, warm Tsk conditions (WC and WW) were associated with significantly higher average HR and lower SV during exercise compared to cool Tsk conditions (CC and CW); 165.2 ± 3.3 vs. 155.1 ± 3.4 beats/min and 140.8 ± 7.8 vs. 146.0 ± 8.7 mL/beat, respectively. Additionally, there was also a trend for Q to be elevated with warm skin (23.0 ± 0.6 vs. 22.4 ± 0.6, P=0.075). Although combined data indicated that warm Tsk conditions significantly lowered average SV by ~6 mL/beat, there was no reduction in SV during exercise by warm Tsk, when Tes was cool (i.e. <37.0°C), as evidenced by identical values for SV in CC and WC, 147.7 ± 9.8 vs. 147.7 ± 9.0 mL/beat, respectively. In contrast, SV was significantly lower in WW compared with CW, 133.9 ± 7.0 vs. 144.4 ± 7.8 mL/beat, respectively. Therefore, the major reduction in SV by warm Tsk occurred during WW, when Tes was elevated (i.e. >38.0°C). Analyzing data independently for precooling and preheating conditions revealed that warm Tsk was associated with greater HR drift from 5 to 20 min of exercise, compared to cool Tsk, when esophageal temperature (Tes) was both cool or warm (23.9 ± 2.2 vs. 17.5 ± 2.3 and 12.3 ± 1.3 vs. 4.6 ± 1.7 beats/min, respectively). These observations demonstrate that both Tes and Tsk can directly influence cardiovascular responses during exercise, as indicated by elevations in HR during exercise with warm Tsk, with both warm and cool Tes. However SV is not compromised by warm Tsk if Tes is below 37.5°C. Furthermore, when both Tes and Tsk are elevated simultaneously, cardiovascular strain (i.e. increased HR and reduced SV) is much greater than when either is elevated alone. This is demonstrated by the finding that average HR was 175.8 ± 3.2 beats/min in WW, compared to 149.8 ± 4.0, 154.7 ± 4.1, and 160.3 ± 3.5 beats/min, in CC, WC, and CW, respectively, and the fact that SV was lowest during exercise in WW. In conclusion, individuals exercising in the heat should take measures to keep skin cool, especially when Tes is 39°C or greater to attenuate the cardiovascular strain that occurs with warm Tsk, when Tes is elevated. / text

Studies conducted for this dissertation utilized a rodent model exposed to single or multiple short duration heat loads in an effort to: 1) elucidate the changes in energy metabolism occurring at the tissue and whole-body level in response to hyperthermia, 2) characterize specific aspects of glucose utilization and hepatic glucose production following a heat load and 3) determine if aspects of mitochondrial function and/or dysfunction might play a role in the metabolic changes that occur in response to heat stress. Study 1 was conducted to determine if rodents exposed to heat stress shared similarities using a bovine heat stress model. Specifically, we were interested in identifying changes in blood metabolites and hormones, as well as gene expression and protein abundance of enzymes associated with energy metabolism in skeletal muscle (type I and type II), liver and adipose tissue. Previous bovine data indicates glucose may be preferentially utilized during heat stress, suggesting alterations in energy metabolism. This study provided evidence that tissue-specific changes occur in response to a heat load and that full glucose oxidation might be reduced, specifically in skeletal muscle where abundance of PDK4 mRNA was increased. Within skeletal muscle, glucose transporters (GLUTs 1 and 4) also tended to be increased in rats exposed to a heat load. Increases in skeletal muscle AMPK-α and PGC-1α as well as increased expression of energy substrate transporters suggests heat stress may impose a cellular energy deficit and/or increased energy demands which subsequently leads to changes in energy metabolism. Few changes were noted in either hepatic or adipose tissue in response to acute heat stress in this pilot study. Study aim of Chapter 3 was to further characterize the effects of heat stress on energy metabolism at the tissue and whole-body level in rats exposed to either 1 or 2 bouts of heat. Rats exposed to a 6 h heat load tended to have higher plasma glucose but reduced insulin levels, compared to thermal neutral controls, suggesting decreased glucose uptake or increased hepatic glucose output. Additionally, although heat stress likely increases whole-body energy demand, plasma NEFA levels were blunted in the early hours following onset of heat, suggesting increased adipocyte insulin sensitivity. Gene expression of enzymes associated with oxidative energy metabolism were increased in the TA (which is comprised primarily of glycolytic muscle fibers) following 2 bouts and in liver following a single bout of heat, while expression of oxidative enzymes were decreased within the soleus (a primarily oxidative muscle type). AMPK mRNA was increased following a single bout of heat in hepatic tissue and after 2 bouts of heat in type I skeletal muscle. AMPK mRNA abundance remained the same following 1 bout but was reduced following 2 bouts of heat within type II skeletal muscle. In the TA, phosphorylated AMPK protein abundance was reduced by HS. Abundance of PGC-1α mRNA was increased in types I and II skeletal muscle but was only numerically increased in liver following heat exposure. These data suggest differences at the transcription level in how heat effects energy metabolism within types I and II skeletal muscle as well as between muscle and hepatic tissue and also suggests a cellular attempt to increase energy production (by all mechanisms) in response to heat exposure. Study 3 (Chapter 4) focused on the effect of a heat load on glucose utilization in skeletal muscle and hepatic glucose production capacity. Similar to study 1, PDK4 expression was increased in types I and II skeletal muscle, while PDK2 expression was increased in hepatic tissue. Within skeletal muscle, increases in PDK expression paralled the increased protein abundance of PDHE1α following heat exposure, implying a decrease in oxidative glucose metabolism. Within the liver, protein abundance of PDH-E1α was reduced following a single heat load, but returned to TN levels after a 2nd heat exposure, suggesting that glucose oxidative metabolism is increased above normal levels after an initial heat exposure, but reduced following multiple heat bouts. Hepatic mRNA abundance for gluconeogenic enzymes were increased, implying an increase in hepatic glucose output capacity. The purpose of Study 4 (Chapter 5) was to determine if heat stress elicits changes on mitochondrial function/dysfunction (i.e. oxidative stress), that may account for changes observed in energy metabolism. Expression of genes associated with antioxidant defense were increased by heat stress, but differed between types I and II skeletal muscle as well as between muscle, hepatic tissue and WBCs. The abundance of mRNA for antioxidant enzymes was increased the greatest, and expression of DNA repair enzymes were also upregulated the most within hepatic tissue due to heat exposure, suggesting either increased damage at the level of hepatocytes or greater defensive capacity following an environmental insult. Taken together, this data provides evidence that heat alters energy metabolism, but these changes are tissue-specific and may be reflective of where damage is occurring, or which tissues are able to adapt and/or compensate for increased energy demands imposed by an environmental insult.

The goal of this dissertation was to evaluate genomic regulation during bovine mammary epithelial cell (BMEC) growth and differentiation. To accomplish this goal, a collagen gel cell culture system was developed that was capable of mimicking the prepartum stages of epithelial development and differentiation. In addition, a 4,600-cDNA bovine microarray was developed in order to profile gene expression. Analysis of BMEC in collagen cultures using various lactogenic conditions highlighted the critical importance of both hormonal and structural signals. The objective of the first study utilizing the microarray was to evaluate the contribution of the two prominent lactogenic factors in vitro, 1) prolactin and 2) gel release. Collectively, lactogenic stimulation appears to turn off genes associated with structural progression and morphogenesis, and turn on genes involved in alveolar MEC differentiation such as cell polarization, milk protein synthesis and ER/Golgi transport. The objective of the second study utilizing these resources was to evaluate the direct effects of thermal stress on BMEC growth and development. The structural response to thermal stress was characterized by morphogenic inhibition and dramatic regression of the ductal branches. Microarray analysis revealed an overall up-regulation of genes associated with stress response, DNA repair, protein degradation and cell death. In contrast, genes associated with cellular and MEC-specific biosynthesis, metabolism, and morphogenesis, were generally down-regulated. Subsequent to the analysis of BMEC differentiation was a targeted effort focusing on two small molecules hypothesized to be involved in regulating the BMEC secretory response: serotonin and prostaglandin E2. A pilot study suggested that serotonin is produced by bovine MEC and a model was proposed that describes serotonin's role as a feedback inhibitor during milk synthesis and secretion. A second pilot study demonstrated that PGE2 had a consistently positive influence on lumen diameter of alveolar structures in vitro. Overall, this dissertation provides new resources for studying bovine functional genomics, particularly within the mammary gland, and it provides a strong foundation for understanding genomic regulation of mammary epithelial structure and function. Furthermore, it establishes potential roles for local regulation of milk production by serotonin and PGE2.

There has been little research into the use of water cooling methods for pigs during transport to slaughter under conditions of high ambient temperature. The aim of this study was to examine the effects of water sprinkling pigs before departure from the farm and before unloading at the plant on behaviour during transport, unloading and lairage using live and remote observations, and on pig gastrointestinal tract temperature (GTT) and trailer micro-climate measured by data loggers. Above 23oC, the use of water sprinkling tended to decrease GTT upon arrival and significantly decreased drinking bouts during lairage. There were no detrimental effects of the water sprinkling on unloading behaviours (e.g. slips and falls) or on trailer micro-climate conditions in terms of temperature, humidity or ammonia. Water sprinkling to wet the skin of pigs can therefore be used to cool pigs during transport and lairage under high ambient temperatures.

This dissertation investigates: (1) the implications of including high-yielding energy sorghum under the Renewable Fuels Standard (RFS2) program; (2) the effects of RFS2 with and without projected climate change scenarios on U.S. agriculture; (3) the spatial distribution of cattle breeders in Texas to quantify how climate factors influence cattle breed selection. In the RFS2 energy sorghum work, the ability of the agriculture sector to meet the fuel requirements of RFS2 is examined with and without energy sorghum being a possibility using an agricultural sector model. The results show that energy sorghum would be a valuable contributor that would be used as a feedstock producing over 13 billion gallons per year of cellulosic ethanol. Without the presence of energy sorghum it is found that switchgrass serves as the major cellulosic ethanol feedstock. Findings also indicate that the presence of high-yielding energy sorghum does relax commodity prices and export reductions except for grain sorghum as energy sorghum competes with grain sorghum production. In addition, the results show that the introduction of energy sorghum has minimal effects on GHG mitigation potential in the agricultural sector. In the RFS2 and climate change research, the analysis shows that climate change eases the burden of meeting the RFS2 mandates increasing consumer welfare while decreasing producer welfare. The results also show that climate change encourages a more diversified use of biofuel feedstocks for cellulosic ethanol production, in particular crop residues. In the cattle breed research, summer heat stress is found to be a significant factor for breed selection: positive for Bos indicus and negative for Bos taurus and composite breeds. The estimation results also indicate a price-driven trade-off between Bos taurus and Bos indicus breeds.

Sixty steers were grazed on toxic tall fescue for 104 days to determine the effects of hair coats and steroidal ear implants on physiological measurements. Steers were stratified by body weight and hair coat color for assignment to six, 3.0-ha pastures of 'Kentucky 31' tall fescue. Main plot treatments of either ten clipped or ten unclipped steers were randomly assigned to pastures. Five steers in each pasture were implanted with Synovex-S (200 mg progesterone-20 mg estradiol) and five were implanted with Compudose (25 mg estradiol) as sub-plot treatments. Hair growth rate averaged 0.29 mm/day. Sweating rate declined (P andlt; 0.001) over the grazing period and was higher (Pandlt; 0.10) with the estradiol implant. Rectal temperatures were lower (P andlt; 0.05) in clipped cattle (39.3 vs. 39.5??C) when the highest ambient temperature (33??C) of the study was recorded. At high environmental temperatures, percentage of steers actively grazing was negatively correlated (P andlt; 0.10) with ambient temperature. Winter hair coat retention, continuous hair growth, and reduced sweating caused impaired thermoregulation and thus decreased grazing frequency resulting in poor animal performance.